Production Of Propanols, Alcohols, And Polyols In Consolidated Bioprocessing Organisms

Abstract

The present in provides for novel metabolic pathways leading to propanol, alcohol or polyol formation in a consolidated bioprocessing system (CBP), where lignocellulosic biomass is efficiently converted to such products. More specifically, the invention provides for a recombinant microorganism, where the microorganism expresses one or more native and/or heterologous enzymes; where the one or more enzymes function in one or more engineered metabolic pathways to achieve: (1) conversion of a carbohydrate source to 1,2-propanediol, isopropropanol, ethanol and/or glycerol; (2) conversion of a carbohydrate source to n-propanol and isopropanol; (3) conversion of a carbohydrate source to isopropanol and methanol; or (4) conversion of a carbohydrate source to propanediol and acetone; wherein the one or more native and/or heterologous enzymes is activated, upregulated or downregulated.

Description

BACKGROUND OF THE INVENTION

Field of the Invention

[0001] Consolidated Bio-Processing (CBP) in essence describes a mode of operation where biocatalysts produce enzymes that can breakdown inexpensive cellulose into usable sugars and then simultaneously ferment them into value added products in a single vessel. CBP, which reduces the number of unit processes, significantly lowers operating and capital costs associated with cellulosic biofuel production. Furthermore, CBP processes reduce or eliminate the need for externally-added, expensive cellulases. See Lynd et al, "Microbial cellulose utilization: Fundamentals and biotechnology," Microbiology and Molecular Biology Reviews 66(3):506-577 (2002); Lynd et al., "Consolidated bioprocessing of cellulosic, biomass: An update," Current Opinion in Biotechnology 16(5):577-583 (2005); "Breaking the Biological Barriers to Cellulosic Ethanol: A joint Research Agenda," December 2005, Rockville, Md. Publication Date: June 2006; DOE/SC-0095. CBP is widely considered to be the "Ultimate low-cost configuration for cellulose, hydrolysis and fermentation." DOE/USA joint Research Agenda. See DOE/SC-0095 Joint Research Agenda. CBP on plant biomass, e.g., lignocellulosic biomass, also reduces the need to rely on petrochemical feedstocks to produce fermentable, value added products, such as propanols, alcohols, and polyols.

[0002] Among forms of plant biomass, lignocellulosic biomass ("biomass") is particularly well-suited for producing fermentable, value added products because of its large-scale availability, low cost, and environmentally benign production. The primary obstacle impeding the processing of biomass feedstocks is the general absence of low-cost technology for overcoming the recalcitrance of these materials to conversion into useful products. Lignocellulosic biomass contains carbohydrate fractions (e.g., cellulose and hemicellulose) that can be converted into propanols, alcohols, and polyols. In order to convert these fractions, the cellulose and hemicellulose must ultimately be converted or hydrolyzed into monosaccharides; it is the hydrolysis that has historically proven to be problematic.

[0003] Lignocellulosic feedstocks are recalcitrant to hydrolysis and subsequent release of sugars. Concentrated acid pre-treatment can release sugars with some associated loss of either pentose or hexose sugars. However, the larger issue with concentrated acid use is the additional capital cost associated with those pre-treatments. The capital cost implications involve using expensive materials of construction, handling corrosive chemicals and dealing with environmental implications. In fact, a group of eminent scholars in the area of lignocellulosic pretreatment have commented that although concentrated mineral acids are effective, they are too expensive to be practical when measured against the value of the resulting sugars. Mosier et al., (2005), Bioresource Technology 96, 673-686.

[0004] More recently some companies have made technology claims where they have demonstrated concentrated acid recycle at laboratory scale as a means of reducing the cost associated with using concentrated acid pretreatments. A recent article on this recycling technology clarifies that they are only able to recycle 42% of the added acids and reiterates that this technology will only be tested in a pilot facility in the second half of 2010. Technology Review, Wednesday, Jun. 10, 2009 (available at http://www.technologyreview.com/energy/22774/). Additionally, the article includes caution by industry experts against the use of concentrated HCl acids for pretreatment as the plant would require expensive materials of construction. CBP provides a viable alternative to the production of fermentable sugars from biomass.

[0005] CBP biomass processing schemes involving enzymatic or microbial hydrolysis commonly involve four biologically mediated transformations: (1) the production of saccharolytic enzymes (cellulases and hemicellulases); (2) the hydrolysis of carbohydrate components present in pretreated biomass to sugars; (3) the fermentation of hexose sugars (e.g., glucose, mannose, and galactose); and (4) the fermentation of pentose sugars (e.g., xylose and arabinose). These four transformations occur in a single step in CBP, which is distinguished from other less highly integrated configurations in that it does not involve a dedicated process step for cellulase and/or hemicellulase production.

[0006] Thus, CBP offers the potential for lower cost and higher efficiency than processes featuring dedicated cellulase production. The benefits result in part from avoided capital costs, substrate and other raw materials, and utilities associated with cellulose production. In addition, several factors support the realization of higher rates of hydrolysis, and hence reduced reactor volume and capital investment using CBP including enzyme-microbe synergy and the use of thermophilic organisms and/or complexed cellulose systems. Moreover, cellulose-adherent cellulolytic microorganisms are likely to compete successfully for products of cellulose hydrolysis with non-adhered microbes, e.g., contaminants, which could increase the stability of industrial processes based on microbial cellulose utilization. Progress in developing CBP-enabling microorganisms is being made through two strategies: engineering naturally occurring cellulolytic microorganisms to improve product-related properties, such as yield and titer and engineering non-cellulolytic organisms that exhibit high product yields and titers to express a heterologous cellulase and hemicellulase system enabling cellulose and hemicellulose utilization.

[0007] Many bacteria have the ability to ferment simple hexose sugars into a mixture of acidic and pH-neutral products via the process of glycolysis. The glycolytic pathway is abundant and comprises a series Of enzymatic steps whereby a six carbon glucose molecule is broken down, via multiple intermediates, into two molecules of the three carbon compounds dihydroxyacetone phosphate and glyceraldehyde 3-phosphate. This process results in the net generation of ATP (biological energy supply) and the reduced cofactor NADH. From these three carbon compounds, a number of downstream vat added products can be made using the metabolic machinery of the CBP organisms, including, e.g., propanols, alcohols, and polyols.

[0008] Industrial chemicals, such as propanols, alcohols, and polyols are traditionally derived from petrochemical feedstocks. Production of such chemicals from petrochemical feedstocks, however, has its problems, not least of which is the use of a non-renewable resource that is subject to price fluctuations and heavy regulation. Thus, there is a need in the art for the production of propanols, alcohols, and polyols from resources that allow for large-scale availability, low cost, and environmentally benign production, all of which are advantages of CBP. In particular, there is a need for engineered organisms capable of converting biomass into propanols, alcohols, and polyols as part of a CBP system.

BRIEF SUMMARY OF THE INVENTION

[0009] The present invention provides for novel metabolic pathways leading to propanol, alcohol or polyol formation in a consolidated bioprocessing system (CBP), where lignocellulosic biomass is efficiently converted to such products.

[0010] The invention therefore provides for a recombinant microorganism, where the microorganism expresses one or more native and/or heterologous enzymes; where the one or more enzymes function in one or more engineered metabolic pathways to achieve: (1) conversion of a carbohydrate source to 1,2-propanediol, isopropropanol, ethanol and/or glycerol; (2) conversion of a carbohydrate source to n-propanol and isopropanol; (3) conversion of a carbohydrate source to isopropanol and methanol; or (4) conversion of a carbohydrate source to propanediol and acetone.

[0011] The engineered metabolic pathways of the invention are outlined in FIGS. 1-5 and 7. The enzymes that function at various steps along the pathways are identified in Tables 2-5. The engineered metabolic pathways of the invention are utilized to achieve high theoretical yields of products, particularly 1,2-propanediol isopropropanol, n-propanol and methanol in bacteria and yeast.

BRIEF DESCRIPTION OF THE DRAWINGS/FIGURES

[0012] FIG. 1 shows a schematic of theoretical metabolic pathways for the production of mixed alcohols in bacterial and yeast CBP platforms. Yeast-specific branch pathways are depicted by EC numbers in dark gray boxes. Bacteria-specific branch pathways are depicted by EC numbers in light gray boxes.

[0013] FIG. 2 shows a schematic of theoretical metabolic pathways for the production of n-propanol and isopropanol in bacterial CBP platforms.

[0014] FIG. 3 shows a schematic of theoretical metabolic pathways for the production of isopropanol and methanol in bacterial CBP platforms.

[0015] FIG. 4 shows a schematic of theoretical metabolic pathways for the anaerobic production of propanediol and acetone in bacterial and yeast CBP platforms. A yeast-specific branch pathway is depicted by EC numbers in dark gray boxes. A bacteria-specific branch pathway is depicted by the EC number in a light gray box.

[0016] FIG. 5 shows a schematic of a theoretical metabolic pathway for the aerobic production of propanediol in yeast CBP platforms.

[0017] FIG. 6 shows a schematic of propanediol utilization (pdu) gene organization in T. saccharolyticum.

[0018] FIG. 7 shows a schematic of theoretical metabolic pathways for the production of n-propanol and isopropanol in bacterial and yeast CBP platforms.

DETAILED DESCRIPTION THE INVENTION

Metabolic Pathway Engineering

[0019] Many bacteria have the ability to ferment simple hexose sugars into a mixture of acidic and pH-neutral products via the process of glycolysis. The glycolytic pathway is abundant and comprises a series of enzymatic steps whereby a six carbon glucose molecule is broken down, via multiple intermediates, into two molecules of the three carbon compound pyruvate. This process results in the net generation of ATP (biological energy supply) and the reduced cofactor NADH.

[0020] Pyruvate is an important intermediary compound of metabolism. For example, under aerobic conditions pyruvate may be oxidized to acetyl coenzyme A (acetyl CoA), which then enters the tricarboxylic acid cycle (TCA), which in turn generates synthetic precursors, CO2 and reduced cofactors. The cofactors are then oxidized by donating hydrogen equivalents, via a series of enzymatic steps, to oxygen resulting in the formation of water and ATP. This process of energy formation is known as oxidative phosphorylation.

[0021] Under anaerobic conditions (no available oxygen), fermentation occurs in which the degradation products of organic compounds serve as hydrogen donors and acceptors. Excess NADH from glycolysis is oxidized in reactions involving the reduction of organic substrates to products, such as lactate and ethanol. In addition, ATP is regenerated from the production of organic acids, such as acetate, in a process known as substrate level phosphorylation. Therefore, the fermentation products of glycolysis and pyruvate metabolism include a variety of organic acids, alcohols and CO.sub.2.

[0022] Most facultative anaerobes metabolize pyruvate aerobically via pyruvate dehydrogenase (PDH) and the tricarboxylic acid cycle (TCA). Under anaerobic conditions, the main energy pathway (or the metabolism of pyruvate is via pyruvate-formate-lyase (PFL) pathway to give formate and acetyl-CoA. Acetyl-CoA is then converted to acetate, via phosphotransacetylase (PTA) and acetate kinase (ACK) with the co-production of ATP, or reduced to ethanol via acetalaldehyde dehydrogenase (AcDH) and alcohol dehydrogenase (ADH). In order to maintain a balance of reducing equivalents, excess NADH produced from glycolysis is re-oxidized to NAD+ by lactate dehydrogenase (LDH) during the reduction of pyruvate to lactate. NADH can also be reoxidized by AcDH and ADH during the reduction of acetyl-CoA to ethanol, but this is a minor reaction in cells with a functional LDH.

[0023] Ethanologenic organisms, including yeast (e.g., Saccharomyces cerevisiae), are capable of a second type of anaerobic fermentation, commonly referred to as alcoholic fermentation, in which pyruvate is metabolized to acetaldehyde and CO.sub.2 by pyruvate decarboxylase (PDC). Acetaldehyde is then reduced to ethanol by ADH regenerating NAD+. Alcoholic fermentation results in the metabolism of one molecule of glucose to two molecules of ethanol and two molecules of CO.sub.2.

[0024] The present invention is directed to the modification of traditional glycolytic pathways in bacteria and yeast, as described above, to engineer novel metabolic pathways capable of generating or increasing the yield of certain products that could not otherwise be generated by the native organism. Such products include n-propanol or isopropanol along with alcohols, propanediol, ethanol, and glycerol.

[0025] In particular embodiments, the present invention is directed to the production of mixed alcohols in CBP yeast and bacterial platforms. In other embodiments, the present invention is directed to the production of n-propanol and isopropanol in a CBP bacterial platform. In additional embodiments, the present invention is directed to production of isopropanol and methanol in a CBP bacterial platform. In certain other embodiments, the present invention is directed to the production of propanediol in a CBP yeast or bacterial platform. In further embodiments, the propanediol could be directly utilized in industrial applications or condensed to propylene or converted via a chemical or microbial based biocatalysis to propanol.

[0026] The present invention is directed to the engineering of such alternative metabolic pathways in various microorganisms, including bacteria and yeast. The term "microorganism," as used herein, refers to an organism of microscopic or submicroscopic size that can be seen only with the aid of a microscope and that typically consists of only a single cell. Microorganisms include bacteria, protozoans, and certain algae and fungi.

[0027] In certain embodiments, the bacterial microorganism is a species of the genera Thermoanaerobacterium, Thermoanaerobacter, Clostridium, Geobacillus, Saccharococcus, Paenibacillus, Bacillus, Caldicellulosiruptor, Anaerocellum, or Anoxybacillus. In certain embodiments, the microorganism is a bacterium selected from the group consisting of: Thermoanaerobacterium thermosulfurigenes, Thermoanaerobacterium aotearoense, Thermoanaerobacterium polysaccharolyticum, Thermoanaerobacterium zeae, Thermoanaerobacterium xylanolyticum, Thermoanaerobacterium saccharolyticum, Thermoanaerobium brockii, Thermoanaerobacterium thermosaccharolyticum, Thermoanaerobacter thermohydrosulfuricus, Thermoanaerobacter ethanolicus, Thermoanaerobacter brocki, Clostridium thermocellum, Clostridium cellulolyticum, Clostridium phytofermentans, Clostridium straminosolvens, Geobacillus thermoglucosidasius, Geobacillus stearothermophilus, Saccharococcus caldoxylosilyticus, Saccharoccus thermophilus, Paenibacillus campinasensis, Bacillus flavothermus, Anoxybacillus kamchatkensis, Anoxybacillus gonensis, Caldicellulosiruptor acetigenus, Caldicellulosiruptor saccharolyticus, Caldicellulosiruptor kristjanssonii, Caldicellulosiruptor owensensis, Caldicellulosiruptor lactoaceticus, and Anaerocellum thermophilum. In particular embodiments, the microorganism is Clostridium thermocellum or Thermoanaerobacterium saccharolyticum.

[0028] In certain other embodiments, the yeast microorganism is selected from the group consisting of Saccharomyces cerevisiae, Kluyveromyces lactis, Kluyveromyces marxianus, Pichia pastoris, Yarrowia lipolytica, Hansenula polymorpha, Phaffia rhodozyma, Candida utilis, Arxula adeninivorans, Pichia stipitis, Debaryomyces hansenii, Debaryomyces polymorphus, Schizosaccharomyces pombe, Candida albicans, and Schwanniomyces occidentalis. In particular embodiments, the yeast microorganism is Saccharomyces cerevisiae.

[0029] In certain instances, the microorganism of the invention is cellulolytic. The term "cellulolytic" means able to hydrolyze glycosidic linkages in oligohexoses and polyhexoses. Cellulolytic activity can also include the ability to depolymerize or debranch cellulose and hemicellulose.

[0030] The term "ethanologenic" is intended to include the ability of a microorganism to produce ethanol from a carbohydrate as a fermentation product. The term is intended to include, but is not limited to, naturally occurring ethanologenic organisms, ethanologenic organisms with naturally occurring or induced mutations, and ethanologenic organisms which have been genetically modified.

[0031] The terms "fermenting" and "fermentation" are intended to include the enzymatic process (e.g., cellular or acellular, e.g., a lysate or purified polypeptide mixture) by which ethanol is produced from a carbohydrate, in particular, as a product of fermentation.

[0032] By "thermophilic" is meant an organism that thrives at a temperature of about 45.degree. C. or higher.

[0033] By "thermophilic" is meant an organism that thrives at a temperature from about 20- about 45.degree. C.

[0034] The term "CBP organism" is intended to include microorganisms of the invention, e.g., microorganisms that have properties suitable for CBP.

[0035] In certain embodiments of the invention, one or more metabolic engineered pathways are utilized for the combined production of propanediol and isopropanol from glucose. The metabolic pathways and the various distinct enzymes (Table 2) required for the combined production of propanediol and isopropanol are shown in FIG. 1 and described further below in Example 1. These metabolic pathways can be subdivided into the following distinct production routes: i) the conversion of dihydroxyacetone phosphate into propanediol; ii) the conversion of pyruvate into isopropanol; iii) the conversion of pyruvate into ethanol (bacterial platform only); and iv) the conversion of dihydroxyacetone phosphate into glycerol (yeast platform only).

[0036] In certain other embodiments of the invention, one or more metabolic engineered pathways are utilized for the production of n-propanol and isopropanol. The metabolic pathways and the various distinct enzymes (Table 3) required for the production of n-propanol and isopropanol are shown in FIG. 2 and described further below in Example 2. The metabolic pathways can be subdivided into two distinct production routes: i) the conversion of dihydroxyacetone phosphate into n-propanol; and ii) the conversion of pyruvate into isopropanol.

[0037] In additional embodiments of the invention, one or more metabolic engineered pathways are utilized for the combined production of isopropanol and methanol from carbohydrates. The metabolic pathways and the various distinct enzymes (Table 4) required for the production of isoproponal and methanol are shown in FIG. 3 and described further below in Example 3. The metabolic pathways can be subdivided into distinct production routes: i) the conversion of pyruvate into isopropanol; and ii) the conversion of formate into CO.sub.2 and methanol.

[0038] In other embodiments of the invention, one or more metabolic engineered pathways are utilized for the co-production of propanediol and acetone from hexose and pentose sugars in thermophilic clostridia and yeast, such as S. cerevisiae. The metabolic pathways and the various distinct enzymes (Table 5) required for the production of propanediol and acetone are shown in FIGS. 4 and 5 and described further below in Examples 4 and 5. The metabolic pathways can be subdivided as follows: i) the production of dihydroxyacetone phosphate and glyceraldehydes-3-phosphate from glucose; ii) the subsequent generation of propanediol from dihydroxyacetone phosphate; and iii) the generation of acetone from glyceraldehyde-3-phosphate.

[0039] A summary of the pathways of the present invention is provided in Table 1 as follows:

TABLE-US-00001 TABLE 1 Summary: Engineering of CBP biocatalysts for production of propanol Required Metabolic Engineering Theoretical yield Gene KO Gene Gene expression Gene expression Pathway hexose/pentose EC# KO function EC# function Critical Step Mixed Alcohol- Hexose: 1.1.1.27 ldh 4.2.3.3 methylglyoxal pathway flux control, Bacterial 0.21 g/g propanediol 1.1.1.-- synthase methylglyoxal to 1,2 2 glucose .fwdarw. 1,2- 0.17 g/g propanol 2.3.1.9 aldo-keto propanediol propandiol + 0.13 g/g ethanol 2.8.1.8 reductase isopropanol + ethanol + Pentose: 4.1.1.4 thiolase 4 CO2 + H.sup.+ + 3 ATP 0.21 g/g propanediol acetyl-CoA 0.17 g/g propanol 1.1.1.80 transferase 0.13 g/g ethanol 1.1.1.202 acetoacetate decarboxylase Isoprop dehyd oxidoreductase Mixed Alcohol - Yeast Hexose: 4.1.1.1 pdc1 4.2.3.3 methylglyoxal pathway flux control 2 glucose .fwdarw. 0.21 g/g propanediol pdc5 1.1.1.6 synthase (without TPI deletion), isopropanol + 0.17 g/g isopropanol pdc6 2.3.1.54 glycerol methylglyoxal to propanol propanediol + 0.26 g/g glycerol 2.7.2.1 dehydrogenase glycerol + 3 CO2 + 1 Pentose: 2.3.1.8 pyruvate formate- ATP (no tpi deletion) lyase 0.21 g/g propanediol 2.8.3.8 acetate kinase 0.17 g/g isopropanol 4.1.1.4 phosphate 0.26 g/g glycerol acetyltransferase 1.1.1.79 acetate CoA- transferase 1.1.1.80 acetoacetate decarboxylase 1.2.1.49 glyoxylate reductase isopropanol dehydrodenese methylglyoxal dehydrogenase n- Hexose: 1.1.1.27 ldh 4.2.3.3 mgs methylglyoxal to n- propanol/isopropanol 0.44 g/g propanols 1.12.7.2 hyd 1.1.1.-- aldo-keto reduct propanol in bacteria Pentose: 1.2.1.10 acdh 2.3.1.9 thiolase 0.44 g/g propanols 2.8.1.8 acetyl-CoA trans 4.1.1.4 adc 1.1.1.80 Isoprop dehyd 1.1.1.202 oxidoreductase Isopropanol/Methanol - Hexose: 1.1.1.27 ldh 1.2.1.2 formate dehyd formate to methanol Bacterial 0.33 g/g propanol 1.2.1.10 Acdh 1.2.1.46 formaldehyde 2 glucose .fwdarw. 2 0.18 g/g methanol 4.2.3.3 mcs 1.1.1.-- dehyd isopropanol + 2 Pentose: 1.2.7.1 pfor 2.8.3.8 methanol dehyd methanol + 4 CO.sub.2 + 0.33 g/g propanol 4.1.1.4 (oxidoreductase) 4 ATP 0.18 g/g ethanol 2.3.1.9 CoA transferase acetoacetate decarb thiolase 1,2- Hexose: 1.1.1.1 adh 2.3.1.9 thiolase methylglyoxal to Propandiol/Acetone - 0.42 g/g propanediol 1.1.1.27 ldh 2.8.3.8 acetate CoA- propanediol Bacterial 0.16 g/g acetone 1.12.7.2 hyd 4.1.1.4 transferase 2 glucose .fwdarw. 2 Pentose: acetoacetate propanediol + acetone + 0.42 g/g propanediol decarboxylase 3 CO.sub.2 + 1 ATP 0.16 g/g acetone propanediol/Acetone - Hexose: 4.1.1.1 pdc1 4.2.3.3 methylglyoxal pathway flux control Yeast 0.42 g/g propanediol 1.1.1.8 pdc5 2.3.1.54 synthase (without TPI deletion), 2 glucose .fwdarw. 2 0.16 g/g acetone 3.1.3.21 pdc6 2.7.2.1 pyruvate formate- methylglyoxal to propanediol + acetone + Pentose: gpd2 2.3.1.8 lyase propanediol 3 CO.sub.2 + 1 ATP (no tpi deletion) gpp1 2.8.3.8 acetate kinase 0.42 g/g propanediol 4.1.1.4 phosphate 0.16 g/g acetone 1.1.1.79 acetyltransferase 1.2.1.49 acetate CoA- transferase acetoacetate decarboxylase glyoxylate reductase methylglyoxal dehydrogenase propanediol Aerobic- Hexose: 4.1.1.1 pdc1 4.2.3.3 methylglyoxal pathway flux control from Yeast 0.42 to 0.61 g/g pdc5 1.1.1.6 synthase glucose 6-phosphate to 17 glucose + 6 O.sub.2 .fwdarw. propanediol pdc6 1.1.1.79 glycerol PPP and propanediol, 24 propanediol + 6 H.sub.2O + Pentose: 1.1.1.8 gpd2 1.2.1.49 dehydrogenase methylglyoxal to 30 CO.sub.2 + 7 ATP 100% xylose could not be 3.1.3.21 gpp1 glyoxylate propanediol converted via this reductase pathway, but methylglyoxal glucose/xylose mixtures dehydrogenase could, with yield similar to glucose alone

Metabolic Enzymes

[0040] As described above, the engineering of metabolic pathways in microorganisms requires certain enzymes to function at particular steps along the pathways, as shown in FIGS. 1-5.

[0041] The enzymes of the invention as described herein can be endogenous to the native strain of the microorganism, and can thus be understood to be referred to as "native" or "endogenous," An organism is in "a native state" if it has not been genetically engineered or otherwise manipulated by the hand of man in a manner that intentionally alters the genetic and/or phenotypic constitution of the organism. For example, wild-type organisms can be considered to be in a native state.

[0042] For example, in certain embodiments, when the host cell is a particular Thermoanaerobacter(ium) strain, one or more metabolic enzymes can be an enzyme derived from that same Thermoanaerobacter(ium) strain. Source libraries with fragments of whole genomic DNA from such a Thermoanaerobacter(ium) strain can be host-modified with promoters, terminators, replication origins, or homologous recombination targeting. Screening of these libraries can identify DNA encoding for enzymes of interest that function in one or more metabolic engineered pathways of the invention.

[0043] In other embodiments, the enzymes of the invention can be non-native or "heterologous" to the organism, and can be introduced into the organism on a vector by transformation or other methods known to one of ordinary skill in the art, as described further below.

[0044] The terms "activity," "activities," "enzymatic activity," and "enzymatic activities" are used interchangeably and are intended to include any functional activity normally attributed to a selected polypeptide. Typically, the activity of a selected polypeptide encompasses the total enzymatic activity associated with the produced polypeptide. The polypeptide produced by a host cell and having enzymatic activity can be located in the intracellular space of the cell, cell-associated, secreted into the extracellular milieu, or a combination thereof.

[0045] In certain embodiments, enzymes that function in the metabolic pathways of the invention are set forth below in Tables 2-5 and include the following: methylglyoxal synthase, aldo-keto reductase, glyoxylate reductase, methylglyoxal dehydrogenase, aldehyde reductase, pyruvate formate lyase, thiolase, CoA transferase, acetoacetate decarboxylase, isoproponal, aldehyde dehydrogenase, alcohol dehydrogenase, diol-hydrolase, dehydrogenase, phosphotransacetylase, oxidoreductase, formate dehydrogenase, formaldehyde dehydrogenase and methanol dehydrogenase.

[0046] As used herein, the term "methylglyoxal synthase" or "mgs" refers to an enzyme that catalyzes the chemical reaction glycerone phosphate methylglyoxal+phosphate

[0047] As used herein, the term "aldo-keto reductase" can refer to any number of related monomeric NADPH-dependent oxidoreductases, such as aldose reductase, prostaglandin F synthase, xylose reductase, and many others.

[0048] As used herein, the term "oxidoreductase" refers to an enzyme that catalyzes the transfer of electrons from one molecule (the reductant, also called the hydrogen or electron donor) to another (the oxidant, also called the hydrogen or electron acceptor).

[0049] As used herein, the term "glyoxylate reductase" refers to an enzyme that catalyzes the chemical reaction glycolate+NAD.sup.+ glyoxylate+NADH+H.sup.+. This enzyme belongs to the family of oxidoreductases, specifically those acting on the CH--OH group of donor with NAD+ or NADP+ as acceptor.

[0050] As used herein, the term "methylglyoxal dehydrogenase" refers to an enzyme that oxidizes methylglyoxal to pyruvate.

[0051] As used herein, the term "CoA transferase" is an enzyme, for example, such as acetyl CoA transferase that catalyzes the chemical reaction acyl-CoA+acetate a fatty acid anion+acetyl-CoA. The term "CoA transferase" also refers an enzyme that catalyzes the chemical reaction acetoacetyl-CoA+acetate acetoacetate+acetyl-CoA.

[0052] As used herein, the term "acetoacetate decarboxylase" or "ADC" refers to an enzyme involved in both the ketone body production pathway in humans and other mammals, and solventogenesis in certain bacteria. Its reaction involves a decarboxylation of acetoacetate, forming acetone and carbon dioxide.

[0053] As used herein, the teen "aldehyde dehydrogenase" refers to an enzyme that catalyzes the oxidation (dehydrogenation) of aldehydes.

[0054] As used herein, the term "dehydrogenase" refers to an enzyme that oxidizes a substrate by transferring one or more hydrides (H.sup.-) to an acceptor, usually NAD.sup.+/NADP.sup.+.

[0055] As used herein, the term "formate dehydrogenase" is an enzyme that catalyzes the oxidation of formate to bicarbonate or carbon dioxide, donating the electrons to a second substrate, such as NAD.sup.+ in formate:NAD.sup.+ oxidoreductase.

[0056] As used herein, the term "formaldehyde dehydrogenase" refers to an enzyme that catalyzes the chemical reaction formaldehyde+NAD.sup.++H.sub.2O formate+NADH+2 H.sup.+. This enzyme belongs to the family of oxidoreductases, specifically those acting on the aldehyde or oxo group of donor with NAD.sup.+ or NADP.sup.+ as acceptor.

[0057] As used herein, the term "methanol dehydrogenase" is an enzyme that catalyzes the chemical reaction methanol+NAD.sup.+ formaldehyde+NADH+H.sup.+. This enzyme also belongs to the family of oxidoreductases, specifically those acting on the aldehyde or oxo group of donor with NAD.sup.+ or NADP.sup.+ as acceptor.

[0058] As used herein, the term "pyruvate formate lyase" or "PFL" is intended to include the enzyme capable of converting pyruvate into Acetyl CoA and formate.

[0059] As used herein the term "alcohol dehydrogenase" or "ADH" is intended to include the enzyme capable of converting aldehydes, such as acetaldehyde and propionaldehyde, and ketones, such as acetone, into an alcohol, such as ethanol, n-propanol, or isopropanol.

[0060] As used herein, the term "phosphotransacetylase" or "PTA" is intended to include the enzyme capable of converting Acetyl CoA into acetyl phosphate.

[0061] As used herein, the term "diol dehydratase" is intended to include the enzyme capable of converting propanediol to propanal.

[0062] The term "upregulated" means increased in activity, e.g., increase in enzymatic activity of the enzyme as compared to activity in a native host.

[0063] The term "downregulated" means decreased in activity, e.g., decrease in enzymatic activity of the enzyme as compared to activity in a native host.

[0064] The term "activated" means expressed or metabolically functional.

[0065] The polypeptide sequences corresponding to certain of the enzymes of the present invention are as follows:

C. Thermocellum Proteins

TABLE-US-00002 [0066] EC 2.3.1.54 (Cthe0505; SEQ ID NO: 1) MDAWRGFNKGNWCQEIDVRDFIIRNYTPYEGDESFLVGPTDRTRKLWEKVSELLK KERENGGVLDVDTHTISTITSHKPGYIDKELEVIVGLQTDEPLKRAIMPFGGIRMVIKGAE AYGHSVDPQVVEIFTKYRKTHNQGVYDVYTPEMRKAKKAGIITGLPDAYGRGRIIGDYR RVALYGVDRLIAEKEKEMASLERDYIDYETVRDREEISEQIKSLKQLKEMALSYGFDISC PAKDAREAFQWLYFAYLAAVKEQNGAAMSIGRISTFLDIYIERDLKEGKLTEELAQELV DQLVIKLRIVRFLRTPEYEKLFSGDPTWVTESIGGMALDGRTLVTKSSFRFLHTLFNLGH APEPNLTVLWSVNLPEGFKKYCAKVSIHSSSIQYESDDIMRKHWGDDYGIACCVSAMRI GKQMQFFGARCNLAKALLYAINGGKDEMTGEQIAPMFAPVETEYLDYEDVMKRFDMV LDWVARLYMNTLNIIHYMHDKYAYEALQMALHDKDVFRTMACGIAGLSVVADSLSAI KYAKVKPIRNENNLVVDYEVEGDYPKFGNNDERVDEIAVQVVKMFMNKLRKQRAYRS ATPTLSILTITSNVVYGKKTGNTPDGRKAGEPLAPGANPMHGRDINGALAVLNSIAKLPY EYAQDGISYTFSIIPKALGRDEETRINNLKSMLDGYFKQGGHHINVNVFEKETLLDAMEH PEKYPQLTIRVSGYAVNFIKLTREQQLDVINRTIHGKI EC 2.3.1.8 (Cthe1029; SEQ ID NO: 2) VIIYSYKYYKYSFYDNSFGIMICGEEFMSFLEQIIERAKSDVKTIVLPESTDLRVIKA ASMIMKKGIAKVVLIGNEKEIKSLAGDIDLEGVMIEDSLNSEKLEDYANTLYELRKSKGM TIEAARETIKDPLYYGVMMVKKGEADGMVAGAVNSTANTLRPALQILKTAPGTKLVSSF FVMVVPNCEYGHNGTFVYADCGLVENPDADQLSEIAISASKSFEMLVGAKPQVAMLSY SSYGSAKSELTEKVIKATQLAKEKAPHLAIDGELQVDAAIVPEVAKSKAKGSSVAGKAN VLIFPDLDAGNIAYKLTQRLAKAEAYGPITQGLARPVNDLSRGCSAEDIVGVAAITAVQA QYVKA EC 2.7.2.1 (Cthe1028; SEQ ID NO: 3) MNILVINTGSSSLKYQLIDMTNESVLAKGVCDRIGLEHSFLKHTKTGGETWIEKD LYNHKLAIQEVISALTDEKIGVIKSMSEISAVGHRIVHGGEKFKESAIIDEDVMKAIRDCV ELAPLHNPSNIIGIEACKQILPDVPMVAVFDTAFHQTMPRHAYIYALPYEIYEKYKLRKY GFHGTSHKYVAHRAAQMLGKPIESLKLITCHLGNGASICAVKGGKSVDTSMGFTPLQGL CMGTRSGNVDPAVITYLMEKEKMNINDINNFLNKKSGVLGISGVSSDFRDVQDAAEKG DDRAQLALDIFCYGVRKYIGKYIAVLNGVDAVVFTAGIGENNAYIRREVLKDMDFFGIKI DLDKNEVKGKEADISAPDAKVKTLVIPTNEELEIARETLRLVKNL EC 1.1.1.80 (Cthe101; SEQ ID NO: 4) MINFVYKNPTKIIFGRGTELKVGEEVRQYSGKVLLHYGGGSIKKTGLYDRVVNSL KQAGVEVVELGGVMPNPRLGLVNEGIKICREKGIDFILAVGGGSAIDSAKAIAVGVPYDG DVWDFFCGKAEPKEALPVGVVLTIPAAGSEASPNSVITREDGLYKRGMYSELIRPVFAIM NPELTYTLPAYQTACGTADIMAHIMERYFTNETHTDLTDRLCEATLKTMIKNVPIALEEP DNYNARAEIMWAGTIAHNGLLGTGRIEDWASHNIEHEISAIYDVAHGAGLAVVFPAWM KYVYKNNLDRFVQFAVRVWNVEMNFDEPERTALEGIERLKKFFKEIGLPVSLKEMNIGD DRLEEMASKCTNGGKATIGNFVKLNREDVY NILKLAV Cthe0394 (SEQ ID NO: 5) MKAFNYYAPTEIIFGCGRVQEIGSITAQYGKKALLVTVPEFPEVKELYEKVKKSLR ENGVEVVHFDGVIPNPTTDVVTEGANMAKAAGVDWIGLGGGSSIDTAKAIAVEATHPG TAWDYNCHTPGPTSATLPIIAIGTTAGTGSQCTQCAVITKTSEKDKSAIWHKNIFPKVAIV DPEVTVTMPKSVTAQTGFDAFAHNFEAYLSVKTSPLVEMMAIEAIKMIKEYLPKALENP NDIEARSKMSLADTLGGLTNSNAGVTLPHGLGMQVGGHAPHVSHGQALAIIYPQFTRYT YAWAIEKFAKVGRIFNPALNELSDEEAAKEACVAIDDFLKKIGLWIGFKDVNVTKEQIRE IADDGQVLGDYLNNPRVATIDEMYELLMNCYERKE Cthe0423 (SEQ ID NO: 6) MTKIANKYEVIDNVEKLEKALKRLREAQSVYATYTQEQVDKIFFEAAMAANKM RIPLAKMAVEETGMGWEDKVIKNHYASEYIYNAYKNTKTCGVIEEDPAPGIKKIAEPLG VIAAVIPTTNPTSTAIFKTLIALKTRNAIIISPHPRAKNSTIEAAKIVLEAAVKAGAPEGIIGW IDVPSLELTNLVMREADVILATGGPGLVKAAYSSGKPAIGVGAGNTPAIIDDSADIVLAV NSIIHSKTFDNGMICASEQSVIVLDGVYKEVKKEFEKRGCYFLNEDETEKVRKTIIINGAL NAKIVGQKAHTIANLAGFEVPETTKILIGEVTSVDISEEFAHEKLCPVLAMYRAKDFDDA LDKAERLVADGGFGHTSSLYIDTVTQKEKLQKFSERMKTCRILVNTPSSQGGIGDLYNFK LAPSLTLGCGSWGGNSVSDNVGVKHLLNIKTVAERRENMLWFRTPEKIYIKRGCLPVAL DELKNVMGKKKAFIVTDNFLYNNGYTKPITDKLDEMGIVHKTFFDVSPDPSLASAKAGA AEMLAFQPDTIIAVGGGSAMDAAKIMWVMYEHPEVDFMDMAMRFMDIRKRVYTFPK MGQKAYFIAIPTSAGTGSEVTPFAVITDEKTGIKYPLADYELLPDMAIVDADMMMNAPK GLTAASGIDALTHALEAYVSMLATDYTDSLALRAIKMIFEYLPRAYENGASDPVAREKM ANAATIAGMAFANAFLGVCHSMAHKLGAFYHLPHGVANALMINEVIRFNSSEAPTKMG TFPQYDHPRTLERYAEIADYIGLKGKNNEEKVENLIKAIDELKEKVGIRKTIKDYDIDEKJE FLDRLDEMVEQAFDDQCTGTNPRYPLMNEIRQMYLNAYYGGAKK Cthe2445 (SEQ ID NO: 7) MKGKMKVCVLTGKEKLEWVERDIPQPGRGELQIKLKHVGVCGSDLHFYKEGRL ANWELDGPLALGHEPGGIVSAIGEGVEGFEIGDKVALEPGVPCGECEDCRKGHYNLCKH IKFMAIPHEKDGVFAEYCVHSASMCYKLPENVDTMEGGLMEPLSVALHATELSNAKIGE TAIVLGSGCIGLCTVMALKARGVSEIYVTDVVDKRLEKALEVGATRVFNSQREDIVEFA KTLPGGGADQVYECAGSRVTTLQTCKLIKRAGKVTLVGVSPEPVLELDIATLNAMEGTV YSVYRYRNMYPIAIAAVSSGVIPLKKIVSHVFDFKDCIEAIEYSTNHKDEVIKSVIKF Cthe2579 (SEQ ID NO: 8) MNFKFKIGTKVFFGKECVKENKAVFKDFRKRALLVTGKNSAKASGAFSDWEVL EEYGIDYEIYDRVANNPSLENVKEGGEAARKFDADFIIGIGGGSPLDASKAVAVLATNDI EPVDLYKNVFENKPLPIIAIPTTAGTGSEVTPYSILTRDDMKTKKSFGNEDTFPAVAFIDA RYTESMSYETTVDTALDAFTHALEGYLGRRSTPVSDILAVEAIRIFGECLENLLNNKFDY DVREKLLYMSMLGGMVISHTGTTIIHGMGYSLTYFKDIPHGRANGMLVREYLKYNYEA AKEKTDNVLRLLKVPSIDAFGEIIDRLIPQKPVLTKEEIELYASLAMKQNSTLSNARTVVK EDMEEIFKNTFGKG EC 4.2.2.3 (Cthe0095; SEQ ID NO: 9) MNIALIAHDKKKELMASFCIAYRSILKNHTLFATGTTGAIIVEATGLNVHRFLPGV MGEQQISARAAYNELDLVIFFRDPISAKSDEPDIHSLLRECDINNIPFATNLGTAEMLIKGL ERGDLDWRELIKK EC 1.1.1 (Cthe0152; SEQ ID NO: 10) LKYCKLGNTGLEVSKLCFGGLIIGPLQANLPPETGAEIILKSFELGVNFIDTAELYG TYSHIGKALKKTNKNIVVATKSYAYSAEGAKESLEKARKEMDIDVIDIFMLHEQESRLTL KGHREALEYYISMKEKGIIKAVGVSTHNVEVVEACCEMPEVDVIHPIVNKAGIGIGDGTI DDMLKAVEKAYSVGKGIYSMKPLGGGNLIKSYKEAMDFVLNIPYIHSIAVGMQSIEEVV MNVCIFEGKEVPQDVQKSLENKKRHLHIDWWCEGCGKCVERCKQKALKLVDGKAKVE EEKCVLCSYCASVCPVFAIKVS Cthe0236 (SEQ ID NO: 11) MQYRGLGKTGVKVSALGFGAMRLPQININGNTRVDEEKSIEMIHRAFELGVNYID TAPGYCNGESEVVVGKALKGWRDKIYLSTKNPIENASGDDWRKRLENSLKKLDTDYID FYHMWGINWETYETKIDVKGGPLEAARKAKEEGLIRHISFSFHDKPENLIKLIDTGNFET VLCQYNLLDRSNEKAIAHAKRKGLGVIIMGPVGGGKLGEPSETIKKLLPKKTVSCAEIAL RFVLANPNVDCALSGMSTIEMVEENVRVASNDTPLTKEELEMIRASMEENICRMEDLYC TGCNYCMPCPVGVNIPLNFQLMNYHRVYKITDYARGQYSQIGKVEWYKGKPAHECIEC GVCETKCPQKLEIRKQLKETARVLSVK Cthe0283 (SEQ ID NO: 12) MKYRKMGRTGLYISEISLGSWLTYGNSTDKETAVKVIDTAYSLGINYFDTANVY ANGRAEVIVGEALKKYPRESYILATKAFWPMGTGPNDKGLSRKHVFEQVHASLKRLNV DYIDIFYCHRYDPETPLEETLRTIDDLLRQGKILYVGVSEWTAAQMAQALHIADRYLLDR IVVNQPQYNMFHRYIEKEIIPFGEKNGISQIVFSPLAQGVLTGKYKPGGNIPRDSRAADPN SNMYIGQFLKEDKLLKVEKLKAVADEMGITLSQLAIAWVLRQPNVTSALIGASKPEQVE ENVKASGINLSDEILNKIEAILQ EC 5.3.1.1. (Cthe0139; SEQ ID NO: 13) MSRKVIAAGNWKMNKTPKEAVEFVQALKGRVADADTEVVVGVPFVCLPGVVE AAKGSNIKVAAQNMHWEEKGAFTGEVSGPMLAELGVDYVIIGHSERRQYFGETDETVN KKVHAAFKYGLKPIICVGESLTQREQGVTAELVRYQVKIALLGLSAEQVKEAVIAYEPIW AIGTGKTATNEQAEEVCGIIRECIKELYGQDVAEAIRIQYGGSVNAANAAELFNMPNIDG GLVGGASLKLDDFEKIAKYNK EC 1.2.7.1 Cthe2390 (SEQ ID NO: 14) MGKVVEIRWHGRGGQGAKTASLLLADAAFNTGKYIQGFPEYGPERMGAPITAY NRISDEKLTIHSNIYEPDYVVVVDDTLLTSVDVTAGLKEDGAHVNTPKTPDEIRPLLKGY KGKVCTIDARKISIETLGKYFPNTPMLGAVVKVSKIMDEEEFLKDMVESFKHKFANKPE VVEGNIKALERSMQEVKGL Cthe2391 (SEQ ID NO: 15) MSKELRDVKPDVTWKEITSOGVIDSPGNAHLFKTODWRSMKPVWNEEKCKQCL LCNPVCPDSSIMVSEEGKMTGIDYDHCKGCGICSKVCPFKAIDFVEEV Cthe2392 (SEQ ID NO: 16) MGIRERLSGNEATAIAMRQINPDVVAAFPITPSTEIPQYFSSYVADGLVDTEFVAV ESEHSAMSACIGAQAAGARAMTATSANGLAYMWEALYIAASMRLPIVLAAVNRALSGP INIHNDHSDTMGARDSGWIQLYSENNQEAYDNMLMAHRIGEHPDVMLPVMVCQDGFIT SHAIENIELVEDEKVKAFVGEYKPTHYLLDRENPISVGPLDLQMHYFEHKRQQAQAMEN AKKVILEVAEEFYKITGRKYGFFEEYKTDDADVAIVVMNSTAGTVKYVIDEYRAKGKK VGLIKPRVFRPFPVDELAQALSKFKAVAVMDKADSFNAAGGPLFTEVTSALFTKGVFGP KVINYKFGLGGRDVKVDDIEVVCEKLLEIASTGKVDSVYNYLGVRE

Cthe2393 (SEQ ID NO: 17) MAYNLKEVAKKPERLTGGHRMCAGCGAPIVVRQVLKALKPEDHAVISAATGCL EVSTFIYPYTAWKDSFIHSAFENTGATISGAEAAYKVLKKKGKIEGETKFIAFGGDGGTY DIGLQALSGAMERGHDMVYVCYDNGAYMNTGIQRSSATPKYADTTTSPVGKKIPGKM QPRKDLTEVLVNHRIPYVAQTAPFGNMKDLYEKAEKAIYTPGPAFLNVLAPCPRGWRY NTPDLMELSKLAVETCFWPLYEVIDGKYIINYKPKEKVPVKEFLKLQGRFKHLFKAGNE YMLEEIQKEVDLRWERLLKLAGEA EC 1.1.1.27 (Cthe1053; SEQ ID NO: 18) MNNNKVIKKVTVVGAGFVGSTTAYTLMLSGLISEIVLIDINAKKADGEVMDLNH GMPFVRPVEIYRGDYKDCAGSDIVIITAGANQKEGETRIDLVKRNTEVFKNIINEIVKYNN DCILLVVTNPVDILTYVTYKLSGFPKNKVIGSGTVLDTARFRYLLSEHVKVDARNVHAYI IGEHGDTEVAAWSLANIAGIPMDRYCDECHQCEEQISRNKIYESVKNAAYEIIRNKGATY YAVALAVRRIVEAIVRNENSILTVSSLLEGQYGLSDVCLSVPTIVGVNGIEEILNVPFNDEE IQLLRKSGNTLKEIIKTLDI EC 1.12.7.2 Cthc425 (SEQ ID NO: 19) MKVSICIGSSCHLKGAKQIVEQLQSLVADYNLKEKVELGGAFCMKNCVNGVSVT VDDKLFSVTPENVKSFFETEILKKLED Cthe426 (SEQ ID NO: 20) MTECLQTKKSNCKNCYKCIRIICPVKSLKFTDGQAHIVRDECVLCGECYWCPQN AKQIRSDVEKAKQLVLKYDVYASIAPSFVAWFHNKSIHDMEQALIKLGFKGADETAKG AYIVKKQYEKMIEEKKSKIIISSCCHTYNTLIQRIIYTGAIQYLADVVSPMLAHAQMLKKE HKGAKVVFIGPCISKKDEAEKYKGYVELVLTFDELDEWLKSENITIESNRGSSKEGRTRS FPVSGGIISSMDKDLGYHYMVVDGMENCINALENIERGEIDNCFIEMSACRGSCINGPPA RRKSNNIVGAILAVNKNTGAKDFSVPMPEPEKLKKEFRFEGVHKIMPGGTAIEEILKKMG KTSIEHELNCGSCGYDTCRDKAVAVLNGKADLTMCLPYLKEKAESFSDAIIKNTPNGVIV LNEDLEIQQINNSAKRILNLSPSTDLLGSPVSRILDPIDYILALREGKNCYYKRKYFAEYKK YVDETIIYDKEYHVIIIIMRDVTEEEKIKALKNKQSEAAIEIADKVVEKQMRVVQEIALLL GETAAETKIALTKLKETMEDE Cthe427 (SEQ ID NO: 21) MNDLCVDLGYKSLNKFGEQLCGDMIQVVKDDDTTILVLADGLGSGVKANILSTL TSKIISTMIAAHMGIEECVNTIMSTLPVCKVRGIAYSTFTIIKITNNTYAEIIQYDNPLVILLR NGKKYDYPTQTKIISGKKIVESKIRLNCDDVFVVMSDGAIYAGVGQTLNYGWQRENIIEF IESHYDKSLSANALTSLLIDTCNNLYANMPGDDTTIAAIKIRKRQWNLMFGPPQNPEDV HNMMSLFFAKQGRHIVCGGTTSTLAAKFLGKELETTIDYIDPRIPPIARIEGVDLVTEGVL TISRVLEYAKDYIGKNILYNEWHSKNDGASIIARMLFEEATDINFYVGKAINPAHQNPNL PIGFNIKMQLVEELSKILKQMGKTINLSYF Cthe428 (SEQ ID NO: 22) MSVTMSEAFDYSMIDNILSEHGTSETAIIAILQSIQEEYHYIPKEVFPYLSKKLKVSE ARIFSVATFYENFSLEPKGKYIIKVCDGTACHVRKSIPIIERLRKELGLSGTKPTTDDLMFT VETVSCLGACGLAPVITVNDKVYAEMTPDKASELIKQLREGDADA Cthe429 (SEQ ID NO: 23) MLKNREELRKAREMYSRYLKAEKRRVLVCAGTGCVSGGSMEIFERLSELVSKRG MDCQVELKEEPHDNTIGMKKSGCHGFCEMGPLVRIEPEGYLYTKVKLEDCEEIVDRTIV AGEHIERLAYKQNGVVYKKQDEIPFYKKQTRLVLEHCGQIDSTSITEYLATGGYYALEK ALFDMTGDEIINEITEANLRGRGGGGFPAGRKWAQVKRQNAKQKYVVCNGDEGDPGAF MDRSIMEGDPHRMIEGMIIAGIACGASEGYIYVRAEYPLAVSRLKRAIEQAKEFGLLGENI LGSNFSFNIHINRGAGAFVCGEGSALTASIEGKRGMPRVKPPRTVEQGLFDMPTVLNNVE TFANVPLIIKNGADWYKSIGTEKSPGTKAFALTGNIENTGLIEIPMGTTLREVIFDIGGGMR NGADFKAVQIGGPSGGCLSEKDLDLPLDFDSLKKAGAMIGSGGLVVMDSNTCMVEVAR FFMNFTQNESCGKCVPCREGTKRMLEILERIVEGNGQDGDIELLLELADTISATALCGLG KAAAFPVVSTIKNFREEYEAHIYDKRCPTGNCQKLKTITIDASLCKGCSKCARSCPVGAIT GKVKEPFVIDQSKCIKCGACIETCAFHAILEG Cthe430 (SEQ ID NO: 24) MDNREYMLIDGIPVEINGEKNLLELIRKAGIKLPTFCYHSELSVYGACRMCMVEN EWGGLDAACSTPPRAGMSIKTNTERLQKYRKMILELLLANHCRDCTTCNNNGKCKLQD LAMRYNISHIRFPNTASNPDVDDSSLCITRDRSKCILCGDCVRVCNEVQNVGAIDFAYRG SKMTISTVFDKPIFESNCVGCGQCALACPTGAIVVKDDTQKVWKEIYDKNTRVSVQIAPA VRVALGKELGLNDGENAIGKIVAALRRMGFDDIFDTSTGADLTVLEESAELLRRIREGKN DMPLFTSCCPAWVNYCEKFYPELLPHVSTCRSPMQMFASIIREEYSTSSKRLVHVAVMP CTAKKFEAARKEFKVNGVPNVDYVLTTQELVRMIKESGIVFSELEPEAIDMPFGTYTGA GVIFGVSGGVTEAVLRRVVSDKSPTSFRSLAYTGVRGMNGVKEASVMYGDRKLKVAV VSGLKNAGDLIERIKAGEHYDLVEVMACPGGCINGGGQPFVQSEEREKRGKGLYSADKL CNIKSSEENPLMMTLYKGILKGRVHELLHVDYASKKEAK Cthe431 SEQ ID NO: 25: MLEIKICVGSSCHLKGSYNVINEFQHLIEEKALHDKIDIKATFCMKQCQKNGVAV EVNNEIFGVLPEAAEEFFKNVILPKV EC 1.12.7.2 Hyd Cthe3019-24 Cthe3019 (SEQ ID NO: 26) MSFFTMTKTLIKSIFHGPYTVRYPLEKKEPFPASRGRIEINIQDCIFCGLCARRCPTG AINVEKPESRWSINRLRCIQCGYCSEVCPKKCLKMNNMYPAPSFENIEDVYQNARVPDN KENNRNIAGAC Cthe3020 (SEQ ID NO: 27) MGKKTVIPFGPQHPVLPEPIHLDLVLEDETVVEAIPSIGYIHRGLEKLVEKKDYQQ FVYVAERICGICSFMHGMGYCMSIENIMGVQIPERAEFLRTIWAELSRIHSHMLWLGLLA DALGFESLFMHSWRLREQILDIFEETTGGRVIFSVCDIGGVRRDIDSEMLKKINSILDGFEK EFSEITKVFLNDSSVKLRTQGLGVLSREEAFELGAVGPMARASGIDIDMRKSGYAAYGK LKIEPVVETAGDCYARTSVRIREVFQSIDLIRQCISLIPDGEIKVKIVGNPSGEYFTRLEQPR GEVLYYVKANGTKFLERFRVRTPTFANIPALLHTLKGCQLADVPVLILTIDPCISCTER Cthe3021 (SEQ ID NO: 28) MAQQTINTISPNELLAYALRLKNANYRLVAISCTNAENGVEMSYSFDSGSDFTNL RITVAPGDEIESISSIYSYSFLYENEIKELFGVNITGISPDYKDKLYRISVKTPFNMKEGDKN G Cthe3022 (SEQ ID NO: 29) MNFSKKSPWILHYDGSSCNGCDIEVLACLTPLYDIERFGVINTGNPKHADILLITGS INEQNKSVVKQLYEQMADPKVVVAVGICAATGGIFSECYNVSGGVDKIIPVDVYVPGCA ARPEAIIDGVVKALGILEERQKYARKKDK Cthe3023 (SEQ ID NO: 30) MSQIIRLVLYIIAIIIVAPLLGGLLTGIDRVITARMQGRKGPSVLQPFYDVLKLFQKE SIEVNTMHRFFVYISLIFVIFTTVIMLLGGDILLALFALTLGSIFFVLGGYASNSPYSTIGSER ELLQMMAFEPMLLLAAIGLYYGDKSFFIKDIVTARIPSIVYLPGVFLGLLYVLTFKLRKSP FDLSMSHHGHQEIVQGITTEYSGKDLAIIQITHWYETIIALALVYLFFAFRSPFSHVIAILAC IIAFLLEIVVDNAFARAKWEFALKSTWIVTGVLASVNLIILSFFR Cthe3024 (SEQ ID NO: 31) MNAILILILFPLLASVTVLSVRKDAIRNIIVRIFAFITGILTLFVVCRYFKDGISLSIEN RNIIDMTISLAEVLIAAYIIFTGIKNKKFIVSIFAAVQTALILWFEFTQKHGINVHSDIVFDRL SAVMVLIVGCIGSLILIYTVGYMICWYHIHHEGYKERKSFFFSVIFLFLFAMFGLIFSNNLI WMYFCWELTTLCSYLLIGYTRTPEAVNNSFHALAINLGGGLAFASAMVYIGTNFKTLEL SALTAMKLELAVLIPVFLLCIAALTKSAQMPFSSWLLGAMVAPTPSSALLHSATMVKAG VYLLIRLAPLLAGTTIGKVIALLGAVTFLASSIIAISKSDAKKILAYSTISNLGLIVTCAA1GT QESLWAAILLLIFHSISKSLLFLTGGSVEHQIGSRNVEDMDILLQVSRRLSVYMIVGIAGM FLAPFGMLISKWVAMKAFIDSKNILTVIILGYGSATTLFYWTKWMGKLVANANRKDHIK HTFH1DEEIPIFIHAVLVVLSCFTFPLVSRYVLVPYLSGLFGPDVPIPIGTSDVNIMLIMLSM LLILPISFIPIYKSDRRRIVPIYMAGENTGDNESFYGAFDEKRKVELHNWYMKNFFSVKKL TFWSNLLCAVVILVGVVLLIGGITK Cthe342 (SEQ ID NO: 32) MQMVNVTIDNCKIQVPANYTVLEAAKQANIDIPTLCFLKDINEVGACRMCWEV KGARSLQAACVYPVSEGLEVYTQTPAVREARKVTLELILSNHEKKCLTCVRSENCELQR LAKDLNVKDIRFEGEMSNLPIDDLSPSVVRDPNKCVLCRRCVSMCKNVQTVGAIDVTER GFRTTVSTAFNKPLSEVPCVNCGQCINVCPVGALREKDDIDKVWEALANPELHVVVQTA PAVRVALGEEFGMPIGSRVTGKMVAALSRLGFKKVFDTDTAADLTIMEEGTELINRIKN GGKLPLITSCSPGWIKFCEHNYPEFLDNLSSCKSPHEMFGAVLKSYYAQICNGIDPSKVFV VSIMPCTAKKFEAQRPELSSTGYPDVDVVLTTRELARMIKETGIDFNSLPDKQFDDPMGE ASGAGVIFGATGGVMEAAIRTVGELLSGKPADKIEYTEVRGLDGIKEASIELDGFTLKAA VAHGLGNARKLLDKIKAGEADYHFIEIMACPGGCINGGGQPIQPSSVRNWKDIRCERAK AIYEEDESLPIRKSHENPKIKMLYEEFFGEPGSHKAHELLHTHYEKRENYPVK Cthe430 (SEQ ID NO: 33) MDNREYMLIDGIPVEINGEKNLLELIRKAGIKLPTFCYHSELSVYGACRMCMVEN EWGGLDAACSTPPRAGMSIKTNTERLQKYRKMILELLLANHCRDCTTCNNNGKCKLQD LAMRYNISHIRFPNTASNPDVDDSSLCITRDRSKCILCGDCVRVCNEVQNVGAIDFAYRG SKMTISTVFDKPIFESNCVGCGQCALACPTGAIVVKDDTQKVWKEIYDKNTRVSVQIAPA VRVALGKELGLNDGENAIGKIVAALRRMGFDDIFDTSTGADLTVLEESAELLRRIREGKN DMPLFTSCCPAWVNYCEKFYPELLPHVSTCRSPMQMFASIIKEEYSTSSKRLVHVAVMP CTAKKFEAARKEFKVNGVPNVDYVLTTQELVRMIKESGIVFSELEPEAIDMPFGTYTOA GVIFGVSGGVTEAVLRRVVSDKSPTSFRSLAYTGVRGMNGVKEASVMYGDRKLKVAV VSGLKNAGDLIERIKAGEHYDLVEVMACPGGCINGGGQPFVQSEEREKRGKGLYSADKL CNIKSSEENPLMMTLYKGILKGRVHELLHVDYASKKEAK Cthe3003 (SEQ ID NO: 34) MDSFLMKGYIKEANIDYSCSRGSMEDLPKWEFREIPKVPRAVMPSLSLEERKNNF NEVELGLSEEVARKEARRCLKCGCSARFTCDLRKEASNHGIVYEEPIHDRPYIPKVDDHP FIVRDHNKCISCGRCIAACAEIEGPGVLTFYMKNGRQLVGTKSGLPLRDTDCVSCGQCVT

ACPCAALDYRRERGKVVRAINDPKKTVVGFVAPAVRSLISNTFGVSYEEASPFMAGLLK KLGFDKVFDFTFAADLTIVEETTEFLSRIQNKGVMPQFTSCCPGWINFVEKRYPEIIPHLST CKSPQMMMGATVKNHYAKLMGINKEDLFVVSIVPCLAKKYEAARPEFIHDGIRDVDAV LTTTEMLEMMELADIKPSEVVPQEFDEPYKQVSGAGILFGASGGVAEAALRMAVEKLTG KVLTDHLEFEEIRGFEGVKESTIDVNGTKVRVAVVSGLKNAEPIIEKILNGVDVGYDLIEV MACPGGCICGAGHPVPEKIDSLEKRQQVLVNIDKVSKYRKSQENPDILRLYNEFYGEPNS PLAHELLHTHYTPKHGDSTCSPERKKGTAAFDVQEFTICMCESCMEKGAENLYNDLSSK IRLFKMDPFVQIKRIRLKETHPGKGVYIALNGKQIEEPMLSGNIPDESESE Cthe3004 (SEQ ID NO: 35) MKTLENHNRIKVTVNGREIEVYDNLTILQALLQEDIHIPHLCYDIRLERSNGNCGL CWTLISPDGERDVKACQTPIKEGMVICTNTPKLENYRKIRLEQLLSDHNADCVAPCVMT CPANIDIQSYLRHVGNGDFEAAIRVIKERNPFPIVCGRVCPHTCESQCRRNLVDAPVAINY VKRFAADWDMARPEPWTPEKKPPTGKKIAIVGAGPSGLSAAYYSAIKGHDVTVFERQPH PGGMMRYGIPEYRLPKAILDKEIEMIKKLGVKIMTEKALGIHIRLEDLSKDFDAVYLAIGS WQATPMHIEGEKLDGVWAGINYLEQVAKNVDIPLGDNVWIGGGNTAIDCARTALRKG AKSVKLVYRCTREEMPAAPYEVEEAIHEGVEMIFLMAPTKIIVKDGKKKLVCIRMQLGE PDRSGRRRPVPIEGSEVEIDADTIIGAIGQSTNTQFLYNDLPVKLNKWGDIEVNGKTLQTS EYNIFAGGDCVTGPATVI Cthe0349 (SEQ ID NO: 36) MPLVTSTEMFKKAYEGKYAIGAFNVNNMEIIQGITEAAKEVNAPLILQVSAGARK YANHTYLVKLVEAAVEETGLPICLHLDHGDSFELCKSCIDGGFTSVMIDGSHLPFEENIKL TKQVVDYAHSKGVVVEGELGRLAGIEDDVNVSEADAAFTDPDQAEEFVKRTGVDSLAI AIGTSHGAYKFKGEAKLRFDILEEIEKRLPGFPIVLHGASSVIPEYVDMINKYGGDMPGA KGVPEDMLRKAASMAVCKINIDSDLRLAMTATIRKYFAENPSHFDPRQYLGPARNAIKE LVKHKIVNVLGCDGKA Cthe1019 (SEQ ID NO: 37) MDIQLKKSGIGVKEKKSKNHLLYSIKQNLFAYAMLIPTFVCMMCIHFIPMLQGIYL SLLDLNQLTMTKFLNAPFIGLKNYYEILFDEKSLIRRGFWFALRNTAIYTWVTFATFALG IILAMLVNREFKGRGIVRTALLMPWVVPSYVVGMTWGFLWRQDSGLINIILCDILHILPE KPYWLVGSNQIWAIIIPTIWRGLPLSMILMLAGLQSISPDYYEAADIDGANGWQKFWHIT LPLLKPILAINVMFSLISNIYSFNIVSMMFGNGAGIPGEWGDLLMTYIQRNTFQMWRFGP GAAALMIVMFFVLGIVALWYTLFKDDLVVK Cthe0390 (SEQ ID NO: 38) VDKFTKLDLNSITSNNRMNIFNCILEAKEINRAVIAKKVGLSIPAVMSITDDLIQKG IIYVIGKGKSSGGKRPELLAVVPDRFFFVGVDVGRTSVRWVMNNCRDVVYKVSKPTES VEPDEUNQITEMTMESINESKFPLDRVVGIGVAMPGLIERGTGRVIFSPNFGWNNLALQD ELKKHLPFNVLVENANRALVIGEIKNTQPNPTSCIVGVNLGYGIGSAIVLPNGLYYGVSG TSGEIGHIIVENHGSYCSCGNYGCIESIASGEAIAREARIAIANKIQSSVFEKCEGDLKKIDA KMVFDAAKEGDHLAQSIVEKAADYIGKGLAITINMLDPEQIILCGGLTLSGDFFIDMIKK AVSKYQMRYAGGNVKIVVGKSGLYATAIGGAWIVANNIDFLSSN Cthe2938 (SEQ ID NO: 39) MYYIGIDLGGTNIAVGLVNEEGKILHKDSVPTLRERPYQEIIKDMAMLTLKVIKD ADVSIDQVKSIGVGSPGTPNCKDGILIYNNNLNFRNVPIRSEIQKYIDLPVYLDNDANCAA LAESVAGAAKGANTSVTITLGTGIGGGVVIDGKIYSGFNYAGGELGHTVLMMDGEPCTC GRKGCWEAYASATALIRQARKAAEANPDSLINKLVGGDLSKIDAKIPFDAAKQGDKTGE MVVQQYIRYIAEGLINMINIFMPEVLVIGGGVCKEGEYLLKPLRELIKQGVYSKEDIPQTE LRTAQMGNDAGIIGAAMLGKEC Cthe0217 (SEQ ID NO: 40) MERIKFDYSKALPFVSEREVAYFENFVRSAHDMLHNKTGAGNDFVGWVDLPVN YDREEFARIKAAAEKIKSDSDALVVIGIGGSYLGARAAIEMLSHSFHNLMPKSKRNAPEI YFVGNNISSTYIADLLEVIEGKEISVNVISKSGTTTEPAIAFRIFKIEYMENKYGKDGASKRI YATTDKEKGALRKLATEEGYETFVVPDDIGGRFSVLTAVGLLPIAVAGIDIDSMMKGAA DARELYSNPNLMENDCYKYAAVRNALYRKNKTIEIMVNYEPSLHYFTEWWKQLYGESE GKDQKGIFPAGVDFTTDLHSMGQYIQDGLRNIFETVIRVEKPRKNIVIKEEKDNLDGLNFI AGKDVDYVNKKAMEGTVLAHTDGGVPNLVVTVPELSAYYFGNMVYFFEKACGISGYL LGVNPFDQPGVEAYKICNMFALLGKPG YEEQRKKLEERL Cthe1261 (SEQ ID NO: 41) MSSVRTIGVLTSGGDAPGMNAAIRSVVRTGLYYGFKVLGIRKGFNGLINGDIEEL TARSVGDIIHRGGTILQTARSPQFKTEEGLfCKAMSMAKVFGIDALVVIGGDGSYRGARDI SKLGLNVIGIPGTIDNDIGCTDYTIGFDTAMNTVQDAIDKIRDTAYSHERCSVLEVMGRH AGYIAVNVSISGGAEAVVLPEKPFDMDTDVIKPIIEGRNRGKKHYLVIVAEGGEGKAIEIA KEITEKTGIEARATILGHIQRGGSPTVYDRVMASQMGAKAVEVLMENKRNRVIVFKDNQ IGDMDLEEALQVKKTISEDLIQLSKILAL T. saccharolyticum proteins Reaction 6b, EC or0411 (SEQ ID NO: 42) MSYIPNENRYEKMIYRRCGRSGIMLPAISLGLWHNFGGYDVFENMREMVKKAFD LGITHFDLANNYGPPPGSAEENFGKILRTDLRGYRDELLISTKAGYTMWPGPYGDWGSR KYLLSSLDQSLKRMGIDYVDIFYSHRRDPNTPLEETMSALAQAVRQGKALYVGISNYNA EDTKKAAEILRQLGTPLLINQPSYSMFNRWIEDGLTDVLEEEGVGSIAFSPLAQGLLTDK YLNGVPDDSRAVRKNTSLRGNLTEENINKVRELKKIADKRGQSIAQMALAWDLRKVTS VIIGASRVSQIEENVKALDNLEFSHEELKQIDEILSK EC4.2.3.4 or2316 (SEQ ID NO: 43) LNIALIAHDMKKSIMVDFAIAYKEILKKCNIYATGATGQLVEEATGIKVNKFLPGP MGGDQQIGAMIAENNMDLVIFLRDPLTAQPHEPDILALLRVCDVHSIPLATNLATAEVLI KGLDAGFLEWRDAVK EC5.3.1.1 or2687 (SEQ ID NO: 44) LRRPIIAGNWKMYMTPSEAVNLVNELKPLVSGAEAEVVVIPPFVDLVDVKKAID ASNIKLGAQNMHWEEKGAFTGEVSPIMLKEIGVEYWIGHSERRQYFAETDETVNKKVK SALSHGLKPIVCVGESLSQREAGEAFNVVREQTKKALDGIKSEDVLNVVIAYEPIWAIGT GKTATSKDANDVIKVIRETIADIYSIDIANEVRIQYGGSVKPDNAKELMSESDIDGALVGG ASLKAQDFAKIVNY Reaction 7 (gldA) or104 (SEQ ID NO: 45) MYMKTNFTYFMPTEIFGPGTLGKLATVKLPGKKALLVIGSGNSMRRHGYLDRW NYLKQNGVDYVVYDKILPNPIAEHVAEGAKVAKDNGCDFVIGLGGGSTIDSSKAIAVMA KNPGDYWDYVSGGSGKGMEVKNGALPIVAIPTTAGTGTESDPWAVVTKTETNEKIGFG CKYTYPTLSIVDPELMVSIPPKFTAYQGMDAFFHSVEGYLATVNQPGSDVLALQSISLITE NLPKAVADGNNMEARTALAWASTAAGIVESLSSCISHHSLEHALSAYHPEIPHGAGLIML SVSYFSFMASKAPERFVDIAKAMGEEFVGNTVEEQAMCFINGLKKLIRNIGMEDLSLSSF GVTEDEATKLAKNAMDTMGGLFNVDPYKLSLDEVVSIYKNCF EC2.3.1.54 (SEQ ID NO: 46) VDDKKVFDHLFILTDDTGMMQHSVGSVPDPKYGYTTDDNGRALIACAMMYEK YKDDAYINLKKYLSFLMYAQEDDGRFRNFMSFDRKFIDEDFSEDCFGRCMWALGYLIN SNIDERVKLPAYKMIEKSLLLVDTLNYIRGKAYTLIGLYYIYNSFKNLDKDFVRKKMDKL AHDIVEEYEKNSSEDWQWFEDVVSYDNGVIPLSLLKYFSIAKDEEVLDIALKTIDFLDSV CFKNGYFKAVGCKGWYRKGKDIAEYDEQPVEAYTMALMYIEAYKLTGDEKYKKRAID CDKWFYGKNSKGLSLYDEDSGGCSDGITEDGVNSNEGAESLISIMISHCAIDQLK EC2.3.1.8 (SEQ ID NO: 47) MKTSELLAMVVEKGASDLHITVGVPPVLRINGQUKLNLPQLTPQDTEEITKDLLS SDELKKLEDMGDIDLSYSVKGLGRFRINAYKQRGTYSLAIRSVALRIPTIDELGLPEVIKE LALKTRGLIIVTGPTGSGKSTTLASMIDLINEERNCHILTLEDPIEYLHKHKKSIVNQREIG HDAASYASALRAALREDPDVILVGEMRDLETIQIAITAAETGHLVLSTLHTIGSAKTIDRII DVFPPHQQQQIKVQLSNVLEGIVSQQLLPKIDNSGRVVAVEVMIATPAIRNLIREGKSFQI QSMVQTGNKFGMVTMDMWISQLLKRNLISMDDALTYCVDRENFSRLVV EC 2.7.2.1 (SEQ ID NO: 48) MIKKKLGDLLVEVGLLDESQLNNAIKIQKKTGEKLGKILVKEGYLTEEQIIEALEF QLGIPHIDMKKVFIDANVAKLIPESMAKRHVAIPIKKENNSIFVAMADPLNIFAIDDIKLVT KLDVKPLIASEDGILKAIDRVFGKEEAERAVQDFKKELSHDSAEDDGNLLRDISEDEINN APAVRLVNSIIEQAVKNRASDVHIEPTENDLRIRFRIDGELHEAMRVFKSTQGPVITRIKIM ANMNIAERRIPQDGKIEMNAGGKNIDIRVSSLPTIYGEKLVLRILDKSGYIITKDKLGLGN DDLKLFDNLLKHPNGIILLTGPTGSGKTTTLYAMLNELNKPDKNIITVEDPVEYTLEGLN QVQVNEKAGLTFASALRSILRQDPDIIMIGEIRDRETAEIAIRSSITGHLVLSTLHTNDSAG AITRLIDMGIEPYLVSSSVVGVIAQRLARKICDNCKIEYDASKREKIILGIDADESLKLYRS KGCAVCNKTGYRGRVPIYEIMMMTPKIKELTNEKAPADVILNEAVSNGMSTLKESAKKL VLSGVTTVDEMLRLTYDDAY EC 1.2.7.1 or0047 (SEQ ID NO: 49) MSKVMKTMDGNTAAAHVAYAFTEVAAIYPITPSSPMAEHVDEWSAHGRKNLFG QEVKVIEMQSEAGAAGAVHGSLAAGALTTTFTASQGLLLMIPNMYKIAGELLPGVFHVS ARALASHALSIFGDHQDVMACRQTGFALLASGSVQEVMDLGSVAHLAAIKGRVPFLHFF DGFRTSHEYQKIEVMDYEDLRKLLDMDAVREFKKRALNPEHPVTRGTAQNPDIYFQERE ASNRYYNAVPEIVEEYMKEISKITGREYKLFNYYGAPDAERIVIAMGSVTETIEETIDYLL KKGEKVGWKVHLYRPFSFKHFMDAIPKTVKKIAVLDRTKEAGAFGEPLYEDVRAAFY DSEMKPIIVGGRYGLGSKDTTPAQIVAVFDNLKSDTPKNNFTIGIVDDVTYTSLPVGEEIE TTAEGTISCKFWGFGSDGTVGANKSAIQIIGDNTDMYAQAYFSYDSKKSGGVTISHLRFG KKPIRSTYLINNADFVACHKQAYVYNYDVLAGLKKGGTFLLNCTWKPEELDEKLPASM KRYIAKNNINFYIINAVDIAKELGLGARINMIMQSAFFKLANIIPIDEAVKHLKDAFVKSYG HKGEKIVNMNYAAVDRGIDALVKVDVPASWANAEDEAKVERNVPDFIKNIADVMNRQ EGDKLPVSAFVGMEDGTFPMGTAAYEKRGIAVDVPEWQIDNCIQCNQCAYVCPHAAIR

PFLLNEEEVKNAPEGFTSKKAIGKGLEGLNFRIQVSVLDCTGCGVCANTCPSKEKSLIMK PLETQLDQAKNWEYAMSLSYKENPLGTDTVKGSQFEKPLLEFSGACAGCGETPYARLV TQLFGDRMLIANATGCSSIWGGSAPSTPYTVNKDGHGPAWANSLFEDNAEFGFGMALA VKQQREKLADIVKEALELDLTQDLKNALKLWLDNFNSSEITKKTANIIVSLIQDYKTDDS KVKELLNEILDRKEYLVKKSQWIFGGDGWAYDIGFGGLDHVLASGEDVNVLVFDTEVY SNTGGQSSKATPVGAIAQFAAAGKGIGKKDLGRIAMSYGYVYVAQIAMGANQAQTIKA LKEAESYPGPSLIIAYAPCINHGIKLGMGCSQIEEKKAVEAGYWHLYRYNPMLKAEGKN PFILDSKAPTASYKEFIMGEVRYSSLAKTFPERAEALFEKAEELAKEKYETYKKLAEQN EC 1.1.1.2 Or180 (SEQ ID NO: 50) MSKVAIIGSGFVGATSAFTLALSGTVTDIVLVDLNKDKAIGDAIDISHGIPLIQPVN VYAGDYKDVKGADVIWTAGAAQKPGETRLDLVKKNTAIFKSMIPELLKYNDKAIYLIV TNPVDILTYVTYKISGLPWGRVFGSGTVIDSSRFRYLLSKHCNIDPRNVHGRIIGEHGDTE FAAWSITNISGISFNEYCSICGRVCNTNFRKEVEEEVYNAAYKIIDKKGATYYAVAVAVR RIVECILRDENSILTVSSPLNGQYGVKDVSLSLPSIVGRNGVARILDLPLSDEEVEKFRHSA SVMADVIKQLDI EC 2.3.1.54 (SEQ ID NO: 51) MINEWRGFQEGKWQKTIDVQDFIQKNYTLYEGDDSFLEGPTEKTIKLWNKVLEL MKEELKKGVLDIDTKTVSSITSHDAGYIDKDLEEIVGLQTDKPLKRAIMPYGGIRMVKKA CEAYGYKVDPKVEEIFTKYRKTHNDGVFDAYTPEIRAARHAGIITGLPDAYGRGRIIGDY RRVALYGIDRLIEEKEKEKLELDYDEFDEATIRLREELTEQIKALNEMKEMALKYGYDIS KPAKNAKEAVQWTYFAFLAAIKEQNGAAMSLGRVSTFLDIYIERDLKEGTLTEKQAQEL MDHFVMKLRMVRFLRTPDYNELFSGDPVWVTESIGGVGVDGRPLVTKNSFRILNTLYN LGPAPEPNLTVLWSKNLPENFKRFCAKVSIDTSSIQYENDDLMRPIYNDDYSIACCVSAM KTGEQMQFFGARANLAKALLYAINGGIDERYKTQVAPKFNPITSEYLDYDEVMAAYDN MLEWLAKVYVKAMNIIHYMHDKYAYERSLMALHDRDIVRTMAFGIAGLSVAADSLSAI KYAKVKAIRDENGIAIDYEVEGDFPKFGNDDDRVDSIAVDIVERFMNKLKICHKTYRNSIP TLSVLTITSNVVYGKKTGATPDGRKAGEPFAPGANPMHGRDTKGAIASMNSSKIPYDSSL DGISYTFTIVPNALGKDDEDKINNLVGLLDGYAFNAGHHININVLNRDMLLDAMEHPEK YPQLTIRVSGYAVNFNKLTREQQLEVISRTFHESM EC1.2.7.1 (following four proteins) Or1545 (SEQ ID NO: S2) MVITVCVGSSCHLKGSYDVINKLKEMIKNYGIEDKVELKADFCMGNCLRAVSVK IDGGACLSIKPNSVERFFKEHVLGELK Or1546 (SEQ ID NO: 53) MSVINFKEANCRNCYKCIRYCPVKAIKVNDEQAEIIEYRCIACGRCLNICPQNAKT VRSDVERVQSFLNKGEKVAFTVAPSYPALVGHDGALNFLKALKSLGAEMIVETSVGAM LISKEYERYYNDLKYDNLITTSCPSVNYLVEKYYPDLIKCLVPVVSPMVAVGRAIKNIHG EGVKVVFIGPCLAKKAEMSDFSCEGAIDAVLTFEEVMNLFNTNKIGVECTKENLEDVDS ESRFKLYPIEGKTMDCMDVDLNLRKFISVSSIENVKDILNDLRAGNLHGYWIEANACDG GCINGPAFGKLESGIAKRKEEVISYSRMKERFSGDFSGITDFSLDLSRKFIDLSDRWKMPS EMEIICEILSKIGKFSVEDELNCGACGYDTCREKAIAVFNGMAEPYMCLPYMRGRAETLS NIIISSTPNAIIAVNNEYEIQDMNRAFEKMFLVNSAMVKGEDLSLIFDISDFVEVIENKKSIF NKKVSFKNYGIIALESIYYLEEYKIAIGIFTDITKMEKQKESFSKLKRENYQLAQQVIDRQ MKVAQEIASLLGETTAETKVILTKMKDMLLNQGDDE or1547 (SEQ ID NO: 54) MSHYIDIAHASLNKYDEELCGDSVQIIRKKDYAMAVMADGLGSGVKANILSTLT TRTVSKMLDMGSELRDVVETVAETLPICKERNIAYSTFTWSIYGDNAHLVEYDNPSVFY FKNGVHKKVDRKCVEIGDKKIFESSFKLDLNDALIVVSDGVIHAGVGGILNLGWQWDN VKQYLSKVLEVYSDASDICSQLITTCNNLYKNRPGDDTTAIVIKVNESKKVTVMVGPPIL KNMDEWVVKICLMKSEGLKVVCGGTAAKIVSRILNKDVITSTEYIDPDIPPYAHIDGIDLV TEGVLTLRKTVEIFKEYMNDKDSNLLRFSKKDAATRLFKILNYATDVNFLVGQAVNSAH QNPDFPSDLRIKVRIVEELISLLERLNKNVEVNYF or1548 (SEQ ID NO: 55) LFKFNTDVQMLKYEVLYNVAKLTLEDRLEDEYDEIPYEIIPGTKPRFRCCVYKER AIIEQRTKVAMGKNLKRTMKHAVDGEEPIIQVLDIACEECPIKRYRVTEACRGCITHRCT EVCPKGAITIINKKANIDYDKCIECGRCKDACPYNAISDNLRPCIRSCSAKAITMDEELKA AINYEKCTSCGACTLACPFGAITDKSYIVDIIRAIKSGKKVYALVAPAIASQFKDVTVGQI KSALKEFGFVDVIEVALGADFVAMEEAKEFSHKIKDIKVMTSSCCPAFVAHIKKSYPELS QNISTTVSPMTAISKYIKKHDPMAVTVFIGPCTAKICSEVMRDDVKGITDFAMTFEEMVA VLDAAKIDMKEQQDVEVDDATLFGRICFARSGGVLEAVVEAVKEIGADVEVNPWCNG LDECNKTLKIMKAGKLPNNFIEGMACIGGCIGGAGVINNNVNQAKLAVNKFGDSSYHKS IKDRISQFDTDDVDFHVDSGEDESSETSFKEA EC 1.2.1.43 or2328 (SEQ ID NO: 56) MDKVRITIDGIPAEVPANYTVLQAAKYAKIEIPTLCYLEEINEIGACRLCVVEIKGV RNLQASCVYPVSDGMEIYTNTPRVREARRSNLELILSAHDRSCLTCVRSGNCELQDLSRK SGIDEIRFMGENIKYQKDESSPSIVRDPNKCVLCRRCVATCNNVQNVFAIGMVNRGFKTI VAPSFGRGLNESPCISCGQCIEACPVGAIYEKDHTKIVYDALLDEKKYVVVQTAPAVRVA LGEEFGMPYGSIVTGKMVSALKRLGFDKVFDTDFAADLTIIEEGNELLKRLNEGGKLPMI TSCSPGWINYCERYYPEFIDNLSTCKSPHMMMGAIIKSYFAEKEGIDPKDIFVVSIMPCTA KKYEIDRPQMIVDGMKDVDAVLTTRELARMIKQSGIDFVNLPDSEYDNPLGESSGAGVIF GATGGVMEAALRTVADIVEGKDIENFEYEEVRGLEGIKEAKIDIGGKEIKIAVANGTGNA KKLLDKIKNGEAEYHFIEVMGCPGGCIMGGGQPIHNPNEKDLVRKSRLKAIYEADKDLPI RKSHKNPMITKLYEEFLISPLGEKSHHLLHTTYSKKDLYPMND EC 4.1.2.13 or0260 (SEQ ID NO: 57) LNDILVKARNNKYAIGGFNFNFYDDALGIISAAYELKSPIILMASEGCVKFLGVKH IVNFVNQLKDEYNIPIILHLDHGKDIEIIKNCIDNKFDSIMYDGSLLNFEENIKNTKFIADLC HDKGMTIEGELGRISGAEENIENSEDVFTDPDSVAEFTERSDVDSLAVAIGNAHGLYKGR PRLDFERLSKINKISKVPLVLHGGTGIPYEDIQKAIQLGISKVNVGTEIKIAYIKSIKKHLETI NDNDIRHLVSMVQNDIKELVKQYLDIFGTANKYSQLQSM or0330 (SEQ ID NO: 58) MLVTGIELLKKANEEGYAVGAFNTSNLEITQAIVEAAEEMRSPAIIQVSEGGLKY AGIETISAIVRTLATKASVPIALHLDHGTDFNNVMKCLRNGWTSVMMDASKLPLEKNIE VTKNVYTIAHGMGVSVEAEIGKIGGTEDNVTVDEREASMTDPDEAFKFAKETGVDYLAI SIGTAHGPYKGEPKLDFDRLVKIKEMLKMPIVLHGASGVPEADIRKAVSLGVNKINIDTDI RQAFAARLRELLKNDEEVYDPRKILGPCKEAMKEVIKNKM RMFGSEGRA or0272 (SEQ ID NO: 59) MITGDQLLIKQINKSIVLNTIRKKGLISRADLANITGLNKSTVSSLVDELIKEGFVEE EGPGESKGGRKPIMLMINSLAGCVIGVDLDVNYILVILTDILANILWQKRINLKLGESKED IISKMLELIDEAIKNSPNTVKGILGIGIGVPGITDYKRGVVLKAPNLNWENVELKKMVEER FNLKVYIDNEANTGAIGEKWFGGGRNAKNFVYVSAGIGIGTGIIINNELYRGSNGLAGEM GHMTIDINDHMCSCGNRGCWENYASEKSLFRYIKERLEAGQEDDFIDSENIDSLDINDIA GYAELGSKLAIDAINEISKNLSVGIVNIVNTFNPDLVLIGNTLSAIGDMLIDAVKEYVREK CLVSRYNDIAIEISKLGMLERAIGAVTLVISEVFSYPGL or1389 (SEQ ID NO: 60) MTNVLNFDYSNALNFVNEHEISYLEKQALLSLDMVLNKTAQGSDFLGWVDLPK DYDKEEFARIKKAAEKIKSDSDALVVIGIGGSYLGARAAIEMLTHSFYNVLPQSVRKAPEI YFAGNSISSTYLQDLLEILEGKDVSINVISKSGTTTEPAIAFRVFRDFLEKKYGKEEAKSRI YVTTDRQKGALKKLADEEGYETFVIPDDVGGRYSVLTAVGLLPIAAAGIDIDEMMKGA YDASIVFKKPDIKENLSMQYAVLRNALYRKGKSVEILVNYEPRLHYFSEWWKQLYGESE GKDHKGIYPASVDFSTDLHSMGQFIQDGSRIMFETVINVEKPLKEITINEDKDNVDGLNFL TGKTVDLVNKKAFEGTVLAHNDGGVPNLIVNVPEISAYNFGYLVYFFEMACGISGYLNG VNPFDQPGVEAYKKNMFALLGKPGYEKEKEELEKRLKR or2875 (SEQ ID NO: 61) MYNIQLDSPNLGDKEKDYLVKCIESGYVSTVGPFVPEFERRFAEFLNVNHCVSVQ SGTAALYMALYELGIKDGDEVIVPAITFVATVNPIVYCGATPVFVDVDKDTWNIDPKEIE KAITPKTKAIIPVHLYGNPCDMDKIMEIAKENNIYVIEDATESLGALYKGRMTGTIGHIGC FSFNGNKVITTGGGGMVASNNEDWSHIRFLWQARDMTQGYFHTEIGFNYRMTNLEA SLGIAQLERLAGFLEKKRMYFEIYKKIFNGIEEISLQTEYEGAKSSDWLSSVKIDCKKVGM TIHQIQDELKRRGIPTRRIFNPIVDLPPYKKYKKGSYSNSYEIYENGLNLPSSTLNTYEDVK YVAKTLLDILSIKKR T. saccharolyticum pdu genes or228-or200 or228 SEQ ID NO: 62: MLAIERRKRIMRLIQENQSVLVPELSKLFNVTEETIRRDLEKLEAEGLLKRTYGGA VINENSSADIPLNIREITNIESKQAISMKVAEYIEDGDTLLLDSSSTVLQVAKQLKFKKKLT VITNSEKIILELANAKDCKVISTGGVLKQNSMSLIGNFAEDMIKNFCVDKAIISSKGFDMT NGITESNEMEAEIKKAMANSAEKVFLLLDHNKFDKSSFVKMFDLDKIDYLFTDRKLSLE WEEFLKKHNIDLIYC SEQ ID NO: 63: ATGCTTGCGATAGAACGAAGGAAGAGGATAATGAGGCTTATACAGGAAAATC AAAGCGTTTGGTGCCTGAGTTAAGTAAATTGTTTAATGTGACAGAGGAAACTATAAG GAGAGATTTAGAGAAACTTGAAGCAGAAGGGCTTTTAAAGAGGACTTATGGTGGTG CTGTTATAAATGAAAATTCAAGTGCTGATATCCCCTTAAATATAAGGGAAATAACGA ATATAGAAAGCAAACAGGCCATAAGTATGAAGGTTGCCGAATACATTGAAGATGGT GATACACTTTTGCTTGATTCAAGCTCTACAGTTCTTCAAGTAGCAAAGCAATTAAAA TTCAAAAAGAAGCTTACAGTCATAACAAATTCGGAAAAGATAATATTAGAATTAGC AAATGCGAAAGATTGCAAAGTCATTTCTACAGGAGGAGTATTGAAGCAAAATTCTAT

GTCGCTAATTGGAAATTTCGCGGAAGATATGATAAAAAATTTCTGTGTAGATAAAGC CATAATATCATCAAAAGGTTTTGACATGACAAATGGCATTACAGAGTCAAACGAAAT GGAAGCTGAAATAAAAAAAGCCATGGCCAACTCGGCAGAAAAAGTGTTTTTACTTC TTGATCACAACAAATTTGACAAGTCATCGTTCGTCAAGATGTTTGACTTAGATAAAA TCGATTATCTATTTACCGATAGAAAGCTGTCTTTAGAATGGGAAGAATTCTTGAAAA AACACAATATTGATTTAATCTATTGTTAG or227 SEQ ID NO: 64: VYSEYEVKKQICEIGKRIYMNGFVAANDGNITVRIGENEIITTPTGVSKGFMTPDM LLNINLNGEVLKSSGDYKPSTEIKMHLRVYRERPDVKSVIHAHPPFGTGFAIVGIPLTKPI MPEAVISLGCVPIAEYGTPSTEELPDAVSKYLQNYDALLLENHGALTYGPDLISAYYKME SLEFYAKLTFISTLLGGPKELSDSQVEKLYEIRRKFGLKGRHPGDLCSTLGCSTNSAKSND DDISELVNVITKKVLEQLKYN SEQ ID NO: 65: GTGTATTCTGAATATGAGGTAAAAAAACAGATCTGCGAAATAGGAAAGAGAA TCTACATGAATGGGTTTGTGGCAGCGAATGACGGCAATATCACCGTTAGGATTGGTG AAAATGAAATAATAACGACGCCTACCGGTGTCAGCAAAGGTTTCATGACTCCAGAC ATGCTATTAAATATTAATTTAAACGGTGAAGTATTAAAATCTTCAGGCGACTACAAA CCGTCCACAGAAATAAAGATGCATCTTAGAGTCTATAGAGAAAGGCCAGATGTCAA ATCAGTCATACATGCACATCCACCATTTGGCACAGGTTTTGCTATTGTAGGGATCCC GCTTACAAAGCCAATAATGCCAGAAGCAGTTATATCTTTAGGCTGTGTGCCOATAGC CGAATACGGGACGCCTTCTACAGAAGAGCTGCCAGATGCCGTCTCTAAATATTTGCA AAATTACGATGCGCTTTTATTAGAAAATCATGGTGCGTTGACATACGGTCCTGATTT AATTAGCGCATACTACAAGATGGAATCACTTGAATTTTACGCAAAATTGACATTTAT TTCTACACTTCTCGGAGGTCCAAAAGAATTATCAGATAGCCAAGTAGAAAAGCTTTA TGAAATTAGGAGAAAATTCGGTTTAAAAGGAAGACATCCAGGCGATTTGTGCAGTA CATTAGGATGCAGCACAAATTCTGCAAAATCGAATGATGATGACATTTCTGAACTTG TGAATGTTATCACTAAGAAAGTATTAGAACAATTGAAATACAATTAA or226 SEQ ID NO: 66: MKHSKRFEVLGKRPVNQDGFINEWPEKGFIAMCSPNDPKPSIKIENDKIVEMDGK RREDFDFIDLFIADHAINIYQAEKSMKMNSLDIAKMLVDINVERKTIIKVVSGLTPAKIME WNHLNVVEMMMAMQKMRARKIPANQSHITNLKDNPVQIAADAAECALRGFREEETT VGVTKYAPFNAIALLIGSQALKRGVLTQCAVEEATELELGMRGFTTYAETISVYGTESVF IDGDDTPYSKAFLASAYASRGLKMRFTSGTGSEVLMGNAEGKSMLYLEIRCIMVTKGAG VQGLQNGAISCIGITSSVPSGIRAVLAENLIASMLDLEVASGNDQTFTHSDIRRTARTMMQ FLPGTDFIFSGYSGTPNYDNMFAGSNFDAEDFDDYNVLQRDLMVDGGLRPVKEEDVVE VRRKAAKALQDVFRELNLGWTDEEVEAAAYAHGSKDMPERDVLSDLESIDEMMKRGI TGIDIVKALYRSGHEDIAENILNMLKQRISGDYLQTSAILDEDFNVISAINCPNDYLGPGT GYRIDKDRWEEIKNIPYTINPDNL SEQ ID NO: 67: ATGAAACATTCTAAGCGATTTGAGGTTCTCGGCAAAAGACCTGTAAATCAGG ATGGATTTATAAATGAATGGCCAGAAAAAGGCTTCATAGCAATGTGTAGTCCCAATG ATCCTAAGCCATCAATAAAGATTGAAAACGACAAGATCGTTGAGATGGATGGGAAG AGAAGAGAAGACTTTGATTTTATAGATTTATTCATAGCTGATCACGCTATAAATATTT ATCAGGCTGAGAAATCCATGAAAATGAACTCGCTTGATATAGCCAAAATGCTTGTAG ATATAAATGTAGAGAGAAAGACTATAATAAAAGTAGTTTCGGGACTTACACCTGCC AAAATAATGGAAGTTGTAAATCATCTTAATGTCGTTGAAATGATGATGGCTATGCAG AAAATGCGAGCAAGAAAGATTCCGGCTAATCAATCACATATTACAAATCTTAAAGA TAATCCTGTGCAGATTGCAGCGGATGCTGCCGAATGTGCTTTAAGAGGTTTTAGGGA AGAAGAGACCACCGTAGGAGTGACAAAATATGCTCCGTTTAATGCAATAGCGTTATT GATAGGGTCTCAGGCATTAAAAAGAGGCGTGCTTACTCAATGTGCTGTTGAGGAGGC GACGGAACTTGAATTAGGCATGAGGGGATTTACCACATACGCTGAGACTATATCTGT TTATGGAACTGAAAGTGTTTTTATAGATGGTGACGATACACCTTACTCCAAAGCATT CCTTGCTTCTGCTTATGCGTCAAGAGGATTGAAAATGAGGTTTACGTCAGGTACAGG TTCAGAAGTTCTTATGGGAAATGCAGAGGGTAAATCGATGTTGTACCTGGAAATCAG GTGCATCATGGTTACAAAAGGTGCAGGAGTGCAGGGGCTTCAAAATGGTGCAATAA GCTGTATAGGCATAACTAGCTCAGTTCCTTCAGGTATAAGGGCGGTGCTGGCTGAAA ACCTTATAGCATCTATGCTTGATTTAGAGGTAGCATCAGGCAATGATCAGACTTTTA CACATTCAGACATAAGAAGGACAGCAAGGACTATGATGCAGTTTTTACCCGGTACTG ATTTCATATTTTCAGGTTACAGTGGAACGCCTAATTATGACAATATGTTTGCAGGTTC CAATTTTGATGCAGAAGATTTTGATGACTACAATGTACTGCAAAGGGATTTAATGGT AGATGGAGGGTTAAGGCCTGTAAAAGAAGAAGATGTGGTAGAAGTGAGGCGAAAG GCAGCTAAAGCTTTGCAGGATGTATTTAGAGAGTTAAATCTTGGAGTAGTTACAGAT GAAGAAGTAGAAGCAGCAGCATATGCACACGGCAGCAAAGATATGCCTGAAAGAG ATGTTTTGTCTGACCTTGAATCAATCGATGAGATGATGAAAAGAGGGATTACAGGCA TTGACATCGTAAAGGCTTTATATAGATCTGGACATGAGGATATAGCGGAAAACATTT TAAACATGTTAAAACAGCGCATATCTGGAGACTATTTGCAGACATCAGCTATTCTTG ATGAAGATTTTAATGTTATAAGCGCCATAAATTGTCCAAATGATTACTTAGGACCTG GAACAGGATATAGGATTGATAAAGATAGATGGGAAGAGATAAAGAATATTCCTTAC ACCATTAATCCTGACAATTTGTAA or225 SEQ ID NO: 68: MYVDEELLKEITKRVIEELNNKHKTDNVPSYFIENGVAYKGKNIEEVVIGVGPAF GKHIKKTINGLDHRDVIBCEIIAGIEEEGMVHRIVRVLKTSDVAFIGKEAALLSGSGIGIGIQ SKGTTVIHQKDLYPLSNLELFPQAPLLNLELYREIGKNAARYAKGMMVKPILIQNDYMV RPKYQVKAAIMHIKETEKILKNAQSIQLTIDL SEQ ID NO: 69: ATGTACGTAGATGAAGAACTGTTAAAAGAAATTACTAAACGTGTTATAGAAG AATTAAATAATAAGCATAAAACTGATAATGTGCCTTCGTATTTTATTGAAAATGGAG TTGCCTATAAGGGTAAAAATATAGAGGAAGTCGTCATTGGTGTTGGGCCTGCATTTG GAAAGCATATAAAAAAGACTATAAATGGCCTTGACCATAGAGATGTCATAAAAGAA ATAATTGCAGGCATCGAAGAAGAAGGTATGGTTCATAGAATTGTAAGAGTTCTAAA GACTTCTGATGTGGCGTTCATAGGCAAAGAAGCTGCTTTATTAAGCGGATCGGGAAT AGGCATAGGCATACAATCAAAAGGTACTACAGTGATTCATCAAAAAGATTTATATCC TTTAAGCAATTTAGAACTGTTTCCACAAGCTCCACTGCTAAATTTAGAATTATACAG GGAAATAGGCAAAAATGCGGCGAGATATGCTAAAGGCATGATGGTAAAGCCTATTT TGATTCAAAATGATTACATGGTGAGACCTAAATACCAAGTGAAAGCTGCTATAATGC ATATAAAAGAGACGGAAAAGATATTGAAAAATGCTCAATCAATCCAATTGACGATA GACTTGTAA or224 SEQ ID NO: 70: MEEYPLSKSAFDKLVTKTGKHLNEINIENVMKGNVKPDDIKISKEVLLMQGQIAE RYGRHQMKENFTRASELTDVPDEKILEIYESLRPFRSTKEELINLAYELRDKYNAINCANL ILEAAEVYEKRNILKT SEQ ID NO: 71: ATGGAAGAATATCCGCTATCAAAAAGTGCTTTTGATAAATTGGTGACAAAAA CAGGCAAACATTTGAATGAAATAAATATTGAAAATGTAATGAAGGGAAACGTAAAA CCCGATGATATCAAGATATCCAAAGAAGTGCTTTTAATGCAAGGGCAAATTGCAGA AAGATACGGCAGGCATCAGATGAAGGAGAATTTCACAAGAGCATCGGAGCTTACAG ATGTTCCAGATGAAAAGATTTTGGAAATATATGAGAGCTTAAGGCCGTTTAGATCTA CAAAGGAAGAGCTTATAAATCTTGCCTATGAATTAAGAGATAAGTACAATGCCATTA ACTGTGCAAACTTGATACTTGAGGCTGCTGAAGTATATGAAAAAAGAAATATTTTGA AAACTTAA or223 SEQ ID NO: 72: MKLIAGVDIGNSTTEVCIAAIKDDNTLEFLSSSLTATTGVKGTVDNVTGVINGLTE ALKKIGKNIRDLSLIRINEAAPVVCGAAMETITETVITGSTMIGHNPSTPGGVGLGVGEIIH INDLADATKGKNYIVVIPKEIGYEEASIMINKSFENDIDVKAAIVQSDEAVLINNRLKKIIPI VDEVRQIEKIPSGWAAVEVAPEGKSISTLSNPYGIATIFDLTPEETKYVIPISKSLMGKKS AVVIKTPRGQVKERIIPAGNLLIMGPTMSSKVSVDSGAEAIMESVEEVGTIDDVEGEENT NVGNMIKNLKNKMANITGQKVDKIKIKDIFAVDTTVPVKVEGGLAGETSMEKAVVLAA MVKTDTLPMIEIAEKLQRKLGVFVKIAGVEAVMATLGALTTPGTKLPLAILDIGGGSTDA ALIDEKGIVKSIHMAGAGELVTMLIDSELGLNDRYLSEEIKRNPIGKVESLFHIRMENREI KFFDKPLNPRYYGRIVILKENDMIPVFKEDLTMEKIIYVRRQAKDKVFVKNAIRALKKIA PENNLRRIPNVVLVGGSALDFEIPEMILSELSKYKIIAGRGNIRKIEGPRNAVATGLVMSY LG SEQ ID NO: 73: ATGAAACTCATAGCAGGTGTTGATATTGGCAATTCTACAACAGAAGTGTGTAT AGCCGCTATTAAAGATGACAATACATTAGAATTTTTAAGCAGTTCCTTGACAGCTAC GACAGGTGTAAAAGGCACTGTGGATAATGTGACAGGGGTTATTAATGGATTGACTG AGGCACTAAAAAAAATTGGCAAGAATATTAGGGATTTAAGCCTCATTAGAATCAAT GAAGCCGCCCCAGTTGTCTGTGGTGCTGCTATGGAGACAATAACGGAAACTGTTATC ACTGGTTCGACTATGATAGGTCATAATCCATCCACGCCGGGTGGTGTCGGACTTGGA GTAGGCGAGATAATACATATAAATGATTTAGCTGATGCTACTAAAGGCAAAAATTAC ATTGTGGTTATACCTAAGGAGATTGGCTATGAAGAAGCTTCAATAATGATAAACAAA TCTTTTGAAAACGATATTGATGTAAAAGCTGCTATAGTTCAAAGCGATGAAGCAGTT TTAATCAACAACAGGCTTAAAAAGATTATACCAATTGTTGACGAAGTAAGGCAGAT AGAAAAGATTCCATCGGGTGTTGTAGCGGCTGTAGAGGTGGCACCAGAAGGCAAGT CCATAAGCACGTTATCAAATCCTTATGGTATCGCAACAATATTTGACTTAACTCCAG AAGAGACAAAGTATGTCATACCGATTTCGAAAAGTTTGATGGGGAAAAAGTCAGCA GTTGTCATAAAAACACCGAGGGGACAAGTGAAAGAAAGAATAATTCCGGCTGGTAA TCTCTTAATCATGGGGCCTACTATGTCATCAAAAGTAAGTGTTGATTCTGGTGCTGAA GCTATAATGGAATCAGTTGAAGAAGTCGGCACAATTGATGACGTAGAAGGTGAAGA

AAATACAAATGTTGGGAATATGATAAAAAATCTAAAAAACAAGATGGCAAATATAA CTGGGCAAAAAGTAGATAAGATAAAGATTAAAGATATCTTCGCTGTTGATACGACA GTCCCTGTTAAAGTAGAGGGCGGACTTGCTGGTGAGACTTCAATGGAAAAAGCAGT CGTGTTGGCGGCTATGGTAAAGACAGATACGCTTCGATGATAGAAATTGCAGAAAA GCTTCAAAGAAAGTTGGGTGTATTTGTAAAAATAGCTGGAGTAGAAGCTGTGATGGC TACATTAGGTGCGCTTACAACTCCAGGCACAAAGTTGCCACTTGCAATACTGGATAT CGGTGGGGGTTCTACAGATGCAGCTTTGATTGATGAAAAAGGCATTGTAAAATCTAT ACACATGGCAGGTGCTGGAGAATTAGTCACAATGCTTATTGATTCAGAATTAGGGTT AAATGATAGATATTTGTCTGAAGAAATAAAGAGAAATCCGATTGGAAAAGTTGAAA GCCTATTTCACATAAGAATGGAAAATAGGGAGATAAAGTTTTTTGACAAACCTTTAA ATCCTCGATATTACGGTAGGATCGTAATTTTAAAAGAAAATGACATGATCCCTGTAT TTAAAGAAGATTTGACAATGGAAAAGATTATTTACGTGCGAAGACAAGCGAAGGAT AAAGTTTTCGTTAAAAATGCTATTAGAGCTTTGAAAAAAATTGCTCCGGAAAATAAT TTAAGGCGAATACCAAATGTAGTCTTGGTTGGCGGTTCTGCTTTGGACTTTGAAATTC CAGAGATGATTTTATCAGAGCTATCAAAATACAAAATCATAGCAGGCAGAGGGAAT ATAAGAAAAATCGAAGGGCCAAGAAATGCTGTAGCGACAGGTCTTGTGATGTCTTA TTTAGGGTGA or222 SEQ ID NO: 74: MEFIKPQIVIFANTENKYIINEVIAGIEEEGALYRLSYNECADVMKMAYDAAKAS VLGIGIGISGDLVCLHSKNLEINTPLILSKTSENFDPRLVGCNAAKYVKGLPLKYLD SEQ ID NO: 75: ATGGAATTTATAAAGCCTCAAATAGTGATTTTTGCAAATACAGAAAACAAAT ATATAATAAACGAGGTTATAGCTGGCATTGAAGAAGAAGGTGCATTATATAGATTAT CTTACAATGAATGTGCTGATGTTATGAAAATGGCTTATGATGCAGCAAAAGCATCTG TATTAGGTATCGGAATAGGCATATCTGGAGATTTAGTGTGTTTGCACTCTAAAAACT TGGAAATCAATACACCTTTGATTCTTTCAAAGACAAGTGAAAACTTTGATCCACGAC TCGTTGGATGCAATGCTGCAAAATATGTAAAGGGTTTGCCACTTAAATACTTAGATT AG or221 SEQ ID NO: 76: MSVYTKTGDDGYTLLLNGERIPKDDLRIETLGNLDELTSYLGFAKAQINDDSIKK R SEQ ID NO: 77: ATGAGTGTTTATACTAAAACTGGTGATGATGGTTACACGTTGCTATTAAATGG AGAAAGAATTCCAAAGGACGATTTGAGAATAGAGACATTGGGAAATTTGGATGAAT TGACAAGCTATTTAGGATTTGCAAAAGCTCAAATAAATGATGATTCCATAAAAAAGA GATAG or220 SEQ ID NO: 78: MVKIKNGFVIPGKNQISALLDIVRTITRKTERSLIKVDKKYPVNINSKVYINRLSDY LFVLARYMEIRTEIEEKVKDVIRKHYGKNKGEIKLNLDIAKNLMAKVEKKAESINLPVAI AIVDMHGNLIAAHFMDGTLLESMNLAINKAYTSVVLKMSTQELSKLAQPGQPLYGINTT DNRIVVFGGGCPIKHQGEIVGGIGVSGGTVEQDIELSIYGADVFEEVIS SEQ ID NO: 79: ATGGTAAAGATTAAAAATGGTTTTGTAATACCTGGTAAAAACCAAATCTCAG CATTATTAGATATTGTAAGGACTATAACGAGAAAAACTGAGAGAAGCTTAATCAAA GTTGACAAGAAATATCCTGTAAATATTAATTCGAAAGTTTACATCAATAGATTGTCT GATTATTTGTTTGTTTTAGCAAGGTATATGGAAATAAGAACGGAAATAGAAGAAAA AGTAAAAGACGTGATAAGAAAGCATTATGGAAAGAACAAAGGCGAAATAAAGCTA AATTTAGATATAGCAAAAAATTTAATGGCTAAGGTAGAAAAGAAGGCAGAAAGCAT TAATCTACCGGTTGCTATTGCAATAGTTGACATGCATGGCAATTTGATAGCGGCTCA TTTTATGGATGGTACACTTCTTGAAAGCATGAATCTAGCTATAAATAAAGCTTATAC ATCAGTGGTGCTTAAAATGTCGACGCAAGAGTTATCAAAACTTGCACAACCAGGGC AGCCTCTTTACGGGATAAATACAACTGATAATAGAATCGTAGTGTTTGGAGGTGGGT GCCCTATAAAACATCAAGGTGAAATAGTTGGTGGAATTGGAGTTAGCGGTGGTACA GTAGAACAAGATATAGAACTTTCTATTTATGGTGCAGATGTATTTGAGGAGOTTATA TCATGA or219 SEQ ID NO: 80: MKVKEEDIEAIVKKVLSEFNFEKNTKSFRDFGVFQDMNDAIRAAKDAQKKLRNM SMESREKIIQNIRKKIMENKKILAEMGVSETGMGKVEHKIIKHELVALKTPGTEDIVTTA WSGDKGLTLVEMGPFGVIGTITPSTNPSETVLCNSIGMIAAGNSVVFNPHPGAVNVSNYA VKLVNEAVMEAGGPENLVASVEKPTLETGNIMFKSPDVSLLVATGGPGVVTSVLSSGKR AIGAGAGNPPVVVDETADIKKAAKDIVDGATFDNNLPCIAEKEVVSVDKITDELIYYMQ QNGCYKIEGREIEKLIELVLDHKGGKITLNRKWVGKDAHLILKAIGIDADESVRCIIFEAE KDNPLVVEELMMPILGIVRAKNVDEAIMIATELEHGNRHSAHMHSKNVDNLTKFGKIID TAIFVKNAPSYAALGYGGEGYCTFTIASRTGEGLTSARTFTKSRRCVLADGLSIR SEQ ID NO: 81: ATGAAAGTTAAAGAGGAAGATATTGAAGCGATCGTCAAAAAAGTCTTATCGG AATTTAATTTTGAAAAAAATACTAAAAGTTTCAGAGATTTTGGCGTATTTCAAGATA TGAATGATGCTATTCGTGCTGCAAAAGATGCCCAGAAAAAATTGAGAAATATGTCCA TGGAGTCGAGAGAAAAGATTATACAGAATATAAGAAAAAAGATTATGGAGAATAAA AAAATACTTGCAGAGATGGGCGTCAGTGAAACTGGCATGGGGAAAGTAGAGCACAA AATAATAAAACATGAGCTTGTAGCACTTAAGACACCTGGTACCGAAGATATAGTGA CAACAGCATGGTCTGGCGATAAGGGACTGACATTGGTTGAAATGGGGCCATTTGGTG TAATAGGTACGATTACTCCTTCGACAAATCCAAGTGAAACCGTCCTTTGCAATAGCA TAGGTATGATAGCCGCAGGTAATTCAGTCGTATTTAATCCACATCCAGGTGCGGTAA ATGTATCTAATTACGCTGTCAAGTTAGTAAATGAAGCGGTGATGGAAGCTGGCGGCC CTGAGAATTTAGTCGCATCTGTTGAAAAACCTACACTTGAAACTGGAAATATTATGT TCAAGAGTCCTGATGTTTCGCTATTAGTAGCGACAGGCGGACCTGGTGTAGTAACAT CGGTTCTCTCATCTGGCAAAAGGGCAATAGGAGCAGGAGCAGGAAATCCACCAGTT GTAGTTGATGAAACGGCAGATATAAAAAAAGCTGCGAAAGATATAGTCGATGGTGC TACATTTGACAACAATTTGCCTTGTATTGCTGAAAAGGAAGTAGTTTCTGTAGATAA AATAACAGATGAACTGATTTACTACATGCAACAGAATGGCTGCTACAAGATTGAGG GGCGAGAAATTGAAAAGCTCATTGAACTTGTATTGGATCACAAAGGTGGCAAGATA ACATTAAACAGGAAATGGGTTGGCAAAGATGCTCATTTAATACTAAAAGCTATAGG CATAGATGCTGATGAAAGCGTAAGGTGCATAATTTTTGAGGCGGAAAAAGACAATC CGTTAGTGGTAGAAGAGCTGATGATGCCTATTTTAGGAATAGTAAGAGCCAAAAAT GTAGATGAAGCGATAATGATTGCGACAGAGTTAGAACATGGCAATAGGCATTCAGC ACATATGCATTCTAAAAACGTTGATAATTTAACAAAGTTTGGAAAAATAATTGACAC TGCTATATTTGTAAAAAATGCTCCATCGTATGCCGCGTTAGGATATGGTGGTGAAGG TTATTGCACATTTACGATTGCAAGCAGAACAGGTGAAGGATTGACATCTGCAAGGAC TTTTACTAAAAGTCGTAGATGTGTCTTGGCAGATGGATTATCAATAAGATAG or218 SEQ ID NO: 82: MEVNQIDIEEIVKKILNDLRNEPKENIKESNSKIPSICRAAVLTDVKKIEVKEFNIPEI NDDEMLVKVEGCGVCGTDVHEYKGDPFGLIPLVLGHEGTGEIVKLGKNVRRDSAGKEI KEGDKIVTSVVPCGECDICLNHPDKTNLCENSKIYGLISDDNYHLNGWFSEYIVIRKGSTF YKVNDINLNLRLLVEPAAVVVHAVERAKSTGLMKFNSKVLVQGCGPIGLLLLSVVKTL GVENIIAVDGDENRLNMAKRLGATALINFTKYSNIDELVDAVKKASDGIGADFAFQCTG VPSAASNIWKFVRRGGGLCEVGFFVNNGDCKINPHYDICNKEITAVGSWTYTPQDYLTT FDFLKRAKEIGLPIEELITHRFSLDKMNEAMEVNMKQEGIKVVYINDRF SEQ ID NO: 83: ATGGAAGTCAATCAGATAGACATTGAGGAGATAGTTAAGAAAATATTAAATG ATTTAAGAAATGAGCCTAAAGAAAACATTAAAGAGAGCAATTCAAAAATACCATCT ATCTGCAGAGCTGCTGTACTTACAGATGTTAAAAAAATAGAAGTAAAAGAATTTAAT ATTCCAGAAATAAATGATGATGAAATGCTTGTCAAGGTGGAAGGCTGTGGCGTTTGC GGTACTGATGTTCATGAATACAAAGGAGATCCTTTTGGACTTATACCATTGGTTTTAG GACACGAAGGTACAGGTGAGATAGTCAAGCTGGGGAAAAACGTGAGACGAGATTCT GCTGGTAAAGAAATCAAAGAAGGCGATAAGATTGTTACATCTGTCGTTCCGTGCGGT GAATGCGATATATGTTTGAATCATCCAGACAAGACAAATTTGTGTGAAAACTCAAAG ATTTACGGCTTAATATCCGATGATAATTACCATTTAAATGGTTGGTTCTCAGAGTACA TCGTCATAAGGAAAGGCTCAACATTTTATAAGGTCAATGATATAAACCTTAATTTGA GGCTTTTGGTAGAACCGGCTGCAGTAGTCGTACATGCAGTAGAGCGCGCAAAATCCA CAGGTCTTATGAAATTCAACAGTAAAGTTCTCGTACAAGGCTGTGGCCCTATAGGAT TACTGCTATTGTCGGTTGTAAAGACGCTTGGAGTAGAAAATATCATAGCCGTCGACG GCGATGAGAATAGACTCAACATGGCTAAAAGATTAGGTGCTACAGCACTCATTAATT TTACTAAATACAGCAATATTGATGAGCTTGTTGATGCTGTTAAAAAAGCAAGCGATG GAATTGGCGCAGATTTTGCATTTCAATGTACAGGCGTTCCTTCTGCAGCGTCTAATAT TTGGAAGTTTGTAAGGCGGGGAGGTGGTTTATGCGAAGTTGGATTTTTTGTAAATAA TGGTGATTGTAAGATAAACCCCCATTATGATATTTGCAATAAGGAGATAACAGCAGT TGGCTCATGGACTTACACTCCTCAAGACTATTTGACAACTTTTGATTTTCTCAAAAGA GCTAAAGAAATAGGACTTCCAATTGAAGAGCTGATAACACATAGATTTTCACTTGAT AAAATGAATGAAGCTATGGAAGTTAATATGAAGCAGGAAGGGATAAAAGTAGTGTA TATAAATGACAGATTTTAG or217 SEQ ID NO: 84: MQAVGLIEVYGLVAAFVAADAACKKANVVIESFDNNKPLNAEALPVPLIIVVKL RGDLEDVKIAVDAAVDAANKISGVVATNIIAKPEEDTEKLLKLNCLK SEQ ID NO: 85: ATGCAGGCTGTTGGATTGATTGAAGTTTATGGATTAGTAGCGGCATTTGTGGC AGCAGATGCTGCATGCAAAAAAGCGAATGTCGTAATAGAGTCTTTTGACAACAATA AGCCATTAAATGCTGAAGCATTGCCAGTTCCATTGATAATAGTCGTTAAGCTCAGAG GAGATCTTGAGGATGTAAAAATAGCGGTAGATGGTGCAGTTGATGCAGCTAATAAA ATATCTGGTGTAGTTGCTACAAATATAATAGCAAAACCAGAAGAAGATACTGAAAA

GCTATTAAAGCTAAATTGTCTTAAATAA or216 SEQ ID NO: 86: MVQEALGMVETRGLVAAIEAADAMVKAADVTLIGTEKIGSGLVTVMVRGDVG AVKAATEVGASAASKLGELVAVHVIPRPHTDVEKILPTIK SEQ ID NO: 87: ATGGTACAAGAAGCATTGGGAATGGTAGAAACGAGAGGATTGGTAGCAGCA ATAGAAGCAGCAGATGCTATGGTAAAGGCTGCGGATGTCACTTTGATAGGAACTGA AAAAATAGGTTCAGGACTTGTAACAGTCATGGTAAGAGGAGATGTCGGTGCAGTAA AAGCAGCGACAGAAGTTGGCGCAAGTGCAGCTTCAAAATTGGGAGAGTTAGTGGCT GTTCACGTAATACCAAGGCCTCATACTGATGTTGAAAAGATACTGCCGACAATTAAA TAA or215 SEQ ID NO: 88: MYAIGLIEVNGFVTAVETLDAMLKTANVEFVTWEKKLGGRLVTinKGDVSAVEE AILTGKIEADKITRTVAYAVIPNPHPETIKMVNISAGKLFKADGGEINEF SEQ ID NO:89: ATGTATGCAATTGGACTTATTGAAGTAAATGGGTTTGTCACAGCGGTTGAAAC ACTGGATGCAATGTTGAAAACAGCCAATGTAGAGTTTGTAACATGGGAGAAAAAAC TTGGAGGCAGACTTGTGACAATCATTATTAAAGGAGATGTTTCAGCAGTTGAAGAAG CAATTTTAACTGGAAAGATTGAAGCTGACAAGATTACACGGACAGTAGCATACGCA GTTATTCCAAATCCACATCCAGAAACTATAAAGATGGTAAATATTAGTGCAGGAAAG CTATTTAAAGCAGATGGTGGTGAAATAAATGAGTTCTGA or214 SEQ ID NO: 90: MSSEEKDTNAKDVKVEKQKNNLTKTSNKEFKEELIMEQQALGMVETRGLVAAIE AADAMVKAANVTLIGTEKIGSGLVTVMVRGDVGAVKAATETGANAAKKLGELVAVH VIPRPHADVEKILPTIK SEQ ID NO: 91: ATGAGTTCTGAAGAAAAGGATACGAATGCAAAAGATGTTAAAGTCGAAAAG CAGAAAAATAATTTAACGAAAACATCAAATAAAGAATTTAAGGAGGAATTGATTAT GGAACAACAAGCATTAGGAATGGTAGAGACGAGAGGATTGGTAGCAGCGATAGAA GCTGCTGATGCAATGGTAAAGGCTGCTAATGTCACGTTAATAGGAACTGAAAAAAT AGGTTCAGGACTTGTAACAGTCATGGTAAGAGGAGATGTTGGTGCAGTAAAAGCAG CGACAGAGACTGGAGCAAATGCAGCTAAAAAGTTAGGGGAGTTAGTAGCTGTTCAC GTAATACCAAGACCTCATGCAGATGTAGAGAAAATACTGCCTACGATAAAGTAG or213 SEQ ID NO: 92: VITVNEKLIEIISKTIADTISERNSLKIPVGVSARHVHLTKEHLDILFGKDYILKKKK ELMGGQFAAEECVTIIGFKLNAIEKVRVLGPLRDKTQVEISKTDAISLGLNPPIRESGDIKG SSPITIVGPRGAISLKEGCIIAKRHIHMSPEDSKRFNVKDDDIISVKINGQRGGILENVQIRV DEKYTLEMHIDTDEANCMGLKSGDFVEIVRDNRS SEQ ID NO: 93: GTGATAACAGTGAACGAAAAATTGATAGAGATTATATCAAAAACTATAGCGG ATACGATTAGTGAAAGGAATTCGCTTAAGATACCAGTAGGCGTATCAGCCCGACATG TACATCTGACTAAAGAACATTTGGATATATTATTTGGAAAAGATTATATCCTTAAAA AGAAAAAGGAATTGATGGGTGGACAGTTCGCAGCAGAGGAATGTGTGACAATTATC GGATTTAAATTAAATGCTATTGAGAAAGTGAGAGTTTTGGGTCCTTTAAGAGATAAA ACGCAGGTAGAAATATCGAAGACCGATGCAATAAGTTTAGGGTTAAACCCTCCTATA CGGGAATCAGGTGATATAAAAGGTTCATCGCCAATTACAATTGTAGGGCCGAGAGG AGCAATATCATTAAAAGAAGGATGTATAATAGCAAAACGACATATTCACATGTCAC CGGAAGATTCCAAAAGATTCAATGTTAAAGACGACGATATAATATCAGTAAAAATA AATGGTCAGCGAGGCGGAATTTTAGAAAATGTACAGATTAGAGTTGACGAAAAGTA TACACTTGAGATGCATATTGACACAGATGAAGCTAATTGCATGGGACTAAAAAGCG GCGATTTTGTTGAAATAGTAAGAGATAATAGGAGTTGA or212 SEQ ID NO: 94: LIIAKVVGTVISTRKNQNLIGNKFUVEPVSEMNYDSKNRVVAIDNVGAGVGEIVL VTFGSSARIGCGMPDSPVDAAIVGIVDSIKDIIIDD SEQ ID NO: 95: TTGATAATAGCTAAAGTTGTTGGTACTGTTATTTCTACCCGCAAGAATCAAAA TTTAATAGGCAATAAATTTTTAATAGTAGAACCAGTAAGTGAAATGAATTATGACAG TAAAAATAGGGTTGTTGCAATAGATAATGTAGGTGCAGGTGTAGGAGAGATAGTAT TAGTTACCTTTGGAAGTTCAGCAAGAATCGGTTGTGGTATGCCAGATTCGCCTGTAG ATGCGGCAATTGTCGGAATTGTTGATAGCATAAAAGATATTATCATTGATGATTAG or211 SEQ ID NO: 96: MMNIDELKNIVFENGIVGAGGAGFPTHAKLTTGIDTIILNGAECEPLLRVDRQLLA IYTDEILMTLSFIVDTLGAKRGIVAIKSAYKTAISSVKNLIGNYKNLELKVLPDVYPAGDE VVLIYETTGRIVPEGSIPISVGTLVMNVETVLNVYNAIYLKFIPVTEKYVTVTGDVKYPSTF KAKVGTSVARLIEKAGGCLEKDCEVIMGGPMTGKIVDVKTPITKTTKAIIVLPKDHPVIT KRKTNIRIGLKRAMSVCSQCQMCTDLCPRHLLGHSIKPHKVMNAVANSIIDDTAAYTMT MLCSECGLCEMYSCHQSLSPRKIISQIKIKLRQNGVKNPFINKRPETANVMRDERLVPME RLISRLSLKKYDVDAPMNFDTVIPSHHVVMQLSQHVGAKAIPVVKVGDIVKEGDLIGDV PNNKLGAKLHASIDGIIIDVTDDSIVIKPRGDFDGQSDRIG SEQ ID NO: 97: ATGATOAATATTGATGAACTTAAAAATATCGTATTTGAAAATGGAATAGTCG GTGCAGGCGGAGCTGGATTTCCTACACATGCAAAACTTACTACAGGTATAGATACAA TCATATTAAATGGCGCTGAATGTGAACCGCTTTTAAGAGTAGATAGGCAGCTACTTG CAATATATACTGATGAAATATTGATGACTTTATCATTCATAGTTGATACTTTAGGAGC CAAACGTGGCATTGTAGCAATAAAATCAGCATACAAAACTGCCATCAGCTCAGTTAA GAATTTGATTGGTAATTATAAAAACTTGGAGTTAAAGGTATTGCCAGACGTTTATCC TGCTGGTGATGAAGTTGTATTAATATATGAAACGACTGGAAGAATTGTGCCAGAAGG TTCTATACCTATTTCTGTTGGCACGTTGGTAATGAATGTGGAAACTGTGCTTAATGTT TATAATGCTATTTATTTAAAACATCCAGTCACAGAAAAGTATGTAACAGTAACGGGA GATGTCAAATATCCCAGCACATTTAAAGCAAAAGTAGGAACATCTGTAGCTCGTCTT ATTGAAAAAGCAGGAGGATGCTTAGAAAAAGATTGTGAAGTGATAATGGGTGGTCC TATGACTGGGAAAATAGTTGATGTAAAGACTCCAATAACAAAAACTACAAAAGCTA TTATCGTTCTCCCAAAAGACCACCCTGTGATAACAAAGAGAAAGACAAACATAAGG ATAGGGTTAAAACGAGCAATGTCTGTTTGCTCTCAATGCCAAATGTGCACAGATCTA TGTCCTAGAAATTTATTAGGTCATTCCATCAAACCTCATAAAGTCATGAATGCAGTT GCAAATAGTATTATTGATGATACCGCTGCATATACGATGACAATGTTATGTTCTGAA TGTGGATTGTGCGAGATGTATTCATGTCATCAAAGTTTGTCGCCGAGAAAGATAATA AGCCAGATAAAGATAAAATTAAGGCAAAATGGTGTAAAAAATCCACACAACAAAAG ACCAGAAACAGCAAATGTCATGCGAGATGAGAGATTAGTGCCGATGGAAAGGCTTA TTTCAAGACTTTCGCTCAAAAAATACGATGTAGATGCTCCGATGAATTTTGATACTGT TATTCCTTCACATCACGTTGTCATGCAACTAAGTCAGCATGTTGGTGCCAAAGCGAT ACCTGTAGTAAAGGTAGGAGATATTGTGAAAGAAGGAGATCTGATAGGCGATGTGC CTAATAATAAGCTGGGTGCTAAATTGCATGCCAGTATTGACGGCATTATAATAGATG TAACTGATGACAGTATTGTTATCAAACCAAGAGGTGATTTTGATGGACAAAGCGATA GGATTGGTTGA or210 SEQ ID NO: 98: MDKAIGLVEYKSVATGITAADDMAKTADVEIIEAYTVCPGKYIVLLAGKLSAVN SAIEKGINQYSENVIDSFILGNPHETIYKAMSGTSVIEDVEALGIIETFSAASIILAADTAAK AAKVNLVEIRIARGMCGKSYLLLTGELAAVEASINAGCKALERTGMLLNKSIIPNPDRAI WDKII SEQ ID NO: 99: ATGGACAAAGCGATAGGATTGGTTGAATACAAATCAGTTGCTACAGGTATAA CTGCTGCTGATGACATGGCTAAAACTGCTGATGTGGAAATAATAGAAGCATATACAG TATGTCCGGGGAAATACATTGTTCTGTTAGCTGGGAAATTAAGTGCAGTTAATTCGG CGATAGAAAAGGGCATAAATCAGTATTCGGAAAATGTCATTGATAGCTTTATATTGG GAAATCCGCATGAAACAATATATAAAGCTATGAGTGGCACGTCTGTAATTGAAGAT GTAGAAGCACTTGGTATCATAGAGACATTTTCTGCAGCATCAATAATACTTGCAGCA GATACGGCTGCAAAAGCTGCAAAAGTGAATCTGGTAGAGATAAGAATAGCCAGAGG TATGTGCGGCAAGTCATATCTACTGCTTACAGGAGAACTTGCTGCTGTTGAAGCATC TATAAATGCAGGATGCAAAGCTTTGGAGAGAACGGGTATGCTTTTAAATAAGTCTAT AATACCCAATCCAGATAGAGCTATTTGGGATAAGATAATTTAA or209 SEQ ID NO: 100: MYEAEKDKILNDYYNAKEIYAKFDIDIDKVLDKMKKIRISLFICWQGDDVTGFEK SANGLSGGGILATGNWPGRARNGEELRQDIEKALSLIPGKHKINLHAIYAETDGEFVDRD EINVEHFRKWIYWAKENGLGLDFNPTFFSHPKANDGYTLSSKDENIRKFWIQHGKRCREI ANEIGRELKTQCVNNVWIPDGSKDLPANRIEHRKILKESLDEIFSVKYDKSNIVDSVESKL FGIGSESYVVGSHEFYMNYASRNDVMLCLDMGHFHPTENIADKISSILTFNDNLLIHVSR GVRWDSDHVVILNEDLLSLAKEIRRCDAYDKVYIALDFFDASINRIMAWVIGARATLKAI LISLLEPVHLLMEEENKGNFGARLALMEEFKTLPFYSVWNKYCMDENVPIGTSWIDDVK EYEKEIVKNRA SEQ ID NO: 101: ATGTATGAAGCAGAAAAAGATAAAATTTTAAATGATTATTATAATGCAAAAG AGATTTATGCAAAGTTTGACATAGATATTGATAAAGTATTAGATAAAATGAAGAAG ATTCGTATTTCACTTCACTGCTGGCAAGGCGATGATGTAACTGGATTCGAAAAAAGT GCCAATGGATTAAGCGGTGGAGGTATTTTGGCGACAGGAAACTGGCCTGGTAGAGC AAGAAATGGTGAAGAATTAAGGCAAGACATTGAAAAAGCCTTAAGCCTTATACCAG GCAAACACAAAATCAATTTACATGCCATTTACGCAGAAACGGATGGTGAATTTGTAG ACAGAGATGAAATAAACGTGGAGCATTTCAGGAAATGGATTTACTGGGCAAAAGAA AATGGCCTTGGCCTTGACTTCAATCCTACGTTTTTTTCGCATCCTAAAGCAAATGATG GCTATACGCTTTCAAGCAAAGATGAAAACATAAGAAAATTTTGGATCCAACATGGTA AAAGATGCCGTGAAATCGCAAATGAAATAGGAAGAGAGCTAAAAACTCAATGTGTG AATAATGTTTGGATTCCTGATGGTTCAAAAGATTTGCCTGCTAATAGGATTGAACAC

AGAAAAATACTTAAAGAATCTTTAGATGAGATATTTTCAGTAAAATATGACAAATCA AATATCGTTGATTCTGTTGAAAGCAAATTATTTGGCATTGGATCTGAAAGCTATGTG GTTGGTTCACATGAGTnTATATGAACTATGCGTCGAGAAATGATGTAATGCTGTGC CTTGATATGGGACATTTTCATCCTACTGAGAATATTGCTGATAAGATATCATCAATAC TTACATTCAATGACAATTTGTTGATTCATGTAAGCCGTGGTGTCCGGTGGGATAGCG ACCATGTAGTCATTTTAAATGAAGATTTGCTTTCATTAGCAAAAGAAATAAGAAGAT GTGATGCTTATGACAAAGTGTATATTGCATTAGATTTCTTTGATGCAAGCATAAATA GGATAATGGCATGGGTAATAGGTGCAAGAGCGACGCTAAAAGCCATATTAATATCA CTATTAGAGCCTGTGCATCTACTTATGGAAGAGGAGAATAAAGGAAATTTTGGTGCA AGACTTGCTTTGATGGAGGAATTCAAAACATTGCCATTTTACTCTGTTTGGAACAAA TACTGCATGGACGAAAATGTGCCTATTGGTACATCGTGGATTGATGATGTTAAAGAA TATGAAAAAGAAATTGTAAAAAATAGGGCTTAA or208 SEQ ID NO: 102: MKDIVYNLAFDFGASSGRLMLSAFDGEKITIEEIYRFPNEPVKLGQSFYWDFLRLF HELKNGLKIASKRKIKISGIGIDTWGVDYGLLDKNDQLISNPFHYRDKRTDGIIKDFENM ALLEEIYNVTGIQFMEFNTIFQLYCDYKKRPELLDNAKTLLFIPDLFNFYLTNEKYNEYTV ASTSQMLDANKKDWANDLIEKLNLPEGIFQKILMPGNTIGYLTKEIQEETGLSEVPVISVG SHDTASAVAGTPIENGSSAYLICGTWSLLGVESEKPIINENTKKYNFTNEGGVEGLIRLLK NINGLWIIQQLKQSWNSNGIKIGFPEISQMASKAEFIEEFIINPDDKLFIAPDDMAEAIRQYC TKTGQGLPQNIGDIARAAYNGIVEQYKNCLNNLEDIVGQEIDNIHMVGGGIQDKFLCKLT ADVTGKKVITGPVEASIYGNVIVQLMALGYIKDLREGRK1IKNSIENDEEMFAK SEQ ID NO: 103: ATGAAAGATATTGTGTATAATCTGGCTTTTGATTTTGGAGCTTCAAGTGGCCG TCTTATGCTATCCGCGTTTGATGGCGAAAAAATCACAATTGAAGAGATTTATAGATT TCCAAATGAGCCAGTCAAGCTGGGACAATCATTTTATTGGGATTTTTTAAGGCTTTTT CACGAATTAAAAAACGGATTAAAAATAGCATCAAAGAGGAAAATCAAAATATCCGG CATTGGTATAGACACTTGGGGTGTCGATTATGGATTGCTTGATAAAAATGATCAATT GATTTCAAATCCTTTTCATTACAGAGATAAAAGAACGGATGGCATAATAAAAGATTT TGAAAATATGGCGTTACTGGAGGAAATCTACAACGTAACTGGTATACAGTTTATGGA ATTTAATACAATATTCCAATTGTATTGCGATTATAAAAAGCGTCCAGAATTATTGGA TAATGCAAAGACATTGTTGTTTATTCCAGATTTATTTAACTTTTATTTGACAAATGAG AAATACAATGAATATACTGTTGCATCCACATCGCAAATGTTGGATGCTAACAAGAAA GATTGGGCAAATGATCTTATAGAAAAGTTAAATTTGCCAGAAGGTATTTTTCAAAAG ATACTGATGCCAGGAAATACAATTGGTTATCTAACAAAAGAAATTCAAGAAGAAAC AGGATTGTCTGAAGTTCCCGTGATTTCTGTTGGCAGCCATGATACGGCATCAGCAGT TGCAGGTACACCTATTGAAAACGGTTCAAGTGCTTATTTGATTTGTGGTACTTGGTCA TTATTAGGTGTTGAAAGTGAAAAACCTATAATAAATGAAAATACAAAGAAGTACAA TTTTACAAATGAAGGCGGTGTCGAAGGCCTTATAAGGCTACTTAAAAATATTAATGG TCTGTGGATAATTCAGCAATTAAAACAAAGTTGGAATTCAAATGGCATTAAAATAGG ATTTCCAGAAATCAGCCAGATGGCATCTAAAGCAGAGCACGAAGAATTTATCATAA ATCCTGATGACAAATTGTTTATAGCTCCAGATGATATGGCTGAGGCGATAAGGCAAT ATTGTACAAAAACAGGACAGGGTTTGCCGCAGAATATTGGCGACATAGCAAGAGCC GCTTACAATGGTATAGTTGAACAATACAAAAATTGCTTAAACAATTTAGAAGATATT GTAGGGCAAGAAATAGATAATATTCACATGGTTGGTGGTGGGATACAGGATAAGTT CCTGTGCAAGCTGACTGCAGATGTTACAGGGAAAAAAGTCATAACAGGCCCTGTAG AAGCTTCAATCTATGGCAATGTGATAGTCCAGCTTATGGCATTGGGATATATAAAAG ACTTGAGAGAAGGAAGAAAGATAATAAAGAATTCTATAGAGAATGATGAAGAGATG TTTGCTAAATAG or207 SEQ ID NO: 104: VSNIYTLWVEDEYEIRTGLVNCFPWNKMGFVVAEEFENGGECFEYLCKNKVDT ILCDIKMPVMSGIELAKKIFESNISTKIVIISGYTDFEYARQALRYGVKDYIVKPTKYNEDD VFSRIKKELDNENTKEILNNSCNNEIDQYSSIISIIEKYVDEHYRDVTLEDVAKVVYMNPY YLSKYFKQKTGMNFSDYITEVRMKKAVEFLKNPLYKTYEISYMIGYKNPKNFTRAFKKY YKKSPREFVNSAINFKE SEQ ID NO: 105: GTGTCTAATATTTATACGCTTGTAGTAGTAGAAGATGAATATGAGATAAGAA CAGGATTAGTTAACTGCTTTCCATGGAACAAAATGGGTTTTGTTGTTGCAGAAGAAT TTGAAAATGGAGGAGAATGTTTTGAGTATTTGTGTAAAAATAAGGTTGATACAATTT TATGTGATATAAAAATGCCAGTTATGTCTGGTATAGAGTTGGCAAAGAAAATTTTTG AAAGTAATATAAGCACTAAAATAGTTATAATCAGTGGTTATACTGATTTTGAATATG CCAGACAGGCGTTAAGATATGGTGTTAAAGATTATATAGTAAAACCTACTAAATATA ATGAAATAATTGATGTTTTCAGCAGAATAAAAAAAGAATTAGACAATGAAAATACA AAGGAAATATTGAATAACTCATGTAACAATGAAATTGATCAGTACAGCAGCATAATT TCAATCATAGAAAAATATGTTGATGAACATTACAGAGATGTGACATTGGAAGATGTA GCTAAAGTAGTTTATATGAATCCGTATTATTTAAGCAAATATTTTAAACAAAAAACC GGTATGAATTTTTCTGATTATATAACTGAGGTCAGAATGAAAAAAGCTGTAGAGTTT CTAAAAAATCCTTTGTATAAAACTTATGAAATAAGTTATATGATTGGATATAAAAAT CCAAAAAATTTTACTAGAGCATTTAAAAAATATTATAAAAAATCCCCAAGAGAATTT GTAAATTCAGCAATAAATTTTAAGGAATGA or206 SEQ ID NO: 106: MRELNNKFFYKNLFVLALPLILIVIVLGSFSILITERYVRDEIYKNSREILKQSSNDL SILFNDINKIYLTFGTNKDVTLYLERILNTNKYSLDDMWHLSMIESLFDSTSFSEPYIQSIY LYFNNPNKNFLVTGNGrNSVTNYIDNKWYDSFLNAPKDEISWIEVRNLKMYSFDKKGIK VLSIYKKIANFNGDKIDGVLVLNIYLDYIENLLNTSTIFPDQKILILDAHDNLICQNINGNFT GKIDLDNYSKANIITKLESPNYNIKYVSIVPKKYLYEVPIKLLKMTLVLLLTSIFFVILITFRI TKRNYENVNKILKIIEAEKTNEIFPEIPVESRDEYSYIIYNIINSYIEKSQLKMELAEKKYKM KAMELLALQSQISPHFLSNALEIIYLRALSYTNGPNDVTKMIENLSQILKYLLSNPNETVT VKEEIENTKAYIQILKVRYRDKFKVNLIYDESILSCLMMKLMLQHLIENSIKHGLKKKNY EGSIKIKIKAVDKKKIKISVIDNGIGMSKERLNYVKRILDSDFDFYEHIGLMNTNERLKLL YGKDCEILIRSKLNIGTAVYIIFPYQLKNQNNDDYNK SEQ ID NO: 107: ATGAGAGAATTAAACAATAAATTTTTTTATAAAAATCTTTTTGTTTTGGCATT GCCATTAATTTTAATTGTTATTGTATTAGGTTCATTTTCAATATTAATAACAGAAAGA TATGTTAGAGATGAAATATACAAAAATAGTAGAGAAATATTAAAGCAAAGCAGTAA TGATTTGTCAATTTTATTTAATGATATAAATAAAATTTATTTAACATTTGGAACAAAC AAAGATGTGACATTGTATTTGGAAAGGATCTTAAATACAAATAAATATTCTTTAGAT GATATGTGGCATCTTAGCATGATAGAAAGTTTATTTGATTCTACGTCGTTTTCAGAAC CITATATACAATCAATTTATTTGTATTTTAACAATCCTAATAAAAATTTTTTAGTGAC AGGAAATGGTATTAATTCTGTAACAAATTATATTGATAATAAATGGTATGACAGCTT TTTAAATGCACCAAAAGATGAGATTTCTTGGATAGAGGTTAGAAATTTAAAAATGTA TAGTTTCGATAAAAAGGGGATAAAAGTCCTAAGTATATACAAAAAAATTGCAAACT TTAACGGGGATAAAATTGATGGTGTGCTTGTACTAAATATATATTTGGACTATATTG AAAATTTGCTAAATACTTCAACAATATTTCCTGACCAAAAAATTCTTATATTAGATGC CCACGACAATTTAATATGTCAAAATATTAATGGGAATTTCACTGGGAAGATAGACTT AGATAATTATAGCAAAGCAAACATCATAACAAAATTAGAATCTCCAAATTATAATAT AAAATATGTATCTATTGTTCCTAAAAAATACCTTTATGAAGTTCCTATAAAGCTTTTA AAGATGACTTTAGTTTTACTTTTGACGTCAATTTTTTTTGTGATATTGATAACATTTAG AATCACTAAACGAAATTACGAAAATGTAAATAAAATATTAAAGATTATAGAGGCAG AAAAGACAAATGAGATATTTCCAGAAATTCCAGTAGAAAGTAGAGATGAGTACAGC TATATAATTTACAACATTATTAATAGTTATATTGAAAAAAGTCAATTOAAAATGGAA TTAGCAGAAAAGAAGTATAAAATGAAAGCAATGGAGTTATTAGCACTGCAATCGCA AATTAGTCCTCATTTTTTGTCTAATGCGTTGGAGATTATTTATCTTAGGGCATTGTCA TACACAAACGGTCCTAATGATGTCACAAAAATGATTGAAAATTTGTCACAGATTTrA AAGTATTTGTTAAGTAATCCAAATGAAACAGTAACTGTAAAAGAAGAAATTGAAAA TACAAAGGCATATATACAAATATTGAAGGTCAGGTATAGAGATAAATTTAAAGTAA ATCTAATTTATGATGAAAGTATTTTATCATGTCTCATGATGAAACTGATGCTGCAACA TTTAATAGAAAATTCTATAAAACATGGGCTTAAGAAGAAAAATTATGAAGGATCAA TAAAAATCAAAATAAAAGCAGTTGATAAAAAGAAAATAAAAATTTCAGTAATCGAT AATGGCATAGGAATGTCCAAAGAGAGGCTAAATTATGTAAAAAGAATTCTTGACTCT GACTTCGATTTTTATGAACATATTGGACTAATGAATACAAATGAACGGTTAAAACTT CTCTATGGGAAAGATTGTGAAATATTAATAAGAAGTAAATTGAATATTGGTACTGCC GTATATATAATTTTTCCATATCAATTAAAAAATCAGAATAATGATGATTATAATAAG TGA or205 SEQ ID NO: 108: MGINRYDLVKRHNVILEKADIENPLSVONGEIAFIADITGMQTFIDDYKSIPLCTM SQWGFHTTPAQNDKGYYTLEDLNLKYYDAFDRKVGYVTSAENQENVFNWLRSNPHRI NLGNIGLNIILDDGTKAELKDIFEIHQVLDLWNGILISDFKVEKVPVHVETFCHPYEDMIN FSVESELLKQNKIYIEVKFPYGAANISGSDWDRNDRHDTNVVDYGRDFVELLRTVDEDV YFVKIEYSKGVYLNRIGENHFALKQKEYNGRIEFSCLFSKQKPLKCLHSFSESKRMCKEY WNSFWRGGGAIDFSKCEDKRAFELERRVILSQYLTAIQCSGSMPPQETGLTCNSWYGKF HLEMHWWHAVHFALWGRMPLLSRSIWWYRSIFNVSRDIARKQGYKGVRWPKMVGPD GRDSPSPIGPLLVWQQPHLIYYSELFFRENPTEETLDMFKDIVINTADFIASFVAYDRKND RYILAPPLIPAQENHDPNVTLNPVFELEYFSFALEIAVKWIERLGLNVNQEWNEIRFKLAN LPSKDGVYISHEKCINTYEKFNFDHPSMLAALGMLPGRKVDKETMRRTLHRVLKEWKF EEMWGWDFPMMAMTATRLGEPETAINILLMDSPKNTYMVNGHNNQIPNKELPVYLPG NGGLLAAMALMTAGWDGNSQSTPGFPKNGMWNVEWEGLKAMI SEQ ID NO: 109

ATGGGAATTAACAGATATGATCTTGTAAAAAGGCATAATGTAATTTTGGAAA AAGCAGATATCGAAAATCCATTGTCAGTAGGTAATGGAGAAATTGCTTTTACAGCTG ATATAACGGGAATGCAAACTTTTATTGATGACTATAAGAGCATTCCTTTATGTACCA TGTCACAGTGGGGGTTTCATACTACGCCGGCACAGAATGATAAGGGCTATTATACTT TGGAAGATTTGAACCTCAAGTATTACGATGCATTTGACCGAAAGGTTGGATATGTAA CATCAGCAGAAAATCAAGAGAATGTATTTAATTGGTTGAGGAGTAATCCTCATAGAA TTAATTTAGGTAATATAGGATTAAATATAATTCTTGATGATGGCACAAAAGCAGAAT TGAAAGATATTTTCGAAATACACCAAGTATTAGATTTGTGGAACGGAATATTGATAA GTGACTTTAAAGTCGAAAAAGTCCCTGTTCACGTTGAGACTTTTTGCCATCCATATGA AGATATGATAAATTTTTCTGTTGAATCAGAACTGCTAAAACAAAATAAAATTTATAT TGAAGTAAAATTTCCATATGGTGCGGCCAATATATCAGGCTCCGATTGGGATAGAAA TGATAGACATGATACAAATGTGGTTGATTATGGCAGAGATTTTGTCGAATTATTGAG AACTGTCGATGAAGATGTTTATTTTGTAAAAATAGAGTACTCAAAAGGCGTTTATTT AAATAGAATCGGGGAAAATCATTTTGCATTAAAGCAAAAAGAGTATAATGGGAGAA TAGAATTTTCGTGCTTGTTTTCGAAGCAAAAACCTCTTAAGTGCTTGCATTCATTTAG TOAAAGCAAAAGGATGTGTAAAGAATATTGGAATAGCTTTTGGAGAGGAGGTGGTG CAATAGATTTTTCAAAGTGTGAGGATAAAAGAGCTTTTGAATTGGAGAGAAGGGTA ATACTTTCGCAATATCTTACAGCTATTCAATGTTCGGGTTCTATGCCGCCGCAAGAAA CAGGGCTCACCTGTAATAGCTGGTATGGTAAATTTCATTTGGAAATGCATTGGTGQC ATGCTGTACATTTTGCTTTATGGGGTAGAATGCCTTTGCTGAGTAGAAGTATATGGTG GTACAGGAGCATTTTCAATGTATCACGTGACATTGCGAGAAAGCAAGGATACAAAG GTGTACGCTGGCCTAAAATGGTTGGACCAGATGGAAGGGATAGCCCTTCTCCGATAG GACCATTGCTTGTTTGGCAGCAGCCTCATCTTATATATTACAGTGAACTGTTTTTTAG AGAAAATCCTACGGAAGAAACATTAGATATGTTTAAAGACATAGTAATTAATACTGC TGATTTTATTGCATCATTTGTTGCATATGATAGAAAAAATGATAGATATATACTTGCG CCACCTTTGATTCCAGCACAAGAAAATCATGATCCTAACGTTACATTAAATCCGGTA TTTGAATTGGAGTATTTTTCGnTGCGCTGGAAATAGCAGTTAAATGGATTGAAAGG TTAGGACTAAATGTGAACCAAGAGTGGAATGAAATACGTTTTAAATTAGCTAATTTA CCTTCAAAAGACGGTGTATATATATCGCATGAAAAATGTATTAACACTTATGAGAAA TTTAATTTTGACCATCCATCTATGCTTGCAGCATTGGGGATGCTACCAGGCCGCAAG GTTGATAAAGAAACTATGAGAAGGACTTTACATAGAGTATTAAAAGAGTGGAAATT TGAGGAAATGTGGGGTTGGGATTTTCCGATGATGGCTATGACTGCAACAAGATTAGG CGAACCGGAGACAGCAATAAATATTCTTTTGATGGATTCACCAAAAAATACTTATAT GGTAAATGGCCATAATAACCAAATACCGAATAAAGAACTACCAGTATATTTGCCTGG AAATGGTGGACTATTGGCGGCAATGGCCCTCATGACAGCTGGTTGGGATGGGAATA GCCAAAGCACACCTGGATTTCCTAAAAATGGGATGTGGAATGTTGAATGGGAAGGG TTAAAAGCGATGATATGA or204 SEQ ID NO: 110: MIKRKDLYIRDPFVVPVPNEKIYYMFGTTDINCWNDEKATGFDYYKSSDLENFEG PFIAFRPDKNFIWDKNFWAPEVHKYNDMYYMFATFFADGRNRGTQILVSEKISGPYRPW SIEPVTPKDWMCLDGTFYVDENGEPWMIFCHEWVQIYDGEICAVRLSKDLKTTIGNPITL FKASSANWTRSIKKIKDHECYVTDGPFIYRSEEGKLYMLWSSFIENNIYAVGISLSRTGKI TGPWVHSENPIFAGDGGHGMIFKTFEGNLTLAVHTPNKRKEERPLFITLEKSVLNDTL SEQ ID NO: 111: ATGATAAAACGAAAGGATCTTTATATACGTGATCCATTTGTAGTTCCAGTACC GAATGAAAAAATATATTATATGTTTGGAACTACTGATATAAATTGCTGGAATGATGA GAAAGCAACTGGATTTGATTACTATAAATCATCTGATTTAGAAAATTTTGAAGGACC TTTTATTGCATTTAGACCAGATAAAAACTTTATTTGGGATAAAAATTTTTGGGCTCCA GAAGTGCACAAATACAATGACATGTATTATATGTTTGCTACATTTTTCGCTGATGGC AGAAATAGAGGAACGCAAATTTTAGTATCTGAAAAAATAAGTGGGCCATATAGACC ATGGAGTATTGAACCGGTGACGCCGAAGGATTGGATGTGTTTAGATGGGACTTTTTA TGTAGATGAGAATGGGGAACCCTGGATGATATTTTGCCATGAATGGGTACAAATATA TGATGGGGAAATTTGTGCTGTAAGATTGTCGAAAGATTTAAAAACAACGATAGGAA ATCCTATTACACTTTTTAAAGCTTCCAGTGCTAATTGGACAAGAAGTATTAAAAAGA TTAAAGATCATGAATGCTACGTTACGGATGGCCCTTTTATTTATAGGTCTGAAGAGG GAAAGCTTTATATGTTGTGGTCCAGTTTTATTGAAAACAATATATACGCTGTTGGTAT ATCATTATCGAGAACAGGCAAAATAACCGGCCCGTGGGTACACAGTGAAAATCCAA TTTTCGCAGGTGATGGTGGGCATGGTATGATATTTAAGACCTTTGAAGGGAATCTAA CATTGGCAGTACACACACCTAATAAAAGGAAAGAAGAACGGCCCCTTTTTATAACTT TAGAAAAATCTGTGCTTAATGATACCTTATAA or203 SEQ ID NO: 112: MFKKITSLLISLLLIISLVTGCSSSSNSSSSSKNSSENNTSPKTVTLRFMWWGGDAR HKATLDAISLYEKEHPNVKINAEYGGVTDYLQKLITQLSSGTAPDLIQIDVTWLQQLFSQ GDFFADLSKLKDINVNAFDQNFLKNYCYVNNKLIGLPTGINNSAMYINKDFFNKFGIDD KTVWTWDNLLQTAKMVHEKDKNAYLLDADSTICDYILVTYVGQKTGNQWVKDDYTL GFDKQTLTEAFKYLNDLFEVGAIEPFSQSAPYEGKPDQNPMWLNGQTGMLWNWSSIYA GVKANIKNLSLALPPIDPNAKQTGIVVRPSQLIAINKDSKNIDEAAKFLNWFFTNTDA1KT LKDVRGVPATADARKILSENNLLDSTLTDNANQAMEKMAPPENGISGNQELEKINTDHQ ELAYKKITPEQAADELINTYKQKLPELKSQQ SEQ ID NO: 113: ATGTTTAAAAAAATTACATCTCTGTTAATATCGCTTCTTTTGATAATTTCATTA GTTACAGGATGTAGCAGTTCTTCGAATTCTTCGAGTTCATCGAAAAATAGTTCTGAA AATAATACCAGCCCAAAAACCGTAACATTAAGATTTATGTGGTGGGGTGGAGATGC CAGACATAAAGCAACACTTGATGCCATAAGTCTTTATGAAAAAGAACATCCCAATGT AAAGATTAATGCTGAATATGGCGGCGTTACTGACTATCTCCAAAAGCTGATAACTCA ATTAAGCAGTGGTACAGCACCTGATCTTATACAAATAGATGTAACATGGTTGCAGCA ACTTTTTAGCCAAGGTGATTTTTTTGCAGATTTAAGTAAGTTAAAAGATATCAATGTG AATGCATTTGATCAAAATTTTCTTAAAAATTATTGCTATGTCAACAATAAGTTGATAG GTTTGCCTAAGGAATAAACAATTCGGCAATGTATATTAACAAAGACTTTTTTAATA AATTTGGCATAGACGATAAGACGGTTTGGACATGGGATAATCTCTTGCAAACCGCTA AGATGGTGCATGAAAAGGATAAAAATGCTTATCTTTTAGATGCTGATTCTACTATTT GTGATTATATATTGGTCACATACGTGGGGCAAAAAACTGGAAATCAGTGGGTGAAA GATGATTACACTTTAGGTTTTGATAAACAAACATTGACAGAGGCATTCAAATATTTA AACGATTTGTTCGAAGTAGGCGCTATAGAGCCATTTTCTCAAAGTGCTCCATACGAA GGAAAACCTGATCAAAATCCTATGTGGCTTAATGGTCAAACGGGTATGCTTTGGAAC TGGTCATCTATATATGCTGGTGTAAAAGCAAACATAAAGAACCTGTCATTGGCATTG CCACCTATTGACCCTAATGCAAAACAGACAGGCATAGTTGTAAGACCATCACAGCTT ATTGCTATTAACAAGGATTCTAAAAATATCGATGAAGCAGCAAAATTTTTAAATTGG TTCTTTACGAATACAGATGCTATAAAAACACTTAAAGATGTCAGAGGAGTTCCAGCT ACCGCAGATGCACGCAAAATTTATCAGAAAATAATTTGTTGGATTCGACTTTAACT GATAATGCAAATCAAGCTATGGAAAAGATGGCACCTCCTGAAAACGGTATAAGTGG TAATCAAGAGTTAGAAAAGATAAATACTGATATCATACAAGAACTGGCTTATAAAA AGATAACGCCAGAGCAGGCTGCTGATGAATTGATAAATACTTATAAACAGAAACTT CCAGAATTAAAAAGCCAGCAATAA or202 SEQ ID NO: 114: MSYNKKRNLMGYLYISPWIIGFLIFTLYPFAMTFIYSFCNYSITKSPVFIGLGNYIT MFTKDMYFWPSLINTIKYVLMTVPLKLCFALFVAMILNIDIKGVNVFRTTYYLPSIFGGS VALSVIWKFLFMDNGIMNKFLSYFHIHGPSWIGNPHISLFTISLLSVWEFGSSMVIFLAAL KQVPNELYEASMLDGASKIRRFFSITLPMISPVLLFNLVMQTINAFQEFTGPYVITGGGPM NSTYVYSMLIYDNAFRYFRMGYSSALSWILFLLILIVTVIIFKSSNTWVYYENGGR SEQ ID NO: 115: ATGAGTTATAATAAAAAGAGAAATTTGATGGGGTATTTATATATTAGTCCATG GATTATAGGCTTTTTAATATTTACTCTGTATCCATTTGCTATGACTTTTATCTATTCAT TTTGTAACTACAGTATTACAAAATCACCTGTATTTATTGGATTAGGCAATTATATAAC TATGTTTACTAAAGATATGTATTTTTGGCCATCTTTAATTAATACTATAAAATATGTA TTAATGACAGTTCCTTTAAAATTATGTTTTGCACTTTTTGTTGCAATGATCTTAAATAT TGATATTAAAGGAGTTAATGTGTTTAGAACAACTTATTATCTGCCTTCTATTTTTGGA GGAAGTGTTGCTTTATCTGTTATATGGAAATTTTTATTCATGGATAATGGTATTATGA ATAAATTTCTTTCATACTTTCATATACACGGGCCAAGTTGGCTTGGAAACCCACACAT ATCATTATTTACTATAAGTTTATTGTCAGTGTGGGAATTTGGGTCTTCTATGGTAATA TTTTTGGCAGCCCTAAAACAGGTCCCGAATGAGTTGTATGAAGCATCTATGTTAGAT GGTGCAAGCAAAATAAGAAGGTTTTTCTCAATAACTTTACCTATGATATCGCCTGTG CTATTATTTAATTTGGTTATGCAGACTATAAATGCTTTTCAGGAATTTACAGGTCCAT ACGTGATAACTGGTGGAGGACCGATGAACTCTACTTATGTGTACAGTATGTTGATTT ATGATAATGCGTTTAGGTATTTTAGGATGGGTTATTCATCTGCCTTGTCTTGGATTTT ATTTTTGTTAATATTGATTGTTACAGTTATAATATTTAAATCTTCAAATACATGGGTG TATTACGAAAATGGAGGTAGATGA or201 SEQ ID NO: 116: MKAKNSQNNDIIRKVFIYVFLVAFGIFMIYPLLWVFASSFKSNDEIFKSISLIPKHIV TNSYFEGWKGTGQYSFGTFILNSITLVVPVVVFTAISSTIVAYGFARFEFPLKTILFTLMIST MMLPGTAVLIPRYILFNWLGWINTYKPFIVPALFGTTPFFIFMMVQFLRGLPKELEESATI DGCNSFQILMKILIPLCKPAIISMCIFQFIWTWNDFFNPLIYINSVEKYTVSLGLNMTIDGTS VVNWNQIMAMTLISMIPSIIIFFSAQKYFVEGIATTGLKN SEQ ID NO: 117: ATGAAAGCAAAGAATAGTCAAAATAACGATATAATCAGAAAAGTATTTATAT ATGTTTTCTTGGTGGCTTTTGGTATTTTCATGATATATCCTTTACTTTGGGTTTTTGCA TCATCATTTAAATCAAATGATGAAATCTTTAAATCGATAAGCCTTATACCAAAACAC ATTGTGACAAATTCATATTTTGAAGGATGGAAAGGTACGGGACAATACTCTTTTGGT

ACATTTATTTTAAACAGCATTACGCTTGTTGTACCTGTTGTTGTATTTACTGCTATATC ATCAACAATTGTAGCCTATGGATTTGCAAGATTTGAGTTTCCGCTTAAAACTATTTTG TTTACTTTGATGATATCTACTATGATGTTGCCGGGCACTGCAGTTTTGATACCAAGAT ATATATTGTTTAATTGGTTAGGCTGGATAAACACTTATAAACCATTTATTGTTCCCGC TTTGTTCGGAACAACGCCTTTTTTCATTTTTATGATGGTTCAATTTTTGAGAGGTCTTC CTAAAGAATTAGAAGAATCGGCTACAATTGATGGTTGCAATTCATTTCAAATACTTA TGAAGATTTTAATACCATTGTGTAAACCTGCAATTATTTCTATGTGTATATTTCAGTT ATATACAGTTTCTCTCGGGCTTAATATGACAATTGATGGGACTTCAGTTGTAAATTGG AACCAAATAATGGCAATGACAATTATTTCAATGATACCGAGCATCATAATATTTTTT TCAGCGCAAAAATACTTCGTTGAAGGTATTGCAACAACTGGATTAAAGAACTAA or200 SEQ ID NO: 118: MRYTDGKVHDITIAYIGGGSRGWAWNLMTDLAKEESISGTVKLYDIDYDAAHD NEIIGNALSMRQDVKGKWLYKACETLEESLKGADFVIISILPGTFDEMESDVHAPEKYGI YQSVGDTVGPGGIVRALRTIPMFVDIANAIKEHCPDAWVINYTNPMTLCVRTLYEIFPQI KAFGCCHEVFGTQKLLSRALQDIEGIENVPREEIKINVLGINHFTWIDNARYKDIDLMYV YKQFVNKYYESGFVSDANNNWMNNSFVSAERVKFDLFLRYGVIAAAGDRHLAEFVPG YWYLKDPETVREWMFGLTTVSWRKEDLKRRLERSKRLKTGEEKFELKETGEEGVRQ1K ALLGLGDLVTNVNMPNHGQIEGIPYGAWETNALFSGNKLKPVLSGKLPDNVNSLVLRQ VYNQETTLKAALKRDFDLAFSAFVNDPLVTISLKDAKKLFKEMLENTKKYLDGWK1KA SEQ ID NO: 119: ATGAGATATACAGATGGAAAGGTTCATGACATTACTATTGCTTATATCGGTGG TGGTTCAAGAGGATGGGCGTGGAATTTAATGACTGACTTAGCAAAAGAGGAAAGTA TTTCTGGTACAGTAAAGTTATACGACATAGATTACGATGCGGCACATGACAATGAGA TAATAGGCAATGCTTTATCAATGAGACAGGATGTTAAAGGCAAATGGCTTTATAAAG CTTGTGAGACGTTAGAAGAGTCACTAAAAGGTGCTGATTTTGTCATAATATCTATTTT GCCAGGTACGTTCGACGAGATGGAATCTGATGTTCATGCACCAGAAAAGTATGGCAT TTATCAGTCAGTAGGTGATACAGTAGGACCTGGTGGAATAGTCAGAGCTTTAAGGAC GATTCCGATGTTTGTGGACATTGCCAATGCGATTAAAGAGCATTGTCCAGATGCATG GGTCATAAATTATACAAATCCTATGACACTTTGTGTAAGGACATTGTATGAAATTTTC CCTCAAATTAAAGCATTTGGATGCTGCCATGAAGTTTTTGGCACACAGAAGCTATTA TCTCGTGCTCTGCAGGATATAGAAGGCATTGAAAATGTTCCGAGGGAAGAGATAAA GATAAATGTTTTAGGTATAAATCATTTTACGTGGATCGACAATGCAAGGTACAAAGA CATAGATTTAATGTATGTTTATAAACAATTTGTGAATAAGTACTATGAAAGCGGATT TGTCAGCGATGCTAACAATAATTGGATGAACAATTCATTTGTATCTGCAGAGAGAGT AAAGTTTGATCTGTTTTTGAGGTATGGAGTAATAGCTGCAGCGGGAGATAGACATCT GGCGGAATTTGTGCCGGGATATTGGTATTTAAAAGATCCAGAGACAGTCAGAGAAT GGATGTTTGGCTTAACGACTGTAAGTTGGAGAAAAGAAGACTTAAAACGCAGGCTT GAAAGAAGTAAAAGGCTTAAGACAGGTGAGGAAAAATTTGAGTTAAAGGAAACAG GCGAAGAAGGTGTTAGGCAAATTAAAGCACTATTAGGCTTAGGCGATTTAGTGACTA ATGTCAACATGCCGAACCATGGACAGATTGAAGGAATACCATACGGTGCGGTAGTT GAAACAAACGCTTTATTTTCAGGTAATAAACTAAAGCCTGTATTATCAGGAAAATTG CCTGACAATGTAAACAGCCTCGTGTTAAGGCAAGTATACAACCAAGAAACGACGTT GAAAGCTGCTTTAAAGAGAGATTTTGATTTGGCTTTTAGTGCTTTTGTAAATGATCCA CTTGTTACAATATCTTTAAAAGATGCAAAAAAATTATTTAAGGAAATGCTTGAAAAT ACGAAGAAATATCTAGATGGATGGAAAATAAAAGCTTGA Non-Native proteins EC 2.3.1.9 C. acetobutylicum ThlA (SEQ ID NO: 120) MKEVVIASAVRTAIGSYGKSLKDVPAVDLGATAIKEAVKKAGIKPEDVNEVILGN VLQAGLGQNPARQASFKAGLPVEIPAMTINKVCGSGLRTVSLAAQIIKAGDADV1IAGGM ENMSRAPYLANNARWGYRMGNAKFVDEMITDGLWDAFNDYHMGITAENIAERWNISR EEQDEFALASQKKAEEAIKSGQFKDEIVPVVIKGRKGETVVDTDEHPRFGSTIEGLAKLK PAFKKDGTVTAGNASGLNDCAAVLVIMSAEKAKELGVKPLAKIVSYGSAGVDPAIMGY GPFYATKAAIEKAGWTVDELDLIESNEAFAAQSLAVAKDLKFDMNKVNVNGGAIALGH PIGASGARILVTLVHAMQKRDAKKGLATLCIGGGQGTAILLEKC EC 2.8.3.8 C. acetobutylicum CtfAB CtfA (SEQ ID NO: 121) MNSKIIRFENLRSFFKDGMTIMIGOFLNCGTPTKLIDFLVNLNIKNLTIISNDTCYPN TGIGKLISNNQVKKLIASYIGSNPDTGKKLFNNELEVELSPQGTLVERIRAGGSGLGGVLT KTGLGTLIEKGKKKISINGTEYLLELPLTADVALIKGSIVDEAGNTFYKGTTKNFNPYMA MAAKTVIVEAENLVSCEKLEKEKAMTPGVLINYIVKEPA CtfB (SEQ ID NO: 122) MINDKNLAKEIIAKRVARELKNGQLVNLGVGLPTMVADYIPKNFKITFQSENGIV GMGASPKINEADKDVVNAGGDYTTVLPDGTFFDSSVSFSLIRGGHVDVTVLGALQVDE KGNIANWIVPGKMLSGMGGAMDLVNGAKKVIIAMRHTNKGQPKILKKCTLPLTAKSQA NLIVTELGVIEVINDGLLLTEINKNTTIDEIRSLTAADLLISNELRPMAV EC 4.1.1.4 C. acetobutylicum Adc, Aad Adc (SEQ ID NO: 123) MLKDEVIKQISTPLTSPAFPRGPYKFHNREYFNIVYRTDMDALRKVVPEPLEIDEP LVRFEIMAMHDTSGLGCYTESGQAIPVSFNGVKGDYLHMMYLDNEPAIAVGRELSAYP KKLGYPKLFVDSDTLVGTLDYGKLRVATATMGYKHKALDANEAKDQICRPNYMLKIIP NYDGSPRICELINAKITDVTVHEAWTGPTRLQLFDHAMAPLNDLPVKEIVSSSHILADIILP RAEVIYDYLK Aad (SEQ ID NO: 124) MLKDEVIKQISTPLTSPAFPRGPYKFHNREYFNIVYRTDMDALRKVVPEPLEIDEP LVRFEIMAMHDTSGLGCYTESGQAIPVSFNGVKGDYLHMMYLDNEPAIAVGRELSAYP KKLGYPKLFVDSDTLVGTLDYGKLRVATATMGYKHKALDANEAKDQICRPNYMLKIIP NYDGSPRICELINAKITDVTVHEAWTGPTRLQLFDHAMAPLNDLPVICEIVSSSHILADIILP RAEVIYDYLK EC 1.2.1.43 Formate dehydrogenase (M. thermoacetica) Moth_2312 (SEQ ID NO: 125) MVNLTIDGQRVTAPEGMTILEVARENGIHIPTLCHHPKLRPLGYCRLCLVDIEGAA KPMTACNTPVAEGMVIRTSTPVIEEMRKGIIEMLLSLHPEDCLTCEKAGNCQLQDCAYT YGVKHGELPVKREELPVLKENPFIVRDYNKCIVCGRCVRACQEVQVQRVVDLVGKGSA ARVGATKAGAEVSLEEGGCVFCGNCVQVCPVGALTEKAGLGQGREWEFKKVRSICSYC GVGCNLTLYVKDGKVVKVRGYENPEVNNGWLCVKGRFGFDYIHNPDRITRPLIREGDR EKGYFREASWEEALALVSQKLTQIKGSYGSEALGFLCSAKCTNEENYLLQKLARGVLGT NNVDHCARLHSSTVAGLATTFGSGAMTNSIADIASADCIFVIGSNTTENHPVIALKVKEA VRRGARUVADPRRIELVNFSYLWLRQKPGTDLALLNGLLHVIIKEELYDKEFIAQRTEGF EALKLAVEEYTPAKVSEVTGVPAGDIIEAARTYARGPSSTILYAMGITQHITGTANVMAL ANLAMACGQVGKEGSGVNPLRGQSNVQGACDMGGLPNVLPGYQPVTDPGVRHKFSEA WGVPDLPGEPGLTLMEMMAAAQEGKLKGMYILGENPVLTDPDVSHVKEALKNLEFLV VQDIFLTETARMADVVLPGASFAEKEGTFTSTERRVQLLHKAIEPPGEARPDWLILNDLL LLMGYPRKYSSPGEIMQEIAGLTPSYAGITYERLEDKGLQWPVLSLEHPGTPVLHREKFS RGYGQFQVVHYRPPAEEPDEEYPFLFTTGRNLYHYHTVTSRKSRGLEEMCPAPVVEINDN DAARLGIREGEMIEIVSRRGKVRVKALVTDRIPRGQVFMNFHFHEAAANLLTIAALDPVA KIPEYKTCAVAIKVKK Proteins sequences for Saccharomyces cerevisae engineering EC 4.2.3.3 Oryza sativa-mgs (SEQ ID NO: 126) MEUTRTIAERKHIALVAHDHRKQALLEWVESHKTILAQHQLYATGTTGNLIQR ASGIPVTSMLSGPMGGDQQVGALIAEGKIDMLIFFWDPLNAVPHDPDVKAILRLATVW NIPVATNRSTADFLIDSPLFKSEVAIAIPDYQRYLQDRLK EC 2.3.1.8 T. saccharolyticum-or1741 (SEQ ID NO: 127) MKTSELLAMVVEKGASDLHITVGVPPVLRINGQLIKLNLPQLTPQDTEEITKDLLS SDELKKLEDMGDIDLSYSVKGLGRFRINAYKQRGTYSLAIRSVALRIPTIDELGLPEVIKE LALKTRGLIIVTGPTGSGKSTTLASMIDLINEERNCHILTLEDPIEYLHKHKKSIVNQREIG HDAASYASALRAALREDPDVILVGEMRDLETIQIAITAAETGHLVLSTLHTIGSAKTIDRII DVFPPHQQQQIKVQLSNVLEGIVSQQLLPKIDNSGRWAVEVMIATPAIRNLIREGKSFQI QSMVQTGNKFGMVTMDMWISQLLKRNLISMDDALTYCVDRENFSRLVV EC 1.1.1.6 Pseudomonas putida gldA (SEQ ID NO: 128) MDRAIQSPGKYVQGADALQRLGDYLKPLADSWLVIADKFVLGFAEDTIRQSLSK AGLAMDIVAFNGECSQGEVDRLCQLATQNGRSAIVGIGGGKTLDTAKAVAFFQKVPVA VAPTIASTDAPCSALSVLYTDEGEFDRYLMLPTNPALVVVDTAIVARAPARLLAAGIGDA LATWFEARAASRSSAATMAGGPATQTALNLARFCYDTLLEEGEKAMLAVQAQWTPA LERIVEANTYLSGVGFESGGVAAAHAVHNGLTAVAETHHFYHGEKVAFGVLVQLALEN ASNAEMQEVMSLCHAVGLPITLAQLDITEDIPTKMRAVAELACAPGETIHNMPGGVTVE QVYGALLVADQLGQHFLEF EC 2.7.2.1 T. saccharolyticum or1742 (SEQ ID NO: 129) MIKKKLGDLLVEVGLLDESQLNNAIKIQKKTGEKXGKILVKEGYLTEEQIIEALEF QLGIPHIDMKKVFIDANVAKLIPESMAKRHVAIPIKKENNSIFVAMADPLNIFAIDDIKLVT KLDVKPLIASEDGILKAIDRVFGKEEAERAVQDFKKELSHDSAEDDGNLLRDISEDEINN APAVRLVNSIIEQAVKNRASDVHIEPTENDLRIRFRIDGELHEAMRVFKSTQGPVITRIKIM ANMNIAERRIPQDGKIEMNAGGKNIDIRVSSLPTIYGEKLVLRILDKSGYIITKDKLGLGN DDLKLFDNLLKHPNGIILLTGPTGSGKTTTLYAMLNELNKPDKNIITVFDPVEYTLEGLN QVQVNEKAGLTFASALRSILRQDPDIIMIGEIRDRETAEIAIRSSITGHLVLSTLHTNDSAG AITRLIDMGIEPYLVSSSVVGVIAQRLARKICDNCKIEYDASKREKHLGIDADESLKLYRS KGCAVCNKTGYRGRVPIYEIMMMTPKIKELTNEKAPADVILNEAVSNGMSTLKESAKKL VLSGVTTVDEMLRLTYDDAY

EC 2.8.3.8 C. acelobulylicum CtfAB CtfA (SEQ ID NO: 130) MNSKIIRFENLRSFFKDGMTIMIGGFLNCGTPTKLIDFLVNLNIKNLTIISNDTCYPN TGIGKLISNNQVKKLIASYIGSNPDTGKKLFNNELEVELSPQGTLVERIRAGGSGLGGVLT KTGLGTLIEKGKKKISINGTEYLLELPLTADVALIKGSIVDEAGNTFYKGTTKNFNPYMA MAAKTVIVEAENLVSCEKLEKEKAMTPGVLINYIVKEPA CtfB (SEQ ID NO: 131) MINDKNLAKEIIAKRVARELKNGQLVNLGVGLPTMVADYIPKNFKITFQSENGIV GMGASPKINEADKDVVNAGGDYTTVLPDGTFFDSSVSFSLIRGGHVDVTVLGALQVDE KGNIANWIVPGKMLSGMGGAMDLVNGAKKVIIAMRHTNKGQPKILKKCTLPLTAKSQA NLIVTELGVIEVINDGLLLTEINKNTTIDEIRSLTAADLLISNELRPMAV EC 4.1.1.4 C. acetobutylicum-Adc (SEQ ID NO: 132) MLKDEVIKQISTPLTSPAFPRGPYKFHNREYFNIVYRTDMDALRKVVPEPLEIDEP LVRFEIMAMHDTSGLGCYTESGQAIPVSFNGVKGDYLHMMYLDNEPAIAVGRELSAYP KKLGYPKLFVDSDTLVGTLDYGKLRVATATMGYKHKALDANEAKDQICRPNYMLKIIP NYDGSPRICELINAKITDVTVHEAWTGPTRLQLFDHAMAPLNDLPVKEIVSSSHILADIILP RAEVIYDYLK EC 2.3.1.54 Escherichia coli-pflA (SEQ ID NO: 133) MSVIGRIHSFESCGTVDGPGIRFITFFQGCLMRCLYCHNRDTWDTHGGKEVTVED LMKEVVTYRHFMNASGGGVTASGGEAILQAEFVRDWFRACKKEGIHTCLDTNGFVRRY DPVIDELLEVTDLVMLDLKQMNDEIHQNLVGVSNHRTLEFAKYLANKNVKVWIRYVVV PGWSDDDDSAHRLGEFTRDMGNVEKIELLPYHELGKHKWVAMGEEYKLDGVKPPKKE TMERVKGILEQYGHKVMF EC 2.3.1.54 Escherichia coli-pflB (SEQ ID NO: 134) MSELNEKLATAWEGFTKGDWQNEVNVRDFIQKNYTPYEGDESFLAGATEATTT LWDKVMEGVKLENRTHAPVDFDTAVASTITSHDAGYINKQLEKIVGLQTEAPLKRALIP FGGIKMIEGSCKAYNRELDPMIKKIFTEYRKTHNQGVFDVYTPDILRCRKSGVLTGLPDA YGRGRIIGDYRRVALYGIDYLMKDKLAQFTSLQADLENGVNLEQTIRLREEIAEQHRAL GQMKEMAAKYGYDISGPATNAQEAIQWTYFGYLAAVKSQNGAAMSFGRTSTFLDVYIE RDLKAGKITEQEAQEMVDHLVMKLRMVRFLRTPEYDELFSGDPIWATESIGGMGLDGR TLVTKNSFRFLNTLYTMGPSPEPNMTILWSEKLPLNFKKFAAKVSIDTSSLQYENDDLMR PDFNNDDYAIACCVSPMIVGKQMQFFGARANLAKTMLYAINGGVDEKLKMQVGPKSEP IKGDVLNYDEVMERMDHFMDWLAKQYITALNIIHYMHDKYSYEASLMALHDRDVIRT MACGIAGLSVAADSLSAIKYAKVKPIRDEDGLAIDFEIEGEYPQFGNNDPRVDDLAVDLV ERFMKKIQKLHTYRDAIPTQSVLTITSNVVYGKKTGNTPDGRRAGAPFGPGANPMHGRD QKGAVASLTSVAKLPFAYAKDGISYTFSIVPNALGKDDEVRKTNLAGLMDGYFHHEASI EGGQHLNVNVMNREMLLDAMENPEKYPQLTIRVSGYAVRFNSLTKEQQQDVITRTFTQ SM EC 2.3.1.9 Saccharomyces cerevisiae ERG10 (SEQ ID NO: 135) MSQNVYIVSTARTPIGSFQGSLSSKTAVELGAVALKGALAKVPELDASKDFDEIIF GNVLSANLGQAPARQVALAAGLSNHIVASTVNKVCASAMKAIILGAQSIKCGNADVVV AGGCESMTNAPYYMPAARAGAKFGQTVLVDGVERDGLNDAYDGLAMGVHAEKCARD WDITREQQDNFAIESYQKSQKSQKEGKFDNEIVPVTIKGFRGKPDTQVTKDEEPARLHVE KLRSARTVFQKENGTVTAANASPINDGAAAVILVSEKVLKEKNLKPLAIIKGWGEAAHQ PADFTWAPSLAVPKALKHAGIEDINSVDYFEFNEAFSVVGLVNTKILKLDPSKVNVYGG AVALGHPLGCSGARVVVTLLSILQQEGGKIGVAAICNGGGGASSIVIEKI EC 1.1.1.1 Saccharomyces cerevisiae ADH1 (SEQ ID NO: 136) MSIPETQKGVIFYESHGKLEYKDIPVPKPKANELLINVKYSGVCHTDLHAWHGD WPLPVKLPLVGGHEGAGVVVGMGENVKGWKIGDYAGIKWLNGSCMACEYCELGNES NCPHADLSGYTHDGSFQQYATADAVQAAHIPQGTDLAQVAPILCAGITVYKALKSANL MAGHWVAISGAAGGLGSLAVQYAKAMGYRVLGIDGGEGKEELFRSIGGEVFIDFTKEK DIVGAVLKATDGGAHGVINVSVSEAAIEASTRYVRANGTTVLVGMPAGAKCCSDVFNQ VVKSISIVGSYVGNRADTREALDFFARGLVKSPIKVVGLSTLPEIYEKMEKGQIVGRYVV DTSK EC 1.1.1.1 Saccharomyces cerevisiae ADI-12 (SEQ ID NO: 137) MSIPETQKAIIFYESNGKLEHKDIPVPKPKPNELLINVKYSGVCHTDLHAWHGDW PLPTKLPLVGGHEGAGVVVGMGENVKGWKIGDYAGIKWLNGSCMACEYCELGNESNC PHADLSGYTHDGSFQEYATADAVQAAHIPQGTDLAEVAPILCAGITVYKALKSANLRAG HWAAISGAAGGLGSLAVQYAKAMGYRVLGIDGGPGKEELFTSLGGEVFIDFTKEKDIVS AVVKATNGGAHGIINVSVSEAAIEASTRYCRANGTVVLVGLPAGAKCSSDVFNHWKSI SIVGSYVGNRADTREALDFFARGLVKSPIKVVGLSSEPEIYEKMEKGQIAGRYVVDTSK EC 1.1.1.1 Saccharomyces cerevisiae ADH3 (SEQ ID NO: 138) MLRTSTLFTRRVQPSLFSRNILRLQSTAAIPKTQKGVIFYENKGKLHYKDIPVPEPK PNEILINVKYSGVCHTDLHAWHGDWPLPVKLPLVGGHEGAGVVVKLGSNVKGWKVGD LAGIKWLNGSCMTCEFCESGHESNCPDADLSGYTHDGSFQQFATADAIQAAKIQQGTDL AEVAPILCAGVTVYKALKEADLKAGDWVAISGAAGGLGSLAVQYATAMGYRVLGIDA GEEKEKLFKKLGGEVF1DFTKTKNMVSDIQEATKGGPHGVTNVSVSEAAISLSTEYVRPC GTVVLVGLPANAYVKSEVFSHVVKSINIKGSYVGNRADTREALDFFSRGLIKSPIKIVGLS ELPKVYDLMEKGKILGRYVVDTSK EC 1.1.1.1 Saccharomyces cerevisiae ADH4 (SEQ ID NO: 139) MSSVTGFYIPPISFFGEGALEETADYIKNKDYKKALIVTDPGIAAIGLSORVQKML EERDLNVAIYDKTQPNPNIANVTAGLKVLKEQNSEIWSIGGGSAHDNAKAIALLATNG GEIGDYEGVNQSKKAALPLFAINTTAGTASEMTRFTIISNEEKKIKMAIIDNNVTPAVAVN DPSTMFGLPPALTAATGLDALTHCIEAYVSTASNPITDACALKGIDLINESLVAAYKDGK DKKARTDMCYAEYLAGMAFNNASLGYVHALAHQLGGFYHLPHGVCNAVLLPHVQEA NMQCPKAKKRLGEIALHFGASQEDPEETIKALHVLNRTMNIPRNLKELGVKTEDFEILAE HAMHDACHLTNPVQFTKEQVVAIIKKAYEY EC 1.1.1.1 Saccharomyces cerevisiae ADH5 (SEQ ID NO: 140) MPSQVIPEKQKAIVFYETDGKLEYKDVTVPEPKPNEILVHVKYSGVCHSDLHAW HGDWPFQLKFPLIGGHEGAGWVKLGSNVKGWKVGDFAGIKWLNGTCMSCEYCEVGN ESQCPYLDGTGFTHDGTFQEYATADAVQAAHIPPNVNLAEVAPILCAGITVYKALKRAN VIPGQWVTISGACGGLGSLAIQYALAMGYRVIGIDGGNAKRKLFEQLGGEIFIDFTEEKDI VGAIIKATNGGSHGVINVSVSEAAIEASTRYCRPNGTVVLVGMPAHAYCNSDVFNQVVK SISIVGSCVGNRADTREALDFFARGLIKSPIHLAGLSDVPEIFAKMEKGEIVGRYVVETSK EC 1.1.1.1 Saccharomyces cerevisiae ADH6 (SEQ ID NO: 141) MSYPEKFEGIAIQSHEDWKNPKKTKYDPKPFYDHDIDIKIEACGVCGSDIHCAAG HWGNMKMPLVVGHEIVGKVVKLGPKSNSGLKVGQRVGVGAQVFSCLECDRCKNDNEP YCTKFVTTYSQPYEDGYVSQGGYANYVRVHEHFVVPIPENIPSHLAAPLLCGGLTVYSPL VRNGCGPGKKVGIVGLGGIGSMGTLISKAMGAETYVISRSSRKREDAMKMGADHYIAT LEEGDWGEKYFDTFDLIVVCASSLTDIDFNIMPKAMKVGGRIVSISIPEQHEMLSLKPYGL KAVSISYSALGSIKELNQLLKLVSEKDIKIWVETLPVGEAGVHEAFERMEKGDVRYRFTL VGYDKEFSD EC 1.1.1.1 Saccharomyces cerevisiae ADH7 (SEQ ID NO: 142) MLYPEKFQGIGISNAKDWKHPKLVSFDPKPFGDHDVDVEIEACGICGSDFHIAVG NWGPVPENQILGHEIIGRVVKVGSKCHTGVKIGDRVGVGAQALACFECERCKSDNEQYC TNDHVLTMWTPYKDGYISQGGFASHVRLHEHFAIQIPENIPSPLAAPLLCGGITVFSPLLR NGCGPGKRVGIVGIGGIGHMGILLAKAMGAEVYAFSRGHSKREDSMKLGADHYIAMLE DKGWTEQYSNALDLLVVCSSSLSKVNFDSIVKIMKIGGSIVSIAAPEVNEKLVLKPLGLM GVSISSSAIGSRKEIEQLLKLVSEKNVKIWVEKLPISEEGVSHAFTRMESGDVKYRFTLVD YDKKFHK EC 1.1.1.1 Saccharomyces cerevisiae BDH2 (SEQ ID NO: 143) MRALAYFGKGNIRFTNHLKEPHIVAPDELVIDIEWCGICGTDLHEYTDGPIFFPED GHTHEISHNPLPQAMGHEMAGTVLEVGPGVKNLKVGDKVVVEPTGTCRDRYRWPLSP NVDKEWCAACKKGYYNICSYLGLCGAGVQSGGFAERVVMNESHCYKVPDFVPLDVAA LIQPLAVCWHAIRVCEFKAGSTALIIGAGPIGLGTILALNAAGCKDIVVSEPAKVRRELAE KMGARVYDPTAHAAKESIDYLRSIADGGDGFDYTFDCSGLEVTLNAAIQCLTFRGTAVN LAMWGHHKIQFSPMDITLHERKYTGSMCYTHHDFEAVIEALEEGRIDIDRARHMITGRV NIEDGLDGAIMKLINEKESTIKIILTPNNHGELNREADNEKKEISELSSRKDQERLRESINE AKLRHT Saccharomyces cerevisiae SFA1 (SEQ ID NO: 144) MSAATVGKPIKCIAAVAYDAKKPLSVEEITVDAPKAHEVRIKIEYTAVCHTDAYT LSGSDPEGLFPCVLGHEGAGIVESVGDDVITVKPGDHVIALYTAECGKCKFCTSGKTNLC GAVRATQGKGVMPDGTTRFHNAKGEDIYHFMGCSTFSEYTVVADVSVVAIDPKAPLDA ACLLGCGVTTGFGAALKTANVQKGDTVAVFGCGTVGLSVIQGAKLRGASKIIAIDINNK KKQYCSQFGATDFVNPKEDLAKDQTIVEKLIEMTDGGLDFTFDCTGNTKIMRDALEACH KGWGQSIIIGVAAAGEEISTRPFQLVTGRVWKGSAFGGIKGRSEMGGLIKDYQKGALKV EEFITHRRPFKEINQAFEDLHNGDCLRTVLKSDEIK EC 1.1.1.1 Saccharomyces cerevisiae YPL088W (SEQ ID NO: 145) MVLVKQVRLGNSGLKISPIVIGCMSYGSKKWADWVIEDKTQIFKIMKHCYDKGL

RTFDTADFYSNGLSERIIKEFLEYYSIKRETVVIMTKIYFPVDETLDLHHNFTLNEFEELDL SNQRGLSRKHIIAGVENSVKRLGTYIDLLQIHRLDHETPMKEIMKALNDVVEAGHVRYIG ASSMLATEFAELQFTADKYGWFQFISSQSYYNLLYREDERELIPFAKRHNIGLLPWSPNA RGMLTRPLNQSTDRIKSDPTFKSLHLDNLEEEQKEIINRVEKVSKDKKVSMAMLSIAWVL HKGCHPIVGLNTTARVDEAIAALQVTITEEEIKYLEEPYKPQRQRC* EC 4.1.2.13 Saccharomyces cerevisiae FBA1 (SEQ ID NO: 146) MGVEQILKRKTGVIVGEDVHNLFTYAKEHKFAIPAINVTSSSTAVAALEAARDSK SPIILQTSNGGAAYFAGKGISNEGQNASIKGAIAAAHYIRSIAPAYGIPWLHSDHCAKKL LPWFDGMLEADEAYFKEHGEPLFSSHMLDLSEETDEENISTCVKYFICRMAAMDQWLEM EIGITGGEEDGVNNENADKEDLYTKPEQVYNVYKLALHPISPNFSIAAAFGNCHGLYAGDI ALRPEILAEHQKYTREQVGCKEEKPLFLVFHGGSGSTVQEFHTGIDNGVVKVNLDTDCQ YAYLTGIRDYVLNKKDYIMSPVGNPEGPEKPNKKFFDPRVWVREGEKTMGAKITKSLET FRTTNTL EC 5.3.1.1 Saccharomyces cerevisiae TPI1 (SEQ ID NO: 147) MARTFFVGGNFKLNGSKQSIKEIVERLNTASIPENVEVVICPPATYLDYSVSLVKK PQVTVGAQNAYLKASGAFTGENSVDQIKDVGAKWVILGHSERRSYFHEDDKFIADKTK FALGQGVGVILCIGETLEEKKAGKTLDVVERQLNAVLEEVKDWTNVWAYEPVWAIGT GLAATPEDAQDIHASIRKFLASKLGDKAASELRILYGGSANGSNAVTFKDKADVDGFLV GGASLKPEFVDIINSRN EC 1.2.1.2 Saccharomyces cerevisiae FDH1 (SEQ ID NO: 148) MSKGKVLLVLYEGGKHAEEQEKLLGCIENELGIRNFIEEQGYELVTTIDKDPEPTS TVDRELKDAEIVITTPFFPAYISRNRIAEAPNLKLCVTAGVGSDHVDLEAANERKITVTEV TGSNVVSVAEHVMATILVLIRNYNGGHQQAINGEWDIAGVAKNEYDLEDKIISTVGAGR IGYRVLERLVAFNPKKLLYYDYQELPAEAINRLNEASKLFNGRGDIVQRVEKLEDMVAQ SDVVTINCPLHKDSRGLFNKKLISHMKDGAYLVNTARGAICVAEDVAEAVKSGKLAGY GGDVWDKQPAPKDHPWRTMDNKDHVGNAMTVHISGTSLDAQKRYAQGVKNILNSYF SKKFDYRPQDIIVQNGSYATRAYGQKK EC 1.1.1.21 Saccharomyces cerevisiae GRE3 (SEQ ID NO: 149) MSSLVTLNNGLKMPLVGLGCWKIDKKVCANQIYEAIKLGYRLFDGACDYGNEK EVGEGIRKAISEGLVSRKDIFWSKLWNNFHHPDHVKLALKKTLSDMGLDYLDLYYIHF PIAFKYVPFEEKYPPGFYTGADDEKKGHITEAHVPIIDTYRALEECVDEGLIKSIGVSNFQ GSLIQDLLRGCRIKPVALQIEHHPYLTQEHLVEFCKLHDIQVVAYSSFGPQSFIEMDLQLA KTTPTLFENDVIKKVSQNHPGSTTSQVLLRWATQRGIAVIPKSSKKERLLGNLEIEKKFTL TEQELKDISALNANIRFNDPWTWLDGKFPTFA EC 1.1.1.79 Saccharomyces cerevisiae GOR1 (SEQ ID NO: 150) MSKKPIVLKLGKDAFGDQAWGELEKIADVITIPESTTREQFLREVKDPQNKLSQV QVITRTARSVKNTGRFDEELALALPSSVVAVCHTGAGYDQIDVEPFKKRHIQVANVPDL VSNATADTHVFLLLGALRNFGIGNRRLIEGNWPEAGPACGSPFGYDPEGKTVGILGLGRI GRCILERLKPFGFENFIYHNRHQLPSEEEHGCEYVGFEEFLKRSDIVSVNVPLNHNTHHLI NAETIEKMKDGVVIVNTARGAVIDEQAMTDALRSGKIRSAGLDVFEYEPKISKELLSMSQ VLGLPHMGTHSVETRKKMEELWENAKNVILTGKVLTIVPELQNEDWPNESKPLV EC 1.1.1.79 Saccharomyces cerevisiae YPL113C (SEQ ID NO: 151) MITSIDIADVTYSAKPRILVPYKTQWEVASHLPEYRKLAERVEFYKYEMSTKDDF VKFLETHRINGFWLTEEFFTVLGNPSSYIEFFPASLKVILVPWVGCDFIDGKLLRSKGITLC NIGPHAADHVTELAIFLAISCFRMTSFWEYCFKYVENGNVEQCKKYISSDSYEIVTDSYH GQEMKFPSRTDKCKPNKDRKVVHLAEKYTVGGKKMESPMNKKVLILGFGSIGQNIGSN LHKVFNMSIEYYKRTGPVQKSLLDYNAKYHSDLDDPNTWKNADLIILALPSTASTNN1IN RKSLAWCKDGVRIVNVGRGTCIDEDVLLDALESGKVASCGLDVFKNEETRVKQELLRR WDVTALPHIGSTVADMVIKQTHTLENVQDIFVEGGDGKYVLN EC 1.2.1.49 Saccharomyces cerevisiae GCY1 (SEQ ID NO: 152) MPATLHDSTKILSLNTGAQIPQIGLGTWQSKENDAYKAVLTALKDGYRHIDTAAI YRNEDQVGQAIKDSGVPREEIFVTTKLWCTQHHEPEVALDQSLKRLGLDYVDLYLMHW PARLDPAYIKNEDILSVPTKKDGSRAVDITNWNFIKTWELMQELPKTGKTKAVGVSNFSI NNLKDLLASQGNKLTPAANQVEIHPLLPQDELINFCKSKGIVVEAYSPLGSTDAPLLKEP VILEIAKKNNVQPGHVVISWHVQRGYVVLPKSVNPDRIKTNRKIFTLSTEDFEAINN1SKE KGEKRVVHPNWSPFEVFK EC 1.2.1.49 Saccharomyces cerevisiae ALD2 (SEQ ID NO: 153) MPTLYTDIEIPQLKISLKQPLGLFINNEFCPSSDGKTIETVNPATGEPITSFQAANEK DVDKAVKAARAAFDNVWSKTSSEQRGIYLSNLLKLIEEEQDTLAALETLDAGKPYSNAK GDLAQILQLTRYFAGSADKFDKGATIPLTFNKFAYTLKVPFGVVAQIVPWNYPLAMAC WKLQGALAAGNTVIIKPAENTSLSLLYFATLIKKAGFPPGWNIVPGYGSLVGQALASH MDIDKISFTGSTKVGGFVLEASGQSNLKDVTLECGGKSPALVFEDADLDKAIDWIAAGIF YNSGQNCTANSRVYVQSSIYDKFVEKFKETAKKEWDVAGKFDPFDEKCIVGPVISSTQY DRIKSYIERGKREEKLDMFQTSEFPIGGAKGYFIPPTIFTDVPQTSKLLQDEIFGPVVVVSK FTNYDDALKLANDTCYGLASAVFTKDVKKAHMFARDIKAGTVWINSSNDEDVTVPFGG FKMSGIGRELGQSGVDTYLQTKAVFIINLSLDN EC 1.2.1.49 Saccharomyces cerevisiae ALD3 (SEQ ID NO: 154) MPTLYTDIEIPQLKISLKQPLGLFINNEFCPSSDGKTIETVNPATGEPITSFQAANEK DVDKAVKAARAAFDNVWSKTSSEQRGIYLSNLLKLIEEEQDTLAALETLDAGKPFHSNA KQDLAQIIELTRYYAGAVDKFNMGETIPLTFNKFAYTLKVPFGVVAQIVPWNYPLAMAC RKMQGALAAGNTVIIKPAENTSLSLLYFATLIKKAGFPPGVVNVIPGYGSVVGKALGTH MDIDKISFTGSTKVGGSVLEASGQSNLKDITLECGGKSPALVFEDADLDKAIEWVANGIF FNSGQICTANSRVYVQSSIYDKFVEKFKETAKKEWDVAGKFDPFDEKCIVGPVISSTQYD RIKSYIERGKKEEKLDMFQTSEFPIGGAKGYFIPPTIFTDVPETSKLLRDEIFGPVWVSKFT NYDDALKLANDTCYGLASAVFTKDVKKAHMFARDIKAGTVWINQTNQEEAKVPFGGF KMSGIGRESGDTGVDNYLQIKSVHVDLSLDK EC 1.2.1.49 Saccharomyces cerevisiae ALD4 (SEQ ID NO: 155) MFSRSTLCLKTSASSIGRLQLRYFSHLPMTVPIKLPNGLEYEQPTGLFINNKFVPSK QNKTFEVINPSTEEEICHIYEGREDDVEEAVQAADRAFSNGSWNGIDPIDRGKALYRLAE LIEQDKDVIASIETLDNGKAISSSRGDVDLVINYLKSSAGFADKIDGRMIDTGRTHFSYTK RQPLGVCGQIIPWNFPLLMWAWKIAPALVTGNTVVLKTAESTPLSALYVSKYIPQAOIPP GVINIVSGFGKIVGEAITNHPKIKKVAFTGSTATGRHIYQSAAAOLKKVTLELGGKSPNIV FADAELKKAVQNIILGIYYNSGEVCCAGSRVYVEESIYDKFIEEFKAASESIKVGDPFDES TFQGAQTSQMQLNKILKYVDIGKNEGATLITGGERLGSKGYFIKPTVFGDVKJEDMRIVK EEIFGPVVTVTKFKSADEVINMANDSEYGLAAGIHTSNINTALKVADRVNAGTVWINTY NDFHHAVPFGGFNASGLGREMSVDALQNYLQVKAVRAKLDE EC 1.2.1.49 Saccharomyces cerevisiae ALD5 (SEQ ID NO: 156) MLSRTRAAAPNSRIFTRSLLRLYSQAPLRVPITLPNGFTYEQPTGLFINGEFVASKQ KKTFDVINPSNEEKITTVYKAMEDDVDEAVAAAKKAFETKWSIVEPEVRAKALFNLADL VEKHQETLAAIESMDNGKSLFCARGDVALVSKYLRSCGGWADKIYGNVIDTGKNHFTY SIKEPLGVCGQIIPWNFPLLMWSWKIGPALATGNTVVEKPAETTPLSALFASQLCQEAGIP AGWNILPGSGRVVGERLSAHPDVKKIAFTGSTATGRHIMKVAADTVKKVTLELGGKSP NIVFADADLDKAVKNIAFGIFYNSGEVCCAGSRIYIQDTVYEEVLQKLKDYTESLKVGDP FDEEVFQGAQTSDKQLHKILDYVDVAKSEGARLVTGGARHGSKGYFVKPTVFADVKGD MRIVKEEVFGPIVTVSKFSTVDEVIAMANDSQYGLAAGIHTNDINKAVDVSKRVKAGTV WINTYNNFHQNVPFGGFGQSGIGREMGEAALSNYTQTKSVRIAIDKPIR EC 1.2.1.49 Saccharomyces cerevisiae ALD6 (SEQ ID NO: 157) MTKLHFDTAEPVKITLPNGLTYEQPTGLFINNKFMKAQDGKTYPVEDPSTENTVC EVSSATTEDVEYAIECADRAFHDTEWATQDPRERGRLLSKLADELESQIDLVSSIEALDN GKTLALARGDVTIAINCLRDAAAYADKVNGRTINTGDGYMNFTTLEPIGVCGQIIPWNFP IMMLAWKIAPALAMGNVCILKPAAVTPLNALYFASLCKKVGIPAGVVNIVPGPGRTVGA ALTNDPRIRKLAFTGSTEVGKSVAVDSSESNLKKITLELGGKSAHLVFDDANIKKTLPNL VNGIFKNAGQICSSGSRIYVQEGIYDELLAAFKAYLETEIKVGNPFDKANFQGAITNRQQF DTIMNYIDIGKKEGAKILTGGEKVGDKGYFIRPTVFYDVNEDMRIVKEEIFGPVVTVAKF KTLEEGVEMANSSEFGLGSGIETESLSTGLKVAKMLKAGTVWINTYNDFDSRVPFGGVK QSGYGREMGEEVYHAYTEVKAVRIKL EC 1.2.1.49 Saccharomyces cerevisiae HFD1 (SEQ ID NO: 158) MSNDGSKILNYTPVSKIDEIVEISRNFFFEKQLKLSHENNPRKKDLEFRQLQLKKL YYAVKDHEEELIDAMYKDFHRNKIESVLNETTKLMNDILHLIEILPKLIKPRRVSDSSPPF MFGKTIVEKISRGSVLIIAPFNFPLLLAFAPLAAALAAGNTIVLKPSELTPHTAWMENLLT TAGFPDGLIQVVQGAIDETTRLLDCGKFDLIFYTGSPRVGSIVAEKAAKSLTPCVLELGGK SPTFITENFKASNIKIALKRIFFGAFGNSGQICVSPDYLLVHKSIYPKVIKECESVLNEFYPS FDEQTDFTRMIHEPAYKKAVASINSTNGSKIVPSKISINSDTEDLCLVPPTIVYNIGWDDPL MKQENFAPVLPIIEYEDLDETINKIIEEHDTPLVQYIFSDSQTEINRILTRLRSGDCVVGDTV IHVGITDAPFGGIGTSGYGNYGGYYGFNTFSHERTIFKQPYWNDFTLFMRYPPNSAQKEK LVRFAMERKPWFDRNGNNKWGLRQYFSLSAAVILISTIYAHCSS EC 2.7.1.2 Saccharomyces cerevisiae GLK1 (SEQ ID NO: 159) MSFDDLHKATERAVIQAVDQICDDFEVTPEKLDELTAYFIEQMEKGLAPPKEGHT

LASDKGLPMIPAFVTGSPNGTERGVLLAADLGGTNFRICSVNLHGDHTFSMEQMKSKIP DDLLDDENVTSDDLFGFLARRTLAFMKICYHPDELAKGKDAKPMKLGFTFSYPVDQTSL NSGTLIRWTKGFRIADTVGKDVVQLYQEQLSAQGMPMIKWALTNDTVGTYLSHCYTS DNTDSMTSGEISEPVIGCIFGTGTNGCYMEEINKITKLPQELRDKLIKEGKTHMIINVEWG SFDNELKHLPTTKYDWIDQKLSTNPGFHLFEKRVSGMFLGEVLRNILVDLHSQGLLLQQ YRSKEQLPRHLTTPFQLSSEVLSHIEIDDSTGLRETELSLLQSLRLPTTPTERVQIQKLVRAI SRRSAYLAAVPLAAILIKTNALNKRYHGEVEIGCDGSWEYYPGFRSMLRHALALSPLG AEGERKVHLKIAKDGSGVGAALCALVA EC 5.3.1.9 Saccharomyces cerevisiae PGI1 (SEQ ID NO: 160) MSNNSFTNFKLATELPAWSKLQKIYESQGKTLSVKQEFQKDAKRFEKLNKTFTN YDGSKILFDYSKNLVNDEHAALIELAKEANVTGLRDAMFKGEHINSTEDRAVYHVALRN RANKPMYVDGVNVAPEVDSVLKHMKEFSEQVRSGEWKGYTGKKITDWNIGIGGSDLG PVMVTEALKHYAGVLDVHFVSNIDGTHIAETLKVVDPETTLFLIASKTFTTAETITNANT AKNWFLSKTGNDPSHIAKHFAALSTNETEVAKFGIDTKNMFGFESWVGGRYSVWSAIGL SVALYIGYDNFEAFLKGAEAVDNHFTQTPLEDNIPLLGGLLSVWYNNFFGAQTHLVAPF DQYLHRFPAYLQQLSMESNGKSVTRGNVFTDYSTGSILFGEPATNAQHSFFQLVHQGTK LIPSDFILAAQSHNPIENKLHQKMLASNFFAQAEALMVGKDEEQVKAEGATGGLVPHKV FSGNRPTTSILAQKITPATLGALIAYYEHVTFTEGAIWNINSFDQWGVELGKVLAKVIGKE LDNSSTISTHDASTNGLINQFKEWM EC 2.7.1.11 Saccharomyces cerevisiae PFK1 (SEQ ID NO: 161) MQSQDSCYGVAFRSIITNDEALFKKTIHFYHTLGFATVKDFNKFKHGENSLLSSGT SQDSLREVWLESFKLSEVDASGFRIPQQEATNKAQSQGALLKIRLVMSAPIDETFDTNET ATITYFSTDLNKIVEKFPKQAEKLSDTLVFLKDPMGNNITFSGLANATDSAPTSKDAFLEA TSEDEIISRASSDASDLLRQTLGSSQKKKKIAVMTSGGDSPGMNAAVRAVVRTGIPIFGCD VFAVYEGYEGLLRGGKYLKKMAWEDVRGWLSEGGTLIGTARSMEFRKREGRRQAAGN LISQGIDALWCGGDGSLTGADLFRHEWPSLVDELVAEGRFTKEEVAPYKNLSIVGLVGS IDNDMSGTDSTIGAYSALERICEMVDYIDATAKSHSRAFVVEVMGRHCGWLALMAGIA TGADY1FIPERAVPHGKWQDELKEVCQRHRSKGRRNNTIIVAEGALDDQLNPVTANDVK DALIELGLDTKVTILGHVQRGGTAVAHDRWLATLQGVDAVKAVLEFTPETPSPLIGILEN KIIRMPLVESVKLTKSVATAIENKDFDKAISLRDTEFIELYENFLSTTVKDDGSELLPVSDR LNIGIVHVGAPSAALNAATRAATLYCLSHGHKPYAIMNGFSGUQTGEVKELSWIDVEN WHNLGGSEIGTNRSVASEDLGTIAYYFQKNKLDGLIILGGFEGFRSLKQLRDGRTQHPIF NIPMCLIPATVSNNVPGTEYSLGVDTCLNALVNYTDDIKQSASATRRRVFVCEVQGGHS GYIASFTGLITGAVSVYTPEKKIDLASIREDITLLKENFRHDKGENRNGKLLVRNEQASSV YSTQLLADIISEASKGKFGVRTAIPGHVQQGGVPSSKDRVTASRFAVKCIKFIEQWNKKN EASPNTDAKVLRFKFDTHGEKVPTVEHEDDSAAVICVNGSHVSFKPIANLWENETNVEL RKGFEVHWAEYNKIGDILSGRLKLRAEVAALAAENK EC 2.7.1.11 Sacchciromyces cerevisiae PFK.2 (SEQ ID NO: 162) MTVTTPFVNGTSYCTVTAYSVQSYKAAIDFYTKFLSLENRSSPDENSTLLSNDSIS LKIIKRPDEKINKNVEAHLKFXNSITKTQDWRSHATQSLVFNTSDILAVKDTLNAMNAPL QGYPTELFPMQLYTLDPLGNVVGVTSTKNAVSTKPTPPPAPEASAESGLSSKVHSYTDLA YRMKTTDTYPSLPKPLNRPQKAIAVMTSGGDAPGMNSNVRAIVRSAIFKGCRAFVVME GYEGLVRGGPEYIKEFHWEDVRGWSAEGGTNIGTARCMEFKICREGRLLGAQHLIEAGV DALIVCGGDGSLTGADLFRSEWPSHEELLKTNRISNEQYERMKHLNICGTVGSIDNDMS TIDATIGAYSALDRICKAIDYVEATANSHSRAFVVEVMGRNCGWLALLAGIATSADYIFI PEKPATSSEWQDQMCDIVSKHRSRGICRTTIVVVAEGAIAADLTPISPSDVHKVLVDRLGL DTRITTLGHVQRGGTAVAYDRILATLQGLEAVNAVLESTPDTPSPLIAVNENKIVRKPLM ESVKLTKAVAEAIQAKDFKRAMSLRDTEFIEHLNNFMAINSADHNEPKLPKDKRLKIAIV NVGAPAGGINSAVYSMATYCMSQGHRPYAIYNGWSGLARHESVRSLNWKDMLGWQS RGGSEIGTNRVTPEEADLGMIAYYFQKYEFDGLIIVGGFEAFESLHQLERARESYPAFRIP MVLIPATLSNNVPGTEYSLGSDTALNALMEYCDWKQSASSTRGRAFVVDCQGGNSGY LATYASLAVGAQVSYVPEEGISLEQLSEDIEYLAQSFEKAEGRGRFGKLILKSTNASKALS ATKLAEVITAEADGRFDAKPAYPGHVQQGGLPSPIDRTRATRMAIKAVGFIKDNQAAIA EARAAEENFNADDKTISDTAAVVGVKGSHVVYNSIRQLYDYETEVSMRMPKVIHWQAT RLIADHLVGRKRVD EC 4.1.1.1 Sacchciromyces cerevisiae PDC1 (SEQ ID NO: 163) MSEITLGKYLFERLKQVNVNTVFGLPGDFNLSLLDKIYEVEGMRWAGNANELNA AYAADGYARIKGMSCITFGVGELSALNGIAGSYAEHVGVLHVVGVPSISAQAKQLLL HHTLGNGDFTVFHRMSANISETTAMITDIATAPAEIDRCIRTTYVTQRPVYLGLPANLVD LNVPAKXLQTPIDMSLKPNDAESEKEVIDTILALVKDAKNPVILADACCSRHDVKAETKK LIDLTQFPAFVTPMGKGSIDEQHPRYGGVYVGTLSKPEVKEAVESADLILSVGALLSDFN TGSFSYSYKTKNIVEFHSDHMKIRNATFPGVQMKFVLQKLLTTIADAAKGYKPVAVPAR TPANAAVPASTPLKQEWMWNQLGNFLQEGDVVIAETGTSAFGINQTTFPNNTYGISQVL WGSIGFTTGATLGAAFAAEEIDPKKRVILFIGDGSLQLTVQEISTMIRWGLKPYLFVLNND GYTIEKLIHGPKAQYNEIQGWDHLSLLPTFGAKDYETHRVATTGEWDKLTQDKSFNDNS KIRMIEIMLPVFDAPQNLVEQAKLTAATNAKQ EC 4.1.1.1 Sacchciromyces cerevisiae PDC5 (SEQ ID NO: 164) MSEITLGKYLFERLSQVNCNTVFGLPGDFNLSLLDKLYEVKGMRWAGNANELN AAYAADGYARIKGMSCTTTTFGVGELSALNGIAGSYAEHVGVLHVVGVPSISSQAKQLLL HHTLGNGDFTVFHRMSANISETTAMITDIANAPAEIDRCIRTTYTTQRPVYLGLPANLVD LNVPAKLLETPIDLSLKPNDAEAEAEVVRTVVELIKDAKNPVILADACASRHDVKAETK KLMDLTQFPVYVTPMGKGAIDEQHPRYGGVYVGTLSRPEVKKAVESADLILSIGALLSD FNTGSFSYSYKTKNIVEFHSDHIKIRNATFPGVQMKFALQKLLDAIPEVVKDYKPVAVPA RVPITKSTPANTPMKQEWMWNHLGNFLREGDIVIAETGTSAFGINQTTFPTDVYAIVQVL WGSIGFTVGALLGATMAAEELDPKKRVILFIGDGSLQLTVQEISTMIRWGLICPYIFVLNN NGYTIEKLIHGPHAEYNEIQGWDHLALLPTFGARNYETHRVATTGEWEKLTQDKDFQD NSKIRMIEVMLPVFDAPQNLVKQAQLTAATNAKQ EC 4.1.1.1 Saccharomyces cerevisiae PDC6 (SEQ ID NO: 165) MSEITLGKYLFERLKQVNVNTIFGLPGDFNLSLLDKIYEVDGLRWAGNANELNA AYAADGYARIKGLSVLVTTFGVGELSALNGIAGSYAEHVGVLHVVGVPSISAQAKQLLL HHTLGNGDFTVFHRMSANISETfSMITDIATAPSEIDRLIRTTFITQRPSYLGLPANLVDLK VPGSLLEKPIDLSLKPNDPEAEKEVIDTVLELIQNSKNPVILSDACASRHNVKKETQKLID LTQFPAFVTPLGKGSIDEQHPRYGGVYVGTLSKQDVKQAVESADLILSVGALLSDFNTGS FSYSYKTKNVVEFHSDYVKVKNATFLGVQMKFALQNLLKVIPDVVKGYKSVPVPTKTP ANKGVPASTPLKQEWLWNELSKFLQEGDVIISETGTSAFGINQTIFPKDAYGISQVLWGSI GFTTGATLGAAFAAEEIDPNKRVILFIGDGSLQLTVQEISTMIRWGLKPYLFVLNNDGYTI EKLIHGPHAEYNEIQTWDHLALLPAFGAKKYENHKIATTGEWDALTTDSEFQKNSVIRLI ELKLPVFDAPESLIKQAQLTAATNAKQ EC 1.1.1.8 Saccharomyces cerevisiae GPD2 (SEQ ID NO: 166) MLAVRRLTRYTFLKRTHPVLYTRRAYKILPSRSTFLRRSLLQTQLHSKMTAHTNI KQHKHCHEDHPIRRSDSAVSIVHLKRAPFKVTVIGSGNWGTTIAKVIAENTELHSHIFEPE VRMWVFDEKIGDENLTDIINTRIIQNVKYLPNIDLPHNLVADPDLLHSIKGADILVFNIPH QFLPNIVKQLQGHVAPHVRAISCLKGFELGSKGVQLLSSYVTDELGIQCGALSGANLAPE VAKEHWSETTVAYQLPKDYQGDGKDVDHKILKLLFHRPYFHVNVIDDVAGISIAGALK NVVALACGFVEGMGWGNNASAAIQRLGLGEIIKFGRMFFPESKVETYYQESAGVADLIT TCSGGRNVKVATYMAKTGKSALEAEKELLNGQSAQGIITCREVHEWLQTCELTQEFPLF EAVYQIVYNNVRMEDLPEMIEELDIDDE EC 3.1.3.21 Saccharomyces cerevisiae GPP1 (SEQ ID NO: 167) MPLTTKPLSLKINAALFDVDGTIIISQPAIAAFWRDFGKDKPYFDAEHVIHISHGW RTYDAIAKFAPDFADEEYVNKLEGEIPEKYGEHSIEVPGAVICLCNALNALPKEKWAVAT SGTRDMAKICWFDILKIKRPEYFITANDVKQGKPHPEPYLKGRNGLGFPINEQDPSKSKVV VFEDAPAGIAAGKAAGCKIVGIATTFDLDFLKEKGCDIIVKNHESIRVGEYNAETDEVELI FDDYLYAKDDLLKW

[0067] In certain embodiments, an enzyme of the present invention includes any enzyme that is at least about 70%, 80%, 90%, 95%, 99% identical, or sharing at least about 60%, 70%, 80%, 90%, 95% sequence identity to any of the enzymes of the metabolic engineered pathways as described above. These enzymes sharing the requisite sequence identity or similarity can be wild-type enzymes from a different organism, or can be artificial, i.e., recombinant, enzymes.

[0068] In certain embodiments, any genes encoding for enzymes with the same activity as any of the enzymes of the metabolicly engineered pathways as described above may be used in place of the enzymes. These enzymes may be wild-type enzymes from a different organism, or may be artificial, recombinant or engineered enzymes.

[0069] Additionally, due to the inherent degeneracy of the genetic code, other nucleic acid sequences which encode substantially the same or a functionally equivalent amino acid sequence can also be used to express the polynucleotide encoding such enzymes. As will be understood by those of skill in the art, it can be advantageous to modify a coding sequence to enhance its expression in a particular host. The codons that are utilized most often in a species are called "optimal codons", and those not utilized very often are classified as "rare or low-usage codons". Codons can be substituted to reflect the preferred codon usage of the host, a process sometimes called "codon optimization" or "controlling for species codon bias." Methodology for optimizing a nucleotide sequence for expression in, e.g. Saccharomyces cerevisiae, are known to one of ordinary skill in the art.

Modified Strains

[0070] The present invention further provides for knockout strains in which the metabolic engineered pathways of the invention are carried out. Such a genetically modified microorganism would have an increased ability to produce lactate or acetate as a fermentation product. "Knock out" of the genes means partial, substantial, or complete deletion, silencing, inactivation, or down-regulation.

[0071] Thus, certain embodiments of the present invention provide for the "inactivation" or "deletion" of certain genes or particular polynucleotide sequences within thermophilic, or mesophilic microorganisms, which "inactivation" or "deletion" of genes or particular pub/nucleotide sequences can be understood to encompass "genetic modification(s)" or "transformation(s)" such that the resulting strains of said thermophilic or mesophilic microorganisms can be understood to be "genetically modified" or "transformed." In certain embodiments, strains can be of bacterial, fungal, or yeast origin.

[0072] A genetically modified strain that is a knockout strain can have the advantage of eliminating the production of certain organic acids or products that interfere with the ability of the strain to generate a high yield of an alternative product, such as isopropanol or propanediol.

[0073] For example, if the conversion of pyruvate to lactate (the salt form of lactic acid) by the action of LDH was not available in the early stages of the glycolytic pathway, then the pyruvate could be more efficiently converted to acetyl CoA by the action of pyruvate dehydrogenase or pyruvate-ferredoxin oxidoreductase.

[0074] Genes to be targeted for knockout for the present invention include lactate dehydrogenase (ldh), hydrogenase (hyd), acetaldehyde dehydrogenase (acdh), acetate kinase (ack), pyruvate-ferredoxin oxidoreductase (por) or pyruvate decarboxylase (pdc).

[0075] As used herein, the term "lactate dehydrogenase" or "LDH" is intended to include the enzyme capable of converting pyruvate into lactate. It is understood that LDH can also catalyze the oxidation of hydroxybutyrate.

[0076] As used herein, the term "acetate kinase" or "ACK" is intended to include the enzyme capable of converting acetyl phosphate into acetate.

[0077] As used herein, the term "pyruvate-ferredoxin oxidoreductase" or "POR" is intended to include the enzyme capable of converting pyruvate into acetyl CoA, carbon dioxide, and reduced ferredoxin.

[0078] The term "pyruvate decarboxylase activity" is intended to include the ability of a polypeptide to enzymatically convert pyruvate into acetaldehyde (e.g., "pyruvate decarboxylase" or "PDC"). Typically, the activity of a selected polypeptide encompasses the total enzymatic activity associated with the produced polypeptide, comprising, e.g., the superior substrate affinity of the enzyme, thermostability, stability at different pHs, or a combination of these attributes.

[0079] Certain embodiments of the present invention, alternatively, provide for the "insertion," (e.g., the addition, integration, incorporation, or introduction) of certain genes or particular polynucleotide sequences within thermophilic or mesophilic microorganisms, which insertion or genes or particular polynucleotide sequences can be understood to encompass "genetic modification(s)" or "transformation(s)" such that the resulting strains of said thermophilic or mesophilic microorganisms can be understood to be "genetically modified" or "transformed." In certain embodiments, strains can be of bacterial, fungal, or yeast origin.

[0080] In one aspect, of the invention, the genes or particular polynucleotide sequences are inserted to activate the activity for which they encode, such as the expression of an enzyme. In certain embodiments, genes encoding enzymes in the metabolic production of ethanol, e.g., enzymes that metabolize pentose and/or hexose sugars, can be added to a mesophilic or thermophilic organism. In certain embodiments of the invention, the enzyme can confer the ability to metabolize a pentose sugar and be involved, for example, in the D-xylose pathway and/or L-arabinose pathway.

[0081] In one aspect of the invention, the genes or particular polynucleotide sequences are partially, substantially, or completely deleted, silenced, inactivated, or down-regulated in order to inactivate the activity for which they encode, such as the expression of an enzyme. Deletions provide maximum stability because there is no opportunity for a reverse mutation to restore function. Alternatively, genes can be partially, substantially, or completely deleted, silenced, inactivated, or down-regulated by insertion of nucleic acid sequences that disrupt the function and/or expression of the gene (e.g., P1 transduction or other methods known in the art). The terms "eliminate," "elimination," and "knockout" are used interchangeably with the terms "deletion," "partial deletion," "substantial deletion," or "complete deletion." In certain embodiments strains of thermophilic or mesophilic microorganisms of interest can be engineered by site directed homologous recombination to knockout the production of organic acids. In still other embodiments, RNAi or antisense DNA (asDNA) can be used to partially, substantially, or completely silence, inactivate, or down-regulate a particular gene of interest.

Vectors and Host Cells

[0082] The present invention also relates to vectors which include genes encoding for enzymes of tile present invention, as described above, as well as host cells which are genetically engineered with vectors of the invention and the production of polypeptides of the invention by recombinant techniques.

[0083] Host cells are genetically engineered (transduced or transformed or transfected) with the vectors of this invention which can be, for example, a cloning vector or an expression vector. The vector can be, for example, in the form of a plasmid, a viral particle, a phage, etc. The engineered host cells can be cultured in conventional nutrient media modified as appropriate for activating promoters, selecting transformants or amplifying the genes of the present invention. The culture conditions, such as temperature, pH and the like, are those previously used with the host cell selected for expression, and will be apparent to the ordinarily skilled artisan.

[0084] The DNA sequence in the expression vector is operatively associated with an appropriate expression control sequence(s) (promoter) to direct mRNA synthesis. Any suitable promoter to drive gene expression in the host cells of the invention can be used. Additionally, promoters known to control expression of genes in prokaryotic or lower eukaryotic cells can be used. The expression vector also contains a ribosome binding site for translation initiation and a transcription terminator. The vector can also include appropriate sequences for amplifying expression, or can include additional regulatory regions.

[0085] The vector containing the appropriate selectable marker sequence as used herein, as well as an appropriate promoter or control sequence, can be employed to transform an appropriate thermophilic host to permit the host to express the protein.

[0086] The terms "promoter" or "surrogate promoter" is intended to include a polynucleotide segment that can transcriptionally control a gene-of-interest that it does not transcriptionally control in nature. In certain embodiments, the transcriptional control of a surrogate promoter results in an increase in expression of the gene-of-interest. In certain embodiments, a surrogate promoter is placed 5' to the gene-of-interest. A surrogate promoter can be used to replace the natural promoter, or can be used in addition to the natural promoter. A surrogate promoter can be endogenous with regard to the host cell in which it is used, or it can be a heterologous polynucleotide sequence introduced into the host cell, e.g., exogenous with regard to the host cell in which it is used.

[0087] The terms "gene(s)" "polynucleotide segment" or "polynucleotide sequence(s)" are intended to include nucleic acid molecules, e.g., polynucleotides which include an open reading frame encoding a polypeptide, and can further include non-coding regulatory sequences, and introns. In addition, the terms are intended to include one or more genes that map to a functional locus. In addition, the terms are intended to include a specific gene for a selected purpose. The gene can be endogenous to the host cell or can be recombinantly introduced into the host cell, e.g., as a plasmid maintained episomally or a plasmid (or fragment thereof) that is stably integrated into the genome. In addition to the plasmid form, a gene can, for example, be in the form of linear DNA. In certain embodiments, the gene encodes a polypeptide, such as an enzyme of the present invention. The term gene is also intended to cover all copies of a particular gene, e.g., all of the DNA sequences in a cell encoding a particular gene product.

[0088] The term "transcriptional control" is intended to include the ability to modulate gene expression at the level of transcription. In certain embodiments, transcription, and thus gene expression, is modulated by replacing or adding a surrogate promoter near the 5' end of the coding region of a gene-of-interest, thereby resulting in altered gene expression. In certain embodiments, the transcriptional control of one or more gene is engineered to result in the optimal expression of such genes, e.g., in a desired ratio. The term also includes inducible transcriptional control as recognized in the art.

[0089] The term "expression" is intended to include the expression of a gene at least at the level of mRNA production.

[0090] The term "expression product" is intended to include the resultant product, e.g., a polypeptide, of an expressed gene.

[0091] The term "increased expression" is intended to include an alteration in gene expression at least at the level of increased mRNA production and, preferably, at the level of polypeptide expression. The term "increased production" is intended to include an increase in the amount of a polypeptide expressed, in the level of the enzymatic activity of the polypeptide, or a combination thereof.

[0092] In certain aspects, the present invention relates to host cells containing the above-described constructs. The host cell can be an anaerobic thermophilic bacterial cell, including an anaerobic xylanolytic and/or cellulolytic host cell. The selection of an appropriate host is deemed to be within the scope of those skilled in the art from the teachings herein.

[0093] The present invention also includes recombinant constructs comprising one or more of the selectable marker sequences as broadly described above. The constructs comprise a vector, such as a plasmid or viral vector, into which a sequence of the invention has been inserted, in a forward or reverse orientation. In one aspect of this embodiment, the construct further comprises regulatory sequences, including, for example, a promoter, operably associated to the sequence. Large numbers of suitable vectors and promoters are known to those of skill in the art, and are commercially available. The following vectors are provided by way of example only.

[0094] The term "derived from" is intended to include the isolation (in whole or in part) of a polynucleotide segment from an indicated source or the purification of a polypeptide from an indicated source. The term is intended to include, for example, direct cloning, PCR amplification, or artificial synthesis from or based on a sequence associated with the indicated polynucleotide source.

[0095] Introduction of the construct in host cells can be done using methods known in the art. Introduction can also be effected by electroporation methods as described in U.S. Prov. Appl. No. 61/109,642, filed Oct. 30, 2008, the contents of which are herein incorporated by reference.

[0096] Furthermore, the use of positive and/or negative selection markers, genetic tools, and homologous recombination-based genome integration adapted for use in, e.g., thermophilic organisms, that can be used to efficiently select modified strains, including modified strains of C. thermocellum and T. saccharolyticum can be done using methods as described in U.S. Prov. Appl. No. 61/232,648, filed Aug. 10, 2009, the contents of which are herein incorporated by reference. Methods for the expression of foreign genes, knockout and overexpression of native genes, and creation of clean industrial strains that do not contain antibiotic markers or other extraneous DNA can be performed, as described in U.S. Prov. Appl. No. 61/232,648.

Biomass

[0097] The terms "lignocellulosic material," "lignocellulosic substrate," and "cellulosic biomass" mean any type of biomass comprising cellulose, hemicellulose, lignin, or combinations thereof, such as but not limited to woody biomass, forage grasses, herbaceous energy crops, non-woody-plant biomass, agricultural wastes and/or agricultural residues, forestry residues and/or forestry wastes, paper-production sludge and/or waste paper sludge, waste-water-treatment sludge, municipal solid waste, corn fiber from wet and dry mill corn ethanol plants, and sugar-processing residues.

[0098] In a non-limiting example, the lignocellulosic material can include, but is not limited to, woody biomass, such as recycled wood pulp fiber, sawdust, hardwood, softwood, and combinations thereof; grasses, such as switch grass, cord grass, rye grass, reed canary grass, miscanthus, or a combination thereof; sugar-processing residues, such as but not limited to sugar cane bagasse; agricultural wastes, such as but not limited to rice straw, rice hulls, barley straw, corn cobs, cereal straw, wheat straw, canola straw, oat straw, oat hulls, and corn fiber; stover, such as but not limited to soybean stover, corn stover; succulent plants, such as but not limited to agave; and forestry wastes, such as but not limited to recycled wood pulp fiber, sawdust, hardwood (e.g., poplar, oak, maple, birch, willow), softwood, or any combination thereof. Lignocellulosic material can comprise one species of fiber; alternatively, lignocellulosic material can comprise a mixture of fibers that originate from different lignocellulosic materials. Particularly advantageous lignocellulosic materials are agricultural wastes, such as cereal straws, including wheat straw, barley straw, canola straw and oat straw; corn fiber; stovers, such as corn stover and soybean stover; grasses, such as switch grass, reed canary grass, cord grass, and miscanthus, or combinations thereof.

[0099] Paper sludge is also a viable feedstock for lactate or acetate production. Paper sludge is solid residue arising from pulping and paper-making, and is typically removed from process wastewater in a primary clarifier. At a disposal cost of $30/wet ton, the cost of sludge disposal equates to $5/ton of paper that is produced for sale. The cost of disposing of wet sludge is a significant incentive to convert the material for other uses, such as conversion to ethanol. Processes provided by the present invention are widely applicable. Moreover, the saccharification and/or fermentation products can be used to produce ethanol or higher value added chemicals, such as organic acids, aromatics, esters, acetone and polymer intermediates. During glycolysis, cells convert simple sugars, such as glucose, into pyruvic acid, with a net production of ATP and NADH. In the absence of a functioning electron transport system for oxidative phosphorylation at least 95% of the pyruvic acid is consumed in short pathways which regenerate NAD.sup.+, an obligate requirement for continued glycolysis and ATP production. The waste products of these NAD regeneration systems are commonly referred to as fermentation products.

EXEMPLIFICATION

Example 1

1.1 Production of Mixed Alcohols in Bacterial and Yeast CBP Platforms

[0100] Production of mixed alcohols in bacteria and yeast makes use of bacterial and yeast CBP platforms, and their available toolboxes, to produce a combination of propanediol, isopropanol, glycerol and ethanol. Trace amounts of microbially produced propanediol were first detected in 1954 during cultivation of Clostridium thermobutyricum. See Enebo L. 1954, "Studies in cellulose decomposition by an anaerobic thermophilic bacterium and two associated non-cellulolytic species," p. 94-96. Viktor Pettersons Bokindustrie Aktiebolag, Stockholm. Since then, reports have indicated native production of propanediol from common sugars during fermentations of C. sphenoides and T. thermosaccharolyticum. See Tran-Din, K., & Gottschalk, G., 1985, Arch. Microbiol. 142, 87-92; Cameron, D. C., Clooney, C., 1986, Bio/Technology 4, 651-654. Recombinant E. coli strains have been developed that produce propanediol from dihydroxyacetone phosphate, an intermediate of sugar metabolism, using multiple recombinant genes. See Altaras, N. E., & Cameron, D. C., 1999, Appl Environ Microbiol. 65(3), 1180-5; U.S. Pat. No. 6,303,352.

[0101] The objective of this example is to provide new pathways for the production of high yields of mixed alcohols in bacteria and yeast. The bacterial CBP platforms comprise microorganisms that are in the same family as C. sphenoides and T. thermosaccharolyticum, which contain native genes for propanediol production and, unlike the literature, do not rely on expression of recombinant activities to convert dihydroxyacetone phosphate to propanediol. For example, T. saccharolyticum is able to ferment L-Rhamnose to equimolar amounts of propanediol and a mixture of ethanol, acetic acid, lactic acid, H.sub.2 and CO.sub.2. See Lee et al., International Journal of Systematic Bacteriology, 43(1): 41-51 (1993). However, in the past, the exploitation of thermophilic clostridia for production of propanediol was not feasible due to a lack of genetically tractable systems required for stable genetic engineering. The successful genetic engineering of thermophilic clostridia and thermoanaerobacter and thermoanaerobacterium strains now makes such exploitation for metabolic engineering possible. See U.S. Prov. Appl. No. 61/232,648, filed Aug. 10, 2009. Further, production of propanedial in yeast has been observed by the expression of a single gene, methylglyoxal synthase (mgs), indicating that additional activities necessary to convert methygloxal to propanediol are endogenous to yeast. See Lee, W., & DaSilva, N. A., 2006, Metabolic Eng. 8, 58-65.

[0102] The 1,2-propandiol produced using these platforms can be used as a valuable intermediate or converted to propionate and propanol using microbes such as Lactobacillus reuteri strain isolated from sourdough that is known to do this reaction. See Sriramulu, D. D., et al., 2008, J. Bacteriol. 190(13):4559-67. Chemical routes might also exist for direct conversion of propanediol to propanol or even propylene.

[0103] Isopropanol can be produced by the addition of a pathway to produce acetone and a dehydrogenase capable of utilizing acetone as a substrate. The best known and studied acetone production route is from the metabolism of Clostridium acetobutylicum. All enzymes in this pathway have been sequenced and cloned into other hosts such as E. coli. See Bermejo, L. L., et al., 1998, Appl Environ Microbiol. 64(3), 1079-85. C. acetobutylicum has been used in industrial fermentations beginning in the early 1900's and the acetone produced was used as a major source for gunpowder during the First World War. The fermentation was widely used until the 1960's when the process was no longer able to compete with the emergent petrochemical process due to rising costs of fermentable sugars. The bacterial and yeast CBP platforms makes the production of isopropanol readily tractable.

1.2 Pathway Definition and Stoichiometric Calculations for Production of Mixed Alcohols

[0104] The combined production of propanediol and isopropanol from glucose is outlined in the pathways of FIG. 1 and requires the activity of several distinct enzymes (Table 2).

TABLE-US-00003 TABLE 2 List of native and non-native gene candidates pertaining to engineering of mixed alcohols in bacteria and yeast CBP platforms. Non-native- Non-native- Activity EC Cthe Tsacch Yeast bacteria yeast methylglyoxal 4.2.3.3 95 or2316 Oryza sativa synthase mgs aldo-keto reductase 1.1.1.-- 152 or1401 (methylglyoxal to 236 or1402 acetol) 283 or785 or414 or2491 aldo-keto reductase 1.1.1.-- 101 or1043 (acetol to 394 or2289 propanediol) 423 or411 2445 or2426 2579 or0286 phosphotransacetylase 2.3.1.8 1029 or1741 Tsacch or1741 acetate kinase 2.7.2.1 1028 or1742 Tsacch or1742 thiolase 2.3.1.9 ERG10 C. acetobutylicum coA transferase 2.8.3.8 C. acetobutylicum C. acetobutylicum acetoacetate 4.1.1.4 C. acetobutylicum C. acetobutylicum decarboxylase isopropanol 1.1.1.80 101 or1411 ADH1 dehydrogenase 394 or1043 ADH2 423 or2426 ADH3 2445 or2289 ADH4 2579 or0286 ADH5 ADH6 ADH7 BDH2 SFA1 YPL088W alcohol 1.1.1.1 423 or411 dehydrogenase PFOR 1.2.7.1 2390-3 or0047 (oxidoreductase) fructose 1,6- 4.1.2.13 0349 or0260 FBA1 biphosphate aldolase 1019 or0330 triose-phophate 5.3.1.1 0139 or2687 TPI1 isomerase glycerol-3-phosphate 1.1.1.8 GPD2 dehydrogenase glycerol-3- 3.1.3.21 GPP1 phosphatase pyruvate formate- 2.3.1.54 E. coli pflA/ lyase pflB formate 1.2.1.2 FDH1 dehydrogenase aldehyde reductase 1.1.1.21 101 or1043 GRE3 394 or2289 423 or411 2445 or2426 2579 or0286 glyoxylate reductase 1.1.1.79 152 or1401 GOR1 236 or1402 YPL113C 283 or785 or414 or2491 methylglyoxal 1.2.1.49 152 or1401 GCY1 dehydrogenase 236 or1402 ALD2 283 or785 ALD3 or414 ALD4 or2491 ALD5 ALD6 HFD1 Genes to KO Activity EC Cthe Tsacch Yeast lactate dehydrogenase 1.1.1.27 1053 or180 pyruvate 4.1.1.1 PDC1 decarboxylase PDC5 PDC6

[0105] The branched metabolic pathways can be subdivided into distinct production rotates as follows:

[0106] (i) the conversion of dihydroxyacetone phosphate into propanediol

[0107] (ii) the conversion of pyruvate into isopropanol

[0108] (iii) the conversion of pyruvate into ethanol (bacterial CBP platform only)

[0109] (iv) the conversion of dihydroxyacetone phosphate into glycerol (yeast CBP platform only).

[0110] The combined production of isopropanol, propanediol, and ethanol (routes (i), (ii), and (iii)) from two glucose molecules during bacterial metabolism is governed by the overall stoichiometric equation with a theoretical yield of one propanol, one propanediol, and one ethanol per two glucose, as follows:

2C.sub.6H.sub.12O.sub.6.fwdarw.C.sub.3H.sub.8O+C.sub.3H.sub.8O.sub.2+C.s- ub.2H.sub.6O+4CO.sub.2+H.sub.2+3ATP

[0111] The theoretical yield of propanediol, propanol, and ethanol on hexose and pentose sugar for the above pathway is:

TABLE-US-00004 Hexose Pentose 0.21 g propanediol/g sugar 0.21 g propanediol/g sugar 0.17 g isopropanol/g sugar 0.17 g isopropanol/g sugar 0.13 g ethanol/g sugar 0.13 g ethanol/g sugar

[0112] The combined production of isopropanol, propanediol, and glycerol in yeast, S. cerevisiae, (routes (i), (ii), and (iv)) results in the net gain of one ATP, and is governed by the overall stoichiometric equation:

2C.sub.6H.sub.12O.sub.6.fwdarw.C.sub.3H.sub.8O+C.sub.3H.sub.8O.sub.2+C.s- ub.3H.sub.8O.sub.3+3CO.sub.2+ATP

[0113] The co-production of isopropanol and propanediol together with the loss of carbon to glycerol and CO.sub.2 are necessary to maintain the redox balance. The theoretical yield of propanediol, propanol, and glycerol on hexose and pentose sugar for the above pathway is:

TABLE-US-00005 Hexose Pentose 0.21 g propanediol/g sugar 0.21 g propanediol/g sugar 0.17 g isopropanol/g sugar 0.17 g isopropanol/g sugar 0.26 g glycerol/g sugar 0.26 g glycerol/g sugar

[0114] The above stoichiometric equations were calculated using a hexose as a carbohydrate source; however, pentose sugars, including but not limited to xylose, can be readily utilized as well. When a pentose sugar is used as the carbohydrate source, six pentose sugars are required as the equivalent for five hexose sugars.

1.3 Production Routes for Mixed Alcohols and Corresponding Enzymology

[0115] Bacterial CBP Platforms

[0116] The combined production of propanediol, isopropanol, and ethanol from glucose in a bacterial CBP platform can be subdivided into the following distinct production routes: (i) the conversion of dihydroxyacetone phosphate into propanediol; (ii) the conversion of pyruvate into isopropanol; and (iii) the conversion of pyruvate into ethanol (FIG. 1). The microbial hosts utilize carbohydrate sources, shown as glucose in FIG. 1, to produce the mixed alcohols, but as mentioned above, pentose sugars such as xylose can be readily utilized as well, requiring six pentose sugars as equivalent for five hexose sugars. The first step in the pathway uses the microbial host's cellular metabolism to metabolize the carbohydrate source, employing, e.g., the Embden-Meyerhof-Parnas (EMP) pathway to produce dihydroxyacetone phosphate and glyceraldehyde 3-phosphate (FIG. 1). These metabolites can be interchanged using triosephosphate isomerase (E.C. 5.3.1.1).

[0117] During route (i), dihydroxyacetone phosphate is converted to methyglyoxal by methylglyoxal synthase (E.C. 4.2.3.3). Methylglyoxal is subsequently converted to either acetol by an oxidoreductase, which is to be identified from EC 1.1.1.- (see Table 2), or lactaldehyde by a keto-reductase (E.C. 1.1.1.79, 1.2.1.49). These intermediates are further reduced to propanediol by, oxidoredutases (E.C. 1.1.1.-) for acetol or (E.C. 1.1.1.2) 1 lactaldehyde.

[0118] For route (ii), glyceraldehyde 3-phosphate is further metabolized to pyruvate through standard glycolysis reactions, producing ATP to power the cellular reactions and the required reducing equivalents needed to reduce the carbon end-products. During bacterial metabolism, pyruvate is metabolized to acetyl-CoA, reduced ferredoxin, and CO.sub.2 by pyruvate ferredoxin oxidoreductase (E.C. 1.2.7.1) (FIG. 1, light gray box). NADH and H.sub.2 are subsequently produced during the oxidation of ferredoxin. Acetyl-CoA is then converted to acetate by phosphate acetytransferse (E.C. 2.3.1.8) and acetate kinase (E.C. 2.7.2.1) in an ATP generating reaction. Two acetyl-CoA molecules are converted to acetoacetyl-CoA by thiolase (E.C. 2.3.1.9). Acetoacetyl-CoA is then converted to acetoacetate by CoA enyzyme transferase (E.C. 2.8.3.8), where the CoA species is transferred from acetoacetyl-CoA to acetate, replenishing the acetyl-CoA consumed during the thiolase reaction. Acetoacetate is then converted to acetone by acetoacetate decarboxylase (E.C. 4.1.1.4). The reduction of acetone to isopropanol can be accomplished by an alcohol dehydrogenase (E.C. 1.1.1.80).

[0119] In route (iii), acetyl-CoA is converted to ethanol by acetaldehyde dehydrogenase (EC 1.2.1.3) and an alcohol dehydrogenase (E.C. 1.1.1.1), or through a bi-functional enzyme catalyzing both steps.

[0120] All the required enzymatic activities have been demonstrated in C. thermosaccharolyticum (see Cameron, D. C. & Clooney, C., 1986, Bio/Technology 4, 651-654) and relevant endogenous enzymes in the bacteria CBP platform production strains that exhibit high levels of homology to the desired enzymatic domains have been identified (see Table 2). The enzymes catalyzing the production of acetone from acetyl-CoA have been identified in the literature, and activities associated with (E.C. 2.3.1.9), (E.C. 2.8.3.8), and (E.C. 4.1.1.4) can be engineered using genes from C. acetobutylicum. See Bermejo, L. L., et al., 1998, Appl Environ Microbiol. 64(3), 1079-85.

[0121] The conversion of acetone to isopropanol has been shown by multiple alcohol dehydrogenases and endogenous enzymes from the microbial CBP hosts can be screened for their capability to accept acetone as a substrate. Additional efforts must be made to readily control the flux through the different metabolic branch points through the modulation of enzyme levels and regulation. To this end, the deletion of ldh (E.C. 1.1.1.27) will prevent flow of carbon from pyruvate to lactic acid (see Table 2, "Genes to KO").

[0122] Yeast CBP Platforms

[0123] The combined production of propanediol, isopropanol, and glycerol from glucose in a yeast CBP platform can be subdivided into the following distinct production routes: (i) the conversion of dihydroxyacetone phosphate into propanediol; (ii) the conversion of pyruvate into isopropanol; and (iv) the conversion of dihydroxyacetone phosphate into glycerol (FIG. 1). As described above, the microbial hosts utilize carbohydrate sources, such as glucose as shown in FIG. 1, or a pentose sugar such as xylose. The first step in the pathway uses the microbial host's cellular metabolism to metabolize the carbohydrate source, employing, e.g., the Embden-Meyerhof-Parnas (EMP) pathway to produce dihydroxyacetone phosphate and glyceraldehyde phosphate (FIG. 1). These metabolites can be interchanged using triosephosphate isomerase (E.C. 5.3.1.1).

[0124] Route (i) is proposed in the yeast CBP platform in a similar manner as route (i) in the bacteria CBP platform, converting dihydroxyacetone phosphate to methyglyoxal and using the two alternate pathways presented to generate propanediol from methyglyoxal. See FIG. 1. However, based on current yeast literature, only a third route might be available, in part, because all result in the same redox change. All three begin with the production of methylglyoxal from dihydroxyacetone phosphate by methylglyoxal synthase, MGS, (E.C. 4.2.3.3) which can be obtained from one of several potential sources. The introduction of the mgs gene alone in yeast has been shown to result in the production of propanediol, but at relatively low titers; subsequent introduction of a glycerol dehydrogenase (E.C. 1.1.1.6) doubled the amount of propanediol formed. See Hoffman, M. L., 1999, Metabolic engineering of 1,2-propanediol production in Saccharomyces cerevisiae. Ph.D. Dissertation, University of Wisconsin--Madison. Alternatively, aldehyde, reductase (E.C. 1.1.1.21) may be capable of converting methylglyoxal to lactaldehyde and then subsequently to propanediol. The native yeast aldehyde reductase, GRE3, can be overexpressed to test this possibility. In addition methyglyoxal could potentially be converted to lactaldehyde by glyoxylate reductase (E.C. 1.1.1.79) or by methylglyoxal dehydrogenase (E.C. 1.2.1.49). These enzymatic activities have not been reported in S. cerevisiae, but there are a number of endogenous genes which may contain these activities: two potential glyoxylate reductases (GOR1 and YPL113C), a glycerol dehydrogenase (GCY1), six aldehyde dehydrogenases (ALD2-6 and HFD1), and the ten alcohol dehydrogenases mentioned below. See Table 2. It might be desirable to engineer in a combination of the two alternate pathways outlined above for producing propanediol from methylglyoxal to reach a desirable titer for propanediol.

[0125] For route (ii), glyceraldehyde 3-phosphate is further metabolized to pyruvate through standard glycolysis reactions, as described above for bacteria CBP platforms. In yeast metabolism, acetyl-CoA and formate is produced from pyruvate by pyruvate formate lyase (E.C. 2.3.1.8) (FIG. 1, dark gray box). Formate is further metabolized to CO.sub.2, NADH, and H.sub.2 by formate dehydrogenase (E.C. 1.2.1.2) (FIG. 1, dark gray box). Production of isopropanol from acetyl-CoA is performed as described above for the bacteria CBP platform.

[0126] Five enzymatic activities can be engineered into yeast for route (ii). The pyruyate formate lyase (PFL) (E.C. 2.3.1.8) is required for the formation of acetyl-CoA in the cytosol, because in a majority of yeast species the endogenously produced acetyl-CoA is sequestered in the mitochondria. Enzymatically active PFL has been expressed in yeast for the production of formate. See Waks, Z., & Silver, P. A., 2009, Appl. Env. Microbiol. 75, 1867-1875. S. cerevisiae has an endogenous formate dehydrogenase (E.C. 1.2.1.2) to convert the formate generated to CO.sub.2 and H.sub.2. The cytosolic acetyl-CoA generated is subsequently converted to acetone by the introduction of the C. acetobutylicum pathway, as described above for the bacteria CBP platform, working together with the yeast acetyl-CoA acetyltransferase, ERG10, (E.C. 2.3.1.9). An alcohol dehydrogenase executes the final reaction in this section acetone to isopropanol. The S. cerevisiae genome encodes for ten alcohol dehydrogenases (ADH1-7, BDH2, SFA1, and YPL088W), which can be assayed for the capability of converting acetone to isopropanol. See Table 2. If necessary an exogenous alcohol dehydrogenase can be engineered into S. cerevisiae. Three pyruvate decarboxylase genes (E.C. 4.1.1.1) can be deleted: PDC1, PDC5, and PDC6. The presence of these three enzymes would result in the loss of significant pyruvate to acetaldehyde.

[0127] In route (iv), dihydroxyacetone phosphate is converted to glycerol by glycerol-3-phosphate dehydrogenase (E.C. 1.1.1.8) and glycerol-3-phosphatase (E.C. 3.1.3.21) (FIG. 1, dark gray boxes). The enzymes required for route (iv) are already present in S. cerevisiae.

Example 2

2.1 Production of n-Propanol and Isopropanol in Bacterial CBP Platforms

[0128] All current native and recombinant propanol producing metabolic pathways have at most a theoretical yield of 0.33 g propanol/g carbohydrate. Yan Y. & Liao J. 2009, J Indus Microbiol and Biotech 36(4):471-479. This yield, corresponding to one mole isopropanol per mole glucose, incorporates into isopropanol only 75% of the free energy available from glucose during anaerobic fermentation. The additional 25% of the free energy, also referred to as available electrons, must be incorporated into a co-product during anaerobic fermentation, or consumed by oxygen during aerobic fermentation.

[0129] The present example proposes a new pathway for propanol production from lignocellulosic carbohydrates at a yield of 0.44 g/g carbohydrate, and incorporates 100% of the free energy available from carbohydrate conversion. In order to produce propanol at this theoretical maximum yield using biochemical pathways found in nature, production of both n- and iso-forms are required. In the metabolic pathway described here, isopropanol production serves in an ATP generating capacity, while n-propanol production serves as an electron sink to balance the anaerobic fermentation. This pathway allows for a balanced fermentation equation that is thermodynamically feasible.

[0130] Both products can be recovered from the fermentation broth via distillation, reducing downstream processing complexity. Isopropanol is a product natively produced by solventogenic Clostridia, and is rapidly produced by Thermoanaerobacter species when fed with acetone, indicating the presence of a native alcohol dehydrogenase with high activity for the desired reaction. See Lamed R J and Zeikus J G, 1981, The Biochemical J 195(1):183-190. Acetone production has been extensively studied, and the Clostridial pathway has been heterologously expressed in E. coli as described above. See Bermejo, L. L., et al., 1998, Appl. Environ. Microbiol. 64(3), 1079-85. n-propanol is a natural product of propanediol degradation, with many microorganisms reported to perform this catalysis under anaerobic conditions. Recently, the genes involved in this conversion have been identified in one species, Listeria innocula, which will facilitate the expression of this pathway in the bacterial CBP organisms. See Xue J. et al., 2008, Applied and Environmental Microbiol. 74(22);7073-7079. Propanediol, a key intermediate of the n-propanol pathway, is a natural fermentation product of thermophilic bacteria. T. thermosaccharolyticum HG-8, the organism reported to produce the highest titer of propanediol, can be engineered for the production of n-propanol.

2.2 Pathway Definition and Stoichiometric Calculations for Production of Propanols

[0131] The combined production of n-propanol and isopropanol from glucose or xylose is outlined in the pathways of FIG. 2 and requires the activity of several distinct enzymes (Table 3).

TABLE-US-00006 TABLE 3 List of native and non-native gene candidates pertaining to engineering of n-propanol and isopropanol in the CBP bacterial platform. Non-native Activity EC C. thermocellum T. saccharolyticum bacteria triose phosphate 5.3.1.1 139 or2687 isomerase methylglyoxal 4.2.3.3 95 or2316 synthase aldo-keto reductase 1.1.1.- 152 or1401 (methylglyoxal to 236 or1402 acetol) 283 or785 or414 or2491 aldo-keto reductase 1.1.1.- 101 or1043 (acetol to propanediol) 394 or2289 423 or411 2445 or2426 2579 or0286 propanediol 4.2.1.28 or0222, T. sacch genes dehydratase or0224-or0226 can be expressed in C. therm propanaldehyde 1.1.1.202 101 0411 dehydrogenase 394 1043 423 2426 2579 2289 0286 phosphotransacetylase 2.3.1.8 1029 or1741 acetate kinase 2.7.2.1 1028 or1742 thiolase 2.3.1.9 C. acetobutylicum coA transferase 2.8.3.8 C. acetobutylicum CtfAB acetoacetate 4.1.1.4 C. acetobutylicum decarboxylase Adc, Aad PFOR 1.2.7.1 2390-93 or0047 (oxidoreductase) Genes to KO alcohol dehydrogenase 1.1.1.1 423 or411 lactate dehydrogenase 1.1.1.27 1053 or180 hydrogenase 1.12.7.2 425-31 or1545-48

[0132] The combined production of n-propanol and isopropanol from 3 glucose molecules during bacterial metabolism is governed by the overall stoichiometric equation:

3C.sub.6H.sub.12O.sub.6.fwdarw.2(n-)C.sub.3 H.sub.8O+2(iso-)C.sub.3H.sub.8O+6CO.sub.2+2H.sub.2O+4 ATP

[0133] The theoretical yield of propanols on a hexose sugar for the above pathway is 0.44 g propanols/g hexose.

[0134] The combined production of n-propanol and isopropanol from 9 xylose molecules during bacterial metabolism is governed by the overall stoichiometric equation:

9C.sub.5H.sub.10O.sub.5.fwdarw.5(n-)C.sub.3H.sub.8O+5(iso-)C.sub.3H.sub.- 2O+15CO.sub.2+5H.sub.2O+12 ATP

[0135] The theoretical yield of propanols on a pentose sugar for the above pathway is 0.44 g propanols/g hexose.

[0136] For this metabolic pathway, product yields are identical for hexose, e.g., glucose, and pentose, e.g., xylose, carbohydrates due to the activity of triosephosphate isomerase (tpi) (E.C. 5.3.1.1). Pentose fermentation produces more of the isomer glyceraldehyde 3-phosphate (GAP) than dihydroxyacetone phosphate (DHAP) compared to hexose fermentation, which produces equimolar ratios of the two compounds. However, tpi allows for the conversion of GAP to DHAP and vice-versa, creating equal product yields for both carbohydrates.

2.3 Production Routes for Propanols and Corresponding Enzymology

[0137] The metabolic pathways for the production of n-propanol and isopropanol can be subdivided into two distinct production routes: (i) the conversion of dihydroxyacetone phosphate into n-propanol: and (ii) the conversion of pyruvate into isopropanol.

[0138] For the n-propanol route, route (i), dihydroxyacetone phosphate is converted to methyglyoxal by methylglyoxal synthase (E.C. 4.2.3.3). Methylglyoxal is subsequently converted to acetol by an oxidoreductase (E.C. 1.1.1.-) or to lactaldehyde by a keto-reductase (1.1.1.79 or 1.2.1.49). These intermediates are then further reduced to propanediol by enzymes from (E.C. 1.1.1.-). Propanediol is then dehydrated to propanal by a diol-hydrolase (E.C. 4.2.1.28) and reduced to n-propanol by a dehydrogenase (E.C. 1.1.1.202). See FIG. 2.

[0139] All the required enzymatic activities for the production of propanediol have been demonstrated in C. thermosaccharolyticum, a strain that can be genetically engineered. Cameron, D. C., et al., 1998, Biotechnol. Prog. 14, 116-125. Relevant endogenous enzymes in the bacterial CBP platform production strains that exhibit high levels of homology to the desired enzymatic domains have also been identified (Table 3). The enzymes leading to propanediol in the bacterial CBP platform production strains can be characterized for implementation in route (i).

[0140] For the isopropanol route, route (ii), glyceraldehyde 3-phosphate is further metabolized to pyruvate through standard glycolysis reactions, producing ATP to power cellular reactions and reducing equivalents needed to balance n-propanol production during anaerobic fermentation. Pyruvate is then metabolized to acetyl-CoA, reduced ferredoxin, and CO.sub.2 by pyruvate ferredoxin oxidoreductase (E.C. 1.2.7.1). NADH and H.sub.2 are subsequently produced during the oxidation of ferredoxin. See FIG. 2.

[0141] Acetyl-CoA is then converted to acetate by phosphate acetytransferse (EC 2.3.1.8) and acetate kinase (E.C. 2.7.2.1) in an ATP generating reaction. Two acetyl-CoA molecules are converted to acetoacetyl-CoA by thiolase (E.C. 2.3.1.9). Acetoacetyl CoA is then converted to acetoacetate by CoA enyzyme transferase (E.C. 2.8.3.8), where the CoA species is transferred from acetoacetyl-CoA to acetate, replenishing the acetyl-CoA consumed during the thiolase reaction. Acetoacetate is then converted to acetone by acetoacetate decarboxylase (E.C. 4.1.1.4). The reduction of acetone to isopropanol can be accomplished by alcohol dehydrogenases E.C.(1.1.1.80).

[0142] The enzymes catalyzing the production of acetone from acetyl-CoA have been identified in the literature from C. acetobutylicum. See Bermejo, L. L., et al., 1998, Appl Environ Microbiol. 64(3), 1079-85. The conversion of acetone to isopropanol has been shown by multiple alcohol dehydrogenases and endogenous bacterial enzymes can be screened for their capability to accept acetone as a substrate.

[0143] Gene deletions will also be required to achieve high yields of propanol production. These include deletion of L-lactate dehydrogeanse ldh (E.C. 1.1.1.27); hydrogenase, hyd (E.C. 1.12.7.2); and acetaldehyde dehydrogenase, acdh (E.C. 1.2.1.10).

Example 3

3.1 Production of Isopropanol and Methanol in Bacterial CBP Platforms

[0144] Co-production of isopropanol and methanol from lignocellulosic carbohydrates allows for a balanced fermentation equation that is thermodynamically feasible. Isopropanol is theoretically produced at 0.33 g/g carbohydrate and incorporates 75% of the electrons available from carbohydrate conversion. Both isopropanol and methanol can be recovered from the fermentation broth via distillation, reducing downstream processing complexity. Further, methanol is a natural product of pectin degradation, and many characterized methylotropic organisms contain genes for methanol metabolism.

3.2 Pathway Definition and Stoichiometric Calculations for Production of Isopropanol and Methanol

[0145] The production of isopropanol and methanol from carbohydrates is outlined in the pathways in FIG. 3 and requires the activity of several distinct enzymes (see Table 4).

TABLE-US-00007 TABLE 4 List of native and non-native gene candidates pertaining to engineering of isopropanol and methanol in the CBP bacterial platform. Non-native- Activity EC C. the T. sacch bacteria pyruvate formate lyase 2.3.1.54 505 or0628 phosphotransacetylase 2.3.1.8 1029 or1741 acetate kinase 2.7.2.1 1028 or1742 formaldehyde 1.2.1.46 218 2445, Pput_0350 dehydrogenase 0388 P. putida methanol 1.1.1.244 101 or1411 dehydrogenase 394 or1043 423 or2476 2445 or2289 2579 or286 formate dehydrogenase 1.2.1.43 342 or2328 Moth_2312 430 M. thermoacetica 3004 3003 thiolase 2.3.1.9 C. acetobutylicum ThlA coA transferase 2.8.3.8 C. acetobutylicum CtfAB acetoacetate 4.1.1.4 C. acetobutylicum decarboxylase Adc, Aad oxidoreductase 1.1.1.80 101 or 1411 394 or1043 423 or2426 2445 or2289 2579 or0286 Genes to KO lactate dehydrogenase 1.1.1.27 1053 or180 alcohol dehydrogenase 1.1.1.1 423 or411 methylglyoxal 4.2.2.3 95 or2316 synthase PFOR 1.2.7.1 2390-93 or0047 (oxidoreductase)

[0146] The combined production of isopropanol and methanol from one glucose molecule during bacterial metabolism is governed by the overall stoichiometric equation, with a theoretical yield of one propanol and one methanol per glucose, as follows:

C.sub.6H.sub.12O.sub.6.fwdarw.C.sub.3H.sub.8O+CH.sub.4O+2CO.sub.2+3ATP

[0147] The theoretical yield of isopropanol and methanol on hexose and pentose sugar for the above pathways (see FIG. 3) are:

Hexose Yield:

[0148] 0.33 g isopropanol/g hexose [0149] 0.18 g methanol/g hexose

Pentose Yield:

[0149] [0150] 0.33 g isopropanol/g pentose [0151] 0.18 g methanol/g pentose

[0152] During cellular metabolism, the microbial hosts can utilize hexose or pentose carbohydrate sources, with six pentose sugars equivalent to five hexose sugars, employing, e.g., the Embden-Meyerhof-Parnas (EMP) pathway to produce dihydroxyacetone phosphate and glyceraldehyde 3-phosphate. These metabolites can be interchanged using the triosephosphate isomerase (E.C. 5.3.1.1).

3.3 Production Routes for isopropanol and Methanol and Corresponding Enzymology

[0153] The branched metabolic pathways for the combined production of isopropanol and methanol from carbohydrates can be subdivided into the following production routes: (i) the conversion of pyruvate into isopropanol: and (ii) the conversion of formate into CO.sub.2 and methanol.

[0154] As described above, glyceraldehyde 3-phosphate is metabolized to pyruvate through standard glycolysis reactions, producing ATP to power the cellular reactions and the required reducing equivalents needed to reduce the carbon end-products. From pyruvate, acetyl-CoA and formate are produced by pyruvate formate lyase (E.C. 2.3.1.54). For isopropanol production, route (i), acetyl-CoA is converted to acetate by phosphate acetytransferse (E.C. 2.3.1.8) and acetate kinase (E.C. 2.7.2.1) in an ATP generating reaction. Two acetyl-CoA molecules are converted to acetoacetyl-CoA by thiolase (E.C. 2.3.1.9). Acetoacetyl-CoA is then converted to acetoacetate by CoA enyzyme transferase (E.C. 2.8.3.8), where the CoA species is transferred from acetoacetyl-CoA to acetate, replenishing the acetyl-CoA consumed during the thiolase reaction. Acetoacetate is then converted to acetone by acetoacetate decarboxylase (E.C. 4.1.1.4). The reduction of acetone to isopropanol can be accomplished by alcohol dehydrogenases (E.C. 1.1.1.80).

[0155] As described above, the enzymes catalyzing the production of acetone from acetyl-CoA have been identified in the literature from C. acetobutylicum. See Bermejo, L. L., et al., 1998, Appl Environ Microbiol. 64(3), 1079-85. The conversion of acetone to isopropanol has been shown by multiple alcohol dehydrogenases and endogenous bacterial enzymes can be screened for their capability to accept acetone as a substrate.

[0156] In route (ii), formate is further metabolized via two pathways in an equimolar ratio first leading to CO.sub.2 and NADPH by formate dehydrogenase (E.C. 1.2.1.43), and the second leading to methanol with the incorporation of two NADH and production of water by the combined action of formaldehyde dehydrogenase (E.C. 1.2.1.46) and methanol dehydrogenase (E.C. 1.1.1.244).

[0157] The production of CO.sub.2 and NADPH via formate is a well characterized pathway with a large body of literature. However, the production of methanol via formate is a less well characterized pathway. The majority of characterized organisms that have methanol metabolism pathways consume methanol, rather than produce it. Methanol production from formate is thermodynamically feasible under anaerobic conditions. The most likely route for engineering a high yielding pathway is to introduce enzymes that natively catalyze the net reaction in the reverse direction and then use evolutionary engineering techniques to select for strains with increased flux towards methanol formation. This strategy for pathway flux improvement has been successfully employed both in the engineering of other metabolic pathways and is anticipated to work for this pathway due to the thermodynamic favorability of the net reaction.

Example 4

4.1 Anaerobic Production of Propanediol and Acetone in Bacterial and Yeast CBP Platforms

[0158] The native microbial production of propanediol has been well documented in Clostridium thermosaccharolyticum by Cameron, D. C., & Clooney, C., 1986 Bio/Technology 4, 651-654, although the endogenous enzymes have yet to be identified and cloned. The native enzymes can be identified from the bacterial CBP platform microbes and utilized in the bacterial CBP platform hosts eliminating the need for "recombinant" genes (e.g., Thermoanerobactor saccharolyticum and Clostridium thermocellum) and/or readily transferred to the yeast CBP platform hosts.

[0159] The theoretical maximum yield for anaerobic propanediol production that includes ATP generation requires the production of a co-fermentation product such as acetate. See U.S. Pat. No. 6,303,352. The pathways presented in this Example achieve the anaerobic maximum theoretical yield and use acetate as art intermediate during the generation of acetone as the co-fermentation product. Acetone was chosen as a co-fermentation product because it is potentially a chemical of value and a less toxic fermentation product to the microorganisms relative to acetate. The simultaneous production of propanediol and acetone represents a novel fermentation process. In addition, relatively little is known about the enzymology converting methygloxal to propanediol, but as described above, can now be ascertained.

4.2 Pathway Definition and Stoichiometric Calculations for Production of Propanediol and Acetone

[0160] The anaerobic production of propanediol and acetone from carbohydrates is outlined in the pathways in FIG. 4 and requires the activity of several distinct enzymes (see Table 5).

TABLE-US-00008 TABLE 5 List of native and non-native gene candidates pertaining to engineering of propanediol and acetone in the CBP bacterial and CBP yeast platforms. Non-native- non-native- Activity EC C. the T. sacch Yeast bacteria yeast methylglyoxal 4.2.3.3 95 or2316 Oryza synthase sativa mgs aldo-keto reductase 1.1.1.-- 152 or1401 P. putida gldA (methylglyoxal to 236 or1402 acetol) 283 or785 or414 or2491 aldo-keto reductase 1.1.1.-- 101 or1043 (acetol to propanediol) 394 or2289 423 or411 2445 or2426 2579 or0286 phosphotransacetylase 2.3.1.8 1029 or1741 Tsacch or1741 acetate kinase 2.7.2.1 1028 or1742 Tsacch or1742 thiolase 2.3.1.9 ERG10 C. acetobutylicum ThlA coA transferase 2.8.3.8 C. acetobutylicum C. acetobutylicum CtfAB CtfAB acetoacetate 4.1.1.4 C. acetobutylicum C. acetobutylicum decarboxylase Adc, Aad Adc, Aad alcohol dehydrogenase 1.1.1.1 ADH1 ADH2 ADH3 ADH4 ADH5 ADH6 ADH7 BDH2 SFA1 YPL088W PFOR 1.2.7.1 2390-3 or0047 (oxidoreductase) fructose 1,6- 4.1.2.13 0349 or0260 FBA1 biphosphate aldolase 1019 or0330 triose-phophate 5.3.1.1 0139 or2687 TPI1 isomerase pyruvate formate- 2.3.1.54 E. coli pflA/ lyase pflB formate 1.2.1.2 FDH1 dehydrogenase aldehyde reductase 1.1.1.21 101 or1043 GRE3 394 or2289 423 or411 2445 or2426 2579 or0286 glyoxylate reductase 1.1.1.79 101 or1043 GOR1 394 or2289 YPL113C 423 or411 2445 or2426 2579 or0286 methylglyoxal 1.2.1.49 152 or1401 GCY1 dehydrogenase 236 or1402 ALD2 283 or785 ALD3 or414 ALD4 or2491 ALD5 ALD6 HFD1 glucokinase 2.7.1.2 0390 or0272 GLK1 2938 glucose 6 phophate 5.3.1.9 0217 or1389 PGI1 isomerase 6-phosphofructokinase 2.7.1.11 1261 or2875 PFK1 PFK2 Genes to KO Activity EC C. the T. sacch Yeast lactate dehydrogenase 1.1.1.27 1053 or180 alcohol dehydrogenase 1.1.1.1 423 or411 pyruvate 4.1.1.1 PDC1 decarboxylase PDC5 PDC6 glycerol-3-phosphate 1.1.1.8 GPD2 dehydrogenase glycerol-3- 3.1.3.21 GPP1 phosphatase

[0161] The combined production of propanediol and acetone from two glucose molecules during bacterial or yeast anaerobic metabolism is governed by the overall stoichiometric equation, resulting in overall redox balance and the net gain of one ATP, as follows:

2C.sub.6H.sub.12O.sub.6.fwdarw.2C.sub.3H.sub.8O.sub.2+C.sub.3H.sub.6O+AT- P+H.sub.2O

[0162] The theoretical yield of propanediol and acetone on hexose and pentose sugar for the above pathway are:

TABLE-US-00009 Hexose Pentose 0.42 g propanediol/g hexose 0.42 g propanediol/g pentose 0.16 g acetone/g hexose 0.16 g acetone/g pentose

[0163] During cellular metabolism, the microbial hosts can utilize hexose or pentose carbohydrate sources, with six pentose sugars equivalent to five hexose sugars, employing the Embden-Meyerhof-Parnas (EMP) pathway to produce dihydroxyacetone phosphate and glyceraldehyde 3-phosphate. These metabolites can be interchanged using the triosephosphate isomerase (EC 5.3.1.1).

4.3 Anaerobic Production Routes for Propanediol and Acetone and Corresponding Enzymology

[0164] The co-production of propanediol and acetone from hexose and pentose sugars in thermophilic clostridia and S. cerevisiae can be broken down into three routes: (i) the production of dihydroxyacetone phosphate and glyceraldehyde 3-phosphate from glucose; (ii) the subsequent generation of propanediol from dihydroxyacetone phosphate; and (iii) the generation of acetone from glyceraldehyde 3 phosphate. See FIG. 4.

[0165] For the bacterial and yeast CBP platforms, the enzyme activities required for route (i), production of dihydroxyacetone phosphate and glyceraldehyde 3-phosphate from glucose, are part of the native glycolytic pathway, e.g., the EMP pathway, as described above. See Table 5.

[0166] For route (ii), the subsequent generation of propanediol from dihydroxyacetone phosphate, two alternative routes are presented, in part because both result in the same redox balance and a priori the best route is not known. Both begin with the production of methylglyoxal from dihydroxyacetone phosphate by methylglyoxal synthase, mgs (E.C. 4.2.3.3). See FIG. 4. This gene is endogenous to the bacterial CBP platform organisms, however for yeast it will have to be obtained from one of several potential sources.

[0167] For the bacterial CBP platform, which comprises thermophilic bacteria, acetol is the likely intermediate from methylglyoxal to propanediol, as has been shown in T. thermosaccarolyticum. See Cameron, D. C., & Clooney, C., 1986, Bio/Technology 4, 651-654. In E. coli, various aldo-keto reductases have been shown to catalyze the conversion of methyglyoxal to acetol (E.C. 1.1.1.-). See Ko, J., et al., 2005, J Bacteriol. 187(16), 5782-9. The list of endogenous aldo-keto reductases for the bacterial platform organisms are shown in Table 5. These genes can be over-expressed and/or deleted to determine their role in propanediol production. It is also possible that lactaldehyde, produced by a glyoxylate reductase (E.C. 1.1.1.79) and a methylglyoxal dehydrogenase (E.C. 1.2.1.49) is an intermediate. To determine if acetol or lactaldehyde is the primary intermediate during conversion of methylglyoxal to propanediol, analytical chemistry procedures such as HPLC can be used to identify these intermediates in fermentation samples. See e.g., Cameron, D. C., &. Clooney, C., 1986, Bio/Technology 4, 651-654; Altaras N. E., Cameron, D. C., 1999, Appl Environ Microbiol. 65(3), 1180-5. Alternatively, cells can be fed acetol or lactaldehyde to determine which intermediate is more effectively converted to propanediol. To determine which genes are responsible for the production of propanediol from acetol or lactaldehyde, the native alcohol dehydrogenases and aldo-keto reductases listed in Table 5 can be deleted and/or over-expressed while propanediol production is monitored.

[0168] For the yeast CBP platform, multiple routes from methylglyoxal to propanediol also exist. See FIG. 4. One route through lactaldehyde involves introduction of a glycerol dehydrogenase (E.C. 1.1.1.-), which doubled the amount of propanediol formed. See Hoffman, M. L., 1999, Metabolic engineering of 1,2-propanediol production in Saccharomyces cerevisiae. Ph.D. Dissertation, University of Wisconsin--Madison. Alternatively, aldehyde reductase (E.C. 1.1.1.21) may be capable of converting methylglyoxal to lactaldehyde and then subsequently to propanediol--the native yeast aldehyde reductase. GRE3, can be overexpressed to test this possibility. In addition, methylglyoxal could potentially be converted to lactaldehyde by glyoxylate reductase (E.C. 1.1.1.79) or to lactaldehyde by methylglyoxal dehydrogenase (E.C. 1.2.1.49). The presence of these alcohol dehydrogenase activities can be screened among the ten native alcohol dehydrogenases. See Table 5. It might be necessary to engineer in a combination of the two pathways outlined above to reach a desirable titer for propanediol.

[0169] The enzymes that convert methylglyoxal to propanediol are oxidoreductases, of which there are examples using either NADH or NADPH as a co-factor. Knowledge of the co-factor is important for producing propanediol in the yeast platform because the compartmentalization of the cell, and the relative difficulty of inter-converting NADH to NADPH, limit the cell's ability to deal with an imbalance in these cofactors. For the anaerobic production of propanediol, an enzyme (or enzymes) that are linked to NADH would be required, since these are the reducing equivalents generated during the production of CO.sub.2 and acetone from glyceraldehyde 3-phosphate. Several of the enzymes identified in bacterial systems have this characteristic.

[0170] For route (iii), the generation of acetone from glyceraldehydes 3-phosphate, the engineering of non-native enzymatic activities into both the bacterial and yeast platforms is required. The bacterial organisms have a native enzyme activity (E.C. 1.2.7.1) that converts pyruvate to acetyl-CoA (FIG. 4, light gray box), while the yeast platform requires the expression of a non-native activity (E.C. 2.3.1.54) to convert pyruvate to acetyl-CoA (FIG. 9a, dark gray box).

[0171] To convert acetyl-CoA to acetone in the bacterial platform, activities associated with (E.C. 2.3.1.9), (E.C. 2.8.3.8), and (E.C. 4.1.1.4) can be engineered using genes from acetobutylicum while activities associated with (E.C. 1.2.7.1), (E.C. 2.3.1.8), and (E.C. 2.7.2.1) are in fact endogenous (FIG. 4). See Bermejo, L. L., et al., 1998, Appl Environ Microbiol. 64(3), 1079-85. Taken together, these activities will allow the formation of acetone from two molecules or pyruvate. For the yeast platform three enzymatic activities can be engineered into yeast. The pyruvate formate lyase, PFL (E.C. 2.3.1.54), is required for the formation of acetyl-CoA in the cytosol, because the majority of yeast endogenously produced acetyl-CoA is sequestered in the mitochondria. Enzymatically active PFL has been expressed in yeast for the production of formate. Waks, Z., & Silver, P. A., 2009, Appl. Env. Microbiol. 75, 1867-1875. S. cerevisiae has an endogenous formate dehydrogenase (E.C. 1.2.1.2) to convert the formate generated to CO.sub.2 and H.sup.+. The cytosolic acetyl-CoA generated can be subsequently converted to acetone by the introduction of the C. acetobutylicum pathway (E.C. 2.8.3.8) and (E.C. 4.1.1.4), as described above, working together with the yeast acetyl-CoA acetyltransferase, ERG10 (E.C. 2.3.1.9).

[0172] The description of the above pathways describes native and non-native genes required to direct carbon flow from sugars to propanediol and acetone. In addition, to prevent decreases in product yield, i.e., carbon from flowing away from desired end products, various genes can be deleted from each platform. For the bacterial CBP system, these genes are shown in Table 5. The deletion of adh (E.C. 1.1.1.1) will prevent flow from acetyl-CoA to acetaldehyde while the deletion of ldh (E.C. 1.1.1.27) will prevent flow of carbon from pyruvate to lactic acid. Deleting the hydrogenase genes (E.C. 1.12.7.2) will ensure that reducing equivalents generated during glycolysis can be used to make reduced end products such as 1,2-propanediol and not the more oxidized couple of H.sub.2 and acetate. For the yeast CBP platform, genes to be deleted are listed in Table 5. Genes encoding activity associated with (E.C. 4.1.1.1) can be deleted to prevent carbon flow from pyruvate to acetaldehyde. In addition, genes associated with (E.C. 1.1.1.8) and (E.C. 3.1.3.21) activity can be deleted to prevent carbon loss from dihydroxyacetone phosphate as glycerol.

Example 5

Aerobic Production of Propanediol in Yeast CBP Platforms

[0173] The purpose of the present Example is to provide a novel pathway for the aerobic production of propanediol in yeast CBP platforms. Aerobic production of propanediol provides some benefits in terms of ATP production. For example, the advantages of aerobic production are discussed in Cameron et al., "Metabolic engineering of propanediol pathways," Biotechnology Progress, 14(1): 116-125 (1998), where a yield of 0.61 g propanediol/g can be achieved in a non-compartmentalized organism. Indeed, the commercial production of 1,3-propanediol is done via an aerobic process. Although not as high as 0.61 g propanediol/g in a non-compartmentalized organism, the present pathway provides for a high yield of propanediol in a compartmentalized organism as discussed below.

[0174] The 1,2-propandiol produced using this platform can be used as a valuable intermediate or converted to propionate and propanol using microbes such as Lactobacillus reuteri strain isolated from sourdough that is known to do this reaction. See Sriramulu, D. D., et al., 2008, J. Bacteriol. 190(13):4559-67. Chemical routes might also exist for direct conversion of propanediol to propanol or even propylene.

Pathway Definition and Stoichiometric Calculations for Production of Propanediol

[0175] The aerobic production of propanediol from carbohydrates is outlined in the pathways in FIG. 5 and requires the activity of several distinct enzymes (see Table 5).

[0176] The production of propanediol, which is the only soluble product of the reaction, from 6 glucose molecules during yeast aerobic metabolism is governed by the overall stoichiometric equation:

6 glucose+12O.sub.2.fwdarw.6 x propanediol+12H.sub.2O+18CO.sub.2+26 ATP

[0177] In order to balance the redox in the cytosol, 1 molecule of glucose 6-phosphate must be completely oxidized by the pentose phosphate pathway (PPP) for every molecule of propanediol produced. In addition, a positive ATP balance is generated via oxidation of the glyceraldehyde 3-phosphate in the TCA cycle and the electron transport chain. See FIG. 4.

[0178] The theoretical yield of propanediol on hexose sugar for the above pathway is 0.42 g propanediol/g hexose, 100% xylose could not be converted via this pathway, but a glucose/xylose mixture could convert with a yield similar to glucose alone. Although not as high of a yield as for a non-compartmentalized organism, the proposed pathway provides a high yield for propanediol. Further, the possibility of shuttling NADH to the cytosol from the mitochondrial matrix cannot be ruled out since such a shuttle has been demonstrated. See Bakker, B. M, et al., 2000, Appl. Env. Micro. 182, 4730-4737. This would potentially allow higher yields in S. cerevisiae. In Kluyveromyces type yeasts, yields might also be increased due to shuttling of reducing equivalents to the cytoplasm, and the enhanced activity of the pentose phosphate pathway in these organisms.

5.3 Aerobic Production Routes for Propanediol and Corresponding Enzymology

[0179] For the production of dihydroxyacetone phosphate and glyceraldehyde 3-phosphate from glucose, the enzyme activities are part of the native glycolytic pathway, e.g., the EMP pathway, as described above. See Table 5 and FIG. 5.

[0180] For the subsequent generation of propanediol from dihydroxyacetone phosphate, two alternative routes are presented as in Example 4 (see FIG. 4), in part because both result in the same redox balance and a priori the best route is not known. Both begin with the production of methylglyoxal from dihydroxyacetone phosphate by methylglyoxal synthase, mgs (E.C. 4.2.3.3). See FIG. 4. For yeast, this gene will have to be obtained from one of several potential sources.

[0181] As described above in Example 4, multiple routes from methylglyoxal to propanediol exist in yeast. See FIG. 4. One route through lactaldehyde involves introduction of a glycerol dehydrogenase (E.C. 1.1.1.-), which doubled the amount of propanediol formed. See Hoffman, M. L., 1999, Metabolic engineering of 1,2-propanediol production in Saccharomyces cerevisiae. Ph.D. Dissertation University of Wisconsin--Madison. Alternatively, aldehyde reductase (E.C. 1.1.1.21) may be capable of converting methylglyoxal to lactaldehyde and then subsequently to propanediol the native yeast aldehyde reductase, GRE3, can be overexpressed to test this possibility. In addition, methylglyoxal could potentially be converted to lactaldehyde by glyoxylate reductase (E.C. 1.1.1.79) or to lactaldehyde by methylglyoxal dehydrogenase (E.C. 1.2.1.49). The presence of these alcohol dehydrogenase activities can be screened among the ten native alcohol dehydrogenases. See Table 5. It might be necessary to engineer in a combination of the two pathways outlined above to reach a desirable titer for propanediol.

[0182] As described above, the enzymes that convert methylglyoxal to propanediol are oxidoreductases, of which there are examples using either NADH or NADPH as a co-factor. Knowledge of the co-factor is important for producing propanediol in the yeast platform because the compartmentalization of the cell, and the relative difficulty of inter-converting NADH to NADPH, limit the cell's ability to deal with an imbalance in these cofactors. In the aerobic production of propanediol, the NADPH linked versions of an enzyme (or enzymes) are required, since the production of reducing equivalents in the form of NADPH is accomplished in the pentose phosphate pathway. The S. cerevisiae gre3 gene is a good example (and candidate) for use in the aerobic system.

[0183] To convert the carbohydrate source to propanediol in yeast using an aerobic process, control of the flux of carbon down particular pathways will be needed. Redox balance is obtained by controlling flux to the PPP and propanediol, while optimal product yield is obtained when the flux to the TCA cycle and electron transport chain is held to a minimal level. Controlling flux to the PPP involves manipulating the expression level of zwfl, which converts glucose 6-phosphate to D-glucono-1,5-lactone 6-phosphate, relative to the activity of pgi, which converts glucose 6-phosphate to fructose 6-phosphate. In order to control the amount of flux to the TCA cycle and the electron transport chain, one of two methods could be used. One would be to down-regulate PDH, and thereby reduce the amount of pyruvate being converted to acetyl-CoA in the mitochondria. The other would be to control the oxygen flux in the fermentation vessel to limit the amount of oxygen available for the electron transport chain. The former genetic approach has an advantage in that it alleviates the necessity of careful process control for aeration at large scale.

Example 6

Identification and Characterization of T. Saccharolyticum pdu Gene Cluster

[0184] Several microorganisms metabolize propanediol to propanol anaerobically. Examples of propanediol utilization can be found among various bacterial species including Thermoanaerobacteria, Salmonella, Listeria, and Clostridia. In some microorganisms, e.g., Listeria spp. and Salmonella spp., the genes required for propanediol utilization (pdu) are clustered on the genome. See generally Scott, K. P., et al. J. Bacteriol. 188(12):4340-49 (2006); Bobik, T. A., et al., J. Bacteriol. 181(19):5967-75; Xue, J., et al., Appl. Env. Microbiol. 74(22):7073-79 (2008).

[0185] Two enzyme activities required for conversion of propanediol propanol include: [0186] 1) diol dehydratase (encoded by pduCDE) and [0187] 2) dehydrogenase (encoded by pduQ). See Table 3 above. In several microorganisms, such as Salmonella spp. or Listeria spp., the first enzyme activity often involves catalysis via a heteromeric diol dehydratase enzyme that is dependent on vitamin B12. The pdu gene dusters are often found to include or be associated with the enzymes required for the synthesis of vitamin B12. Some of the pdu gene clusters include genes for 1) B12 synthesis, 2) AraC type transcription activator, 3) two-component response regulator, 4) an alcohol and aldehyde dehydrogenase, or 5) mfC homolog. See Scott, K. P., et al., J. Bacteriol. 188(12):4340-49 (2006); Bobik, T. A., et al., J. Bacteriol. 181(19):5967-75; Xue, J., et al., Appl. Env. Microbiol. 74(22):7073-79 (2008).

[0188] Thus far, no pdu gene clusters have been identified in thermophilic anaerobic bacteria. This Example provides the identification and characterization of the T. saccharolyticum pdu gene cluster for its use in conversion of propanediol to propanol, following, e.g., the scheme described in Example 2.

[0189] The pdu gene organization in T. saccharolyticum is shown in FIG. 6 and includes several of the genes found in pdu gene clusters from other microorganisms. The T. saccharolyticum pdu genes include ABC-sugar transporter components (or201, or202, or203), a two-component response regulator (or206 or207), rhamnose isomerase (or209) rhamulokinase (or208), rhamnulose-1-phosphate lactaldehyde lyase (or227), a putative propanediol:NAD+ oxidoreductase (or211), micro-compartment proteins (pduJ, pduL, etc. or212, or214, or215, or216, or217), an aldehyde dehydrogenase (or219), an alcohol dehydrogenase (or218), a phosphotransacetylase (or213), B12 accessory enzymes (or223, or222, or221, or220), a B-12 dependent diol dehydratase pduCDE (or226, or225, or224), and a transcriptional regulator (or228). The activities of these genes can be characterized, e.g., through various gene deletion studies, growth on rhamnose, and/or expression into heterologous systems such as T. thermosaccharolyticum and C. thermocellum.

[0190] The ability of T. saccharolyticum which harbors the above-identified pdu gene cluster, to produce detectable levels of n-propanol was determined. The wild-type T. saccharolyticum YS485 strain was grown in TSC1 medium (Table 6) with 10 g/L CaCO.sub.3 and a starting pH of 5.8 at 55.degree. C. and 200 rpm under anaerobic conditions. The medium was supplemented with 0.001 g/L, vitamin B12.

TABLE-US-00010 TABLE 6 Composition of TSC1 medium. Concentration Components (g/L) (NH.sub.4).sub.2SO.sub.4 1.85 FeSO.sub.4 * 7H.sub.2O 0.05 KH.sub.2PO.sub.4 0.5 MgSO.sub.4 1 CaCl.sub.2 * 2H.sub.2O 0.05 Trisodium citrate * 2H.sub.2O 2 Yeast Extract 8.5 CaCO.sub.3 10 L-rhamnose 18

[0191] Batch fermentation was done and samples were drawn at various time points shown in Table 7. The samples were analyzed by HPLC to detect remaining L-rhamnose and end products, including lactic acid (LA), acetic acid (AA), ethanol (Etoh), 1,2-propanediol (1,2 PD), and n-propanol. The results are depicted in Table 7.

TABLE-US-00011 TABLE 7 Production of 1,2-Propanediol and n-Propanol in T. saccharolyticum Grown on L-rhamnose Time L-rhamnose LA AA Etoh 1,2 PD n-propanol (hr) (g/L) (g/L) (g/L) (g/L) (g/L) (g/L) 0 17.820 0.000 0.118 0.000 0.277 0.000 17.5 11.440 0.242 2.054 0.315 2.248 0.363 24.5 2.522 0.346 4.289 0.437 4.623 1.072 41.25 0.679 0.384 5.024 0.527 5.073 1.525 69.5 0.427 0.407 5.135 0.567 5.134 1.638

[0192] These results demonstrate that T. saccharolyticum has the native ability to produce 1,2-propanediol (up to 5.1 g/L) and n-propanol (1.6 g/L) when grown on L-rhamnose. The pdu gene cluster includes some rhamnose utilization and sugar uptake genes indicating that those are likely to be involved in this process. This provides the first example of a thermophilic anaerobic bacterium shown to be capable of producing n-propanol.

Example 7

Production of Propanol Via Propanediol Using a B12-Independent Diol Dehydratase in Yeast

[0193] As described above, one of the two enzyme activities required for conversion of propanediol to propanol includes a diol dehydratase enzyme, which in several microorganisms is dependent on vitamin B12. Yeast lack the metabolic machinery to synthesize vitamin B12, and thus, it is not possible to engineer a vitamin B12-dependent enzyme in yeast without also providing, e.g., the enzyme activities to synthesize vitamin B12. There have been a few reports of propanediol dehydratase enzymes that do not require vitamin B12. See Raynaud, C., et al., PNAS (USA) 100(9):5010-15 (2003); Scott, K. P., et al., J. Bacteriol. 188(12):4340-49 (2006); Hartmanis, M. G., and Stadtman, T. C., Arch. Biochem. Biophys. 245(1)144-52 (1986).

[0194] Because of the requirement for vitamin B12, the anaerobic conversion of propanediol to propanol was thought to be impossible due to the requirement of a vitamin B12-dependent enzyme. Recent reports describing the B12-independent diol dehydratase provide a source and incentive to screen for existing B12-independent diol dehydratases in nature and express them into yeast. See Raynaud, C., et al., PNAS (USA) 100(9):5010-15 (2003); Scott, K. P., et al., J. Bacteriol. 188(12):4340-49 (2006); Hartmanis, M. G., and Stadtman, T. C., Arch. Biochem. Biophys. 245(1)144-52 (1986). If successfully done, this would be the first n-propanol producing yeast engineered so far. The purpose of this Example is to identify and engineer a vitamin B12-independent diol dehydratase, as well as other necessary enzymes, in yeast, e.g., Saccharomyces cerevisiae, to anaerobically convert propanediol to propanol.

[0195] The metabolic pathway for generating propanol from, e.g., a carbohydrate source, in yeast is similar to the route described above in Example 2 and as shown in FIG. 2. In order to successfully achieve this conversion of glucose, several enzyme activities need to be engineered in yeast. Conversion of glucose to pyruvate and dihydroxyacetone-P are achieved via the endogenous enzyme activities in yeast. Those activities which need to be engineered are highlighted in FIG. 7 and are as follows:

[0196] 1) The conversion of pyruvate to acetyl-CoA and formate via pyruvate-formate lyase (PFL) (E.C. 2.3.1.8) has been successfully engineered and demonstrated. See Waks, Z. and Silver, P. A., Appl. Env. Microbiol. 75(7):1867-75 (2009). This is an important step to generate a pool of acetyl-CoA in the yeast cytosol for its subsequent conversion into isopropanol. Simultaneously, the flux of pyruvate to acetyl-CoA via pyruvate decarboxylase (PDC) needs to be avoided liar which the PDC1, PDC5 and PDC6 need to be knocked out. The conversion of formate to carbon dioxide is catalyzed by an endogenous enzyme, formate dehydrogenase (E.C. 1.2.1.2).

[0197] 2) Acetyl-CoA is further converted to acetate by phosphate acetyltransferase (E.C. 2.3.1.8) and acetate kinase (E.C. 2.7.2.1) in an ATP generating reaction. Two acetyl-CoA molecules are converted to acetoacetyl-CoA by thiolase (E.C. 2.3.1.9). Acetoacetyl-CoA is then converted to acetoacetate by CoA enzyme transferase (E.C. 2.8.3.8), where the CoA species is transferred from acetoacetyl-CoA to acetate, replenishing the acetyl-CoA consumed during the thiolase reaction. Acetoacetate is then converted to acetone by acetoacetate decarboxylase (E.C. 4.1.1.4). The reduction of acetone to isopropanol can be accomplished by alcohol dehydrogenases (E.C. 1.1.1.80).

[0198] 3) Synthesis of methylglyoxal from dihydroxyacetone-P can be achieved by expression of heterologous methylglyoxal synthase (mgs) and glycerol dehydrogenase (gldA) as has been previously demonstrated. See Lee, W. and DaSilva, N. A., Metabolic Eng. 8(1):58-65 (2006).

[0199] 4) The conversion of propanediol to propanol requires two enzyme activities as described above, involving a diol dehydratase and a dehydrogenase. Although several microorganisms can convert 1,2-propandiol to propanol using a vitamin B12-dependent diol dehydratase reaction via a vitamin B12-dependent diol dehydratase is not feasible in yeast due to the B12 dependency. The few recently discovered examples of vitamin B12-independent diol dehydratase include those identified from Clostridium butyricum, Roseburia inulinivorans, Clostridium glycolicum and Klebsiella spp. The C. butyricum enzyme is extensively characterized and shown to be functional independent of B12 and in a heterologous system (E. coli). See Tang, X., et al., Appl. Env. Microbiol. 75(6):1628-34 (2009). The results obtained with the C. butyricum B12-independent diol dehydratase activity suggest that the enzyme can be engineered into a heterologous system such as yeast.

[0200] In addition to the incorporation of these enzymatic activities, the flux of carbon from pyruvate to ethanol must be disrupted in yeast. This can be accomplished via the deletion of pdc1, pdc5, and pdc6. PDC deletion strains are slow growing and require a small amount of added ethanol or acetate to be viable; however, these issues can be overcome via an evolutionary based approach. See, e.g., van Maris, A. J. A., et al., Appl. Env. Microbiol. 70(1):159-66 (2004). The fact that such strains produce pyruvate at high levels indicates that this compound would be available for subsequent conversion to propanol via the proposed pathway above.

[0201] In order to identify additional B12-independent diol dehydratases for engineering in part 4 above, other B12-independent diol dehydratase enzymes existing in nature can be identified. Suitable methods for identifying can include, but are not limited to, alignment searches based on homology to known B12-independent diol dehydratases, an enzymatic activity assay combined with protein purification and protein sequencing, and whole-genome transcriptional analysis of 1,2 propanediol utilizing organisms. See, e.g., Scott, K. P. et al., J. Bact 188(12):4340-4349 (2006), and Raynaud, C. et al., PNAS 100(9):5010-5015 (2003).

[0202] Once identified and isolated., the gene responsible for the activity is cloned into yeast along with other enzyme activities as described above. Optimization of expression of the B12-independent diol dehydratase and analytical assays for production of propanol is subsequently followed.

INCORPORATION BY REFERENCE

[0203] All of the U.S. patents and U.S. published patent applications cited herein are hereby incorporated by reference.

EQUIVALENTS

[0204] Those skilled in the art will recognize, or be able to ascertain using no more than routine experimentation, many equivalents to the specific embodiments of the invention described herein. Such equivalents are intended to be encompassed by the following claims.

Sequence CWU 1

1

1671742PRTC. thermocellum 1Met Asp Ala Trp Arg Gly Phe Asn Lys Gly Asn Trp Cys Gln Glu Ile 1 5 10 15 Asp Val Arg Asp Phe Ile Ile Arg Asn Tyr Thr Pro Tyr Glu Gly Asp 20 25 30 Glu Ser Phe Leu Val Gly Pro Thr Asp Arg Thr Arg Lys Leu Trp Glu 35 40 45 Lys Val Ser Glu Leu Leu Lys Lys Glu Arg Glu Asn Gly Gly Val Leu 50 55 60 Asp Val Asp Thr His Thr Ile Ser Thr Ile Thr Ser His Lys Pro Gly 65 70 75 80 Tyr Ile Asp Lys Glu Leu Glu Val Ile Val Gly Leu Gln Thr Asp Glu 85 90 95 Pro Leu Lys Arg Ala Ile Met Pro Phe Gly Gly Ile Arg Met Val Ile 100 105 110 Lys Gly Ala Glu Ala Tyr Gly His Ser Val Asp Pro Gln Val Val Glu 115 120 125 Ile Phe Thr Lys Tyr Arg Lys Thr His Asn Gln Gly Val Tyr Asp Val 130 135 140 Tyr Thr Pro Glu Met Arg Lys Ala Lys Lys Ala Gly Ile Ile Thr Gly 145 150 155 160 Leu Pro Asp Ala Tyr Gly Arg Gly Arg Ile Ile Gly Asp Tyr Arg Arg 165 170 175 Val Ala Leu Tyr Gly Val Asp Arg Leu Ile Ala Glu Lys Glu Lys Glu 180 185 190 Met Ala Ser Leu Glu Arg Asp Tyr Ile Asp Tyr Glu Thr Val Arg Asp 195 200 205 Arg Glu Glu Ile Ser Glu Gln Ile Lys Ser Leu Lys Gln Leu Lys Glu 210 215 220 Met Ala Leu Ser Tyr Gly Phe Asp Ile Ser Cys Pro Ala Lys Asp Ala 225 230 235 240 Arg Glu Ala Phe Gln Trp Leu Tyr Phe Ala Tyr Leu Ala Ala Val Lys 245 250 255 Glu Gln Asn Gly Ala Ala Met Ser Ile Gly Arg Ile Ser Thr Phe Leu 260 265 270 Asp Ile Tyr Ile Glu Arg Asp Leu Lys Glu Gly Lys Leu Thr Glu Glu 275 280 285 Leu Ala Gln Glu Leu Val Asp Gln Leu Val Ile Lys Leu Arg Ile Val 290 295 300 Arg Phe Leu Arg Thr Pro Glu Tyr Glu Lys Leu Phe Ser Gly Asp Pro 305 310 315 320 Thr Trp Val Thr Glu Ser Ile Gly Gly Met Ala Leu Asp Gly Arg Thr 325 330 335 Leu Val Thr Lys Ser Ser Phe Arg Phe Leu His Thr Leu Phe Asn Leu 340 345 350 Gly His Ala Pro Glu Pro Asn Leu Thr Val Leu Trp Ser Val Asn Leu 355 360 365 Pro Glu Gly Phe Lys Lys Tyr Cys Ala Lys Val Ser Ile His Ser Ser 370 375 380 Ser Ile Gln Tyr Glu Ser Asp Asp Ile Met Arg Lys His Trp Gly Asp 385 390 395 400 Asp Tyr Gly Ile Ala Cys Cys Val Ser Ala Met Arg Ile Gly Lys Gln 405 410 415 Met Gln Phe Phe Gly Ala Arg Cys Asn Leu Ala Lys Ala Leu Leu Tyr 420 425 430 Ala Ile Asn Gly Gly Lys Asp Glu Met Thr Gly Glu Gln Ile Ala Pro 435 440 445 Met Phe Ala Pro Val Glu Thr Glu Tyr Leu Asp Tyr Glu Asp Val Met 450 455 460 Lys Arg Phe Asp Met Val Leu Asp Trp Val Ala Arg Leu Tyr Met Asn 465 470 475 480 Thr Leu Asn Ile Ile His Tyr Met His Asp Lys Tyr Ala Tyr Glu Ala 485 490 495 Leu Gln Met Ala Leu His Asp Lys Asp Val Phe Arg Thr Met Ala Cys 500 505 510 Gly Ile Ala Gly Leu Ser Val Val Ala Asp Ser Leu Ser Ala Ile Lys 515 520 525 Tyr Ala Lys Val Lys Pro Ile Arg Asn Glu Asn Asn Leu Val Val Asp 530 535 540 Tyr Glu Val Glu Gly Asp Tyr Pro Lys Phe Gly Asn Asn Asp Glu Arg 545 550 555 560 Val Asp Glu Ile Ala Val Gln Val Val Lys Met Phe Met Asn Lys Leu 565 570 575 Arg Lys Gln Arg Ala Tyr Arg Ser Ala Thr Pro Thr Leu Ser Ile Leu 580 585 590 Thr Ile Thr Ser Asn Val Val Tyr Gly Lys Lys Thr Gly Asn Thr Pro 595 600 605 Asp Gly Arg Lys Ala Gly Glu Pro Leu Ala Pro Gly Ala Asn Pro Met 610 615 620 His Gly Arg Asp Ile Asn Gly Ala Leu Ala Val Leu Asn Ser Ile Ala 625 630 635 640 Lys Leu Pro Tyr Glu Tyr Ala Gln Asp Gly Ile Ser Tyr Thr Phe Ser 645 650 655 Ile Ile Pro Lys Ala Leu Gly Arg Asp Glu Glu Thr Arg Ile Asn Asn 660 665 670 Leu Lys Ser Met Leu Asp Gly Tyr Phe Lys Gln Gly Gly His His Ile 675 680 685 Asn Val Asn Val Phe Glu Lys Glu Thr Leu Leu Asp Ala Met Glu His 690 695 700 Pro Glu Lys Tyr Pro Gln Leu Thr Ile Arg Val Ser Gly Tyr Ala Val 705 710 715 720 Asn Phe Ile Lys Leu Thr Arg Glu Gln Gln Leu Asp Val Ile Asn Arg 725 730 735 Thr Ile His Gly Lys Ile 740 2358PRTC. thermocellum 2Val Ile Ile Tyr Ser Tyr Lys Tyr Tyr Lys Tyr Ser Phe Tyr Asp Asn 1 5 10 15 Ser Phe Gly Ile Met Lys Gly Glu Glu Phe Met Ser Phe Leu Glu Gln 20 25 30 Ile Ile Glu Arg Ala Lys Ser Asp Val Lys Thr Ile Val Leu Pro Glu 35 40 45 Ser Thr Asp Leu Arg Val Ile Lys Ala Ala Ser Met Ile Met Lys Lys 50 55 60 Gly Ile Ala Lys Val Val Leu Ile Gly Asn Glu Lys Glu Ile Lys Ser 65 70 75 80 Leu Ala Gly Asp Ile Asp Leu Glu Gly Val Met Ile Glu Asp Ser Leu 85 90 95 Asn Ser Glu Lys Leu Glu Asp Tyr Ala Asn Thr Leu Tyr Glu Leu Arg 100 105 110 Lys Ser Lys Gly Met Thr Ile Glu Ala Ala Arg Glu Thr Ile Lys Asp 115 120 125 Pro Leu Tyr Tyr Gly Val Met Met Val Lys Lys Gly Glu Ala Asp Gly 130 135 140 Met Val Ala Gly Ala Val Asn Ser Thr Ala Asn Thr Leu Arg Pro Ala 145 150 155 160 Leu Gln Ile Leu Lys Thr Ala Pro Gly Thr Lys Leu Val Ser Ser Phe 165 170 175 Phe Val Met Val Val Pro Asn Cys Glu Tyr Gly His Asn Gly Thr Phe 180 185 190 Val Tyr Ala Asp Cys Gly Leu Val Glu Asn Pro Asp Ala Asp Gln Leu 195 200 205 Ser Glu Ile Ala Ile Ser Ala Ser Lys Ser Phe Glu Met Leu Val Gly 210 215 220 Ala Lys Pro Gln Val Ala Met Leu Ser Tyr Ser Ser Tyr Gly Ser Ala 225 230 235 240 Lys Ser Glu Leu Thr Glu Lys Val Ile Lys Ala Thr Gln Leu Ala Lys 245 250 255 Glu Lys Ala Pro His Leu Ala Ile Asp Gly Glu Leu Gln Val Asp Ala 260 265 270 Ala Ile Val Pro Glu Val Ala Lys Ser Lys Ala Lys Gly Ser Ser Val 275 280 285 Ala Gly Lys Ala Asn Val Leu Ile Phe Pro Asp Leu Asp Ala Gly Asn 290 295 300 Ile Ala Tyr Lys Leu Thr Gln Arg Leu Ala Lys Ala Glu Ala Tyr Gly 305 310 315 320 Pro Ile Thr Gln Gly Leu Ala Arg Pro Val Asn Asp Leu Ser Arg Gly 325 330 335 Cys Ser Ala Glu Asp Ile Val Gly Val Ala Ala Ile Thr Ala Val Gln 340 345 350 Ala Gln Tyr Val Lys Ala 355 3399PRTC. thermocellum 3Met Asn Ile Leu Val Ile Asn Thr Gly Ser Ser Ser Leu Lys Tyr Gln 1 5 10 15 Leu Ile Asp Met Thr Asn Glu Ser Val Leu Ala Lys Gly Val Cys Asp 20 25 30 Arg Ile Gly Leu Glu His Ser Phe Leu Lys His Thr Lys Thr Gly Gly 35 40 45 Glu Thr Val Val Ile Glu Lys Asp Leu Tyr Asn His Lys Leu Ala Ile 50 55 60 Gln Glu Val Ile Ser Ala Leu Thr Asp Glu Lys Ile Gly Val Ile Lys 65 70 75 80 Ser Met Ser Glu Ile Ser Ala Val Gly His Arg Ile Val His Gly Gly 85 90 95 Glu Lys Phe Lys Glu Ser Ala Ile Ile Asp Glu Asp Val Met Lys Ala 100 105 110 Ile Arg Asp Cys Val Glu Leu Ala Pro Leu His Asn Pro Ser Asn Ile 115 120 125 Ile Gly Ile Glu Ala Cys Lys Gln Ile Leu Pro Asp Val Pro Met Val 130 135 140 Ala Val Phe Asp Thr Ala Phe His Gln Thr Met Pro Arg His Ala Tyr 145 150 155 160 Ile Tyr Ala Leu Pro Tyr Glu Ile Tyr Glu Lys Tyr Lys Leu Arg Lys 165 170 175 Tyr Gly Phe His Gly Thr Ser His Lys Tyr Val Ala His Arg Ala Ala 180 185 190 Gln Met Leu Gly Lys Pro Ile Glu Ser Leu Lys Leu Ile Thr Cys His 195 200 205 Leu Gly Asn Gly Ala Ser Ile Cys Ala Val Lys Gly Gly Lys Ser Val 210 215 220 Asp Thr Ser Met Gly Phe Thr Pro Leu Gln Gly Leu Cys Met Gly Thr 225 230 235 240 Arg Ser Gly Asn Val Asp Pro Ala Val Ile Thr Tyr Leu Met Glu Lys 245 250 255 Glu Lys Met Asn Ile Asn Asp Ile Asn Asn Phe Leu Asn Lys Lys Ser 260 265 270 Gly Val Leu Gly Ile Ser Gly Val Ser Ser Asp Phe Arg Asp Val Gln 275 280 285 Asp Ala Ala Glu Lys Gly Asp Asp Arg Ala Gln Leu Ala Leu Asp Ile 290 295 300 Phe Cys Tyr Gly Val Arg Lys Tyr Ile Gly Lys Tyr Ile Ala Val Leu 305 310 315 320 Asn Gly Val Asp Ala Val Val Phe Thr Ala Gly Ile Gly Glu Asn Asn 325 330 335 Ala Tyr Ile Arg Arg Glu Val Leu Lys Asp Met Asp Phe Phe Gly Ile 340 345 350 Lys Ile Asp Leu Asp Lys Asn Glu Val Lys Gly Lys Glu Ala Asp Ile 355 360 365 Ser Ala Pro Asp Ala Lys Val Lys Thr Leu Val Ile Pro Thr Asn Glu 370 375 380 Glu Leu Glu Ile Ala Arg Glu Thr Leu Arg Leu Val Lys Asn Leu 385 390 395 4389PRTC. thermocellum 4Met Ile Asn Phe Val Tyr Lys Asn Pro Thr Lys Ile Ile Phe Gly Arg 1 5 10 15 Gly Thr Glu Leu Lys Val Gly Glu Glu Val Arg Gln Tyr Ser Gly Lys 20 25 30 Val Leu Leu His Tyr Gly Gly Gly Ser Ile Lys Lys Thr Gly Leu Tyr 35 40 45 Asp Arg Val Val Asn Ser Leu Lys Gln Ala Gly Val Glu Val Val Glu 50 55 60 Leu Gly Gly Val Met Pro Asn Pro Arg Leu Gly Leu Val Asn Glu Gly 65 70 75 80 Ile Lys Ile Cys Arg Glu Lys Gly Ile Asp Phe Ile Leu Ala Val Gly 85 90 95 Gly Gly Ser Ala Ile Asp Ser Ala Lys Ala Ile Ala Val Gly Val Pro 100 105 110 Tyr Asp Gly Asp Val Trp Asp Phe Phe Cys Gly Lys Ala Glu Pro Lys 115 120 125 Glu Ala Leu Pro Val Gly Val Val Leu Thr Ile Pro Ala Ala Gly Ser 130 135 140 Glu Ala Ser Pro Asn Ser Val Ile Thr Arg Glu Asp Gly Leu Tyr Lys 145 150 155 160 Arg Gly Met Tyr Ser Glu Leu Ile Arg Pro Val Phe Ala Ile Met Asn 165 170 175 Pro Glu Leu Thr Tyr Thr Leu Pro Ala Tyr Gln Thr Ala Cys Gly Thr 180 185 190 Ala Asp Ile Met Ala His Ile Met Glu Arg Tyr Phe Thr Asn Glu Thr 195 200 205 His Thr Asp Leu Thr Asp Arg Leu Cys Glu Ala Thr Leu Lys Thr Met 210 215 220 Ile Lys Asn Val Pro Ile Ala Leu Glu Glu Pro Asp Asn Tyr Asn Ala 225 230 235 240 Arg Ala Glu Ile Met Trp Ala Gly Thr Ile Ala His Asn Gly Leu Leu 245 250 255 Gly Thr Gly Arg Ile Glu Asp Trp Ala Ser His Asn Ile Glu His Glu 260 265 270 Ile Ser Ala Ile Tyr Asp Val Ala His Gly Ala Gly Leu Ala Val Val 275 280 285 Phe Pro Ala Trp Met Lys Tyr Val Tyr Lys Asn Asn Leu Asp Arg Phe 290 295 300 Val Gln Phe Ala Val Arg Val Trp Asn Val Glu Met Asn Phe Asp Glu 305 310 315 320 Pro Glu Arg Thr Ala Leu Glu Gly Ile Glu Arg Leu Lys Lys Phe Phe 325 330 335 Lys Glu Ile Gly Leu Pro Val Ser Leu Lys Glu Met Asn Ile Gly Asp 340 345 350 Asp Arg Leu Glu Glu Met Ala Ser Lys Cys Thr Asn Gly Gly Lys Ala 355 360 365 Thr Ile Gly Asn Phe Val Lys Leu Asn Arg Glu Asp Val Tyr Asn Ile 370 375 380 Leu Lys Leu Ala Val 385 5389PRTC. thermocellum 5Met Lys Ala Phe Asn Tyr Tyr Ala Pro Thr Glu Ile Ile Phe Gly Cys 1 5 10 15 Gly Arg Val Gln Glu Ile Gly Ser Ile Thr Ala Gln Tyr Gly Lys Lys 20 25 30 Ala Leu Leu Val Thr Val Pro Glu Phe Pro Glu Val Lys Glu Leu Tyr 35 40 45 Glu Lys Val Lys Lys Ser Leu Arg Glu Asn Gly Val Glu Val Val His 50 55 60 Phe Asp Gly Val Ile Pro Asn Pro Thr Thr Asp Val Val Thr Glu Gly 65 70 75 80 Ala Asn Met Ala Lys Ala Ala Gly Val Asp Val Val Ile Gly Leu Gly 85 90 95 Gly Gly Ser Ser Ile Asp Thr Ala Lys Ala Ile Ala Val Glu Ala Thr 100 105 110 His Pro Gly Thr Ala Trp Asp Tyr Asn Cys His Thr Pro Gly Pro Thr 115 120 125 Ser Ala Thr Leu Pro Ile Ile Ala Ile Gly Thr Thr Ala Gly Thr Gly 130 135 140 Ser Gln Cys Thr Gln Cys Ala Val Ile Thr Lys Thr Ser Glu Lys Asp 145 150 155 160 Lys Ser Ala Ile Trp His Lys Asn Ile Phe Pro Lys Val Ala Ile Val 165 170 175 Asp Pro Glu Val Thr Val Thr Met Pro Lys Ser Val Thr Ala Gln Thr 180 185 190 Gly Phe Asp Ala Phe Ala His Asn Phe Glu Ala Tyr Leu Ser Val Lys 195 200 205 Thr Ser Pro Leu Val Glu Met Met Ala Ile Glu Ala Ile Lys Met Ile 210 215 220 Lys Glu Tyr Leu Pro Lys Ala Leu Glu Asn Pro Asn Asp Ile Glu Ala 225 230 235 240 Arg Ser Lys Met Ser Leu Ala Asp Thr Leu Gly Gly Leu Thr Asn Ser 245 250 255 Asn Ala Gly Val Thr Leu Pro His Gly Leu Gly Met Gln Val Gly Gly 260 265 270 His Ala Pro His Val Ser His Gly Gln Ala Leu Ala Ile Ile Tyr Pro 275 280 285 Gln Phe Thr Arg Tyr Thr Tyr Ala Trp Ala Ile Glu Lys Phe Ala Lys 290 295 300 Val Gly Arg Ile Phe Asn Pro Ala Leu Asn Glu Leu Ser Asp Glu Glu 305 310 315 320 Ala Ala Lys Glu Ala Cys Val Ala Ile Asp Asp Phe Leu Lys Lys Ile 325 330 335 Gly Leu Trp Ile Gly Phe Lys Asp Val Asn Val Thr Lys Glu Gln Ile 340 345 350 Arg Glu Ile Ala Asp Asp Gly Gln Val Leu Gly Asp Tyr Leu Asn Asn 355 360 365 Pro Arg Val Ala Thr Ile Asp Glu Met Tyr Glu Leu Leu Met Asn Cys 370 375 380 Tyr Glu Arg Lys Glu 385 6873PRTC. thermocellum 6Met Thr Lys Ile Ala Asn Lys Tyr Glu Val Ile Asp Asn Val Glu Lys 1 5

10 15 Leu Glu Lys Ala Leu Lys Arg Leu Arg Glu Ala Gln Ser Val Tyr Ala 20 25 30 Thr Tyr Thr Gln Glu Gln Val Asp Lys Ile Phe Phe Glu Ala Ala Met 35 40 45 Ala Ala Asn Lys Met Arg Ile Pro Leu Ala Lys Met Ala Val Glu Glu 50 55 60 Thr Gly Met Gly Val Val Glu Asp Lys Val Ile Lys Asn His Tyr Ala 65 70 75 80 Ser Glu Tyr Ile Tyr Asn Ala Tyr Lys Asn Thr Lys Thr Cys Gly Val 85 90 95 Ile Glu Glu Asp Pro Ala Phe Gly Ile Lys Lys Ile Ala Glu Pro Leu 100 105 110 Gly Val Ile Ala Ala Val Ile Pro Thr Thr Asn Pro Thr Ser Thr Ala 115 120 125 Ile Phe Lys Thr Leu Ile Ala Leu Lys Thr Arg Asn Ala Ile Ile Ile 130 135 140 Ser Pro His Pro Arg Ala Lys Asn Ser Thr Ile Glu Ala Ala Lys Ile 145 150 155 160 Val Leu Glu Ala Ala Val Lys Ala Gly Ala Pro Glu Gly Ile Ile Gly 165 170 175 Trp Ile Asp Val Pro Ser Leu Glu Leu Thr Asn Leu Val Met Arg Glu 180 185 190 Ala Asp Val Ile Leu Ala Thr Gly Gly Pro Gly Leu Val Lys Ala Ala 195 200 205 Tyr Ser Ser Gly Lys Pro Ala Ile Gly Val Gly Ala Gly Asn Thr Pro 210 215 220 Ala Ile Ile Asp Asp Ser Ala Asp Ile Val Leu Ala Val Asn Ser Ile 225 230 235 240 Ile His Ser Lys Thr Phe Asp Asn Gly Met Ile Cys Ala Ser Glu Gln 245 250 255 Ser Val Ile Val Leu Asp Gly Val Tyr Lys Glu Val Lys Lys Glu Phe 260 265 270 Glu Lys Arg Gly Cys Tyr Phe Leu Asn Glu Asp Glu Thr Glu Lys Val 275 280 285 Arg Lys Thr Ile Ile Ile Asn Gly Ala Leu Asn Ala Lys Ile Val Gly 290 295 300 Gln Lys Ala His Thr Ile Ala Asn Leu Ala Gly Phe Glu Val Pro Glu 305 310 315 320 Thr Thr Lys Ile Leu Ile Gly Glu Val Thr Ser Val Asp Ile Ser Glu 325 330 335 Glu Phe Ala His Glu Lys Leu Cys Pro Val Leu Ala Met Tyr Arg Ala 340 345 350 Lys Asp Phe Asp Asp Ala Leu Asp Lys Ala Glu Arg Leu Val Ala Asp 355 360 365 Gly Gly Phe Gly His Thr Ser Ser Leu Tyr Ile Asp Thr Val Thr Gln 370 375 380 Lys Glu Lys Leu Gln Lys Phe Ser Glu Arg Met Lys Thr Cys Arg Ile 385 390 395 400 Leu Val Asn Thr Pro Ser Ser Gln Gly Gly Ile Gly Asp Leu Tyr Asn 405 410 415 Phe Lys Leu Ala Pro Ser Leu Thr Leu Gly Cys Gly Ser Trp Gly Gly 420 425 430 Asn Ser Val Ser Asp Asn Val Gly Val Lys His Leu Leu Asn Ile Lys 435 440 445 Thr Val Ala Glu Arg Arg Glu Asn Met Leu Trp Phe Arg Thr Pro Glu 450 455 460 Lys Ile Tyr Ile Lys Arg Gly Cys Leu Pro Val Ala Leu Asp Glu Leu 465 470 475 480 Lys Asn Val Met Gly Lys Lys Lys Ala Phe Ile Val Thr Asp Asn Phe 485 490 495 Leu Tyr Asn Asn Gly Tyr Thr Lys Pro Ile Thr Asp Lys Leu Asp Glu 500 505 510 Met Gly Ile Val His Lys Thr Phe Phe Asp Val Ser Pro Asp Pro Ser 515 520 525 Leu Ala Ser Ala Lys Ala Gly Ala Ala Glu Met Leu Ala Phe Gln Pro 530 535 540 Asp Thr Ile Ile Ala Val Gly Gly Gly Ser Ala Met Asp Ala Ala Lys 545 550 555 560 Ile Met Trp Val Met Tyr Glu His Pro Glu Val Asp Phe Met Asp Met 565 570 575 Ala Met Arg Phe Met Asp Ile Arg Lys Arg Val Tyr Thr Phe Pro Lys 580 585 590 Met Gly Gln Lys Ala Tyr Phe Ile Ala Ile Pro Thr Ser Ala Gly Thr 595 600 605 Gly Ser Glu Val Thr Pro Phe Ala Val Ile Thr Asp Glu Lys Thr Gly 610 615 620 Ile Lys Tyr Pro Leu Ala Asp Tyr Glu Leu Leu Pro Asp Met Ala Ile 625 630 635 640 Val Asp Ala Asp Met Met Met Asn Ala Pro Lys Gly Leu Thr Ala Ala 645 650 655 Ser Gly Ile Asp Ala Leu Thr His Ala Leu Glu Ala Tyr Val Ser Met 660 665 670 Leu Ala Thr Asp Tyr Thr Asp Ser Leu Ala Leu Arg Ala Ile Lys Met 675 680 685 Ile Phe Glu Tyr Leu Pro Arg Ala Tyr Glu Asn Gly Ala Ser Asp Pro 690 695 700 Val Ala Arg Glu Lys Met Ala Asn Ala Ala Thr Ile Ala Gly Met Ala 705 710 715 720 Phe Ala Asn Ala Phe Leu Gly Val Cys His Ser Met Ala His Lys Leu 725 730 735 Gly Ala Phe Tyr His Leu Pro His Gly Val Ala Asn Ala Leu Met Ile 740 745 750 Asn Glu Val Ile Arg Phe Asn Ser Ser Glu Ala Pro Thr Lys Met Gly 755 760 765 Thr Phe Pro Gln Tyr Asp His Pro Arg Thr Leu Glu Arg Tyr Ala Glu 770 775 780 Ile Ala Asp Tyr Ile Gly Leu Lys Gly Lys Asn Asn Glu Glu Lys Val 785 790 795 800 Glu Asn Leu Ile Lys Ala Ile Asp Glu Leu Lys Glu Lys Val Gly Ile 805 810 815 Arg Lys Thr Ile Lys Asp Tyr Asp Ile Asp Glu Lys Glu Phe Leu Asp 820 825 830 Arg Leu Asp Glu Met Val Glu Gln Ala Phe Asp Asp Gln Cys Thr Gly 835 840 845 Thr Asn Pro Arg Tyr Pro Leu Met Asn Glu Ile Arg Gln Met Tyr Leu 850 855 860 Asn Ala Tyr Tyr Gly Gly Ala Lys Lys 865 870 7347PRTC. thermocellum 7Met Lys Gly Lys Met Lys Val Cys Val Leu Thr Gly Lys Glu Lys Leu 1 5 10 15 Glu Trp Val Glu Arg Asp Ile Pro Gln Pro Gly Arg Gly Glu Leu Gln 20 25 30 Ile Lys Leu Lys His Val Gly Val Cys Gly Ser Asp Leu His Phe Tyr 35 40 45 Lys Glu Gly Arg Leu Ala Asn Trp Glu Leu Asp Gly Pro Leu Ala Leu 50 55 60 Gly His Glu Pro Gly Gly Ile Val Ser Ala Ile Gly Glu Gly Val Glu 65 70 75 80 Gly Phe Glu Ile Gly Asp Lys Val Ala Leu Glu Pro Gly Val Pro Cys 85 90 95 Gly Glu Cys Glu Asp Cys Arg Lys Gly His Tyr Asn Leu Cys Lys His 100 105 110 Ile Lys Phe Met Ala Ile Pro His Glu Lys Asp Gly Val Phe Ala Glu 115 120 125 Tyr Cys Val His Ser Ala Ser Met Cys Tyr Lys Leu Pro Glu Asn Val 130 135 140 Asp Thr Met Glu Gly Gly Leu Met Glu Pro Leu Ser Val Ala Leu His 145 150 155 160 Ala Thr Glu Leu Ser Asn Ala Lys Ile Gly Glu Thr Ala Ile Val Leu 165 170 175 Gly Ser Gly Cys Ile Gly Leu Cys Thr Val Met Ala Leu Lys Ala Arg 180 185 190 Gly Val Ser Glu Ile Tyr Val Thr Asp Val Val Asp Lys Arg Leu Glu 195 200 205 Lys Ala Leu Glu Val Gly Ala Thr Arg Val Phe Asn Ser Gln Arg Glu 210 215 220 Asp Ile Val Glu Phe Ala Lys Thr Leu Pro Gly Gly Gly Ala Asp Gln 225 230 235 240 Val Tyr Glu Cys Ala Gly Ser Arg Val Thr Thr Leu Gln Thr Cys Lys 245 250 255 Leu Ile Lys Arg Ala Gly Lys Val Thr Leu Val Gly Val Ser Pro Glu 260 265 270 Pro Val Leu Glu Leu Asp Ile Ala Thr Leu Asn Ala Met Glu Gly Thr 275 280 285 Val Tyr Ser Val Tyr Arg Tyr Arg Asn Met Tyr Pro Ile Ala Ile Ala 290 295 300 Ala Val Ser Ser Gly Val Ile Pro Leu Lys Lys Ile Val Ser His Val 305 310 315 320 Phe Asp Phe Lys Asp Cys Ile Glu Ala Ile Glu Tyr Ser Thr Asn His 325 330 335 Lys Asp Glu Val Ile Lys Ser Val Ile Lys Phe 340 345 8368PRTC. thermocellum 8Met Asn Phe Lys Phe Lys Ile Gly Thr Lys Val Phe Phe Gly Lys Glu 1 5 10 15 Cys Val Lys Glu Asn Lys Ala Val Phe Lys Asp Phe Arg Lys Arg Ala 20 25 30 Leu Leu Val Thr Gly Lys Asn Ser Ala Lys Ala Ser Gly Ala Phe Ser 35 40 45 Asp Val Val Glu Val Leu Glu Glu Tyr Gly Ile Asp Tyr Glu Ile Tyr 50 55 60 Asp Arg Val Ala Asn Asn Pro Ser Leu Glu Asn Val Lys Glu Gly Gly 65 70 75 80 Glu Ala Ala Arg Lys Phe Asp Ala Asp Phe Ile Ile Gly Ile Gly Gly 85 90 95 Gly Ser Pro Leu Asp Ala Ser Lys Ala Val Ala Val Leu Ala Thr Asn 100 105 110 Asp Ile Glu Pro Val Asp Leu Tyr Lys Asn Val Phe Glu Asn Lys Pro 115 120 125 Leu Pro Ile Ile Ala Ile Pro Thr Thr Ala Gly Thr Gly Ser Glu Val 130 135 140 Thr Pro Tyr Ser Ile Leu Thr Arg Asp Asp Met Lys Thr Lys Lys Ser 145 150 155 160 Phe Gly Asn Glu Asp Thr Phe Pro Ala Val Ala Phe Ile Asp Ala Arg 165 170 175 Tyr Thr Glu Ser Met Ser Tyr Glu Thr Thr Val Asp Thr Ala Leu Asp 180 185 190 Ala Phe Thr His Ala Leu Glu Gly Tyr Leu Gly Arg Arg Ser Thr Pro 195 200 205 Val Ser Asp Ile Leu Ala Val Glu Ala Ile Arg Ile Phe Gly Glu Cys 210 215 220 Leu Glu Asn Leu Leu Asn Asn Lys Phe Asp Tyr Asp Val Arg Glu Lys 225 230 235 240 Leu Leu Tyr Met Ser Met Leu Gly Gly Met Val Ile Ser His Thr Gly 245 250 255 Thr Thr Ile Ile His Gly Met Gly Tyr Ser Leu Thr Tyr Phe Lys Asp 260 265 270 Ile Pro His Gly Arg Ala Asn Gly Met Leu Val Arg Glu Tyr Leu Lys 275 280 285 Tyr Asn Tyr Glu Ala Ala Lys Glu Lys Thr Asp Asn Val Leu Arg Leu 290 295 300 Leu Lys Val Pro Ser Ile Asp Ala Phe Gly Glu Ile Ile Asp Arg Leu 305 310 315 320 Ile Pro Gln Lys Pro Val Leu Thr Lys Glu Glu Ile Glu Leu Tyr Ala 325 330 335 Ser Leu Ala Met Lys Gln Asn Ser Thr Leu Ser Asn Ala Arg Thr Val 340 345 350 Val Lys Glu Asp Met Glu Glu Ile Phe Lys Asn Thr Phe Gly Lys Gly 355 360 365 9131PRTC. thermocellum 9Met Asn Ile Ala Leu Ile Ala His Asp Lys Lys Lys Glu Leu Met Ala 1 5 10 15 Ser Phe Cys Ile Ala Tyr Arg Ser Ile Leu Lys Asn His Thr Leu Phe 20 25 30 Ala Thr Gly Thr Thr Gly Ala Ile Ile Val Glu Ala Thr Gly Leu Asn 35 40 45 Val His Arg Phe Leu Pro Gly Val Met Gly Glu Gln Gln Ile Ser Ala 50 55 60 Arg Ala Ala Tyr Asn Glu Leu Asp Leu Val Ile Phe Phe Arg Asp Pro 65 70 75 80 Ile Ser Ala Lys Ser Asp Glu Pro Asp Ile His Ser Leu Leu Arg Glu 85 90 95 Cys Asp Ile Asn Asn Ile Pro Phe Ala Thr Asn Leu Gly Thr Ala Glu 100 105 110 Met Leu Ile Lys Gly Leu Glu Arg Gly Asp Leu Asp Trp Arg Glu Leu 115 120 125 Ile Lys Lys 130 10315PRTC. thermocellum 10Leu Lys Tyr Cys Lys Leu Gly Asn Thr Gly Leu Glu Val Ser Lys Leu 1 5 10 15 Cys Phe Gly Gly Leu Ile Ile Gly Pro Leu Gln Ala Asn Leu Pro Pro 20 25 30 Glu Thr Gly Ala Glu Ile Ile Leu Lys Ser Phe Glu Leu Gly Val Asn 35 40 45 Phe Ile Asp Thr Ala Glu Leu Tyr Gly Thr Tyr Ser His Ile Gly Lys 50 55 60 Ala Leu Lys Lys Thr Asn Lys Asn Ile Val Val Ala Thr Lys Ser Tyr 65 70 75 80 Ala Tyr Ser Ala Glu Gly Ala Lys Glu Ser Leu Glu Lys Ala Arg Lys 85 90 95 Glu Met Asp Ile Asp Val Ile Asp Ile Phe Met Leu His Glu Gln Glu 100 105 110 Ser Arg Leu Thr Leu Lys Gly His Arg Glu Ala Leu Glu Tyr Tyr Ile 115 120 125 Ser Met Lys Glu Lys Gly Ile Ile Lys Ala Val Gly Val Ser Thr His 130 135 140 Asn Val Glu Val Val Glu Ala Cys Cys Glu Met Pro Glu Val Asp Val 145 150 155 160 Ile His Pro Ile Val Asn Lys Ala Gly Ile Gly Ile Gly Asp Gly Thr 165 170 175 Ile Asp Asp Met Leu Lys Ala Val Glu Lys Ala Tyr Ser Val Gly Lys 180 185 190 Gly Ile Tyr Ser Met Lys Pro Leu Gly Gly Gly Asn Leu Ile Lys Ser 195 200 205 Tyr Lys Glu Ala Met Asp Phe Val Leu Asn Ile Pro Tyr Ile His Ser 210 215 220 Ile Ala Val Gly Met Gln Ser Ile Glu Glu Val Val Met Asn Val Cys 225 230 235 240 Ile Phe Glu Gly Lys Glu Val Pro Gln Asp Val Gln Lys Ser Leu Glu 245 250 255 Asn Lys Lys Arg His Leu His Ile Asp Trp Trp Cys Glu Gly Cys Gly 260 265 270 Lys Cys Val Glu Arg Cys Lys Gln Lys Ala Leu Lys Leu Val Asp Gly 275 280 285 Lys Ala Lys Val Glu Glu Glu Lys Cys Val Leu Cys Ser Tyr Cys Ala 290 295 300 Ser Val Cys Pro Val Phe Ala Ile Lys Val Ser 305 310 315 11376PRTC. thermocellum 11Met Gln Tyr Arg Gly Leu Gly Lys Thr Gly Val Lys Val Ser Ala Leu 1 5 10 15 Gly Phe Gly Ala Met Arg Leu Pro Gln Ile Asn Ile Asn Gly Asn Thr 20 25 30 Arg Val Asp Glu Glu Lys Ser Ile Glu Met Ile His Arg Ala Phe Glu 35 40 45 Leu Gly Val Asn Tyr Ile Asp Thr Ala Pro Gly Tyr Cys Asn Gly Glu 50 55 60 Ser Glu Val Val Val Gly Lys Ala Leu Lys Gly Trp Arg Asp Lys Ile 65 70 75 80 Tyr Leu Ser Thr Lys Asn Pro Ile Glu Asn Ala Ser Gly Asp Asp Trp 85 90 95 Arg Lys Arg Leu Glu Asn Ser Leu Lys Lys Leu Asp Thr Asp Tyr Ile 100 105 110 Asp Phe Tyr His Met Trp Gly Ile Asn Trp Glu Thr Tyr Glu Thr Lys 115 120 125 Ile Asp Val Lys Gly Gly Pro Leu Glu Ala Ala Arg Lys Ala Lys Glu 130 135 140 Glu Gly Leu Ile Arg His Ile Ser Phe Ser Phe His Asp Lys Pro Glu 145 150 155 160 Asn Leu Ile Lys Leu Ile Asp Thr Gly Asn Phe Glu Thr Val Leu Cys 165 170 175 Gln Tyr Asn Leu Leu Asp Arg Ser Asn Glu Lys Ala Ile Ala His Ala 180 185 190 Lys Arg Lys Gly Leu Gly Val Ile Ile Met Gly Pro Val Gly Gly Gly 195 200 205 Lys Leu Gly Glu Pro Ser Glu Thr Ile Lys Lys Leu Leu Pro Lys Lys 210 215 220 Thr Val Ser Cys Ala Glu Ile Ala Leu Arg Phe Val Leu Ala Asn Pro 225 230 235 240 Asn Val Asp Cys Ala Leu Ser Gly Met Ser Thr Ile Glu Met Val Glu 245 250 255 Glu Asn Val Arg Val Ala Ser Asn Asp Thr Pro Leu Thr Lys Glu Glu 260

265 270 Leu Glu Met Ile Arg Ala Ser Met Glu Glu Asn Lys Arg Met Glu Asp 275 280 285 Leu Tyr Cys Thr Gly Cys Asn Tyr Cys Met Pro Cys Pro Val Gly Val 290 295 300 Asn Ile Pro Leu Asn Phe Gln Leu Met Asn Tyr His Arg Val Tyr Lys 305 310 315 320 Ile Thr Asp Tyr Ala Arg Gly Gln Tyr Ser Gln Ile Gly Lys Val Glu 325 330 335 Trp Tyr Lys Gly Lys Pro Ala His Glu Cys Ile Glu Cys Gly Val Cys 340 345 350 Glu Thr Lys Cys Pro Gln Lys Leu Glu Ile Arg Lys Gln Leu Lys Glu 355 360 365 Thr Ala Arg Val Leu Ser Val Lys 370 375 12315PRTC. thermocellum 12Met Lys Tyr Arg Lys Met Gly Arg Thr Gly Leu Tyr Ile Ser Glu Ile 1 5 10 15 Ser Leu Gly Ser Trp Leu Thr Tyr Gly Asn Ser Thr Asp Lys Glu Thr 20 25 30 Ala Val Lys Val Ile Asp Thr Ala Tyr Ser Leu Gly Ile Asn Tyr Phe 35 40 45 Asp Thr Ala Asn Val Tyr Ala Asn Gly Arg Ala Glu Val Ile Val Gly 50 55 60 Glu Ala Leu Lys Lys Tyr Pro Arg Glu Ser Tyr Ile Leu Ala Thr Lys 65 70 75 80 Ala Phe Trp Pro Met Gly Thr Gly Pro Asn Asp Lys Gly Leu Ser Arg 85 90 95 Lys His Val Phe Glu Gln Val His Ala Ser Leu Lys Arg Leu Asn Val 100 105 110 Asp Tyr Ile Asp Ile Phe Tyr Cys His Arg Tyr Asp Pro Glu Thr Pro 115 120 125 Leu Glu Glu Thr Leu Arg Thr Ile Asp Asp Leu Leu Arg Gln Gly Lys 130 135 140 Ile Leu Tyr Val Gly Val Ser Glu Trp Thr Ala Ala Gln Met Ala Gln 145 150 155 160 Ala Leu His Ile Ala Asp Arg Tyr Leu Leu Asp Arg Ile Val Val Asn 165 170 175 Gln Pro Gln Tyr Asn Met Phe His Arg Tyr Ile Glu Lys Glu Ile Ile 180 185 190 Pro Phe Gly Glu Lys Asn Gly Ile Ser Gln Ile Val Phe Ser Pro Leu 195 200 205 Ala Gln Gly Val Leu Thr Gly Lys Tyr Lys Pro Gly Gly Asn Ile Pro 210 215 220 Arg Asp Ser Arg Ala Ala Asp Pro Asn Ser Asn Met Tyr Ile Gly Gln 225 230 235 240 Phe Leu Lys Glu Asp Lys Leu Leu Lys Val Glu Lys Leu Lys Ala Val 245 250 255 Ala Asp Glu Met Gly Ile Thr Leu Ser Gln Leu Ala Ile Ala Trp Val 260 265 270 Leu Arg Gln Pro Asn Val Thr Ser Ala Leu Ile Gly Ala Ser Lys Pro 275 280 285 Glu Gln Val Glu Glu Asn Val Lys Ala Ser Gly Ile Asn Leu Ser Asp 290 295 300 Glu Ile Leu Asn Lys Ile Glu Ala Ile Leu Gln 305 310 315 13251PRTC. thermocellum 13Met Ser Arg Lys Val Ile Ala Ala Gly Asn Trp Lys Met Asn Lys Thr 1 5 10 15 Pro Lys Glu Ala Val Glu Phe Val Gln Ala Leu Lys Gly Arg Val Ala 20 25 30 Asp Ala Asp Thr Glu Val Val Val Gly Val Pro Phe Val Cys Leu Pro 35 40 45 Gly Val Val Glu Ala Ala Lys Gly Ser Asn Ile Lys Val Ala Ala Gln 50 55 60 Asn Met His Trp Glu Glu Lys Gly Ala Phe Thr Gly Glu Val Ser Gly 65 70 75 80 Pro Met Leu Ala Glu Leu Gly Val Asp Tyr Val Ile Ile Gly His Ser 85 90 95 Glu Arg Arg Gln Tyr Phe Gly Glu Thr Asp Glu Thr Val Asn Lys Lys 100 105 110 Val His Ala Ala Phe Lys Tyr Gly Leu Lys Pro Ile Ile Cys Val Gly 115 120 125 Glu Ser Leu Thr Gln Arg Glu Gln Gly Val Thr Ala Glu Leu Val Arg 130 135 140 Tyr Gln Val Lys Ile Ala Leu Leu Gly Leu Ser Ala Glu Gln Val Lys 145 150 155 160 Glu Ala Val Ile Ala Tyr Glu Pro Ile Trp Ala Ile Gly Thr Gly Lys 165 170 175 Thr Ala Thr Asn Glu Gln Ala Glu Glu Val Cys Gly Ile Ile Arg Glu 180 185 190 Cys Ile Lys Glu Leu Tyr Gly Gln Asp Val Ala Glu Ala Ile Arg Ile 195 200 205 Gln Tyr Gly Gly Ser Val Asn Ala Ala Asn Ala Ala Glu Leu Phe Asn 210 215 220 Met Pro Asn Ile Asp Gly Gly Leu Val Gly Gly Ala Ser Leu Lys Leu 225 230 235 240 Asp Asp Phe Glu Lys Ile Ala Lys Tyr Asn Lys 245 250 14192PRTC. thermocellum 14Met Gly Lys Val Val Glu Ile Arg Trp His Gly Arg Gly Gly Gln Gly 1 5 10 15 Ala Lys Thr Ala Ser Leu Leu Leu Ala Asp Ala Ala Phe Asn Thr Gly 20 25 30 Lys Tyr Ile Gln Gly Phe Pro Glu Tyr Gly Pro Glu Arg Met Gly Ala 35 40 45 Pro Ile Thr Ala Tyr Asn Arg Ile Ser Asp Glu Lys Leu Thr Ile His 50 55 60 Ser Asn Ile Tyr Glu Pro Asp Tyr Val Val Val Val Asp Asp Thr Leu 65 70 75 80 Leu Thr Ser Val Asp Val Thr Ala Gly Leu Lys Glu Asp Gly Ala Ile 85 90 95 Ile Val Asn Thr Pro Lys Thr Pro Asp Glu Ile Arg Pro Leu Leu Lys 100 105 110 Gly Tyr Lys Gly Lys Val Cys Thr Ile Asp Ala Arg Lys Ile Ser Ile 115 120 125 Glu Thr Leu Gly Lys Tyr Phe Pro Asn Thr Pro Met Leu Gly Ala Val 130 135 140 Val Lys Val Ser Lys Ile Met Asp Glu Glu Glu Phe Leu Lys Asp Met 145 150 155 160 Val Glu Ser Phe Lys His Lys Phe Ala Asn Lys Pro Glu Val Val Glu 165 170 175 Gly Asn Ile Lys Ala Leu Glu Arg Ser Met Gln Glu Val Lys Gly Leu 180 185 190 15101PRTC. thermocellum 15Met Ser Lys Glu Leu Arg Asp Val Lys Pro Asp Val Thr Trp Lys Glu 1 5 10 15 Ile Thr Ser Gly Gly Val Ile Asp Ser Pro Gly Asn Ala His Leu Phe 20 25 30 Lys Thr Gly Asp Trp Arg Ser Met Lys Pro Val Trp Asn Glu Glu Lys 35 40 45 Cys Lys Gln Cys Leu Leu Cys Asn Pro Val Cys Pro Asp Ser Ser Ile 50 55 60 Met Val Ser Glu Glu Gly Lys Met Thr Gly Ile Asp Tyr Asp His Cys 65 70 75 80 Lys Gly Cys Gly Ile Cys Ser Lys Val Cys Pro Phe Lys Ala Ile Asp 85 90 95 Phe Val Glu Glu Val 100 16394PRTC. thermocellum 16Met Gly Ile Arg Glu Arg Leu Ser Gly Asn Glu Ala Thr Ala Ile Ala 1 5 10 15 Met Arg Gln Ile Asn Pro Asp Val Val Ala Ala Phe Pro Ile Thr Pro 20 25 30 Ser Thr Glu Ile Pro Gln Tyr Phe Ser Ser Tyr Val Ala Asp Gly Leu 35 40 45 Val Asp Thr Glu Phe Val Ala Val Glu Ser Glu His Ser Ala Met Ser 50 55 60 Ala Cys Ile Gly Ala Gln Ala Ala Gly Ala Arg Ala Met Thr Ala Thr 65 70 75 80 Ser Ala Asn Gly Leu Ala Tyr Met Trp Glu Ala Leu Tyr Ile Ala Ala 85 90 95 Ser Met Arg Leu Pro Ile Val Leu Ala Ala Val Asn Arg Ala Leu Ser 100 105 110 Gly Pro Ile Asn Ile His Asn Asp His Ser Asp Thr Met Gly Ala Arg 115 120 125 Asp Ser Gly Trp Ile Gln Leu Tyr Ser Glu Asn Asn Gln Glu Ala Tyr 130 135 140 Asp Asn Met Leu Met Ala His Arg Ile Gly Glu His Pro Asp Val Met 145 150 155 160 Leu Pro Val Met Val Cys Gln Asp Gly Phe Ile Thr Ser His Ala Ile 165 170 175 Glu Asn Ile Glu Leu Val Glu Asp Glu Lys Val Lys Ala Phe Val Gly 180 185 190 Glu Tyr Lys Pro Thr His Tyr Leu Leu Asp Arg Glu Asn Pro Ile Ser 195 200 205 Val Gly Pro Leu Asp Leu Gln Met His Tyr Phe Glu His Lys Arg Gln 210 215 220 Gln Ala Gln Ala Met Glu Asn Ala Lys Lys Val Ile Leu Glu Val Ala 225 230 235 240 Glu Glu Phe Tyr Lys Leu Thr Gly Arg Lys Tyr Gly Phe Phe Glu Glu 245 250 255 Tyr Lys Thr Asp Asp Ala Asp Val Ala Ile Val Val Met Asn Ser Thr 260 265 270 Ala Gly Thr Val Lys Tyr Val Ile Asp Glu Tyr Arg Ala Lys Gly Lys 275 280 285 Lys Val Gly Leu Ile Lys Pro Arg Val Phe Arg Pro Phe Pro Val Asp 290 295 300 Glu Leu Ala Gln Ala Leu Ser Lys Phe Lys Ala Val Ala Val Met Asp 305 310 315 320 Lys Ala Asp Ser Phe Asn Ala Ala Gly Gly Pro Leu Phe Thr Glu Val 325 330 335 Thr Ser Ala Leu Phe Thr Lys Gly Val Phe Gly Pro Lys Val Ile Asn 340 345 350 Tyr Lys Phe Gly Leu Gly Gly Arg Asp Val Lys Val Asp Asp Ile Glu 355 360 365 Val Val Cys Glu Lys Leu Leu Glu Ile Ala Ser Thr Gly Lys Val Asp 370 375 380 Ser Val Tyr Asn Tyr Leu Gly Val Arg Glu 385 390 17311PRTC. thermocellum 17Met Ala Tyr Asn Leu Lys Glu Val Ala Lys Lys Pro Glu Arg Leu Thr 1 5 10 15 Gly Gly His Arg Met Cys Ala Gly Cys Gly Ala Pro Ile Val Val Arg 20 25 30 Gln Val Leu Lys Ala Leu Lys Pro Glu Asp His Ala Val Ile Ser Ala 35 40 45 Ala Thr Gly Cys Leu Glu Val Ser Thr Phe Ile Tyr Pro Tyr Thr Ala 50 55 60 Trp Lys Asp Ser Phe Ile His Ser Ala Phe Glu Asn Thr Gly Ala Thr 65 70 75 80 Ile Ser Gly Ala Glu Ala Ala Tyr Lys Val Leu Lys Lys Lys Gly Lys 85 90 95 Ile Glu Gly Glu Thr Lys Phe Ile Ala Phe Gly Gly Asp Gly Gly Thr 100 105 110 Tyr Asp Ile Gly Leu Gln Ala Leu Ser Gly Ala Met Glu Arg Gly His 115 120 125 Asp Met Val Tyr Val Cys Tyr Asp Asn Gly Ala Tyr Met Asn Thr Gly 130 135 140 Ile Gln Arg Ser Ser Ala Thr Pro Lys Tyr Ala Asp Thr Thr Thr Ser 145 150 155 160 Pro Val Gly Lys Lys Ile Pro Gly Lys Met Gln Pro Arg Lys Asp Leu 165 170 175 Thr Glu Val Leu Val Asn His Arg Ile Pro Tyr Val Ala Gln Thr Ala 180 185 190 Pro Phe Gly Asn Met Lys Asp Leu Tyr Glu Lys Ala Glu Lys Ala Ile 195 200 205 Tyr Thr Pro Gly Pro Ala Phe Leu Asn Val Leu Ala Pro Cys Pro Arg 210 215 220 Gly Trp Arg Tyr Asn Thr Pro Asp Leu Met Glu Leu Ser Lys Leu Ala 225 230 235 240 Val Glu Thr Cys Phe Trp Pro Leu Tyr Glu Val Ile Asp Gly Lys Tyr 245 250 255 Ile Ile Asn Tyr Lys Pro Lys Glu Lys Val Pro Val Lys Glu Phe Leu 260 265 270 Lys Leu Gln Gly Arg Phe Lys His Leu Phe Lys Ala Gly Asn Glu Tyr 275 280 285 Met Leu Glu Glu Ile Gln Lys Glu Val Asp Leu Arg Trp Glu Arg Leu 290 295 300 Leu Lys Leu Ala Gly Glu Ala 305 310 18317PRTC. thermocellum 18Met Asn Asn Asn Lys Val Ile Lys Lys Val Thr Val Val Gly Ala Gly 1 5 10 15 Phe Val Gly Ser Thr Thr Ala Tyr Thr Leu Met Leu Ser Gly Leu Ile 20 25 30 Ser Glu Ile Val Leu Ile Asp Ile Asn Ala Lys Lys Ala Asp Gly Glu 35 40 45 Val Met Asp Leu Asn His Gly Met Pro Phe Val Arg Pro Val Glu Ile 50 55 60 Tyr Arg Gly Asp Tyr Lys Asp Cys Ala Gly Ser Asp Ile Val Ile Ile 65 70 75 80 Thr Ala Gly Ala Asn Gln Lys Glu Gly Glu Thr Arg Ile Asp Leu Val 85 90 95 Lys Arg Asn Thr Glu Val Phe Lys Asn Ile Ile Asn Glu Ile Val Lys 100 105 110 Tyr Asn Asn Asp Cys Ile Leu Leu Val Val Thr Asn Pro Val Asp Ile 115 120 125 Leu Thr Tyr Val Thr Tyr Lys Leu Ser Gly Phe Pro Lys Asn Lys Val 130 135 140 Ile Gly Ser Gly Thr Val Leu Asp Thr Ala Arg Phe Arg Tyr Leu Leu 145 150 155 160 Ser Glu His Val Lys Val Asp Ala Arg Asn Val His Ala Tyr Ile Ile 165 170 175 Gly Glu His Gly Asp Thr Glu Val Ala Ala Trp Ser Leu Ala Asn Ile 180 185 190 Ala Gly Ile Pro Met Asp Arg Tyr Cys Asp Glu Cys His Gln Cys Glu 195 200 205 Glu Gln Ile Ser Arg Asn Lys Ile Tyr Glu Ser Val Lys Asn Ala Ala 210 215 220 Tyr Glu Ile Ile Arg Asn Lys Gly Ala Thr Tyr Tyr Ala Val Ala Leu 225 230 235 240 Ala Val Arg Arg Ile Val Glu Ala Ile Val Arg Asn Glu Asn Ser Ile 245 250 255 Leu Thr Val Ser Ser Leu Leu Glu Gly Gln Tyr Gly Leu Ser Asp Val 260 265 270 Cys Leu Ser Val Pro Thr Ile Val Gly Val Asn Gly Ile Glu Glu Ile 275 280 285 Leu Asn Val Pro Phe Asn Asp Glu Glu Ile Gln Leu Leu Arg Lys Ser 290 295 300 Gly Asn Thr Leu Lys Glu Ile Ile Lys Thr Leu Asp Ile 305 310 315 1981PRTC. thermocellum 19Met Lys Val Ser Ile Cys Ile Gly Ser Ser Cys His Leu Lys Gly Ala 1 5 10 15 Lys Gln Ile Val Glu Gln Leu Gln Ser Leu Val Ala Asp Tyr Asn Leu 20 25 30 Lys Glu Lys Val Glu Leu Gly Gly Ala Phe Cys Met Lys Asn Cys Val 35 40 45 Asn Gly Val Ser Val Thr Val Asp Asp Lys Leu Phe Ser Val Thr Pro 50 55 60 Glu Asn Val Lys Ser Phe Phe Glu Thr Glu Ile Leu Lys Lys Leu Glu 65 70 75 80 Asp 20556PRTC. thermocellum 20Met Thr Glu Cys Leu Gln Thr Lys Lys Ser Asn Cys Lys Asn Cys Tyr 1 5 10 15 Lys Cys Ile Arg His Cys Pro Val Lys Ser Leu Lys Phe Thr Asp Gly 20 25 30 Gln Ala His Ile Val Arg Asp Glu Cys Val Leu Cys Gly Glu Cys Tyr 35 40 45 Val Val Cys Pro Gln Asn Ala Lys Gln Ile Arg Ser Asp Val Glu Lys 50 55 60 Ala Lys Gln Leu Val Leu Lys Tyr Asp Val Tyr Ala Ser Ile Ala Pro 65 70 75 80 Ser Phe Val Ala Trp Phe His Asn Lys Ser Ile His Asp Met Glu Gln 85 90 95 Ala Leu Ile Lys Leu Gly Phe Lys Gly Ala Asp Glu Thr Ala Lys Gly 100 105 110 Ala Tyr Ile Val Lys Lys Gln Tyr Glu Lys Met Ile Glu Glu Lys Lys 115 120 125 Ser Lys Ile Ile Ile Ser Ser Cys Cys His Thr Val Asn Thr Leu Ile 130 135 140 Gln Arg His Tyr Thr Gly Ala Ile Gln Tyr Leu Ala Asp Val Val Ser 145 150 155 160 Pro Met Leu Ala His Ala Gln Met Leu Lys Lys Glu His Lys Gly Ala 165 170 175 Lys Val Val Phe Ile Gly Pro Cys Ile Ser Lys Lys Asp Glu Ala Glu 180 185 190 Lys Tyr Lys Gly Tyr Val Glu Leu Val Leu Thr Phe Asp Glu Leu Asp 195 200

205 Glu Trp Leu Lys Ser Glu Asn Ile Thr Ile Glu Ser Asn Arg Gly Ser 210 215 220 Ser Lys Glu Gly Arg Thr Arg Ser Phe Pro Val Ser Gly Gly Ile Ile 225 230 235 240 Ser Ser Met Asp Lys Asp Leu Gly Tyr His Tyr Met Val Val Asp Gly 245 250 255 Met Glu Asn Cys Ile Asn Ala Leu Glu Asn Ile Glu Arg Gly Glu Ile 260 265 270 Asp Asn Cys Phe Ile Glu Met Ser Ala Cys Arg Gly Ser Cys Ile Asn 275 280 285 Gly Pro Pro Ala Arg Arg Lys Ser Asn Asn Ile Val Gly Ala Ile Leu 290 295 300 Ala Val Asn Lys Asn Thr Gly Ala Lys Asp Phe Ser Val Pro Met Pro 305 310 315 320 Glu Pro Glu Lys Leu Lys Lys Glu Phe Arg Phe Glu Gly Val His Lys 325 330 335 Ile Met Pro Gly Gly Thr Ala Ile Glu Glu Ile Leu Lys Lys Met Gly 340 345 350 Lys Thr Ser Ile Glu His Glu Leu Asn Cys Gly Ser Cys Gly Tyr Asp 355 360 365 Thr Cys Arg Asp Lys Ala Val Ala Val Leu Asn Gly Lys Ala Asp Leu 370 375 380 Thr Met Cys Leu Pro Tyr Leu Lys Glu Lys Ala Glu Ser Phe Ser Asp 385 390 395 400 Ala Ile Ile Lys Asn Thr Pro Asn Gly Val Ile Val Leu Asn Glu Asp 405 410 415 Leu Glu Ile Gln Gln Ile Asn Asn Ser Ala Lys Arg Ile Leu Asn Leu 420 425 430 Ser Pro Ser Thr Asp Leu Leu Gly Ser Pro Val Ser Arg Ile Leu Asp 435 440 445 Pro Ile Asp Tyr Ile Leu Ala Leu Arg Glu Gly Lys Asn Cys Tyr Tyr 450 455 460 Lys Arg Lys Tyr Phe Ala Glu Tyr Lys Lys Tyr Val Asp Glu Thr Ile 465 470 475 480 Ile Tyr Asp Lys Glu Tyr His Val Ile Ile Ile Ile Met Arg Asp Val 485 490 495 Thr Glu Glu Glu Lys Ile Lys Ala Leu Lys Asn Lys Gln Ser Glu Ala 500 505 510 Ala Ile Glu Ile Ala Asp Lys Val Val Glu Lys Gln Met Arg Val Val 515 520 525 Gln Glu Ile Ala Leu Leu Leu Gly Glu Thr Ala Ala Glu Thr Lys Ile 530 535 540 Ala Leu Thr Lys Leu Lys Glu Thr Met Glu Asp Glu 545 550 555 21389PRTC. thermocellum 21Met Asn Asp Leu Cys Val Asp Leu Gly Tyr Lys Ser Leu Asn Lys Phe 1 5 10 15 Gly Glu Gln Leu Cys Gly Asp Met Ile Gln Val Val Lys Asp Asp Asp 20 25 30 Thr Thr Ile Leu Val Leu Ala Asp Gly Leu Gly Ser Gly Val Lys Ala 35 40 45 Asn Ile Leu Ser Thr Leu Thr Ser Lys Ile Ile Ser Thr Met Ile Ala 50 55 60 Ala His Met Gly Ile Glu Glu Cys Val Asn Thr Ile Met Ser Thr Leu 65 70 75 80 Pro Val Cys Lys Val Arg Gly Ile Ala Tyr Ser Thr Phe Thr Ile Ile 85 90 95 Lys Ile Thr Asn Asn Thr Tyr Ala Glu Ile Ile Gln Tyr Asp Asn Pro 100 105 110 Leu Val Ile Leu Leu Arg Asn Gly Lys Lys Tyr Asp Tyr Pro Thr Gln 115 120 125 Thr Lys Ile Ile Ser Gly Lys Lys Ile Val Glu Ser Lys Ile Arg Leu 130 135 140 Asn Cys Asp Asp Val Phe Val Val Met Ser Asp Gly Ala Ile Tyr Ala 145 150 155 160 Gly Val Gly Gln Thr Leu Asn Tyr Gly Trp Gln Arg Glu Asn Ile Ile 165 170 175 Glu Phe Ile Glu Ser His Tyr Asp Lys Ser Leu Ser Ala Asn Ala Leu 180 185 190 Thr Ser Leu Leu Ile Asp Thr Cys Asn Asn Leu Tyr Ala Asn Met Pro 195 200 205 Gly Asp Asp Thr Thr Ile Ala Ala Ile Lys Ile Arg Lys Arg Gln Val 210 215 220 Val Asn Leu Met Phe Gly Pro Pro Gln Asn Pro Glu Asp Val His Asn 225 230 235 240 Met Met Ser Leu Phe Phe Ala Lys Gln Gly Arg His Ile Val Cys Gly 245 250 255 Gly Thr Thr Ser Thr Leu Ala Ala Lys Phe Leu Gly Lys Glu Leu Glu 260 265 270 Thr Thr Ile Asp Tyr Ile Asp Pro Arg Ile Pro Pro Ile Ala Arg Ile 275 280 285 Glu Gly Val Asp Leu Val Thr Glu Gly Val Leu Thr Ile Ser Arg Val 290 295 300 Leu Glu Tyr Ala Lys Asp Tyr Ile Gly Lys Asn Ile Leu Tyr Asn Glu 305 310 315 320 Trp His Ser Lys Asn Asp Gly Ala Ser Ile Ile Ala Arg Met Leu Phe 325 330 335 Glu Glu Ala Thr Asp Ile Asn Phe Tyr Val Gly Lys Ala Ile Asn Pro 340 345 350 Ala His Gln Asn Pro Asn Leu Pro Ile Gly Phe Asn Ile Lys Met Gln 355 360 365 Leu Val Glu Glu Leu Ser Lys Ile Leu Lys Gln Met Gly Lys Thr Ile 370 375 380 Asn Leu Ser Tyr Phe 385 22165PRTC. thermocellum 22Met Ser Val Thr Met Ser Glu Ala Phe Asp Tyr Ser Met Ile Asp Asn 1 5 10 15 Ile Leu Ser Glu His Gly Thr Ser Glu Thr Ala Ile Ile Ala Ile Leu 20 25 30 Gln Ser Ile Gln Glu Glu Tyr His Tyr Ile Pro Lys Glu Val Phe Pro 35 40 45 Tyr Leu Ser Lys Lys Leu Lys Val Ser Glu Ala Arg Ile Phe Ser Val 50 55 60 Ala Thr Phe Tyr Glu Asn Phe Ser Leu Glu Pro Lys Gly Lys Tyr Ile 65 70 75 80 Ile Lys Val Cys Asp Gly Thr Ala Cys His Val Arg Lys Ser Ile Pro 85 90 95 Ile Ile Glu Arg Leu Arg Lys Glu Leu Gly Leu Ser Gly Thr Lys Pro 100 105 110 Thr Thr Asp Asp Leu Met Phe Thr Val Glu Thr Val Ser Cys Leu Gly 115 120 125 Ala Cys Gly Leu Ala Pro Val Ile Thr Val Asn Asp Lys Val Tyr Ala 130 135 140 Glu Met Thr Pro Asp Lys Ala Ser Glu Leu Ile Lys Gln Leu Arg Glu 145 150 155 160 Gly Asp Ala Asp Ala 165 23624PRTC. thermocellum 23Met Leu Lys Asn Arg Glu Glu Leu Arg Lys Ala Arg Glu Met Tyr Ser 1 5 10 15 Arg Tyr Leu Lys Ala Glu Lys Arg Arg Val Leu Val Cys Ala Gly Thr 20 25 30 Gly Cys Val Ser Gly Gly Ser Met Glu Ile Phe Glu Arg Leu Ser Glu 35 40 45 Leu Val Ser Lys Arg Gly Met Asp Cys Gln Val Glu Leu Lys Glu Glu 50 55 60 Pro His Asp Asn Thr Ile Gly Met Lys Lys Ser Gly Cys His Gly Phe 65 70 75 80 Cys Glu Met Gly Pro Leu Val Arg Ile Glu Pro Glu Gly Tyr Leu Tyr 85 90 95 Thr Lys Val Lys Leu Glu Asp Cys Glu Glu Ile Val Asp Arg Thr Ile 100 105 110 Val Ala Gly Glu His Ile Glu Arg Leu Ala Tyr Lys Gln Asn Gly Val 115 120 125 Val Tyr Lys Lys Gln Asp Glu Ile Pro Phe Tyr Lys Lys Gln Thr Arg 130 135 140 Leu Val Leu Glu His Cys Gly Gln Ile Asp Ser Thr Ser Ile Thr Glu 145 150 155 160 Tyr Leu Ala Thr Gly Gly Tyr Tyr Ala Leu Glu Lys Ala Leu Phe Asp 165 170 175 Met Thr Gly Asp Glu Ile Ile Asn Glu Ile Thr Glu Ala Asn Leu Arg 180 185 190 Gly Arg Gly Gly Gly Gly Phe Pro Ala Gly Arg Lys Trp Ala Gln Val 195 200 205 Lys Arg Gln Asn Ala Lys Gln Lys Tyr Val Val Cys Asn Gly Asp Glu 210 215 220 Gly Asp Pro Gly Ala Phe Met Asp Arg Ser Ile Met Glu Gly Asp Pro 225 230 235 240 His Arg Met Ile Glu Gly Met Ile Ile Ala Gly Ile Ala Cys Gly Ala 245 250 255 Ser Glu Gly Tyr Ile Tyr Val Arg Ala Glu Tyr Pro Leu Ala Val Ser 260 265 270 Arg Leu Lys Arg Ala Ile Glu Gln Ala Lys Glu Phe Gly Leu Leu Gly 275 280 285 Glu Asn Ile Leu Gly Ser Asn Phe Ser Phe Asn Ile His Ile Asn Arg 290 295 300 Gly Ala Gly Ala Phe Val Cys Gly Glu Gly Ser Ala Leu Thr Ala Ser 305 310 315 320 Ile Glu Gly Lys Arg Gly Met Pro Arg Val Lys Pro Pro Arg Thr Val 325 330 335 Glu Gln Gly Leu Phe Asp Met Pro Thr Val Leu Asn Asn Val Glu Thr 340 345 350 Phe Ala Asn Val Pro Leu Ile Ile Lys Asn Gly Ala Asp Trp Tyr Lys 355 360 365 Ser Ile Gly Thr Glu Lys Ser Pro Gly Thr Lys Ala Phe Ala Leu Thr 370 375 380 Gly Asn Ile Glu Asn Thr Gly Leu Ile Glu Ile Pro Met Gly Thr Thr 385 390 395 400 Leu Arg Glu Val Ile Phe Asp Ile Gly Gly Gly Met Arg Asn Gly Ala 405 410 415 Asp Phe Lys Ala Val Gln Ile Gly Gly Pro Ser Gly Gly Cys Leu Ser 420 425 430 Glu Lys Asp Leu Asp Leu Pro Leu Asp Phe Asp Ser Leu Lys Lys Ala 435 440 445 Gly Ala Met Ile Gly Ser Gly Gly Leu Val Val Met Asp Ser Asn Thr 450 455 460 Cys Met Val Glu Val Ala Arg Phe Phe Met Asn Phe Thr Gln Asn Glu 465 470 475 480 Ser Cys Gly Lys Cys Val Pro Cys Arg Glu Gly Thr Lys Arg Met Leu 485 490 495 Glu Ile Leu Glu Arg Ile Val Glu Gly Asn Gly Gln Asp Gly Asp Ile 500 505 510 Glu Leu Leu Leu Glu Leu Ala Asp Thr Ile Ser Ala Thr Ala Leu Cys 515 520 525 Gly Leu Gly Lys Ala Ala Ala Phe Pro Val Val Ser Thr Ile Lys Asn 530 535 540 Phe Arg Glu Glu Tyr Glu Ala His Ile Tyr Asp Lys Arg Cys Pro Thr 545 550 555 560 Gly Asn Cys Gln Lys Leu Lys Thr Ile Thr Ile Asp Ala Ser Leu Cys 565 570 575 Lys Gly Cys Ser Lys Cys Ala Arg Ser Cys Pro Val Gly Ala Ile Thr 580 585 590 Gly Lys Val Lys Glu Pro Phe Val Ile Asp Gln Ser Lys Cys Ile Lys 595 600 605 Cys Gly Ala Cys Ile Glu Thr Cys Ala Phe His Ala Ile Leu Glu Gly 610 615 620 24566PRTC. thermocellum 24Met Asp Asn Arg Glu Tyr Met Leu Ile Asp Gly Ile Pro Val Glu Ile 1 5 10 15 Asn Gly Glu Lys Asn Leu Leu Glu Leu Ile Arg Lys Ala Gly Ile Lys 20 25 30 Leu Pro Thr Phe Cys Tyr His Ser Glu Leu Ser Val Tyr Gly Ala Cys 35 40 45 Arg Met Cys Met Val Glu Asn Glu Trp Gly Gly Leu Asp Ala Ala Cys 50 55 60 Ser Thr Pro Pro Arg Ala Gly Met Ser Ile Lys Thr Asn Thr Glu Arg 65 70 75 80 Leu Gln Lys Tyr Arg Lys Met Ile Leu Glu Leu Leu Leu Ala Asn His 85 90 95 Cys Arg Asp Cys Thr Thr Cys Asn Asn Asn Gly Lys Cys Lys Leu Gln 100 105 110 Asp Leu Ala Met Arg Tyr Asn Ile Ser His Ile Arg Phe Pro Asn Thr 115 120 125 Ala Ser Asn Pro Asp Val Asp Asp Ser Ser Leu Cys Ile Thr Arg Asp 130 135 140 Arg Ser Lys Cys Ile Leu Cys Gly Asp Cys Val Arg Val Cys Asn Glu 145 150 155 160 Val Gln Asn Val Gly Ala Ile Asp Phe Ala Tyr Arg Gly Ser Lys Met 165 170 175 Thr Ile Ser Thr Val Phe Asp Lys Pro Ile Phe Glu Ser Asn Cys Val 180 185 190 Gly Cys Gly Gln Cys Ala Leu Ala Cys Pro Thr Gly Ala Ile Val Val 195 200 205 Lys Asp Asp Thr Gln Lys Val Trp Lys Glu Ile Tyr Asp Lys Asn Thr 210 215 220 Arg Val Ser Val Gln Ile Ala Pro Ala Val Arg Val Ala Leu Gly Lys 225 230 235 240 Glu Leu Gly Leu Asn Asp Gly Glu Asn Ala Ile Gly Lys Ile Val Ala 245 250 255 Ala Leu Arg Arg Met Gly Phe Asp Asp Ile Phe Asp Thr Ser Thr Gly 260 265 270 Ala Asp Leu Thr Val Leu Glu Glu Ser Ala Glu Leu Leu Arg Arg Ile 275 280 285 Arg Glu Gly Lys Asn Asp Met Pro Leu Phe Thr Ser Cys Cys Pro Ala 290 295 300 Trp Val Asn Tyr Cys Glu Lys Phe Tyr Pro Glu Leu Leu Pro His Val 305 310 315 320 Ser Thr Cys Arg Ser Pro Met Gln Met Phe Ala Ser Ile Ile Lys Glu 325 330 335 Glu Tyr Ser Thr Ser Ser Lys Arg Leu Val His Val Ala Val Met Pro 340 345 350 Cys Thr Ala Lys Lys Phe Glu Ala Ala Arg Lys Glu Phe Lys Val Asn 355 360 365 Gly Val Pro Asn Val Asp Tyr Val Leu Thr Thr Gln Glu Leu Val Arg 370 375 380 Met Ile Lys Glu Ser Gly Ile Val Phe Ser Glu Leu Glu Pro Glu Ala 385 390 395 400 Ile Asp Met Pro Phe Gly Thr Tyr Thr Gly Ala Gly Val Ile Phe Gly 405 410 415 Val Ser Gly Gly Val Thr Glu Ala Val Leu Arg Arg Val Val Ser Asp 420 425 430 Lys Ser Pro Thr Ser Phe Arg Ser Leu Ala Tyr Thr Gly Val Arg Gly 435 440 445 Met Asn Gly Val Lys Glu Ala Ser Val Met Tyr Gly Asp Arg Lys Leu 450 455 460 Lys Val Ala Val Val Ser Gly Leu Lys Asn Ala Gly Asp Leu Ile Glu 465 470 475 480 Arg Ile Lys Ala Gly Glu His Tyr Asp Leu Val Glu Val Met Ala Cys 485 490 495 Pro Gly Gly Cys Ile Asn Gly Gly Gly Gln Pro Phe Val Gln Ser Glu 500 505 510 Glu Arg Glu Lys Arg Gly Lys Gly Leu Tyr Ser Ala Asp Lys Leu Cys 515 520 525 Asn Ile Lys Ser Ser Glu Glu Asn Pro Leu Met Met Thr Leu Tyr Lys 530 535 540 Gly Ile Leu Lys Gly Arg Val His Glu Leu Leu His Val Asp Tyr Ala 545 550 555 560 Ser Lys Lys Glu Ala Lys 565 2581PRTC. thermocellum 25Met Leu Glu Ile Lys Ile Cys Val Gly Ser Ser Cys His Leu Lys Gly 1 5 10 15 Ser Tyr Asn Val Ile Asn Glu Phe Gln His Leu Ile Glu Glu Lys Ala 20 25 30 Leu His Asp Lys Ile Asp Ile Lys Ala Thr Phe Cys Met Lys Gln Cys 35 40 45 Gln Lys Asn Gly Val Ala Val Glu Val Asn Asn Glu Ile Phe Gly Val 50 55 60 Leu Pro Glu Ala Ala Glu Glu Phe Phe Lys Asn Val Ile Leu Pro Lys 65 70 75 80 Val 26128PRTC. thermocellum 26Met Ser Phe Phe Thr Met Thr Lys Thr Leu Ile Lys Ser Ile Phe His 1 5 10 15 Gly Pro Tyr Thr Val Arg Tyr Pro Leu Glu Lys Lys Glu Pro Phe Pro 20 25 30 Ala Ser Arg Gly Arg Ile Glu Ile Asn Ile Gln Asp Cys Ile Phe Cys 35 40 45 Gly Leu Cys Ala Arg Arg Cys Pro Thr Gly Ala Ile Asn Val Glu Lys 50 55 60 Pro Glu Ser Arg Trp Ser Ile Asn Arg Leu Arg Cys Ile Gln Cys Gly 65 70 75 80 Tyr Cys Ser Glu Val Cys Pro Lys Lys Cys Leu Lys Met Asn Asn Met 85 90 95 Tyr Pro Ala Pro Ser Phe Glu Asn Ile Glu Asp Val Tyr Gln Asn Ala 100 105

110 Arg Val Pro Asp Asn Lys Glu Asn Asn Arg Asn Ile Ala Gly Ala Cys 115 120 125 27359PRTC. thermocellum 27Met Gly Lys Lys Thr Val Ile Pro Phe Gly Pro Gln His Pro Val Leu 1 5 10 15 Pro Glu Pro Ile His Leu Asp Leu Val Leu Glu Asp Glu Thr Val Val 20 25 30 Glu Ala Ile Pro Ser Ile Gly Tyr Ile His Arg Gly Leu Glu Lys Leu 35 40 45 Val Glu Lys Lys Asp Tyr Gln Gln Phe Val Tyr Val Ala Glu Arg Ile 50 55 60 Cys Gly Ile Cys Ser Phe Met His Gly Met Gly Tyr Cys Met Ser Ile 65 70 75 80 Glu Asn Ile Met Gly Val Gln Ile Pro Glu Arg Ala Glu Phe Leu Arg 85 90 95 Thr Ile Trp Ala Glu Leu Ser Arg Ile His Ser His Met Leu Trp Leu 100 105 110 Gly Leu Leu Ala Asp Ala Leu Gly Phe Glu Ser Leu Phe Met His Ser 115 120 125 Trp Arg Leu Arg Glu Gln Ile Leu Asp Ile Phe Glu Glu Thr Thr Gly 130 135 140 Gly Arg Val Ile Phe Ser Val Cys Asp Ile Gly Gly Val Arg Arg Asp 145 150 155 160 Ile Asp Ser Glu Met Leu Lys Lys Ile Asn Ser Ile Leu Asp Gly Phe 165 170 175 Glu Lys Glu Phe Ser Glu Ile Thr Lys Val Phe Leu Asn Asp Ser Ser 180 185 190 Val Lys Leu Arg Thr Gln Gly Leu Gly Val Leu Ser Arg Glu Glu Ala 195 200 205 Phe Glu Leu Gly Ala Val Gly Pro Met Ala Arg Ala Ser Gly Ile Asp 210 215 220 Ile Asp Met Arg Lys Ser Gly Tyr Ala Ala Tyr Gly Lys Leu Lys Ile 225 230 235 240 Glu Pro Val Val Glu Thr Ala Gly Asp Cys Tyr Ala Arg Thr Ser Val 245 250 255 Arg Ile Arg Glu Val Phe Gln Ser Ile Asp Leu Ile Arg Gln Cys Ile 260 265 270 Ser Leu Ile Pro Asp Gly Glu Ile Lys Val Lys Ile Val Gly Asn Pro 275 280 285 Ser Gly Glu Tyr Phe Thr Arg Leu Glu Gln Pro Arg Gly Glu Val Leu 290 295 300 Tyr Tyr Val Lys Ala Asn Gly Thr Lys Phe Leu Glu Arg Phe Arg Val 305 310 315 320 Arg Thr Pro Thr Phe Ala Asn Ile Pro Ala Leu Leu His Thr Leu Lys 325 330 335 Gly Cys Gln Leu Ala Asp Val Pro Val Leu Ile Leu Thr Ile Asp Pro 340 345 350 Cys Ile Ser Cys Thr Glu Arg 355 28119PRTC. thermocellum 28Met Ala Gln Gln Thr Ile Asn Thr Ile Ser Pro Asn Glu Leu Leu Ala 1 5 10 15 Tyr Ala Leu Arg Leu Lys Asn Ala Asn Tyr Arg Leu Val Ala Ile Ser 20 25 30 Cys Thr Asn Ala Glu Asn Gly Val Glu Met Ser Tyr Ser Phe Asp Ser 35 40 45 Gly Ser Asp Phe Thr Asn Leu Arg Ile Thr Val Ala Pro Gly Asp Glu 50 55 60 Ile Glu Ser Ile Ser Ser Ile Tyr Ser Tyr Ser Phe Leu Tyr Glu Asn 65 70 75 80 Glu Ile Lys Glu Leu Phe Gly Val Asn Ile Thr Gly Ile Ser Pro Asp 85 90 95 Tyr Lys Asp Lys Leu Tyr Arg Ile Ser Val Lys Thr Pro Phe Asn Met 100 105 110 Lys Glu Gly Asp Lys Asn Gly 115 29145PRTC. thermocellum 29Met Asn Phe Ser Lys Lys Ser Pro Trp Ile Leu His Tyr Asp Gly Ser 1 5 10 15 Ser Cys Asn Gly Cys Asp Ile Glu Val Leu Ala Cys Leu Thr Pro Leu 20 25 30 Tyr Asp Ile Glu Arg Phe Gly Val Ile Asn Thr Gly Asn Pro Lys His 35 40 45 Ala Asp Ile Leu Leu Ile Thr Gly Ser Ile Asn Glu Gln Asn Lys Ser 50 55 60 Val Val Lys Gln Leu Tyr Glu Gln Met Ala Asp Pro Lys Val Val Val 65 70 75 80 Ala Val Gly Ile Cys Ala Ala Thr Gly Gly Ile Phe Ser Glu Cys Tyr 85 90 95 Asn Val Ser Gly Gly Val Asp Lys Ile Ile Pro Val Asp Val Tyr Val 100 105 110 Pro Gly Cys Ala Ala Arg Pro Glu Ala Ile Ile Asp Gly Val Val Lys 115 120 125 Ala Leu Gly Ile Leu Glu Glu Arg Gln Lys Tyr Ala Arg Lys Lys Asp 130 135 140 Lys 145 30291PRTC. thermocellum 30Met Ser Gln Ile Ile Arg Leu Val Leu Tyr Ile Ile Ala Ile Ile Ile 1 5 10 15 Val Ala Pro Leu Leu Gly Gly Leu Leu Thr Gly Ile Asp Arg Val Ile 20 25 30 Thr Ala Arg Met Gln Gly Arg Lys Gly Pro Ser Val Leu Gln Pro Phe 35 40 45 Tyr Asp Val Leu Lys Leu Phe Gln Lys Glu Ser Ile Glu Val Asn Thr 50 55 60 Met His Arg Phe Phe Val Tyr Ile Ser Leu Ile Phe Val Ile Phe Thr 65 70 75 80 Thr Val Ile Met Leu Leu Gly Gly Asp Ile Leu Leu Ala Leu Phe Ala 85 90 95 Leu Thr Leu Gly Ser Ile Phe Phe Val Leu Gly Gly Tyr Ala Ser Asn 100 105 110 Ser Pro Tyr Ser Thr Ile Gly Ser Glu Arg Glu Leu Leu Gln Met Met 115 120 125 Ala Phe Glu Pro Met Leu Leu Leu Ala Ala Ile Gly Leu Tyr Tyr Gly 130 135 140 Asp Lys Ser Phe Phe Ile Lys Asp Ile Val Thr Ala Arg Ile Pro Ser 145 150 155 160 Ile Val Tyr Leu Pro Gly Val Phe Leu Gly Leu Leu Tyr Val Leu Thr 165 170 175 Phe Lys Leu Arg Lys Ser Pro Phe Asp Leu Ser Met Ser His His Gly 180 185 190 His Gln Glu Ile Val Gln Gly Ile Thr Thr Glu Tyr Ser Gly Lys Asp 195 200 205 Leu Ala Ile Ile Gln Ile Thr His Trp Tyr Glu Thr Ile Ile Ala Leu 210 215 220 Ala Leu Val Tyr Leu Phe Phe Ala Phe Arg Ser Pro Phe Ser His Val 225 230 235 240 Ile Ala Ile Leu Ala Cys Ile Ile Ala Phe Leu Leu Glu Ile Val Val 245 250 255 Asp Asn Ala Phe Ala Arg Ala Lys Trp Glu Phe Ala Leu Lys Ser Thr 260 265 270 Trp Ile Val Thr Gly Val Leu Ala Ser Val Asn Leu Ile Ile Leu Ser 275 280 285 Phe Phe Arg 290 31636PRTC. thermocellum 31Met Asn Ala Ile Leu Ile Leu Ile Leu Phe Pro Leu Leu Ala Ser Val 1 5 10 15 Thr Val Leu Ser Val Arg Lys Asp Ala Ile Arg Asn Ile Ile Val Arg 20 25 30 Ile Phe Ala Phe Ile Thr Gly Ile Leu Thr Leu Phe Val Val Cys Arg 35 40 45 Tyr Phe Lys Asp Gly Ile Ser Leu Ser Ile Glu Asn Arg Asn Ile Ile 50 55 60 Asp Met Thr Ile Ser Leu Ala Glu Val Leu Ile Ala Ala Tyr Ile Ile 65 70 75 80 Phe Thr Gly Ile Lys Asn Lys Lys Phe Ile Val Ser Ile Phe Ala Ala 85 90 95 Val Gln Thr Ala Leu Ile Leu Trp Phe Glu Phe Thr Gln Lys His Gly 100 105 110 Ile Asn Val His Ser Asp Ile Val Phe Asp Arg Leu Ser Ala Val Met 115 120 125 Val Leu Ile Val Gly Cys Ile Gly Ser Leu Ile Leu Ile Tyr Thr Val 130 135 140 Gly Tyr Met Lys Trp Tyr His Ile His His Glu Gly Tyr Lys Glu Arg 145 150 155 160 Lys Ser Phe Phe Phe Ser Val Ile Phe Leu Phe Leu Phe Ala Met Phe 165 170 175 Gly Leu Ile Phe Ser Asn Asn Leu Ile Trp Met Tyr Phe Cys Trp Glu 180 185 190 Leu Thr Thr Leu Cys Ser Tyr Leu Leu Ile Gly Tyr Thr Arg Thr Pro 195 200 205 Glu Ala Val Asn Asn Ser Phe His Ala Leu Ala Ile Asn Leu Gly Gly 210 215 220 Gly Leu Ala Phe Ala Ser Ala Met Val Tyr Ile Gly Thr Asn Phe Lys 225 230 235 240 Thr Leu Glu Leu Ser Ala Leu Thr Ala Met Lys Leu Glu Leu Ala Val 245 250 255 Leu Ile Pro Val Phe Leu Leu Cys Ile Ala Ala Leu Thr Lys Ser Ala 260 265 270 Gln Met Pro Phe Ser Ser Trp Leu Leu Gly Ala Met Val Ala Pro Thr 275 280 285 Pro Ser Ser Ala Leu Leu His Ser Ala Thr Met Val Lys Ala Gly Val 290 295 300 Tyr Leu Leu Ile Arg Leu Ala Pro Leu Leu Ala Gly Thr Thr Ile Gly 305 310 315 320 Lys Val Ile Ala Leu Leu Gly Ala Val Thr Phe Leu Ala Ser Ser Ile 325 330 335 Ile Ala Ile Ser Lys Ser Asp Ala Lys Lys Ile Leu Ala Tyr Ser Thr 340 345 350 Ile Ser Asn Leu Gly Leu Ile Val Thr Cys Ala Ala Ile Gly Thr Gln 355 360 365 Glu Ser Leu Trp Ala Ala Ile Leu Leu Leu Ile Phe His Ser Ile Ser 370 375 380 Lys Ser Leu Leu Phe Leu Thr Gly Gly Ser Val Glu His Gln Ile Gly 385 390 395 400 Ser Arg Asn Val Glu Asp Met Asp Ile Leu Leu Gln Val Ser Arg Arg 405 410 415 Leu Ser Val Tyr Met Ile Val Gly Ile Ala Gly Met Phe Leu Ala Pro 420 425 430 Phe Gly Met Leu Ile Ser Lys Trp Val Ala Met Lys Ala Phe Ile Asp 435 440 445 Ser Lys Asn Ile Leu Thr Val Ile Ile Leu Gly Tyr Gly Ser Ala Thr 450 455 460 Thr Leu Phe Tyr Trp Thr Lys Trp Met Gly Lys Leu Val Ala Asn Ala 465 470 475 480 Asn Arg Lys Asp His Ile Lys His Thr Phe His Ile Asp Glu Glu Ile 485 490 495 Pro Ile Phe Ile His Ala Val Leu Val Val Leu Ser Cys Phe Thr Phe 500 505 510 Pro Leu Val Ser Arg Tyr Val Leu Val Pro Tyr Leu Ser Gly Leu Phe 515 520 525 Gly Pro Asp Val Pro Ile Pro Ile Gly Thr Ser Asp Val Asn Ile Met 530 535 540 Leu Ile Met Leu Ser Met Leu Leu Ile Leu Pro Ile Ser Phe Ile Pro 545 550 555 560 Ile Tyr Lys Ser Asp Arg Arg Arg Ile Val Pro Ile Tyr Met Ala Gly 565 570 575 Glu Asn Thr Gly Asp Asn Glu Ser Phe Tyr Gly Ala Phe Asp Glu Lys 580 585 590 Arg Lys Val Glu Leu His Asn Trp Tyr Met Lys Asn Phe Phe Ser Val 595 600 605 Lys Lys Leu Thr Phe Trp Ser Asn Leu Leu Cys Ala Val Val Ile Leu 610 615 620 Val Gly Val Val Leu Leu Ile Gly Gly Ile Thr Lys 625 630 635 32582PRTC. thermocellum 32Met Gln Met Val Asn Val Thr Ile Asp Asn Cys Lys Ile Gln Val Pro 1 5 10 15 Ala Asn Tyr Thr Val Leu Glu Ala Ala Lys Gln Ala Asn Ile Asp Ile 20 25 30 Pro Thr Leu Cys Phe Leu Lys Asp Ile Asn Glu Val Gly Ala Cys Arg 35 40 45 Met Cys Val Val Glu Val Lys Gly Ala Arg Ser Leu Gln Ala Ala Cys 50 55 60 Val Tyr Pro Val Ser Glu Gly Leu Glu Val Tyr Thr Gln Thr Pro Ala 65 70 75 80 Val Arg Glu Ala Arg Lys Val Thr Leu Glu Leu Ile Leu Ser Asn His 85 90 95 Glu Lys Lys Cys Leu Thr Cys Val Arg Ser Glu Asn Cys Glu Leu Gln 100 105 110 Arg Leu Ala Lys Asp Leu Asn Val Lys Asp Ile Arg Phe Glu Gly Glu 115 120 125 Met Ser Asn Leu Pro Ile Asp Asp Leu Ser Pro Ser Val Val Arg Asp 130 135 140 Pro Asn Lys Cys Val Leu Cys Arg Arg Cys Val Ser Met Cys Lys Asn 145 150 155 160 Val Gln Thr Val Gly Ala Ile Asp Val Thr Glu Arg Gly Phe Arg Thr 165 170 175 Thr Val Ser Thr Ala Phe Asn Lys Pro Leu Ser Glu Val Pro Cys Val 180 185 190 Asn Cys Gly Gln Cys Ile Asn Val Cys Pro Val Gly Ala Leu Arg Glu 195 200 205 Lys Asp Asp Ile Asp Lys Val Trp Glu Ala Leu Ala Asn Pro Glu Leu 210 215 220 His Val Val Val Gln Thr Ala Pro Ala Val Arg Val Ala Leu Gly Glu 225 230 235 240 Glu Phe Gly Met Pro Ile Gly Ser Arg Val Thr Gly Lys Met Val Ala 245 250 255 Ala Leu Ser Arg Leu Gly Phe Lys Lys Val Phe Asp Thr Asp Thr Ala 260 265 270 Ala Asp Leu Thr Ile Met Glu Glu Gly Thr Glu Leu Ile Asn Arg Ile 275 280 285 Lys Asn Gly Gly Lys Leu Pro Leu Ile Thr Ser Cys Ser Pro Gly Trp 290 295 300 Ile Lys Phe Cys Glu His Asn Tyr Pro Glu Phe Leu Asp Asn Leu Ser 305 310 315 320 Ser Cys Lys Ser Pro His Glu Met Phe Gly Ala Val Leu Lys Ser Tyr 325 330 335 Tyr Ala Gln Lys Asn Gly Ile Asp Pro Ser Lys Val Phe Val Val Ser 340 345 350 Ile Met Pro Cys Thr Ala Lys Lys Phe Glu Ala Gln Arg Pro Glu Leu 355 360 365 Ser Ser Thr Gly Tyr Pro Asp Val Asp Val Val Leu Thr Thr Arg Glu 370 375 380 Leu Ala Arg Met Ile Lys Glu Thr Gly Ile Asp Phe Asn Ser Leu Pro 385 390 395 400 Asp Lys Gln Phe Asp Asp Pro Met Gly Glu Ala Ser Gly Ala Gly Val 405 410 415 Ile Phe Gly Ala Thr Gly Gly Val Met Glu Ala Ala Ile Arg Thr Val 420 425 430 Gly Glu Leu Leu Ser Gly Lys Pro Ala Asp Lys Ile Glu Tyr Thr Glu 435 440 445 Val Arg Gly Leu Asp Gly Ile Lys Glu Ala Ser Ile Glu Leu Asp Gly 450 455 460 Phe Thr Leu Lys Ala Ala Val Ala His Gly Leu Gly Asn Ala Arg Lys 465 470 475 480 Leu Leu Asp Lys Ile Lys Ala Gly Glu Ala Asp Tyr His Phe Ile Glu 485 490 495 Ile Met Ala Cys Pro Gly Gly Cys Ile Asn Gly Gly Gly Gln Pro Ile 500 505 510 Gln Pro Ser Ser Val Arg Asn Trp Lys Asp Ile Arg Cys Glu Arg Ala 515 520 525 Lys Ala Ile Tyr Glu Glu Asp Glu Ser Leu Pro Ile Arg Lys Ser His 530 535 540 Glu Asn Pro Lys Ile Lys Met Leu Tyr Glu Glu Phe Phe Gly Glu Pro 545 550 555 560 Gly Ser His Lys Ala His Glu Leu Leu His Thr His Tyr Glu Lys Arg 565 570 575 Glu Asn Tyr Pro Val Lys 580 33566PRTC. thermocellum 33Met Asp Asn Arg Glu Tyr Met Leu Ile Asp Gly Ile Pro Val Glu Ile 1 5 10 15 Asn Gly Glu Lys Asn Leu Leu Glu Leu Ile Arg Lys Ala Gly Ile Lys 20 25 30 Leu Pro Thr Phe Cys Tyr His Ser Glu Leu Ser Val Tyr Gly Ala Cys 35 40 45 Arg Met Cys Met Val Glu Asn Glu Trp Gly Gly Leu Asp Ala Ala Cys 50 55 60 Ser Thr Pro Pro Arg Ala Gly Met Ser Ile Lys Thr Asn Thr Glu Arg 65 70 75 80 Leu Gln Lys Tyr Arg Lys Met Ile Leu Glu Leu Leu Leu Ala Asn His 85 90 95 Cys Arg Asp Cys Thr Thr Cys Asn Asn Asn Gly Lys Cys Lys Leu Gln 100 105 110 Asp Leu Ala Met Arg Tyr Asn Ile Ser His Ile Arg Phe Pro Asn Thr 115 120 125 Ala Ser Asn Pro Asp Val Asp Asp Ser Ser Leu Cys Ile Thr

Arg Asp 130 135 140 Arg Ser Lys Cys Ile Leu Cys Gly Asp Cys Val Arg Val Cys Asn Glu 145 150 155 160 Val Gln Asn Val Gly Ala Ile Asp Phe Ala Tyr Arg Gly Ser Lys Met 165 170 175 Thr Ile Ser Thr Val Phe Asp Lys Pro Ile Phe Glu Ser Asn Cys Val 180 185 190 Gly Cys Gly Gln Cys Ala Leu Ala Cys Pro Thr Gly Ala Ile Val Val 195 200 205 Lys Asp Asp Thr Gln Lys Val Trp Lys Glu Ile Tyr Asp Lys Asn Thr 210 215 220 Arg Val Ser Val Gln Ile Ala Pro Ala Val Arg Val Ala Leu Gly Lys 225 230 235 240 Glu Leu Gly Leu Asn Asp Gly Glu Asn Ala Ile Gly Lys Ile Val Ala 245 250 255 Ala Leu Arg Arg Met Gly Phe Asp Asp Ile Phe Asp Thr Ser Thr Gly 260 265 270 Ala Asp Leu Thr Val Leu Glu Glu Ser Ala Glu Leu Leu Arg Arg Ile 275 280 285 Arg Glu Gly Lys Asn Asp Met Pro Leu Phe Thr Ser Cys Cys Pro Ala 290 295 300 Trp Val Asn Tyr Cys Glu Lys Phe Tyr Pro Glu Leu Leu Pro His Val 305 310 315 320 Ser Thr Cys Arg Ser Pro Met Gln Met Phe Ala Ser Ile Ile Lys Glu 325 330 335 Glu Tyr Ser Thr Ser Ser Lys Arg Leu Val His Val Ala Val Met Pro 340 345 350 Cys Thr Ala Lys Lys Phe Glu Ala Ala Arg Lys Glu Phe Lys Val Asn 355 360 365 Gly Val Pro Asn Val Asp Tyr Val Leu Thr Thr Gln Glu Leu Val Arg 370 375 380 Met Ile Lys Glu Ser Gly Ile Val Phe Ser Glu Leu Glu Pro Glu Ala 385 390 395 400 Ile Asp Met Pro Phe Gly Thr Tyr Thr Gly Ala Gly Val Ile Phe Gly 405 410 415 Val Ser Gly Gly Val Thr Glu Ala Val Leu Arg Arg Val Val Ser Asp 420 425 430 Lys Ser Pro Thr Ser Phe Arg Ser Leu Ala Tyr Thr Gly Val Arg Gly 435 440 445 Met Asn Gly Val Lys Glu Ala Ser Val Met Tyr Gly Asp Arg Lys Leu 450 455 460 Lys Val Ala Val Val Ser Gly Leu Lys Asn Ala Gly Asp Leu Ile Glu 465 470 475 480 Arg Ile Lys Ala Gly Glu His Tyr Asp Leu Val Glu Val Met Ala Cys 485 490 495 Pro Gly Gly Cys Ile Asn Gly Gly Gly Gln Pro Phe Val Gln Ser Glu 500 505 510 Glu Arg Glu Lys Arg Gly Lys Gly Leu Tyr Ser Ala Asp Lys Leu Cys 515 520 525 Asn Ile Lys Ser Ser Glu Glu Asn Pro Leu Met Met Thr Leu Tyr Lys 530 535 540 Gly Ile Leu Lys Gly Arg Val His Glu Leu Leu His Val Asp Tyr Ala 545 550 555 560 Ser Lys Lys Glu Ala Lys 565 34644PRTC. thermocellum 34Met Asp Ser Phe Leu Met Lys Gly Tyr Ile Lys Glu Ala Asn Ile Asp 1 5 10 15 Tyr Ser Cys Ser Arg Gly Ser Met Glu Asp Leu Pro Lys Trp Glu Phe 20 25 30 Arg Glu Ile Pro Lys Val Pro Arg Ala Val Met Pro Ser Leu Ser Leu 35 40 45 Glu Glu Arg Lys Asn Asn Phe Asn Glu Val Glu Leu Gly Leu Ser Glu 50 55 60 Glu Val Ala Arg Lys Glu Ala Arg Arg Cys Leu Lys Cys Gly Cys Ser 65 70 75 80 Ala Arg Phe Thr Cys Asp Leu Arg Lys Glu Ala Ser Asn His Gly Ile 85 90 95 Val Tyr Glu Glu Pro Ile His Asp Arg Pro Tyr Ile Pro Lys Val Asp 100 105 110 Asp His Pro Phe Ile Val Arg Asp His Asn Lys Cys Ile Ser Cys Gly 115 120 125 Arg Cys Ile Ala Ala Cys Ala Glu Ile Glu Gly Pro Gly Val Leu Thr 130 135 140 Phe Tyr Met Lys Asn Gly Arg Gln Leu Val Gly Thr Lys Ser Gly Leu 145 150 155 160 Pro Leu Arg Asp Thr Asp Cys Val Ser Cys Gly Gln Cys Val Thr Ala 165 170 175 Cys Pro Cys Ala Ala Leu Asp Tyr Arg Arg Glu Arg Gly Lys Val Val 180 185 190 Arg Ala Ile Asn Asp Pro Lys Lys Thr Val Val Gly Phe Val Ala Pro 195 200 205 Ala Val Arg Ser Leu Ile Ser Asn Thr Phe Gly Val Ser Tyr Glu Glu 210 215 220 Ala Ser Pro Phe Met Ala Gly Leu Leu Lys Lys Leu Gly Phe Asp Lys 225 230 235 240 Val Phe Asp Phe Thr Phe Ala Ala Asp Leu Thr Ile Val Glu Glu Thr 245 250 255 Thr Glu Phe Leu Ser Arg Ile Gln Asn Lys Gly Val Met Pro Gln Phe 260 265 270 Thr Ser Cys Cys Pro Gly Trp Ile Asn Phe Val Glu Lys Arg Tyr Pro 275 280 285 Glu Ile Ile Pro His Leu Ser Thr Cys Lys Ser Pro Gln Met Met Met 290 295 300 Gly Ala Thr Val Lys Asn His Tyr Ala Lys Leu Met Gly Ile Asn Lys 305 310 315 320 Glu Asp Leu Phe Val Val Ser Ile Val Pro Cys Leu Ala Lys Lys Tyr 325 330 335 Glu Ala Ala Arg Pro Glu Phe Ile His Asp Gly Ile Arg Asp Val Asp 340 345 350 Ala Val Leu Thr Thr Thr Glu Met Leu Glu Met Met Glu Leu Ala Asp 355 360 365 Ile Lys Pro Ser Glu Val Val Pro Gln Glu Phe Asp Glu Pro Tyr Lys 370 375 380 Gln Val Ser Gly Ala Gly Ile Leu Phe Gly Ala Ser Gly Gly Val Ala 385 390 395 400 Glu Ala Ala Leu Arg Met Ala Val Glu Lys Leu Thr Gly Lys Val Leu 405 410 415 Thr Asp His Leu Glu Phe Glu Glu Ile Arg Gly Phe Glu Gly Val Lys 420 425 430 Glu Ser Thr Ile Asp Val Asn Gly Thr Lys Val Arg Val Ala Val Val 435 440 445 Ser Gly Leu Lys Asn Ala Glu Pro Ile Ile Glu Lys Ile Leu Asn Gly 450 455 460 Val Asp Val Gly Tyr Asp Leu Ile Glu Val Met Ala Cys Pro Gly Gly 465 470 475 480 Cys Ile Cys Gly Ala Gly His Pro Val Pro Glu Lys Ile Asp Ser Leu 485 490 495 Glu Lys Arg Gln Gln Val Leu Val Asn Ile Asp Lys Val Ser Lys Tyr 500 505 510 Arg Lys Ser Gln Glu Asn Pro Asp Ile Leu Arg Leu Tyr Asn Glu Phe 515 520 525 Tyr Gly Glu Pro Asn Ser Pro Leu Ala His Glu Leu Leu His Thr His 530 535 540 Tyr Thr Pro Lys His Gly Asp Ser Thr Cys Ser Pro Glu Arg Lys Lys 545 550 555 560 Gly Thr Ala Ala Phe Asp Val Gln Glu Phe Thr Ile Cys Met Cys Glu 565 570 575 Ser Cys Met Glu Lys Gly Ala Glu Asn Leu Tyr Asn Asp Leu Ser Ser 580 585 590 Lys Ile Arg Leu Phe Lys Met Asp Pro Phe Val Gln Ile Lys Arg Ile 595 600 605 Arg Leu Lys Glu Thr His Pro Gly Lys Gly Val Tyr Ile Ala Leu Asn 610 615 620 Gly Lys Gln Ile Glu Glu Pro Met Leu Ser Gly Asn Ile Pro Asp Glu 625 630 635 640 Ser Glu Ser Glu 35493PRTC. thermocellum 35Met Lys Thr Leu Glu Asn His Asn Arg Ile Lys Val Thr Val Asn Gly 1 5 10 15 Arg Glu Ile Glu Val Tyr Asp Asn Leu Thr Ile Leu Gln Ala Leu Leu 20 25 30 Gln Glu Asp Ile His Ile Pro His Leu Cys Tyr Asp Ile Arg Leu Glu 35 40 45 Arg Ser Asn Gly Asn Cys Gly Leu Cys Val Val Thr Leu Ile Ser Pro 50 55 60 Asp Gly Glu Arg Asp Val Lys Ala Cys Gln Thr Pro Ile Lys Glu Gly 65 70 75 80 Met Val Ile Cys Thr Asn Thr Pro Lys Leu Glu Asn Tyr Arg Lys Ile 85 90 95 Arg Leu Glu Gln Leu Leu Ser Asp His Asn Ala Asp Cys Val Ala Pro 100 105 110 Cys Val Met Thr Cys Pro Ala Asn Ile Asp Ile Gln Ser Tyr Leu Arg 115 120 125 His Val Gly Asn Gly Asp Phe Glu Ala Ala Ile Arg Val Ile Lys Glu 130 135 140 Arg Asn Pro Phe Pro Ile Val Cys Gly Arg Val Cys Pro His Thr Cys 145 150 155 160 Glu Ser Gln Cys Arg Arg Asn Leu Val Asp Ala Pro Val Ala Ile Asn 165 170 175 Tyr Val Lys Arg Phe Ala Ala Asp Trp Asp Met Ala Arg Pro Glu Pro 180 185 190 Trp Thr Pro Glu Lys Lys Pro Pro Thr Gly Lys Lys Ile Ala Ile Val 195 200 205 Gly Ala Gly Pro Ser Gly Leu Ser Ala Ala Tyr Tyr Ser Ala Ile Lys 210 215 220 Gly His Asp Val Thr Val Phe Glu Arg Gln Pro His Pro Gly Gly Met 225 230 235 240 Met Arg Tyr Gly Ile Pro Glu Tyr Arg Leu Pro Lys Ala Ile Leu Asp 245 250 255 Lys Glu Ile Glu Met Ile Lys Lys Leu Gly Val Lys Ile Met Thr Glu 260 265 270 Lys Ala Leu Gly Ile His Ile Arg Leu Glu Asp Leu Ser Lys Asp Phe 275 280 285 Asp Ala Val Tyr Leu Ala Ile Gly Ser Trp Gln Ala Thr Pro Met His 290 295 300 Ile Glu Gly Glu Lys Leu Asp Gly Val Trp Ala Gly Ile Asn Tyr Leu 305 310 315 320 Glu Gln Val Ala Lys Asn Val Asp Ile Pro Leu Gly Asp Asn Val Val 325 330 335 Val Ile Gly Gly Gly Asn Thr Ala Ile Asp Cys Ala Arg Thr Ala Leu 340 345 350 Arg Lys Gly Ala Lys Ser Val Lys Leu Val Tyr Arg Cys Thr Arg Glu 355 360 365 Glu Met Pro Ala Ala Pro Tyr Glu Val Glu Glu Ala Ile His Glu Gly 370 375 380 Val Glu Met Ile Phe Leu Met Ala Pro Thr Lys Ile Ile Val Lys Asp 385 390 395 400 Gly Lys Lys Lys Leu Val Cys Ile Arg Met Gln Leu Gly Glu Pro Asp 405 410 415 Arg Ser Gly Arg Arg Arg Pro Val Pro Ile Glu Gly Ser Glu Val Glu 420 425 430 Ile Asp Ala Asp Thr Ile Ile Gly Ala Ile Gly Gln Ser Thr Asn Thr 435 440 445 Gln Phe Leu Tyr Asn Asp Leu Pro Val Lys Leu Asn Lys Trp Gly Asp 450 455 460 Ile Glu Val Asn Gly Lys Thr Leu Gln Thr Ser Glu Tyr Asn Ile Phe 465 470 475 480 Ala Gly Gly Asp Cys Val Thr Gly Pro Ala Thr Val Ile 485 490 36309PRTC. thermocellum 36Met Pro Leu Val Thr Ser Thr Glu Met Phe Lys Lys Ala Tyr Glu Gly 1 5 10 15 Lys Tyr Ala Ile Gly Ala Phe Asn Val Asn Asn Met Glu Ile Ile Gln 20 25 30 Gly Ile Thr Glu Ala Ala Lys Glu Val Asn Ala Pro Leu Ile Leu Gln 35 40 45 Val Ser Ala Gly Ala Arg Lys Tyr Ala Asn His Thr Tyr Leu Val Lys 50 55 60 Leu Val Glu Ala Ala Val Glu Glu Thr Gly Leu Pro Ile Cys Leu His 65 70 75 80 Leu Asp His Gly Asp Ser Phe Glu Leu Cys Lys Ser Cys Ile Asp Gly 85 90 95 Gly Phe Thr Ser Val Met Ile Asp Gly Ser His Leu Pro Phe Glu Glu 100 105 110 Asn Ile Lys Leu Thr Lys Gln Val Val Asp Tyr Ala His Ser Lys Gly 115 120 125 Val Val Val Glu Gly Glu Leu Gly Arg Leu Ala Gly Ile Glu Asp Asp 130 135 140 Val Asn Val Ser Glu Ala Asp Ala Ala Phe Thr Asp Pro Asp Gln Ala 145 150 155 160 Glu Glu Phe Val Lys Arg Thr Gly Val Asp Ser Leu Ala Ile Ala Ile 165 170 175 Gly Thr Ser His Gly Ala Tyr Lys Phe Lys Gly Glu Ala Lys Leu Arg 180 185 190 Phe Asp Ile Leu Glu Glu Ile Glu Lys Arg Leu Pro Gly Phe Pro Ile 195 200 205 Val Leu His Gly Ala Ser Ser Val Ile Pro Glu Tyr Val Asp Met Ile 210 215 220 Asn Lys Tyr Gly Gly Asp Met Pro Gly Ala Lys Gly Val Pro Glu Asp 225 230 235 240 Met Leu Arg Lys Ala Ala Ser Met Ala Val Cys Lys Ile Asn Ile Asp 245 250 255 Ser Asp Leu Arg Leu Ala Met Thr Ala Thr Ile Arg Lys Tyr Phe Ala 260 265 270 Glu Asn Pro Ser His Phe Asp Pro Arg Gln Tyr Leu Gly Pro Ala Arg 275 280 285 Asn Ala Ile Lys Glu Leu Val Lys His Lys Ile Val Asn Val Leu Gly 290 295 300 Cys Asp Gly Lys Ala 305 37327PRTC. thermocellum 37Met Asp Ile Gln Leu Lys Lys Ser Gly Ile Gly Val Lys Glu Lys Lys 1 5 10 15 Ser Lys Asn His Leu Leu Tyr Ser Ile Lys Gln Asn Leu Phe Ala Tyr 20 25 30 Ala Met Leu Ile Pro Thr Phe Val Cys Met Met Cys Ile His Phe Ile 35 40 45 Pro Met Leu Gln Gly Ile Tyr Leu Ser Leu Leu Asp Leu Asn Gln Leu 50 55 60 Thr Met Thr Lys Phe Leu Asn Ala Pro Phe Ile Gly Leu Lys Asn Tyr 65 70 75 80 Tyr Glu Ile Leu Phe Asp Glu Lys Ser Leu Ile Arg Arg Gly Phe Trp 85 90 95 Phe Ala Leu Arg Asn Thr Ala Ile Tyr Thr Val Val Val Thr Phe Ala 100 105 110 Thr Phe Ala Leu Gly Ile Ile Leu Ala Met Leu Val Asn Arg Glu Phe 115 120 125 Lys Gly Arg Gly Ile Val Arg Thr Ala Leu Leu Met Pro Trp Val Val 130 135 140 Pro Ser Tyr Val Val Gly Met Thr Trp Gly Phe Leu Trp Arg Gln Asp 145 150 155 160 Ser Gly Leu Ile Asn Ile Ile Leu Cys Asp Ile Leu His Ile Leu Pro 165 170 175 Glu Lys Pro Tyr Trp Leu Val Gly Ser Asn Gln Ile Trp Ala Ile Ile 180 185 190 Ile Pro Thr Ile Trp Arg Gly Leu Pro Leu Ser Met Ile Leu Met Leu 195 200 205 Ala Gly Leu Gln Ser Ile Ser Pro Asp Tyr Tyr Glu Ala Ala Asp Ile 210 215 220 Asp Gly Ala Asn Gly Trp Gln Lys Phe Trp His Ile Thr Leu Pro Leu 225 230 235 240 Leu Lys Pro Ile Leu Ala Ile Asn Val Met Phe Ser Leu Ile Ser Asn 245 250 255 Ile Tyr Ser Phe Asn Ile Val Ser Met Met Phe Gly Asn Gly Ala Gly 260 265 270 Ile Pro Gly Glu Trp Gly Asp Leu Leu Met Thr Tyr Ile Gln Arg Asn 275 280 285 Thr Phe Gln Met Trp Arg Phe Gly Pro Gly Ala Ala Ala Leu Met Ile 290 295 300 Val Met Phe Phe Val Leu Gly Ile Val Ala Leu Trp Tyr Thr Leu Phe 305 310 315 320 Lys Asp Asp Leu Val Val Lys 325 38404PRTC. thermocellum 38Val Asp Lys Phe Thr Lys Leu Asp Leu Asn Ser Ile Thr Ser Asn Asn 1 5 10 15 Arg Met Asn Ile Phe Asn Cys Ile Leu Glu Ala Lys Glu Ile Asn Arg 20 25 30 Ala Val Ile Ala Lys Lys Val Gly Leu Ser Ile Pro Ala Val Met Ser 35 40 45 Ile Thr Asp Asp Leu Ile Gln Lys Gly Ile Ile Tyr Val Ile Gly Lys 50 55 60 Gly Lys Ser Ser Gly Gly Lys Arg Pro Glu Leu Leu Ala Val Val Pro 65 70 75 80 Asp Arg Phe Phe Phe Val Gly Val Asp Val Gly Arg Thr Ser Val Arg

85 90 95 Val Val Val Met Asn Asn Cys Arg Asp Val Val Tyr Lys Val Ser Lys 100 105 110 Pro Thr Glu Ser Val Glu Pro Asp Glu Leu Ile Asn Gln Ile Thr Glu 115 120 125 Met Thr Met Glu Ser Ile Asn Glu Ser Lys Phe Pro Leu Asp Arg Val 130 135 140 Val Gly Ile Gly Val Ala Met Pro Gly Leu Ile Glu Arg Gly Thr Gly 145 150 155 160 Arg Val Ile Phe Ser Pro Asn Phe Gly Trp Asn Asn Ile Ala Leu Gln 165 170 175 Asp Glu Leu Lys Lys His Leu Pro Phe Asn Val Leu Val Glu Asn Ala 180 185 190 Asn Arg Ala Leu Val Ile Gly Glu Ile Lys Asn Thr Gln Pro Asn Pro 195 200 205 Thr Ser Cys Ile Val Gly Val Asn Leu Gly Tyr Gly Ile Gly Ser Ala 210 215 220 Ile Val Leu Pro Asn Gly Leu Tyr Tyr Gly Val Ser Gly Thr Ser Gly 225 230 235 240 Glu Ile Gly His Ile Ile Val Glu Asn His Gly Ser Tyr Cys Ser Cys 245 250 255 Gly Asn Tyr Gly Cys Ile Glu Ser Ile Ala Ser Gly Glu Ala Ile Ala 260 265 270 Arg Glu Ala Arg Ile Ala Ile Ala Asn Lys Ile Gln Ser Ser Val Phe 275 280 285 Glu Lys Cys Glu Gly Asp Leu Lys Lys Ile Asp Ala Lys Met Val Phe 290 295 300 Asp Ala Ala Lys Glu Gly Asp His Leu Ala Gln Ser Ile Val Glu Lys 305 310 315 320 Ala Ala Asp Tyr Ile Gly Lys Gly Leu Ala Ile Thr Ile Asn Met Leu 325 330 335 Asp Pro Glu Gln Ile Ile Leu Cys Gly Gly Leu Thr Leu Ser Gly Asp 340 345 350 Phe Phe Ile Asp Met Ile Lys Lys Ala Val Ser Lys Tyr Gln Met Arg 355 360 365 Tyr Ala Gly Gly Asn Val Lys Ile Val Val Gly Lys Ser Gly Leu Tyr 370 375 380 Ala Thr Ala Ile Gly Gly Ala Trp Ile Val Ala Asn Asn Ile Asp Phe 385 390 395 400 Leu Ser Ser Asn 39317PRTC. thermocellum 39Met Tyr Tyr Ile Gly Ile Asp Leu Gly Gly Thr Asn Ile Ala Val Gly 1 5 10 15 Leu Val Asn Glu Glu Gly Lys Ile Leu His Lys Asp Ser Val Pro Thr 20 25 30 Leu Arg Glu Arg Pro Tyr Gln Glu Ile Ile Lys Asp Met Ala Met Leu 35 40 45 Thr Leu Lys Val Ile Lys Asp Ala Asp Val Ser Ile Asp Gln Val Lys 50 55 60 Ser Ile Gly Val Gly Ser Pro Gly Thr Pro Asn Cys Lys Asp Gly Ile 65 70 75 80 Leu Ile Tyr Asn Asn Asn Leu Asn Phe Arg Asn Val Pro Ile Arg Ser 85 90 95 Glu Ile Gln Lys Tyr Ile Asp Leu Pro Val Tyr Leu Asp Asn Asp Ala 100 105 110 Asn Cys Ala Ala Leu Ala Glu Ser Val Ala Gly Ala Ala Lys Gly Ala 115 120 125 Asn Thr Ser Val Thr Ile Thr Leu Gly Thr Gly Ile Gly Gly Gly Val 130 135 140 Val Ile Asp Gly Lys Ile Tyr Ser Gly Phe Asn Tyr Ala Gly Gly Glu 145 150 155 160 Leu Gly His Thr Val Leu Met Met Asp Gly Glu Pro Cys Thr Cys Gly 165 170 175 Arg Lys Gly Cys Trp Glu Ala Tyr Ala Ser Ala Thr Ala Leu Ile Arg 180 185 190 Gln Ala Arg Lys Ala Ala Glu Ala Asn Pro Asp Ser Leu Ile Asn Lys 195 200 205 Leu Val Gly Gly Asp Leu Ser Lys Ile Asp Ala Lys Ile Pro Phe Asp 210 215 220 Ala Ala Lys Gln Gly Asp Lys Thr Gly Glu Met Val Val Gln Gln Tyr 225 230 235 240 Ile Arg Tyr Ile Ala Glu Gly Leu Ile Asn Met Ile Asn Ile Phe Met 245 250 255 Pro Glu Val Leu Val Ile Gly Gly Gly Val Cys Lys Glu Gly Glu Tyr 260 265 270 Leu Leu Lys Pro Leu Arg Glu Leu Ile Lys Gln Gly Val Tyr Ser Lys 275 280 285 Glu Asp Ile Pro Gln Thr Glu Leu Arg Thr Ala Gln Met Gly Asn Asp 290 295 300 Ala Gly Ile Ile Gly Ala Ala Met Leu Gly Lys Glu Cys 305 310 315 40448PRTC. thermocellum 40Met Glu Arg Ile Lys Phe Asp Tyr Ser Lys Ala Leu Pro Phe Val Ser 1 5 10 15 Glu Arg Glu Val Ala Tyr Phe Glu Asn Phe Val Arg Ser Ala His Asp 20 25 30 Met Leu His Asn Lys Thr Gly Ala Gly Asn Asp Phe Val Gly Trp Val 35 40 45 Asp Leu Pro Val Asn Tyr Asp Arg Glu Glu Phe Ala Arg Ile Lys Ala 50 55 60 Ala Ala Glu Lys Ile Lys Ser Asp Ser Asp Ala Leu Val Val Ile Gly 65 70 75 80 Ile Gly Gly Ser Tyr Leu Gly Ala Arg Ala Ala Ile Glu Met Leu Ser 85 90 95 His Ser Phe His Asn Leu Met Pro Lys Ser Lys Arg Asn Ala Pro Glu 100 105 110 Ile Tyr Phe Val Gly Asn Asn Ile Ser Ser Thr Tyr Ile Ala Asp Leu 115 120 125 Leu Glu Val Ile Glu Gly Lys Glu Ile Ser Val Asn Val Ile Ser Lys 130 135 140 Ser Gly Thr Thr Thr Glu Pro Ala Ile Ala Phe Arg Ile Phe Lys Glu 145 150 155 160 Tyr Met Glu Asn Lys Tyr Gly Lys Asp Gly Ala Ser Lys Arg Ile Tyr 165 170 175 Ala Thr Thr Asp Lys Glu Lys Gly Ala Leu Arg Lys Leu Ala Thr Glu 180 185 190 Glu Gly Tyr Glu Thr Phe Val Val Pro Asp Asp Ile Gly Gly Arg Phe 195 200 205 Ser Val Leu Thr Ala Val Gly Leu Leu Pro Ile Ala Val Ala Gly Ile 210 215 220 Asp Ile Asp Ser Met Met Lys Gly Ala Ala Asp Ala Arg Glu Leu Tyr 225 230 235 240 Ser Asn Pro Asn Leu Met Glu Asn Asp Cys Tyr Lys Tyr Ala Ala Val 245 250 255 Arg Asn Ala Leu Tyr Arg Lys Asn Lys Thr Ile Glu Ile Met Val Asn 260 265 270 Tyr Glu Pro Ser Leu His Tyr Phe Thr Glu Trp Trp Lys Gln Leu Tyr 275 280 285 Gly Glu Ser Glu Gly Lys Asp Gln Lys Gly Ile Phe Pro Ala Gly Val 290 295 300 Asp Phe Thr Thr Asp Leu His Ser Met Gly Gln Tyr Ile Gln Asp Gly 305 310 315 320 Leu Arg Asn Ile Phe Glu Thr Val Ile Arg Val Glu Lys Pro Arg Lys 325 330 335 Asn Ile Val Ile Lys Glu Glu Lys Asp Asn Leu Asp Gly Leu Asn Phe 340 345 350 Ile Ala Gly Lys Asp Val Asp Tyr Val Asn Lys Lys Ala Met Glu Gly 355 360 365 Thr Val Leu Ala His Thr Asp Gly Gly Val Pro Asn Leu Val Val Thr 370 375 380 Val Pro Glu Leu Ser Ala Tyr Tyr Phe Gly Asn Met Val Tyr Phe Phe 385 390 395 400 Glu Lys Ala Cys Gly Ile Ser Gly Tyr Leu Leu Gly Val Asn Pro Phe 405 410 415 Asp Gln Pro Gly Val Glu Ala Tyr Lys Lys Asn Met Phe Ala Leu Leu 420 425 430 Gly Lys Pro Gly Tyr Glu Glu Gln Arg Lys Lys Leu Glu Glu Arg Leu 435 440 445 41324PRTC. thermocellum 41Met Ser Ser Val Arg Thr Ile Gly Val Leu Thr Ser Gly Gly Asp Ala 1 5 10 15 Pro Gly Met Asn Ala Ala Ile Arg Ser Val Val Arg Thr Gly Leu Tyr 20 25 30 Tyr Gly Phe Lys Val Leu Gly Ile Arg Lys Gly Phe Asn Gly Leu Ile 35 40 45 Asn Gly Asp Ile Glu Glu Leu Thr Ala Arg Ser Val Gly Asp Ile Ile 50 55 60 His Arg Gly Gly Thr Ile Leu Gln Thr Ala Arg Ser Pro Gln Phe Lys 65 70 75 80 Thr Glu Glu Gly Leu Lys Lys Ala Met Ser Met Ala Lys Val Phe Gly 85 90 95 Ile Asp Ala Leu Val Val Ile Gly Gly Asp Gly Ser Tyr Arg Gly Ala 100 105 110 Arg Asp Ile Ser Lys Leu Gly Leu Asn Val Ile Gly Ile Pro Gly Thr 115 120 125 Ile Asp Asn Asp Ile Gly Cys Thr Asp Tyr Thr Ile Gly Phe Asp Thr 130 135 140 Ala Met Asn Thr Val Gln Asp Ala Ile Asp Lys Ile Arg Asp Thr Ala 145 150 155 160 Tyr Ser His Glu Arg Cys Ser Val Leu Glu Val Met Gly Arg His Ala 165 170 175 Gly Tyr Ile Ala Val Asn Val Ser Ile Ser Gly Gly Ala Glu Ala Val 180 185 190 Val Leu Pro Glu Lys Pro Phe Asp Met Asp Thr Asp Val Ile Lys Pro 195 200 205 Ile Ile Glu Gly Arg Asn Arg Gly Lys Lys His Tyr Leu Val Ile Val 210 215 220 Ala Glu Gly Gly Glu Gly Lys Ala Ile Glu Ile Ala Lys Glu Ile Thr 225 230 235 240 Glu Lys Thr Gly Ile Glu Ala Arg Ala Thr Ile Leu Gly His Ile Gln 245 250 255 Arg Gly Gly Ser Pro Thr Val Tyr Asp Arg Val Met Ala Ser Gln Met 260 265 270 Gly Ala Lys Ala Val Glu Val Leu Met Glu Asn Lys Arg Asn Arg Val 275 280 285 Ile Val Phe Lys Asp Asn Gln Ile Gly Asp Met Asp Leu Glu Glu Ala 290 295 300 Leu Gln Val Lys Lys Thr Ile Ser Glu Asp Leu Ile Gln Leu Ser Lys 305 310 315 320 Ile Leu Ala Leu 42327PRTT. saccharolyticum 42Met Ser Tyr Ile Pro Asn Glu Asn Arg Tyr Glu Lys Met Ile Tyr Arg 1 5 10 15 Arg Cys Gly Arg Ser Gly Ile Met Leu Pro Ala Ile Ser Leu Gly Leu 20 25 30 Trp His Asn Phe Gly Gly Tyr Asp Val Phe Glu Asn Met Arg Glu Met 35 40 45 Val Lys Lys Ala Phe Asp Leu Gly Ile Thr His Phe Asp Leu Ala Asn 50 55 60 Asn Tyr Gly Pro Pro Pro Gly Ser Ala Glu Glu Asn Phe Gly Lys Ile 65 70 75 80 Leu Arg Thr Asp Leu Arg Gly Tyr Arg Asp Glu Leu Leu Ile Ser Thr 85 90 95 Lys Ala Gly Tyr Thr Met Trp Pro Gly Pro Tyr Gly Asp Trp Gly Ser 100 105 110 Arg Lys Tyr Leu Leu Ser Ser Leu Asp Gln Ser Leu Lys Arg Met Gly 115 120 125 Ile Asp Tyr Val Asp Ile Phe Tyr Ser His Arg Arg Asp Pro Asn Thr 130 135 140 Pro Leu Glu Glu Thr Met Ser Ala Leu Ala Gln Ala Val Arg Gln Gly 145 150 155 160 Lys Ala Leu Tyr Val Gly Ile Ser Asn Tyr Asn Ala Glu Asp Thr Lys 165 170 175 Lys Ala Ala Glu Ile Leu Arg Gln Leu Gly Thr Pro Leu Leu Ile Asn 180 185 190 Gln Pro Ser Tyr Ser Met Phe Asn Arg Trp Ile Glu Asp Gly Leu Thr 195 200 205 Asp Val Leu Glu Glu Glu Gly Val Gly Ser Ile Ala Phe Ser Pro Leu 210 215 220 Ala Gln Gly Leu Leu Thr Asp Lys Tyr Leu Asn Gly Val Pro Asp Asp 225 230 235 240 Ser Arg Ala Val Arg Lys Asn Thr Ser Leu Arg Gly Asn Leu Thr Glu 245 250 255 Glu Asn Ile Asn Lys Val Arg Glu Leu Lys Lys Ile Ala Asp Lys Arg 260 265 270 Gly Gln Ser Ile Ala Gln Met Ala Leu Ala Trp Asp Leu Arg Lys Val 275 280 285 Thr Ser Val Ile Ile Gly Ala Ser Arg Val Ser Gln Ile Glu Glu Asn 290 295 300 Val Lys Ala Leu Asp Asn Leu Glu Phe Ser His Glu Glu Leu Lys Gln 305 310 315 320 Ile Asp Glu Ile Leu Ser Lys 325 43131PRTT. saccharolyticum 43Leu Asn Ile Ala Leu Ile Ala His Asp Met Lys Lys Ser Ile Met Val 1 5 10 15 Asp Phe Ala Ile Ala Tyr Lys Glu Ile Leu Lys Lys Cys Asn Ile Tyr 20 25 30 Ala Thr Gly Ala Thr Gly Gln Leu Val Glu Glu Ala Thr Gly Ile Lys 35 40 45 Val Asn Lys Phe Leu Pro Gly Pro Met Gly Gly Asp Gln Gln Ile Gly 50 55 60 Ala Met Ile Ala Glu Asn Asn Met Asp Leu Val Ile Phe Leu Arg Asp 65 70 75 80 Pro Leu Thr Ala Gln Pro His Glu Pro Asp Ile Leu Ala Leu Leu Arg 85 90 95 Val Cys Asp Val His Ser Ile Pro Leu Ala Thr Asn Leu Ala Thr Ala 100 105 110 Glu Val Leu Ile Lys Gly Leu Asp Ala Gly Phe Leu Glu Trp Arg Asp 115 120 125 Ala Val Lys 130 44248PRTT. saccharolyticum 44Leu Arg Arg Pro Ile Ile Ala Gly Asn Trp Lys Met Tyr Met Thr Pro 1 5 10 15 Ser Glu Ala Val Asn Leu Val Asn Glu Leu Lys Pro Leu Val Ser Gly 20 25 30 Ala Glu Ala Glu Val Val Val Ile Pro Pro Phe Val Asp Leu Val Asp 35 40 45 Val Lys Lys Ala Ile Asp Ala Ser Asn Ile Lys Leu Gly Ala Gln Asn 50 55 60 Met His Trp Glu Glu Lys Gly Ala Phe Thr Gly Glu Val Ser Pro Ile 65 70 75 80 Met Leu Lys Glu Ile Gly Val Glu Tyr Val Val Ile Gly His Ser Glu 85 90 95 Arg Arg Gln Tyr Phe Ala Glu Thr Asp Glu Thr Val Asn Lys Lys Val 100 105 110 Lys Ser Ala Leu Ser His Gly Leu Lys Pro Ile Val Cys Val Gly Glu 115 120 125 Ser Leu Ser Gln Arg Glu Ala Gly Glu Ala Phe Asn Val Val Arg Glu 130 135 140 Gln Thr Lys Lys Ala Leu Asp Gly Ile Lys Ser Glu Asp Val Leu Asn 145 150 155 160 Val Val Ile Ala Tyr Glu Pro Ile Trp Ala Ile Gly Thr Gly Lys Thr 165 170 175 Ala Thr Ser Lys Asp Ala Asn Asp Val Ile Lys Val Ile Arg Glu Thr 180 185 190 Ile Ala Asp Ile Tyr Ser Ile Asp Ile Ala Asn Glu Val Arg Ile Gln 195 200 205 Tyr Gly Gly Ser Val Lys Pro Asp Asn Ala Lys Glu Leu Met Ser Glu 210 215 220 Ser Asp Ile Asp Gly Ala Leu Val Gly Gly Ala Ser Leu Lys Ala Gln 225 230 235 240 Asp Phe Ala Lys Ile Val Asn Tyr 245 45394PRTT. saccharolyticum 45Met Tyr Met Lys Thr Asn Phe Thr Tyr Phe Met Pro Thr Glu Ile Phe 1 5 10 15 Gly Pro Gly Thr Leu Gly Lys Leu Ala Thr Val Lys Leu Pro Gly Lys 20 25 30 Lys Ala Leu Leu Val Ile Gly Ser Gly Asn Ser Met Arg Arg His Gly 35 40 45 Tyr Leu Asp Arg Val Val Asn Tyr Leu Lys Gln Asn Gly Val Asp Tyr 50 55 60 Val Val Tyr Asp Lys Ile Leu Pro Asn Pro Ile Ala Glu His Val Ala 65 70 75 80 Glu Gly Ala Lys Val Ala Lys Asp Asn Gly Cys Asp Phe Val Ile Gly 85 90 95 Leu Gly Gly Gly Ser Thr Ile Asp Ser Ser Lys Ala Ile Ala Val Met 100 105 110 Ala Lys Asn Pro Gly Asp Tyr Trp Asp Tyr Val Ser Gly Gly Ser Gly 115 120 125 Lys Gly Met Glu Val Lys Asn Gly Ala Leu Pro Ile Val Ala Ile Pro 130 135 140 Thr Thr Ala Gly Thr Gly Thr Glu Ser Asp Pro Trp Ala Val Val Thr 145 150 155 160 Lys Thr Glu Thr Asn Glu Lys Ile Gly Phe Gly Cys Lys Tyr Thr Tyr 165 170 175 Pro Thr Leu

Ser Ile Val Asp Pro Glu Leu Met Val Ser Ile Pro Pro 180 185 190 Lys Phe Thr Ala Tyr Gln Gly Met Asp Ala Phe Phe His Ser Val Glu 195 200 205 Gly Tyr Leu Ala Thr Val Asn Gln Pro Gly Ser Asp Val Leu Ala Leu 210 215 220 Gln Ser Ile Ser Leu Ile Thr Glu Asn Leu Pro Lys Ala Val Ala Asp 225 230 235 240 Gly Asn Asn Met Glu Ala Arg Thr Ala Leu Ala Trp Ala Ser Thr Ala 245 250 255 Ala Gly Ile Val Glu Ser Leu Ser Ser Cys Ile Ser His His Ser Leu 260 265 270 Glu His Ala Leu Ser Ala Tyr His Pro Glu Ile Pro His Gly Ala Gly 275 280 285 Leu Ile Met Leu Ser Val Ser Tyr Phe Ser Phe Met Ala Ser Lys Ala 290 295 300 Pro Glu Arg Phe Val Asp Ile Ala Lys Ala Met Gly Glu Glu Ile Val 305 310 315 320 Gly Asn Thr Val Glu Glu Gln Ala Met Cys Phe Ile Asn Gly Leu Lys 325 330 335 Lys Leu Ile Arg Asn Ile Gly Met Glu Asp Leu Ser Leu Ser Ser Phe 340 345 350 Gly Val Thr Glu Asp Glu Ala Thr Lys Leu Ala Lys Asn Ala Met Asp 355 360 365 Thr Met Gly Gly Leu Phe Asn Val Asp Pro Tyr Lys Leu Ser Leu Asp 370 375 380 Glu Val Val Ser Ile Tyr Lys Asn Cys Phe 385 390 46343PRTT. saccharolyticum 46Val Asp Asp Lys Lys Val Phe Asp His Leu Phe Ile Leu Thr Asp Asp 1 5 10 15 Thr Gly Met Met Gln His Ser Val Gly Ser Val Pro Asp Pro Lys Tyr 20 25 30 Gly Tyr Thr Thr Asp Asp Asn Gly Arg Ala Leu Ile Ala Cys Ala Met 35 40 45 Met Tyr Glu Lys Tyr Lys Asp Asp Ala Tyr Ile Asn Leu Ile Lys Lys 50 55 60 Tyr Leu Ser Phe Leu Met Tyr Ala Gln Glu Asp Asp Gly Arg Phe Arg 65 70 75 80 Asn Phe Met Ser Phe Asp Arg Lys Phe Ile Asp Glu Asp Phe Ser Glu 85 90 95 Asp Cys Phe Gly Arg Cys Met Trp Ala Leu Gly Tyr Leu Ile Asn Ser 100 105 110 Asn Ile Asp Glu Arg Val Lys Leu Pro Ala Tyr Lys Met Ile Glu Lys 115 120 125 Ser Leu Leu Leu Val Asp Thr Leu Asn Tyr Ile Arg Gly Lys Ala Tyr 130 135 140 Thr Leu Ile Gly Leu Tyr Tyr Ile Tyr Asn Ser Phe Lys Asn Leu Asp 145 150 155 160 Lys Asp Phe Val Arg Lys Lys Met Asp Lys Leu Ala His Asp Ile Val 165 170 175 Glu Glu Tyr Glu Lys Asn Ser Ser Glu Asp Trp Gln Trp Phe Glu Asp 180 185 190 Val Val Ser Tyr Asp Asn Gly Val Ile Pro Leu Ser Leu Leu Lys Tyr 195 200 205 Phe Ser Ile Ala Lys Asp Glu Glu Val Leu Asp Ile Ala Leu Lys Thr 210 215 220 Ile Asp Phe Leu Asp Ser Val Cys Phe Lys Asn Gly Tyr Phe Lys Ala 225 230 235 240 Val Gly Cys Lys Gly Trp Tyr Arg Lys Gly Lys Asp Ile Ala Glu Tyr 245 250 255 Asp Glu Gln Pro Val Glu Ala Tyr Thr Met Ala Leu Met Tyr Ile Glu 260 265 270 Ala Tyr Lys Leu Thr Gly Asp Glu Lys Tyr Lys Lys Arg Ala Ile Asp 275 280 285 Cys Asp Lys Trp Phe Tyr Gly Lys Asn Ser Lys Gly Leu Ser Leu Tyr 290 295 300 Asp Glu Asp Ser Gly Gly Cys Ser Asp Gly Ile Thr Glu Asp Gly Val 305 310 315 320 Asn Ser Asn Glu Gly Ala Glu Ser Leu Ile Ser Ile Met Ile Ser His 325 330 335 Cys Ala Ile Asp Gln Leu Lys 340 47350PRTT. saccharolyticum 47Met Lys Thr Ser Glu Leu Leu Ala Met Val Val Glu Lys Gly Ala Ser 1 5 10 15 Asp Leu His Ile Thr Val Gly Val Pro Pro Val Leu Arg Ile Asn Gly 20 25 30 Gln Leu Ile Lys Leu Asn Leu Pro Gln Leu Thr Pro Gln Asp Thr Glu 35 40 45 Glu Ile Thr Lys Asp Leu Leu Ser Ser Asp Glu Leu Lys Lys Leu Glu 50 55 60 Asp Met Gly Asp Ile Asp Leu Ser Tyr Ser Val Lys Gly Leu Gly Arg 65 70 75 80 Phe Arg Ile Asn Ala Tyr Lys Gln Arg Gly Thr Tyr Ser Leu Ala Ile 85 90 95 Arg Ser Val Ala Leu Arg Ile Pro Thr Ile Asp Glu Leu Gly Leu Pro 100 105 110 Glu Val Ile Lys Glu Leu Ala Leu Lys Thr Arg Gly Leu Ile Ile Val 115 120 125 Thr Gly Pro Thr Gly Ser Gly Lys Ser Thr Thr Leu Ala Ser Met Ile 130 135 140 Asp Leu Ile Asn Glu Glu Arg Asn Cys His Ile Leu Thr Leu Glu Asp 145 150 155 160 Pro Ile Glu Tyr Leu His Lys His Lys Lys Ser Ile Val Asn Gln Arg 165 170 175 Glu Ile Gly His Asp Ala Ala Ser Tyr Ala Ser Ala Leu Arg Ala Ala 180 185 190 Leu Arg Glu Asp Pro Asp Val Ile Leu Val Gly Glu Met Arg Asp Leu 195 200 205 Glu Thr Ile Gln Ile Ala Ile Thr Ala Ala Glu Thr Gly His Leu Val 210 215 220 Leu Ser Thr Leu His Thr Ile Gly Ser Ala Lys Thr Ile Asp Arg Ile 225 230 235 240 Ile Asp Val Phe Pro Pro His Gln Gln Gln Gln Ile Lys Val Gln Leu 245 250 255 Ser Asn Val Leu Glu Gly Ile Val Ser Gln Gln Leu Leu Pro Lys Ile 260 265 270 Asp Asn Ser Gly Arg Val Val Ala Val Glu Val Met Ile Ala Thr Pro 275 280 285 Ala Ile Arg Asn Leu Ile Arg Glu Gly Lys Ser Phe Gln Ile Gln Ser 290 295 300 Met Val Gln Thr Gly Asn Lys Phe Gly Met Val Thr Met Asp Met Trp 305 310 315 320 Ile Ser Gln Leu Leu Lys Arg Asn Leu Ile Ser Met Asp Asp Ala Leu 325 330 335 Thr Tyr Cys Val Asp Arg Glu Asn Phe Ser Arg Leu Val Val 340 345 350 48564PRTT. saccharolyticum 48Met Ile Lys Lys Lys Leu Gly Asp Leu Leu Val Glu Val Gly Leu Leu 1 5 10 15 Asp Glu Ser Gln Leu Asn Asn Ala Ile Lys Ile Gln Lys Lys Thr Gly 20 25 30 Glu Lys Leu Gly Lys Ile Leu Val Lys Glu Gly Tyr Leu Thr Glu Glu 35 40 45 Gln Ile Ile Glu Ala Leu Glu Phe Gln Leu Gly Ile Pro His Ile Asp 50 55 60 Met Lys Lys Val Phe Ile Asp Ala Asn Val Ala Lys Leu Ile Pro Glu 65 70 75 80 Ser Met Ala Lys Arg His Val Ala Ile Pro Ile Lys Lys Glu Asn Asn 85 90 95 Ser Ile Phe Val Ala Met Ala Asp Pro Leu Asn Ile Phe Ala Ile Asp 100 105 110 Asp Ile Lys Leu Val Thr Lys Leu Asp Val Lys Pro Leu Ile Ala Ser 115 120 125 Glu Asp Gly Ile Leu Lys Ala Ile Asp Arg Val Phe Gly Lys Glu Glu 130 135 140 Ala Glu Arg Ala Val Gln Asp Phe Lys Lys Glu Leu Ser His Asp Ser 145 150 155 160 Ala Glu Asp Asp Gly Asn Leu Leu Arg Asp Ile Ser Glu Asp Glu Ile 165 170 175 Asn Asn Ala Pro Ala Val Arg Leu Val Asn Ser Ile Ile Glu Gln Ala 180 185 190 Val Lys Asn Arg Ala Ser Asp Val His Ile Glu Pro Thr Glu Asn Asp 195 200 205 Leu Arg Ile Arg Phe Arg Ile Asp Gly Glu Leu His Glu Ala Met Arg 210 215 220 Val Phe Lys Ser Thr Gln Gly Pro Val Ile Thr Arg Ile Lys Ile Met 225 230 235 240 Ala Asn Met Asn Ile Ala Glu Arg Arg Ile Pro Gln Asp Gly Lys Ile 245 250 255 Glu Met Asn Ala Gly Gly Lys Asn Ile Asp Ile Arg Val Ser Ser Leu 260 265 270 Pro Thr Ile Tyr Gly Glu Lys Leu Val Leu Arg Ile Leu Asp Lys Ser 275 280 285 Gly Tyr Ile Ile Thr Lys Asp Lys Leu Gly Leu Gly Asn Asp Asp Leu 290 295 300 Lys Leu Phe Asp Asn Leu Leu Lys His Pro Asn Gly Ile Ile Leu Leu 305 310 315 320 Thr Gly Pro Thr Gly Ser Gly Lys Thr Thr Thr Leu Tyr Ala Met Leu 325 330 335 Asn Glu Leu Asn Lys Pro Asp Lys Asn Ile Ile Thr Val Glu Asp Pro 340 345 350 Val Glu Tyr Thr Leu Glu Gly Leu Asn Gln Val Gln Val Asn Glu Lys 355 360 365 Ala Gly Leu Thr Phe Ala Ser Ala Leu Arg Ser Ile Leu Arg Gln Asp 370 375 380 Pro Asp Ile Ile Met Ile Gly Glu Ile Arg Asp Arg Glu Thr Ala Glu 385 390 395 400 Ile Ala Ile Arg Ser Ser Ile Thr Gly His Leu Val Leu Ser Thr Leu 405 410 415 His Thr Asn Asp Ser Ala Gly Ala Ile Thr Arg Leu Ile Asp Met Gly 420 425 430 Ile Glu Pro Tyr Leu Val Ser Ser Ser Val Val Gly Val Ile Ala Gln 435 440 445 Arg Leu Ala Arg Lys Ile Cys Asp Asn Cys Lys Ile Glu Tyr Asp Ala 450 455 460 Ser Lys Arg Glu Lys Ile Ile Leu Gly Ile Asp Ala Asp Glu Ser Leu 465 470 475 480 Lys Leu Tyr Arg Ser Lys Gly Cys Ala Val Cys Asn Lys Thr Gly Tyr 485 490 495 Arg Gly Arg Val Pro Ile Tyr Glu Ile Met Met Met Thr Pro Lys Ile 500 505 510 Lys Glu Leu Thr Asn Glu Lys Ala Pro Ala Asp Val Ile Leu Asn Glu 515 520 525 Ala Val Ser Asn Gly Met Ser Thr Leu Lys Glu Ser Ala Lys Lys Leu 530 535 540 Val Leu Ser Gly Val Thr Thr Val Asp Glu Met Leu Arg Leu Thr Tyr 545 550 555 560 Asp Asp Ala Tyr 491175PRTT. saccharolyticum 49Met Ser Lys Val Met Lys Thr Met Asp Gly Asn Thr Ala Ala Ala His 1 5 10 15 Val Ala Tyr Ala Phe Thr Glu Val Ala Ala Ile Tyr Pro Ile Thr Pro 20 25 30 Ser Ser Pro Met Ala Glu His Val Asp Glu Trp Ser Ala His Gly Arg 35 40 45 Lys Asn Leu Phe Gly Gln Glu Val Lys Val Ile Glu Met Gln Ser Glu 50 55 60 Ala Gly Ala Ala Gly Ala Val His Gly Ser Leu Ala Ala Gly Ala Leu 65 70 75 80 Thr Thr Thr Phe Thr Ala Ser Gln Gly Leu Leu Leu Met Ile Pro Asn 85 90 95 Met Tyr Lys Ile Ala Gly Glu Leu Leu Pro Gly Val Phe His Val Ser 100 105 110 Ala Arg Ala Leu Ala Ser His Ala Leu Ser Ile Phe Gly Asp His Gln 115 120 125 Asp Val Met Ala Cys Arg Gln Thr Gly Phe Ala Leu Leu Ala Ser Gly 130 135 140 Ser Val Gln Glu Val Met Asp Leu Gly Ser Val Ala His Leu Ala Ala 145 150 155 160 Ile Lys Gly Arg Val Pro Phe Leu His Phe Phe Asp Gly Phe Arg Thr 165 170 175 Ser His Glu Tyr Gln Lys Ile Glu Val Met Asp Tyr Glu Asp Leu Arg 180 185 190 Lys Leu Leu Asp Met Asp Ala Val Arg Glu Phe Lys Lys Arg Ala Leu 195 200 205 Asn Pro Glu His Pro Val Thr Arg Gly Thr Ala Gln Asn Pro Asp Ile 210 215 220 Tyr Phe Gln Glu Arg Glu Ala Ser Asn Arg Tyr Tyr Asn Ala Val Pro 225 230 235 240 Glu Ile Val Glu Glu Tyr Met Lys Glu Ile Ser Lys Ile Thr Gly Arg 245 250 255 Glu Tyr Lys Leu Phe Asn Tyr Tyr Gly Ala Pro Asp Ala Glu Arg Ile 260 265 270 Val Ile Ala Met Gly Ser Val Thr Glu Thr Ile Glu Glu Thr Ile Asp 275 280 285 Tyr Leu Leu Lys Lys Gly Glu Lys Val Gly Val Val Lys Val His Leu 290 295 300 Tyr Arg Pro Phe Ser Phe Lys His Phe Met Asp Ala Ile Pro Lys Thr 305 310 315 320 Val Lys Lys Ile Ala Val Leu Asp Arg Thr Lys Glu Ala Gly Ala Phe 325 330 335 Gly Glu Pro Leu Tyr Glu Asp Val Arg Ala Ala Phe Tyr Asp Ser Glu 340 345 350 Met Lys Pro Ile Ile Val Gly Gly Arg Tyr Gly Leu Gly Ser Lys Asp 355 360 365 Thr Thr Pro Ala Gln Ile Val Ala Val Phe Asp Asn Leu Lys Ser Asp 370 375 380 Thr Pro Lys Asn Asn Phe Thr Ile Gly Ile Val Asp Asp Val Thr Tyr 385 390 395 400 Thr Ser Leu Pro Val Gly Glu Glu Ile Glu Thr Thr Ala Glu Gly Thr 405 410 415 Ile Ser Cys Lys Phe Trp Gly Phe Gly Ser Asp Gly Thr Val Gly Ala 420 425 430 Asn Lys Ser Ala Ile Gln Ile Ile Gly Asp Asn Thr Asp Met Tyr Ala 435 440 445 Gln Ala Tyr Phe Ser Tyr Asp Ser Lys Lys Ser Gly Gly Val Thr Ile 450 455 460 Ser His Leu Arg Phe Gly Lys Lys Pro Ile Arg Ser Thr Tyr Leu Ile 465 470 475 480 Asn Asn Ala Asp Phe Val Ala Cys His Lys Gln Ala Tyr Val Tyr Asn 485 490 495 Tyr Asp Val Leu Ala Gly Leu Lys Lys Gly Gly Thr Phe Leu Leu Asn 500 505 510 Cys Thr Trp Lys Pro Glu Glu Leu Asp Glu Lys Leu Pro Ala Ser Met 515 520 525 Lys Arg Tyr Ile Ala Lys Asn Asn Ile Asn Phe Tyr Ile Ile Asn Ala 530 535 540 Val Asp Ile Ala Lys Glu Leu Gly Leu Gly Ala Arg Ile Asn Met Ile 545 550 555 560 Met Gln Ser Ala Phe Phe Lys Leu Ala Asn Ile Ile Pro Ile Asp Glu 565 570 575 Ala Val Lys His Leu Lys Asp Ala Ile Val Lys Ser Tyr Gly His Lys 580 585 590 Gly Glu Lys Ile Val Asn Met Asn Tyr Ala Ala Val Asp Arg Gly Ile 595 600 605 Asp Ala Leu Val Lys Val Asp Val Pro Ala Ser Trp Ala Asn Ala Glu 610 615 620 Asp Glu Ala Lys Val Glu Arg Asn Val Pro Asp Phe Ile Lys Asn Ile 625 630 635 640 Ala Asp Val Met Asn Arg Gln Glu Gly Asp Lys Leu Pro Val Ser Ala 645 650 655 Phe Val Gly Met Glu Asp Gly Thr Phe Pro Met Gly Thr Ala Ala Tyr 660 665 670 Glu Lys Arg Gly Ile Ala Val Asp Val Pro Glu Trp Gln Ile Asp Asn 675 680 685 Cys Ile Gln Cys Asn Gln Cys Ala Tyr Val Cys Pro His Ala Ala Ile 690 695 700 Arg Pro Phe Leu Leu Asn Glu Glu Glu Val Lys Asn Ala Pro Glu Gly 705 710 715 720 Phe Thr Ser Lys Lys Ala Ile Gly Lys Gly Leu Glu Gly Leu Asn Phe 725 730 735 Arg Ile Gln Val Ser Val Leu Asp Cys Thr Gly Cys Gly Val Cys Ala 740 745 750 Asn Thr Cys Pro Ser Lys Glu Lys Ser Leu Ile Met Lys Pro Leu Glu 755 760 765 Thr Gln Leu Asp Gln Ala Lys Asn Trp Glu Tyr Ala Met Ser Leu Ser 770 775 780 Tyr Lys Glu Asn Pro Leu Gly Thr Asp Thr Val Lys Gly Ser Gln Phe 785 790 795 800 Glu Lys Pro Leu Leu Glu Phe Ser Gly Ala Cys Ala Gly Cys Gly Glu 805 810 815 Thr Pro Tyr Ala Arg Leu Val Thr Gln

Leu Phe Gly Asp Arg Met Leu 820 825 830 Ile Ala Asn Ala Thr Gly Cys Ser Ser Ile Trp Gly Gly Ser Ala Pro 835 840 845 Ser Thr Pro Tyr Thr Val Asn Lys Asp Gly His Gly Pro Ala Trp Ala 850 855 860 Asn Ser Leu Phe Glu Asp Asn Ala Glu Phe Gly Phe Gly Met Ala Leu 865 870 875 880 Ala Val Lys Gln Gln Arg Glu Lys Leu Ala Asp Ile Val Lys Glu Ala 885 890 895 Leu Glu Leu Asp Leu Thr Gln Asp Leu Lys Asn Ala Leu Lys Leu Trp 900 905 910 Leu Asp Asn Phe Asn Ser Ser Glu Ile Thr Lys Lys Thr Ala Asn Ile 915 920 925 Ile Val Ser Leu Ile Gln Asp Tyr Lys Thr Asp Asp Ser Lys Val Lys 930 935 940 Glu Leu Leu Asn Glu Ile Leu Asp Arg Lys Glu Tyr Leu Val Lys Lys 945 950 955 960 Ser Gln Trp Ile Phe Gly Gly Asp Gly Trp Ala Tyr Asp Ile Gly Phe 965 970 975 Gly Gly Leu Asp His Val Leu Ala Ser Gly Glu Asp Val Asn Val Leu 980 985 990 Val Phe Asp Thr Glu Val Tyr Ser Asn Thr Gly Gly Gln Ser Ser Lys 995 1000 1005 Ala Thr Pro Val Gly Ala Ile Ala Gln Phe Ala Ala Ala Gly Lys 1010 1015 1020 Gly Ile Gly Lys Lys Asp Leu Gly Arg Ile Ala Met Ser Tyr Gly 1025 1030 1035 Tyr Val Tyr Val Ala Gln Ile Ala Met Gly Ala Asn Gln Ala Gln 1040 1045 1050 Thr Ile Lys Ala Leu Lys Glu Ala Glu Ser Tyr Pro Gly Pro Ser 1055 1060 1065 Leu Ile Ile Ala Tyr Ala Pro Cys Ile Asn His Gly Ile Lys Leu 1070 1075 1080 Gly Met Gly Cys Ser Gln Ile Glu Glu Lys Lys Ala Val Glu Ala 1085 1090 1095 Gly Tyr Trp His Leu Tyr Arg Tyr Asn Pro Met Leu Lys Ala Glu 1100 1105 1110 Gly Lys Asn Pro Phe Ile Leu Asp Ser Lys Ala Pro Thr Ala Ser 1115 1120 1125 Tyr Lys Glu Phe Ile Met Gly Glu Val Arg Tyr Ser Ser Leu Ala 1130 1135 1140 Lys Thr Phe Pro Glu Arg Ala Glu Ala Leu Phe Glu Lys Ala Glu 1145 1150 1155 Glu Leu Ala Lys Glu Lys Tyr Glu Thr Tyr Lys Lys Leu Ala Glu 1160 1165 1170 Gln Asn 1175 50311PRTT. saccharolyticum 50Met Ser Lys Val Ala Ile Ile Gly Ser Gly Phe Val Gly Ala Thr Ser 1 5 10 15 Ala Phe Thr Leu Ala Leu Ser Gly Thr Val Thr Asp Ile Val Leu Val 20 25 30 Asp Leu Asn Lys Asp Lys Ala Ile Gly Asp Ala Leu Asp Ile Ser His 35 40 45 Gly Ile Pro Leu Ile Gln Pro Val Asn Val Tyr Ala Gly Asp Tyr Lys 50 55 60 Asp Val Lys Gly Ala Asp Val Ile Val Val Thr Ala Gly Ala Ala Gln 65 70 75 80 Lys Pro Gly Glu Thr Arg Leu Asp Leu Val Lys Lys Asn Thr Ala Ile 85 90 95 Phe Lys Ser Met Ile Pro Glu Leu Leu Lys Tyr Asn Asp Lys Ala Ile 100 105 110 Tyr Leu Ile Val Thr Asn Pro Val Asp Ile Leu Thr Tyr Val Thr Tyr 115 120 125 Lys Ile Ser Gly Leu Pro Trp Gly Arg Val Phe Gly Ser Gly Thr Val 130 135 140 Leu Asp Ser Ser Arg Phe Arg Tyr Leu Leu Ser Lys His Cys Asn Ile 145 150 155 160 Asp Pro Arg Asn Val His Gly Arg Ile Ile Gly Glu His Gly Asp Thr 165 170 175 Glu Phe Ala Ala Trp Ser Ile Thr Asn Ile Ser Gly Ile Ser Phe Asn 180 185 190 Glu Tyr Cys Ser Ile Cys Gly Arg Val Cys Asn Thr Asn Phe Arg Lys 195 200 205 Glu Val Glu Glu Glu Val Val Asn Ala Ala Tyr Lys Ile Ile Asp Lys 210 215 220 Lys Gly Ala Thr Tyr Tyr Ala Val Ala Val Ala Val Arg Arg Ile Val 225 230 235 240 Glu Cys Ile Leu Arg Asp Glu Asn Ser Ile Leu Thr Val Ser Ser Pro 245 250 255 Leu Asn Gly Gln Tyr Gly Val Lys Asp Val Ser Leu Ser Leu Pro Ser 260 265 270 Ile Val Gly Arg Asn Gly Val Ala Arg Ile Leu Asp Leu Pro Leu Ser 275 280 285 Asp Glu Glu Val Glu Lys Phe Arg His Ser Ala Ser Val Met Ala Asp 290 295 300 Val Ile Lys Gln Leu Asp Ile 305 310 51741PRTT. saccharolyticum 51Met Ile Asn Glu Trp Arg Gly Phe Gln Glu Gly Lys Trp Gln Lys Thr 1 5 10 15 Ile Asp Val Gln Asp Phe Ile Gln Lys Asn Tyr Thr Leu Tyr Glu Gly 20 25 30 Asp Asp Ser Phe Leu Glu Gly Pro Thr Glu Lys Thr Ile Lys Leu Trp 35 40 45 Asn Lys Val Leu Glu Leu Met Lys Glu Glu Leu Lys Lys Gly Val Leu 50 55 60 Asp Ile Asp Thr Lys Thr Val Ser Ser Ile Thr Ser His Asp Ala Gly 65 70 75 80 Tyr Ile Asp Lys Asp Leu Glu Glu Ile Val Gly Leu Gln Thr Asp Lys 85 90 95 Pro Leu Lys Arg Ala Ile Met Pro Tyr Gly Gly Ile Arg Met Val Lys 100 105 110 Lys Ala Cys Glu Ala Tyr Gly Tyr Lys Val Asp Pro Lys Val Glu Glu 115 120 125 Ile Phe Thr Lys Tyr Arg Lys Thr His Asn Asp Gly Val Phe Asp Ala 130 135 140 Tyr Thr Pro Glu Ile Arg Ala Ala Arg His Ala Gly Ile Ile Thr Gly 145 150 155 160 Leu Pro Asp Ala Tyr Gly Arg Gly Arg Ile Ile Gly Asp Tyr Arg Arg 165 170 175 Val Ala Leu Tyr Gly Ile Asp Arg Leu Ile Glu Glu Lys Glu Lys Glu 180 185 190 Lys Leu Glu Leu Asp Tyr Asp Glu Phe Asp Glu Ala Thr Ile Arg Leu 195 200 205 Arg Glu Glu Leu Thr Glu Gln Ile Lys Ala Leu Asn Glu Met Lys Glu 210 215 220 Met Ala Leu Lys Tyr Gly Tyr Asp Ile Ser Lys Pro Ala Lys Asn Ala 225 230 235 240 Lys Glu Ala Val Gln Trp Thr Tyr Phe Ala Phe Leu Ala Ala Ile Lys 245 250 255 Glu Gln Asn Gly Ala Ala Met Ser Leu Gly Arg Val Ser Thr Phe Leu 260 265 270 Asp Ile Tyr Ile Glu Arg Asp Leu Lys Glu Gly Thr Leu Thr Glu Lys 275 280 285 Gln Ala Gln Glu Leu Met Asp His Phe Val Met Lys Leu Arg Met Val 290 295 300 Arg Phe Leu Arg Thr Pro Asp Tyr Asn Glu Leu Phe Ser Gly Asp Pro 305 310 315 320 Val Trp Val Thr Glu Ser Ile Gly Gly Val Gly Val Asp Gly Arg Pro 325 330 335 Leu Val Thr Lys Asn Ser Phe Arg Ile Leu Asn Thr Leu Tyr Asn Leu 340 345 350 Gly Pro Ala Pro Glu Pro Asn Leu Thr Val Leu Trp Ser Lys Asn Leu 355 360 365 Pro Glu Asn Phe Lys Arg Phe Cys Ala Lys Val Ser Ile Asp Thr Ser 370 375 380 Ser Ile Gln Tyr Glu Asn Asp Asp Leu Met Arg Pro Ile Tyr Asn Asp 385 390 395 400 Asp Tyr Ser Ile Ala Cys Cys Val Ser Ala Met Lys Thr Gly Glu Gln 405 410 415 Met Gln Phe Phe Gly Ala Arg Ala Asn Leu Ala Lys Ala Leu Leu Tyr 420 425 430 Ala Ile Asn Gly Gly Ile Asp Glu Arg Tyr Lys Thr Gln Val Ala Pro 435 440 445 Lys Phe Asn Pro Ile Thr Ser Glu Tyr Leu Asp Tyr Asp Glu Val Met 450 455 460 Ala Ala Tyr Asp Asn Met Leu Glu Trp Leu Ala Lys Val Tyr Val Lys 465 470 475 480 Ala Met Asn Ile Ile His Tyr Met His Asp Lys Tyr Ala Tyr Glu Arg 485 490 495 Ser Leu Met Ala Leu His Asp Arg Asp Ile Val Arg Thr Met Ala Phe 500 505 510 Gly Ile Ala Gly Leu Ser Val Ala Ala Asp Ser Leu Ser Ala Ile Lys 515 520 525 Tyr Ala Lys Val Lys Ala Ile Arg Asp Glu Asn Gly Ile Ala Ile Asp 530 535 540 Tyr Glu Val Glu Gly Asp Phe Pro Lys Phe Gly Asn Asp Asp Asp Arg 545 550 555 560 Val Asp Ser Ile Ala Val Asp Ile Val Glu Arg Phe Met Asn Lys Leu 565 570 575 Lys Lys His Lys Thr Tyr Arg Asn Ser Ile Pro Thr Leu Ser Val Leu 580 585 590 Thr Ile Thr Ser Asn Val Val Tyr Gly Lys Lys Thr Gly Ala Thr Pro 595 600 605 Asp Gly Arg Lys Ala Gly Glu Pro Phe Ala Pro Gly Ala Asn Pro Met 610 615 620 His Gly Arg Asp Thr Lys Gly Ala Ile Ala Ser Met Asn Ser Ser Lys 625 630 635 640 Ile Pro Tyr Asp Ser Ser Leu Asp Gly Ile Ser Tyr Thr Phe Thr Ile 645 650 655 Val Pro Asn Ala Leu Gly Lys Asp Asp Glu Asp Lys Ile Asn Asn Leu 660 665 670 Val Gly Leu Leu Asp Gly Tyr Ala Phe Asn Ala Gly His His Ile Asn 675 680 685 Ile Asn Val Leu Asn Arg Asp Met Leu Leu Asp Ala Met Glu His Pro 690 695 700 Glu Lys Tyr Pro Gln Leu Thr Ile Arg Val Ser Gly Tyr Ala Val Asn 705 710 715 720 Phe Asn Lys Leu Thr Arg Glu Gln Gln Leu Glu Val Ile Ser Arg Thr 725 730 735 Phe His Glu Ser Met 740 5281PRTT. saccharolyticum 52Met Val Ile Thr Val Cys Val Gly Ser Ser Cys His Leu Lys Gly Ser 1 5 10 15 Tyr Asp Val Ile Asn Lys Leu Lys Glu Met Ile Lys Asn Tyr Gly Ile 20 25 30 Glu Asp Lys Val Glu Leu Lys Ala Asp Phe Cys Met Gly Asn Cys Leu 35 40 45 Arg Ala Val Ser Val Lys Ile Asp Gly Gly Ala Cys Leu Ser Ile Lys 50 55 60 Pro Asn Ser Val Glu Arg Phe Phe Lys Glu His Val Leu Gly Glu Leu 65 70 75 80 Lys 53572PRTT. saccharolyticum 53Met Ser Val Ile Asn Phe Lys Glu Ala Asn Cys Arg Asn Cys Tyr Lys 1 5 10 15 Cys Ile Arg Tyr Cys Pro Val Lys Ala Ile Lys Val Asn Asp Glu Gln 20 25 30 Ala Glu Ile Ile Glu Tyr Arg Cys Ile Ala Cys Gly Arg Cys Leu Asn 35 40 45 Ile Cys Pro Gln Asn Ala Lys Thr Val Arg Ser Asp Val Glu Arg Val 50 55 60 Gln Ser Phe Leu Asn Lys Gly Glu Lys Val Ala Phe Thr Val Ala Pro 65 70 75 80 Ser Tyr Pro Ala Leu Val Gly His Asp Gly Ala Leu Asn Phe Leu Lys 85 90 95 Ala Leu Lys Ser Leu Gly Ala Glu Met Ile Val Glu Thr Ser Val Gly 100 105 110 Ala Met Leu Ile Ser Lys Glu Tyr Glu Arg Tyr Tyr Asn Asp Leu Lys 115 120 125 Tyr Asp Asn Leu Ile Thr Thr Ser Cys Pro Ser Val Asn Tyr Leu Val 130 135 140 Glu Lys Tyr Tyr Pro Asp Leu Ile Lys Cys Leu Val Pro Val Val Ser 145 150 155 160 Pro Met Val Ala Val Gly Arg Ala Ile Lys Asn Ile His Gly Glu Gly 165 170 175 Val Lys Val Val Phe Ile Gly Pro Cys Leu Ala Lys Lys Ala Glu Met 180 185 190 Ser Asp Phe Ser Cys Glu Gly Ala Ile Asp Ala Val Leu Thr Phe Glu 195 200 205 Glu Val Met Asn Leu Phe Asn Thr Asn Lys Ile Gly Val Glu Cys Thr 210 215 220 Lys Glu Asn Leu Glu Asp Val Asp Ser Glu Ser Arg Phe Lys Leu Tyr 225 230 235 240 Pro Ile Glu Gly Lys Thr Met Asp Cys Met Asp Val Asp Leu Asn Leu 245 250 255 Arg Lys Phe Ile Ser Val Ser Ser Ile Glu Asn Val Lys Asp Ile Leu 260 265 270 Asn Asp Leu Arg Ala Gly Asn Leu His Gly Tyr Trp Ile Glu Ala Asn 275 280 285 Ala Cys Asp Gly Gly Cys Ile Asn Gly Pro Ala Phe Gly Lys Leu Glu 290 295 300 Ser Gly Ile Ala Lys Arg Lys Glu Glu Val Ile Ser Tyr Ser Arg Met 305 310 315 320 Lys Glu Arg Phe Ser Gly Asp Phe Ser Gly Ile Thr Asp Phe Ser Leu 325 330 335 Asp Leu Ser Arg Lys Phe Ile Asp Leu Ser Asp Arg Trp Lys Met Pro 340 345 350 Ser Glu Met Glu Ile Lys Glu Ile Leu Ser Lys Ile Gly Lys Phe Ser 355 360 365 Val Glu Asp Glu Leu Asn Cys Gly Ala Cys Gly Tyr Asp Thr Cys Arg 370 375 380 Glu Lys Ala Ile Ala Val Phe Asn Gly Met Ala Glu Pro Tyr Met Cys 385 390 395 400 Leu Pro Tyr Met Arg Gly Arg Ala Glu Thr Leu Ser Asn Ile Ile Ile 405 410 415 Ser Ser Thr Pro Asn Ala Ile Ile Ala Val Asn Asn Glu Tyr Glu Ile 420 425 430 Gln Asp Met Asn Arg Ala Phe Glu Lys Met Phe Leu Val Asn Ser Ala 435 440 445 Met Val Lys Gly Glu Asp Leu Ser Leu Ile Phe Asp Ile Ser Asp Phe 450 455 460 Val Glu Val Ile Glu Asn Lys Lys Ser Ile Phe Asn Lys Lys Val Ser 465 470 475 480 Phe Lys Asn Tyr Gly Ile Ile Ala Leu Glu Ser Ile Tyr Tyr Leu Glu 485 490 495 Glu Tyr Lys Ile Ala Ile Gly Ile Phe Thr Asp Ile Thr Lys Met Glu 500 505 510 Lys Gln Lys Glu Ser Phe Ser Lys Leu Lys Arg Glu Asn Tyr Gln Leu 515 520 525 Ala Gln Gln Val Ile Asp Arg Gln Met Lys Val Ala Gln Glu Ile Ala 530 535 540 Ser Leu Leu Gly Glu Thr Thr Ala Glu Thr Lys Val Ile Leu Thr Lys 545 550 555 560 Met Lys Asp Met Leu Leu Asn Gln Gly Asp Asp Glu 565 570 54386PRTT. saccharolyticum 54Met Ser His Tyr Ile Asp Ile Ala His Ala Ser Leu Asn Lys Tyr Asp 1 5 10 15 Glu Glu Leu Cys Gly Asp Ser Val Gln Ile Ile Arg Lys Lys Asp Tyr 20 25 30 Ala Met Ala Val Met Ala Asp Gly Leu Gly Ser Gly Val Lys Ala Asn 35 40 45 Ile Leu Ser Thr Leu Thr Thr Arg Ile Val Ser Lys Met Leu Asp Met 50 55 60 Gly Ser Glu Leu Arg Asp Val Val Glu Thr Val Ala Glu Thr Leu Pro 65 70 75 80 Ile Cys Lys Glu Arg Asn Ile Ala Tyr Ser Thr Phe Thr Val Val Ser 85 90 95 Ile Tyr Gly Asp Asn Ala His Leu Val Glu Tyr Asp Asn Pro Ser Val 100 105 110 Phe Tyr Phe Lys Asn Gly Val His Lys Lys Val Asp Arg Lys Cys Val 115 120 125 Glu Ile Gly Asp Lys Lys Ile Phe Glu Ser Ser Phe Lys Leu Asp Leu 130 135 140 Asn Asp Ala Leu Ile Val Val Ser Asp Gly Val Ile His Ala Gly Val 145 150 155 160 Gly Gly Ile Leu Asn Leu Gly Trp Gln Trp Asp Asn Val Lys Gln Tyr 165 170 175 Leu Ser Lys Val Leu Glu Val Tyr Ser Asp Ala Ser Asp Ile Cys Ser 180 185 190 Gln Leu Ile Thr Thr Cys Asn Asn Leu Tyr Lys Asn Arg Pro Gly Asp 195 200 205 Asp Thr Thr Ala Ile Val Ile Lys Val Asn Glu Ser Lys Lys Val Thr 210

215 220 Val Met Val Gly Pro Pro Ile Leu Lys Asn Met Asp Glu Trp Val Val 225 230 235 240 Lys Lys Leu Met Lys Ser Glu Gly Leu Lys Val Val Cys Gly Gly Thr 245 250 255 Ala Ala Lys Ile Val Ser Arg Ile Leu Asn Lys Asp Val Ile Thr Ser 260 265 270 Thr Glu Tyr Ile Asp Pro Asp Ile Pro Pro Tyr Ala His Ile Asp Gly 275 280 285 Ile Asp Leu Val Thr Glu Gly Val Leu Thr Leu Arg Lys Thr Val Glu 290 295 300 Ile Phe Lys Glu Tyr Met Asn Asp Lys Asp Ser Asn Leu Leu Arg Phe 305 310 315 320 Ser Lys Lys Asp Ala Ala Thr Arg Leu Phe Lys Ile Leu Asn Tyr Ala 325 330 335 Thr Asp Val Asn Phe Leu Val Gly Gln Ala Val Asn Ser Ala His Gln 340 345 350 Asn Pro Asp Phe Pro Ser Asp Leu Arg Ile Lys Val Arg Ile Val Glu 355 360 365 Glu Leu Ile Ser Leu Leu Glu Arg Leu Asn Lys Asn Val Glu Val Asn 370 375 380 Tyr Phe 385 55504PRTT. saccharolyticum 55Leu Phe Lys Phe Asn Thr Asp Val Gln Met Leu Lys Tyr Glu Val Leu 1 5 10 15 Tyr Asn Val Ala Lys Leu Thr Leu Glu Asp Arg Leu Glu Asp Glu Tyr 20 25 30 Asp Glu Ile Pro Tyr Glu Ile Ile Pro Gly Thr Lys Pro Arg Phe Arg 35 40 45 Cys Cys Val Tyr Lys Glu Arg Ala Ile Ile Glu Gln Arg Thr Lys Val 50 55 60 Ala Met Gly Lys Asn Leu Lys Arg Thr Met Lys His Ala Val Asp Gly 65 70 75 80 Glu Glu Pro Ile Ile Gln Val Leu Asp Ile Ala Cys Glu Glu Cys Pro 85 90 95 Ile Lys Arg Tyr Arg Val Thr Glu Ala Cys Arg Gly Cys Ile Thr His 100 105 110 Arg Cys Thr Glu Val Cys Pro Lys Gly Ala Ile Thr Ile Ile Asn Lys 115 120 125 Lys Ala Asn Ile Asp Tyr Asp Lys Cys Ile Glu Cys Gly Arg Cys Lys 130 135 140 Asp Ala Cys Pro Tyr Asn Ala Ile Ser Asp Asn Leu Arg Pro Cys Ile 145 150 155 160 Arg Ser Cys Ser Ala Lys Ala Ile Thr Met Asp Glu Glu Leu Lys Ala 165 170 175 Ala Ile Asn Tyr Glu Lys Cys Thr Ser Cys Gly Ala Cys Thr Leu Ala 180 185 190 Cys Pro Phe Gly Ala Ile Thr Asp Lys Ser Tyr Ile Val Asp Ile Ile 195 200 205 Arg Ala Ile Lys Ser Gly Lys Lys Val Tyr Ala Leu Val Ala Pro Ala 210 215 220 Ile Ala Ser Gln Phe Lys Asp Val Thr Val Gly Gln Ile Lys Ser Ala 225 230 235 240 Leu Lys Glu Phe Gly Phe Val Asp Val Ile Glu Val Ala Leu Gly Ala 245 250 255 Asp Phe Val Ala Met Glu Glu Ala Lys Glu Phe Ser His Lys Ile Lys 260 265 270 Asp Ile Lys Val Met Thr Ser Ser Cys Cys Pro Ala Phe Val Ala His 275 280 285 Ile Lys Lys Ser Tyr Pro Glu Leu Ser Gln Asn Ile Ser Thr Thr Val 290 295 300 Ser Pro Met Thr Ala Ile Ser Lys Tyr Ile Lys Lys His Asp Pro Met 305 310 315 320 Ala Val Thr Val Phe Ile Gly Pro Cys Thr Ala Lys Lys Ser Glu Val 325 330 335 Met Arg Asp Asp Val Lys Gly Ile Thr Asp Phe Ala Met Thr Phe Glu 340 345 350 Glu Met Val Ala Val Leu Asp Ala Ala Lys Ile Asp Met Lys Glu Gln 355 360 365 Gln Asp Val Glu Val Asp Asp Ala Thr Leu Phe Gly Arg Lys Phe Ala 370 375 380 Arg Ser Gly Gly Val Leu Glu Ala Val Val Glu Ala Val Lys Glu Ile 385 390 395 400 Gly Ala Asp Val Glu Val Asn Pro Val Val Cys Asn Gly Leu Asp Glu 405 410 415 Cys Asn Lys Thr Leu Lys Ile Met Lys Ala Gly Lys Leu Pro Asn Asn 420 425 430 Phe Ile Glu Gly Met Ala Cys Ile Gly Gly Cys Ile Gly Gly Ala Gly 435 440 445 Val Ile Asn Asn Asn Val Asn Gln Ala Lys Leu Ala Val Asn Lys Phe 450 455 460 Gly Asp Ser Ser Tyr His Lys Ser Ile Lys Asp Arg Ile Ser Gln Phe 465 470 475 480 Asp Thr Asp Asp Val Asp Phe His Val Asp Ser Gly Glu Asp Glu Ser 485 490 495 Ser Glu Thr Ser Phe Lys Glu Ala 500 56581PRTT. saccharolyticum 56Met Asp Lys Val Arg Ile Thr Ile Asp Gly Ile Pro Ala Glu Val Pro 1 5 10 15 Ala Asn Tyr Thr Val Leu Gln Ala Ala Lys Tyr Ala Lys Ile Glu Ile 20 25 30 Pro Thr Leu Cys Tyr Leu Glu Glu Ile Asn Glu Ile Gly Ala Cys Arg 35 40 45 Leu Cys Val Val Glu Ile Lys Gly Val Arg Asn Leu Gln Ala Ser Cys 50 55 60 Val Tyr Pro Val Ser Asp Gly Met Glu Ile Tyr Thr Asn Thr Pro Arg 65 70 75 80 Val Arg Glu Ala Arg Arg Ser Asn Leu Glu Leu Ile Leu Ser Ala His 85 90 95 Asp Arg Ser Cys Leu Thr Cys Val Arg Ser Gly Asn Cys Glu Leu Gln 100 105 110 Asp Leu Ser Arg Lys Ser Gly Ile Asp Glu Ile Arg Phe Met Gly Glu 115 120 125 Asn Ile Lys Tyr Gln Lys Asp Glu Ser Ser Pro Ser Ile Val Arg Asp 130 135 140 Pro Asn Lys Cys Val Leu Cys Arg Arg Cys Val Ala Thr Cys Asn Asn 145 150 155 160 Val Gln Asn Val Phe Ala Ile Gly Met Val Asn Arg Gly Phe Lys Thr 165 170 175 Ile Val Ala Pro Ser Phe Gly Arg Gly Leu Asn Glu Ser Pro Cys Ile 180 185 190 Ser Cys Gly Gln Cys Ile Glu Ala Cys Pro Val Gly Ala Ile Tyr Glu 195 200 205 Lys Asp His Thr Lys Ile Val Tyr Asp Ala Leu Leu Asp Glu Lys Lys 210 215 220 Tyr Val Val Val Gln Thr Ala Pro Ala Val Arg Val Ala Leu Gly Glu 225 230 235 240 Glu Phe Gly Met Pro Tyr Gly Ser Ile Val Thr Gly Lys Met Val Ser 245 250 255 Ala Leu Lys Arg Leu Gly Phe Asp Lys Val Phe Asp Thr Asp Phe Ala 260 265 270 Ala Asp Leu Thr Ile Ile Glu Glu Gly Asn Glu Leu Leu Lys Arg Leu 275 280 285 Asn Glu Gly Gly Lys Leu Pro Met Ile Thr Ser Cys Ser Pro Gly Trp 290 295 300 Ile Asn Tyr Cys Glu Arg Tyr Tyr Pro Glu Phe Ile Asp Asn Leu Ser 305 310 315 320 Thr Cys Lys Ser Pro His Met Met Met Gly Ala Ile Ile Lys Ser Tyr 325 330 335 Phe Ala Glu Lys Glu Gly Ile Asp Pro Lys Asp Ile Phe Val Val Ser 340 345 350 Ile Met Pro Cys Thr Ala Lys Lys Tyr Glu Ile Asp Arg Pro Gln Met 355 360 365 Ile Val Asp Gly Met Lys Asp Val Asp Ala Val Leu Thr Thr Arg Glu 370 375 380 Leu Ala Arg Met Ile Lys Gln Ser Gly Ile Asp Phe Val Asn Leu Pro 385 390 395 400 Asp Ser Glu Tyr Asp Asn Pro Leu Gly Glu Ser Ser Gly Ala Gly Val 405 410 415 Ile Phe Gly Ala Thr Gly Gly Val Met Glu Ala Ala Leu Arg Thr Val 420 425 430 Ala Asp Ile Val Glu Gly Lys Asp Ile Glu Asn Phe Glu Tyr Glu Glu 435 440 445 Val Arg Gly Leu Glu Gly Ile Lys Glu Ala Lys Ile Asp Ile Gly Gly 450 455 460 Lys Glu Ile Lys Ile Ala Val Ala Asn Gly Thr Gly Asn Ala Lys Lys 465 470 475 480 Leu Leu Asp Lys Ile Lys Asn Gly Glu Ala Glu Tyr His Phe Ile Glu 485 490 495 Val Met Gly Cys Pro Gly Gly Cys Ile Met Gly Gly Gly Gln Pro Ile 500 505 510 His Asn Pro Asn Glu Lys Asp Leu Val Arg Lys Ser Arg Leu Lys Ala 515 520 525 Ile Tyr Glu Ala Asp Lys Asp Leu Pro Ile Arg Lys Ser His Lys Asn 530 535 540 Pro Met Ile Thr Lys Leu Tyr Glu Glu Phe Leu Ile Ser Pro Leu Gly 545 550 555 560 Glu Lys Ser His His Leu Leu His Thr Thr Tyr Ser Lys Lys Asp Leu 565 570 575 Tyr Pro Met Asn Asp 580 57282PRTT. saccharolyticum 57Leu Asn Asp Ile Leu Val Lys Ala Arg Asn Asn Lys Tyr Ala Ile Gly 1 5 10 15 Gly Phe Asn Phe Asn Phe Tyr Asp Asp Ala Leu Gly Ile Ile Ser Ala 20 25 30 Ala Tyr Glu Leu Lys Ser Pro Ile Ile Leu Met Ala Ser Glu Gly Cys 35 40 45 Val Lys Phe Leu Gly Val Lys His Ile Val Asn Phe Val Asn Gln Leu 50 55 60 Lys Asp Glu Tyr Asn Ile Pro Ile Ile Leu His Leu Asp His Gly Lys 65 70 75 80 Asp Ile Glu Ile Ile Lys Asn Cys Ile Asp Asn Lys Phe Asp Ser Ile 85 90 95 Met Tyr Asp Gly Ser Leu Leu Asn Phe Glu Glu Asn Ile Lys Asn Thr 100 105 110 Lys Phe Ile Ala Asp Leu Cys His Asp Lys Gly Met Thr Ile Glu Gly 115 120 125 Glu Leu Gly Arg Ile Ser Gly Ala Glu Glu Asn Ile Glu Asn Ser Glu 130 135 140 Asp Val Phe Thr Asp Pro Asp Ser Val Ala Glu Phe Thr Glu Arg Ser 145 150 155 160 Asp Val Asp Ser Leu Ala Val Ala Ile Gly Asn Ala His Gly Leu Tyr 165 170 175 Lys Gly Arg Pro Arg Leu Asp Phe Glu Arg Leu Ser Lys Ile Asn Lys 180 185 190 Ile Ser Lys Val Pro Leu Val Leu His Gly Gly Thr Gly Ile Pro Tyr 195 200 205 Glu Asp Ile Gln Lys Ala Ile Gln Leu Gly Ile Ser Lys Val Asn Val 210 215 220 Gly Thr Glu Ile Lys Ile Ala Tyr Ile Lys Ser Ile Lys Lys His Leu 225 230 235 240 Glu Thr Ile Asn Asp Asn Asp Ile Arg His Leu Val Ser Met Val Gln 245 250 255 Asn Asp Ile Lys Glu Leu Val Lys Gln Tyr Leu Asp Ile Phe Gly Thr 260 265 270 Ala Asn Lys Tyr Ser Gln Leu Gln Ser Met 275 280 58283PRTT. saccharolyticum 58Met Leu Val Thr Gly Ile Glu Leu Leu Lys Lys Ala Asn Glu Glu Gly 1 5 10 15 Tyr Ala Val Gly Ala Phe Asn Thr Ser Asn Leu Glu Ile Thr Gln Ala 20 25 30 Ile Val Glu Ala Ala Glu Glu Met Arg Ser Pro Ala Ile Ile Gln Val 35 40 45 Ser Glu Gly Gly Leu Lys Tyr Ala Gly Ile Glu Thr Ile Ser Ala Ile 50 55 60 Val Arg Thr Leu Ala Thr Lys Ala Ser Val Pro Ile Ala Leu His Leu 65 70 75 80 Asp His Gly Thr Asp Phe Asn Asn Val Met Lys Cys Leu Arg Asn Gly 85 90 95 Trp Thr Ser Val Met Met Asp Ala Ser Lys Leu Pro Leu Glu Lys Asn 100 105 110 Ile Glu Val Thr Lys Asn Val Val Thr Ile Ala His Gly Met Gly Val 115 120 125 Ser Val Glu Ala Glu Ile Gly Lys Ile Gly Gly Thr Glu Asp Asn Val 130 135 140 Thr Val Asp Glu Arg Glu Ala Ser Met Thr Asp Pro Asp Glu Ala Phe 145 150 155 160 Lys Phe Ala Lys Glu Thr Gly Val Asp Tyr Leu Ala Ile Ser Ile Gly 165 170 175 Thr Ala His Gly Pro Tyr Lys Gly Glu Pro Lys Leu Asp Phe Asp Arg 180 185 190 Leu Val Lys Ile Lys Glu Met Leu Lys Met Pro Ile Val Leu His Gly 195 200 205 Ala Ser Gly Val Pro Glu Ala Asp Ile Arg Lys Ala Val Ser Leu Gly 210 215 220 Val Asn Lys Ile Asn Ile Asp Thr Asp Ile Arg Gln Ala Phe Ala Ala 225 230 235 240 Arg Leu Arg Glu Leu Leu Lys Asn Asp Glu Glu Val Tyr Asp Pro Arg 245 250 255 Lys Ile Leu Gly Pro Cys Lys Glu Ala Met Lys Glu Val Ile Lys Asn 260 265 270 Lys Met Arg Met Phe Gly Ser Glu Gly Arg Ala 275 280 59400PRTT. saccharolyticum 59Met Ile Thr Gly Asp Gln Leu Leu Ile Lys Gln Ile Asn Lys Ser Ile 1 5 10 15 Val Leu Asn Thr Ile Arg Lys Lys Gly Leu Ile Ser Arg Ala Asp Leu 20 25 30 Ala Asn Ile Thr Gly Leu Asn Lys Ser Thr Val Ser Ser Leu Val Asp 35 40 45 Glu Leu Ile Lys Glu Gly Phe Val Glu Glu Glu Gly Pro Gly Glu Ser 50 55 60 Lys Gly Gly Arg Lys Pro Ile Met Leu Met Ile Asn Ser Leu Ala Gly 65 70 75 80 Cys Val Ile Gly Val Asp Leu Asp Val Asn Tyr Ile Leu Val Ile Leu 85 90 95 Thr Asp Ile Leu Ala Asn Ile Leu Trp Gln Lys Arg Ile Asn Leu Lys 100 105 110 Leu Gly Glu Ser Lys Glu Asp Ile Ile Ser Lys Met Leu Glu Leu Ile 115 120 125 Asp Glu Ala Ile Lys Asn Ser Pro Asn Thr Val Lys Gly Ile Leu Gly 130 135 140 Ile Gly Ile Gly Val Pro Gly Ile Thr Asp Tyr Lys Arg Gly Val Val 145 150 155 160 Leu Lys Ala Pro Asn Leu Asn Trp Glu Asn Val Glu Leu Lys Lys Met 165 170 175 Val Glu Glu Arg Phe Asn Leu Lys Val Tyr Ile Asp Asn Glu Ala Asn 180 185 190 Thr Gly Ala Ile Gly Glu Lys Trp Phe Gly Gly Gly Arg Asn Ala Lys 195 200 205 Asn Phe Val Tyr Val Ser Ala Gly Ile Gly Ile Gly Thr Gly Ile Ile 210 215 220 Ile Asn Asn Glu Leu Tyr Arg Gly Ser Asn Gly Leu Ala Gly Glu Met 225 230 235 240 Gly His Met Thr Ile Asp Ile Asn Asp His Met Cys Ser Cys Gly Asn 245 250 255 Arg Gly Cys Trp Glu Asn Tyr Ala Ser Glu Lys Ser Leu Phe Arg Tyr 260 265 270 Ile Lys Glu Arg Leu Glu Ala Gly Gln Glu Asp Asp Phe Ile Asp Ser 275 280 285 Glu Asn Ile Asp Ser Leu Asp Ile Asn Asp Ile Ala Gly Tyr Ala Glu 290 295 300 Leu Gly Ser Lys Leu Ala Ile Asp Ala Ile Asn Glu Ile Ser Lys Asn 305 310 315 320 Leu Ser Val Gly Ile Val Asn Ile Val Asn Thr Phe Asn Pro Asp Leu 325 330 335 Val Leu Ile Gly Asn Thr Leu Ser Ala Ile Gly Asp Met Leu Ile Asp 340 345 350 Ala Val Lys Glu Tyr Val Arg Glu Lys Cys Leu Val Ser Arg Tyr Asn 355 360 365 Asp Ile Ala Ile Glu Ile Ser Lys Leu Gly Met Leu Glu Arg Ala Ile 370 375 380 Gly Ala Val Thr Leu Val Ile Ser Glu Val Phe Ser Tyr Pro Gly Leu 385 390 395 400 60451PRTT. saccharolyticum 60Met Thr Asn Val Leu Asn Phe Asp Tyr Ser Asn Ala Leu Asn Phe Val 1 5 10 15 Asn Glu His Glu Ile Ser Tyr Leu Glu Lys Gln Ala Leu Leu Ser Leu 20 25 30 Asp Met Val Leu Asn Lys Thr Ala Gln Gly Ser Asp Phe Leu Gly Trp 35 40 45 Val Asp Leu Pro Lys Asp Tyr Asp Lys Glu Glu Phe Ala Arg Ile Lys 50 55 60 Lys Ala Ala Glu Lys Ile

Lys Ser Asp Ser Asp Ala Leu Val Val Ile 65 70 75 80 Gly Ile Gly Gly Ser Tyr Leu Gly Ala Arg Ala Ala Ile Glu Met Leu 85 90 95 Thr His Ser Phe Tyr Asn Val Leu Pro Gln Ser Val Arg Lys Ala Pro 100 105 110 Glu Ile Tyr Phe Ala Gly Asn Ser Ile Ser Ser Thr Tyr Leu Gln Asp 115 120 125 Leu Leu Glu Ile Leu Glu Gly Lys Asp Val Ser Ile Asn Val Ile Ser 130 135 140 Lys Ser Gly Thr Thr Thr Glu Pro Ala Ile Ala Phe Arg Val Phe Arg 145 150 155 160 Asp Phe Leu Glu Lys Lys Tyr Gly Lys Glu Glu Ala Lys Ser Arg Ile 165 170 175 Tyr Val Thr Thr Asp Arg Gln Lys Gly Ala Leu Lys Lys Leu Ala Asp 180 185 190 Glu Glu Gly Tyr Glu Thr Phe Val Ile Pro Asp Asp Val Gly Gly Arg 195 200 205 Tyr Ser Val Leu Thr Ala Val Gly Leu Leu Pro Ile Ala Ala Ala Gly 210 215 220 Ile Asp Ile Asp Glu Met Met Lys Gly Ala Tyr Asp Ala Ser Ile Val 225 230 235 240 Phe Lys Lys Pro Asp Ile Lys Glu Asn Leu Ser Met Gln Tyr Ala Val 245 250 255 Leu Arg Asn Ala Leu Tyr Arg Lys Gly Lys Ser Val Glu Ile Leu Val 260 265 270 Asn Tyr Glu Pro Arg Leu His Tyr Phe Ser Glu Trp Trp Lys Gln Leu 275 280 285 Tyr Gly Glu Ser Glu Gly Lys Asp His Lys Gly Ile Tyr Pro Ala Ser 290 295 300 Val Asp Phe Ser Thr Asp Leu His Ser Met Gly Gln Phe Ile Gln Asp 305 310 315 320 Gly Ser Arg Ile Met Phe Glu Thr Val Ile Asn Val Glu Lys Pro Leu 325 330 335 Lys Glu Ile Thr Ile Asn Glu Asp Lys Asp Asn Val Asp Gly Leu Asn 340 345 350 Phe Leu Thr Gly Lys Thr Val Asp Leu Val Asn Lys Lys Ala Phe Glu 355 360 365 Gly Thr Val Leu Ala His Asn Asp Gly Gly Val Pro Asn Leu Ile Val 370 375 380 Asn Val Pro Glu Ile Ser Ala Tyr Asn Phe Gly Tyr Leu Val Tyr Phe 385 390 395 400 Phe Glu Met Ala Cys Gly Ile Ser Gly Tyr Leu Asn Gly Val Asn Pro 405 410 415 Phe Asp Gln Pro Gly Val Glu Ala Tyr Lys Lys Asn Met Phe Ala Leu 420 425 430 Leu Gly Lys Pro Gly Tyr Glu Lys Glu Lys Glu Glu Leu Glu Lys Arg 435 440 445 Leu Lys Arg 450 61372PRTT. saccharolyticum 61Met Tyr Asn Ile Gln Leu Asp Ser Pro Asn Leu Gly Asp Lys Glu Lys 1 5 10 15 Asp Tyr Leu Val Lys Cys Ile Glu Ser Gly Tyr Val Ser Thr Val Gly 20 25 30 Pro Phe Val Pro Glu Phe Glu Arg Arg Phe Ala Glu Phe Leu Asn Val 35 40 45 Asn His Cys Val Ser Val Gln Ser Gly Thr Ala Ala Leu Tyr Met Ala 50 55 60 Leu Tyr Glu Leu Gly Ile Lys Asp Gly Asp Glu Val Ile Val Pro Ala 65 70 75 80 Ile Thr Phe Val Ala Thr Val Asn Pro Ile Val Tyr Cys Gly Ala Thr 85 90 95 Pro Val Phe Val Asp Val Asp Lys Asp Thr Trp Asn Ile Asp Pro Lys 100 105 110 Glu Ile Glu Lys Ala Ile Thr Pro Lys Thr Lys Ala Ile Ile Pro Val 115 120 125 His Leu Tyr Gly Asn Pro Cys Asp Met Asp Lys Ile Met Glu Ile Ala 130 135 140 Lys Glu Asn Asn Ile Tyr Val Ile Glu Asp Ala Thr Glu Ser Leu Gly 145 150 155 160 Ala Leu Tyr Lys Gly Arg Met Thr Gly Thr Ile Gly His Ile Gly Cys 165 170 175 Phe Ser Phe Asn Gly Asn Lys Val Ile Thr Thr Gly Gly Gly Gly Met 180 185 190 Val Ala Ser Asn Asn Glu Asp Trp Val Ser His Ile Arg Phe Leu Val 195 200 205 Asn Gln Ala Arg Asp Met Thr Gln Gly Tyr Phe His Thr Glu Ile Gly 210 215 220 Phe Asn Tyr Arg Met Thr Asn Leu Glu Ala Ser Leu Gly Ile Ala Gln 225 230 235 240 Leu Glu Arg Leu Ala Gly Phe Leu Glu Lys Lys Arg Met Tyr Phe Glu 245 250 255 Ile Tyr Lys Lys Ile Phe Asn Gly Ile Glu Glu Ile Ser Leu Gln Thr 260 265 270 Glu Tyr Glu Gly Ala Lys Ser Ser Asp Trp Leu Ser Ser Val Lys Ile 275 280 285 Asp Cys Lys Lys Val Gly Met Thr Ile His Gln Ile Gln Asp Glu Leu 290 295 300 Lys Arg Arg Gly Ile Pro Thr Arg Arg Ile Phe Asn Pro Ile Val Asp 305 310 315 320 Leu Pro Pro Tyr Lys Lys Tyr Lys Lys Gly Ser Tyr Ser Asn Ser Tyr 325 330 335 Glu Ile Tyr Glu Asn Gly Leu Asn Leu Pro Ser Ser Thr Leu Asn Thr 340 345 350 Tyr Glu Asp Val Lys Tyr Val Ala Lys Thr Leu Leu Asp Ile Leu Ser 355 360 365 Ile Lys Lys Arg 370 62253PRTT. saccharolyticum 62Met Leu Ala Ile Glu Arg Arg Lys Arg Ile Met Arg Leu Ile Gln Glu 1 5 10 15 Asn Gln Ser Val Leu Val Pro Glu Leu Ser Lys Leu Phe Asn Val Thr 20 25 30 Glu Glu Thr Ile Arg Arg Asp Leu Glu Lys Leu Glu Ala Glu Gly Leu 35 40 45 Leu Lys Arg Thr Tyr Gly Gly Ala Val Ile Asn Glu Asn Ser Ser Ala 50 55 60 Asp Ile Pro Leu Asn Ile Arg Glu Ile Thr Asn Ile Glu Ser Lys Gln 65 70 75 80 Ala Ile Ser Met Lys Val Ala Glu Tyr Ile Glu Asp Gly Asp Thr Leu 85 90 95 Leu Leu Asp Ser Ser Ser Thr Val Leu Gln Val Ala Lys Gln Leu Lys 100 105 110 Phe Lys Lys Lys Leu Thr Val Ile Thr Asn Ser Glu Lys Ile Ile Leu 115 120 125 Glu Leu Ala Asn Ala Lys Asp Cys Lys Val Ile Ser Thr Gly Gly Val 130 135 140 Leu Lys Gln Asn Ser Met Ser Leu Ile Gly Asn Phe Ala Glu Asp Met 145 150 155 160 Ile Lys Asn Phe Cys Val Asp Lys Ala Ile Ile Ser Ser Lys Gly Phe 165 170 175 Asp Met Thr Asn Gly Ile Thr Glu Ser Asn Glu Met Glu Ala Glu Ile 180 185 190 Lys Lys Ala Met Ala Asn Ser Ala Glu Lys Val Phe Leu Leu Leu Asp 195 200 205 His Asn Lys Phe Asp Lys Ser Ser Phe Val Lys Met Phe Asp Leu Asp 210 215 220 Lys Ile Asp Tyr Leu Phe Thr Asp Arg Lys Leu Ser Leu Glu Trp Glu 225 230 235 240 Glu Phe Leu Lys Lys His Asn Ile Asp Leu Ile Tyr Cys 245 250 63761DNAT. saccharolyticum 63atgcttgcga tagaacgaag gaagaggata atgaggctta tacaggaaaa tcaaagcgtt 60tggtgcctga gttaagtaaa ttgtttaatg tgacagagga aactataagg agagatttag 120agaaacttga agcagaaggg cttttaaaga ggacttatgg tggtgctgtt ataaatgaaa 180attcaagtgc tgatatcccc ttaaatataa gggaaataac gaatatagaa agcaaacagg 240ccataagtat gaaggttgcc gaatacattg aagatggtga tacacttttg cttgattcaa 300gctctacagt tcttcaagta gcaaagcaat taaaattcaa aaagaagctt acagtcataa 360caaattcgga aaagataata ttagaattag caaatgcgaa agattgcaaa gtcatttcta 420caggaggagt attgaagcaa aattctatgt cgctaattgg aaatttcgcg gaagatatga 480taaaaaattt ctgtgtagat aaagccataa tatcatcaaa aggttttgac atgacaaatg 540gcattacaga gtcaaacgaa atggaagctg aaataaaaaa agccatggcc aactcggcag 600aaaaagtgtt tttacttctt gatcacaaca aatttgacaa gtcatcgttc gtcaagatgt 660ttgacttaga taaaatcgat tatctattta ccgatagaaa gctgtcttta gaatgggaag 720aattcttgaa aaaacacaat attgatttaa tctattgtta g 76164259PRTT. saccharolyticum 64Val Tyr Ser Glu Tyr Glu Val Lys Lys Gln Ile Cys Glu Ile Gly Lys 1 5 10 15 Arg Ile Tyr Met Asn Gly Phe Val Ala Ala Asn Asp Gly Asn Ile Thr 20 25 30 Val Arg Ile Gly Glu Asn Glu Ile Ile Thr Thr Pro Thr Gly Val Ser 35 40 45 Lys Gly Phe Met Thr Pro Asp Met Leu Leu Asn Ile Asn Leu Asn Gly 50 55 60 Glu Val Leu Lys Ser Ser Gly Asp Tyr Lys Pro Ser Thr Glu Ile Lys 65 70 75 80 Met His Leu Arg Val Tyr Arg Glu Arg Pro Asp Val Lys Ser Val Ile 85 90 95 His Ala His Pro Pro Phe Gly Thr Gly Phe Ala Ile Val Gly Ile Pro 100 105 110 Leu Thr Lys Pro Ile Met Pro Glu Ala Val Ile Ser Leu Gly Cys Val 115 120 125 Pro Ile Ala Glu Tyr Gly Thr Pro Ser Thr Glu Glu Leu Pro Asp Ala 130 135 140 Val Ser Lys Tyr Leu Gln Asn Tyr Asp Ala Leu Leu Leu Glu Asn His 145 150 155 160 Gly Ala Leu Thr Tyr Gly Pro Asp Leu Ile Ser Ala Tyr Tyr Lys Met 165 170 175 Glu Ser Leu Glu Phe Tyr Ala Lys Leu Thr Phe Ile Ser Thr Leu Leu 180 185 190 Gly Gly Pro Lys Glu Leu Ser Asp Ser Gln Val Glu Lys Leu Tyr Glu 195 200 205 Ile Arg Arg Lys Phe Gly Leu Lys Gly Arg His Pro Gly Asp Leu Cys 210 215 220 Ser Thr Leu Gly Cys Ser Thr Asn Ser Ala Lys Ser Asn Asp Asp Asp 225 230 235 240 Ile Ser Glu Leu Val Asn Val Ile Thr Lys Lys Val Leu Glu Gln Leu 245 250 255 Lys Tyr Asn 65780DNAT. saccharolyticum 65gtgtattctg aatatgaggt aaaaaaacag atctgcgaaa taggaaagag aatctacatg 60aatgggtttg tggcagcgaa tgacggcaat atcaccgtta ggattggtga aaatgaaata 120ataacgacgc ctaccggtgt cagcaaaggt ttcatgactc cagacatgct attaaatatt 180aatttaaacg gtgaagtatt aaaatcttca ggcgactaca aaccgtccac agaaataaag 240atgcatctta gagtctatag agaaaggcca gatgtcaaat cagtcataca tgcacatcca 300ccatttggca caggttttgc tattgtaggg atcccgctta caaagccaat aatgccagaa 360gcagttatat ctttaggctg tgtgccgata gccgaatacg ggacgccttc tacagaagag 420ctgccagatg ccgtctctaa atatttgcaa aattacgatg cgcttttatt agaaaatcat 480ggtgcgttga catacggtcc tgatttaatt agcgcatact acaagatgga atcacttgaa 540ttttacgcaa aattgacatt tatttctaca cttctcggag gtccaaaaga attatcagat 600agccaagtag aaaagcttta tgaaattagg agaaaattcg gtttaaaagg aagacatcca 660ggcgatttgt gcagtacatt aggatgcagc acaaattctg caaaatcgaa tgatgatgac 720atttctgaac ttgtgaatgt tatcactaag aaagtattag aacaattgaa atacaattaa 78066554PRTT. saccharolyticum 66Met Lys His Ser Lys Arg Phe Glu Val Leu Gly Lys Arg Pro Val Asn 1 5 10 15 Gln Asp Gly Phe Ile Asn Glu Trp Pro Glu Lys Gly Phe Ile Ala Met 20 25 30 Cys Ser Pro Asn Asp Pro Lys Pro Ser Ile Lys Ile Glu Asn Asp Lys 35 40 45 Ile Val Glu Met Asp Gly Lys Arg Arg Glu Asp Phe Asp Phe Ile Asp 50 55 60 Leu Phe Ile Ala Asp His Ala Ile Asn Ile Tyr Gln Ala Glu Lys Ser 65 70 75 80 Met Lys Met Asn Ser Leu Asp Ile Ala Lys Met Leu Val Asp Ile Asn 85 90 95 Val Glu Arg Lys Thr Ile Ile Lys Val Val Ser Gly Leu Thr Pro Ala 100 105 110 Lys Ile Met Glu Val Val Asn His Leu Asn Val Val Glu Met Met Met 115 120 125 Ala Met Gln Lys Met Arg Ala Arg Lys Ile Pro Ala Asn Gln Ser His 130 135 140 Ile Thr Asn Leu Lys Asp Asn Pro Val Gln Ile Ala Ala Asp Ala Ala 145 150 155 160 Glu Cys Ala Leu Arg Gly Phe Arg Glu Glu Glu Thr Thr Val Gly Val 165 170 175 Thr Lys Tyr Ala Pro Phe Asn Ala Ile Ala Leu Leu Ile Gly Ser Gln 180 185 190 Ala Leu Lys Arg Gly Val Leu Thr Gln Cys Ala Val Glu Glu Ala Thr 195 200 205 Glu Leu Glu Leu Gly Met Arg Gly Phe Thr Thr Tyr Ala Glu Thr Ile 210 215 220 Ser Val Tyr Gly Thr Glu Ser Val Phe Ile Asp Gly Asp Asp Thr Pro 225 230 235 240 Tyr Ser Lys Ala Phe Leu Ala Ser Ala Tyr Ala Ser Arg Gly Leu Lys 245 250 255 Met Arg Phe Thr Ser Gly Thr Gly Ser Glu Val Leu Met Gly Asn Ala 260 265 270 Glu Gly Lys Ser Met Leu Tyr Leu Glu Ile Arg Cys Ile Met Val Thr 275 280 285 Lys Gly Ala Gly Val Gln Gly Leu Gln Asn Gly Ala Ile Ser Cys Ile 290 295 300 Gly Ile Thr Ser Ser Val Pro Ser Gly Ile Arg Ala Val Leu Ala Glu 305 310 315 320 Asn Leu Ile Ala Ser Met Leu Asp Leu Glu Val Ala Ser Gly Asn Asp 325 330 335 Gln Thr Phe Thr His Ser Asp Ile Arg Arg Thr Ala Arg Thr Met Met 340 345 350 Gln Phe Leu Pro Gly Thr Asp Phe Ile Phe Ser Gly Tyr Ser Gly Thr 355 360 365 Pro Asn Tyr Asp Asn Met Phe Ala Gly Ser Asn Phe Asp Ala Glu Asp 370 375 380 Phe Asp Asp Tyr Asn Val Leu Gln Arg Asp Leu Met Val Asp Gly Gly 385 390 395 400 Leu Arg Pro Val Lys Glu Glu Asp Val Val Glu Val Arg Arg Lys Ala 405 410 415 Ala Lys Ala Leu Gln Asp Val Phe Arg Glu Leu Asn Leu Gly Val Val 420 425 430 Thr Asp Glu Glu Val Glu Ala Ala Ala Tyr Ala His Gly Ser Lys Asp 435 440 445 Met Pro Glu Arg Asp Val Leu Ser Asp Leu Glu Ser Ile Asp Glu Met 450 455 460 Met Lys Arg Gly Ile Thr Gly Ile Asp Ile Val Lys Ala Leu Tyr Arg 465 470 475 480 Ser Gly His Glu Asp Ile Ala Glu Asn Ile Leu Asn Met Leu Lys Gln 485 490 495 Arg Ile Ser Gly Asp Tyr Leu Gln Thr Ser Ala Ile Leu Asp Glu Asp 500 505 510 Phe Asn Val Ile Ser Ala Ile Asn Cys Pro Asn Asp Tyr Leu Gly Pro 515 520 525 Gly Thr Gly Tyr Arg Ile Asp Lys Asp Arg Trp Glu Glu Ile Lys Asn 530 535 540 Ile Pro Tyr Thr Ile Asn Pro Asp Asn Leu 545 550 671665DNAT. saccharolyticum 67atgaaacatt ctaagcgatt tgaggttctc ggcaaaagac ctgtaaatca ggatggattt 60ataaatgaat ggccagaaaa aggcttcata gcaatgtgta gtcccaatga tcctaagcca 120tcaataaaga ttgaaaacga caagatcgtt gagatggatg ggaagagaag agaagacttt 180gattttatag atttattcat agctgatcac gctataaata tttatcaggc tgagaaatcc 240atgaaaatga actcgcttga tatagccaaa atgcttgtag atataaatgt agagagaaag 300actataataa aagtagtttc gggacttaca cctgccaaaa taatggaagt tgtaaatcat 360cttaatgtcg ttgaaatgat gatggctatg cagaaaatgc gagcaagaaa gattccggct 420aatcaatcac atattacaaa tcttaaagat aatcctgtgc agattgcagc ggatgctgcc 480gaatgtgctt taagaggttt tagggaagaa gagaccaccg taggagtgac aaaatatgct 540ccgtttaatg caatagcgtt attgataggg tctcaggcat taaaaagagg cgtgcttact 600caatgtgctg ttgaggaggc gacggaactt gaattaggca tgaggggatt taccacatac 660gctgagacta tatctgttta tggaactgaa agtgttttta tagatggtga cgatacacct 720tactccaaag cattccttgc ttctgcttat gcgtcaagag gattgaaaat gaggtttacg 780tcaggtacag gttcagaagt tcttatggga aatgcagagg gtaaatcgat gttgtacctg 840gaaatcaggt gcatcatggt tacaaaaggt gcaggagtgc aggggcttca aaatggtgca 900ataagctgta taggcataac tagctcagtt ccttcaggta taagggcggt gctggctgaa 960aaccttatag catctatgct tgatttagag gtagcatcag gcaatgatca gacttttaca 1020cattcagaca taagaaggac agcaaggact atgatgcagt ttttacccgg tactgatttc 1080atattttcag gttacagtgg aacgcctaat tatgacaata tgtttgcagg ttccaatttt 1140gatgcagaag attttgatga ctacaatgta ctgcaaaggg atttaatggt agatggaggg 1200ttaaggcctg taaaagaaga agatgtggta gaagtgaggc gaaaggcagc taaagctttg 1260caggatgtat ttagagagtt aaatcttgga gtagttacag atgaagaagt agaagcagca 1320gcatatgcac acggcagcaa agatatgcct gaaagagatg ttttgtctga ccttgaatca 1380atcgatgaga tgatgaaaag agggattaca ggcattgaca

tcgtaaaggc tttatataga 1440tctggacatg aggatatagc ggaaaacatt ttaaacatgt taaaacagcg catatctgga 1500gactatttgc agacatcagc tattcttgat gaagatttta atgttataag cgccataaat 1560tgtccaaatg attacttagg acctggaaca ggatatagga ttgataaaga tagatgggaa 1620gagataaaga atattcctta caccattaat cctgacaatt tgtaa 166568208PRTT. saccharolyticum 68Met Tyr Val Asp Glu Glu Leu Leu Lys Glu Ile Thr Lys Arg Val Ile 1 5 10 15 Glu Glu Leu Asn Asn Lys His Lys Thr Asp Asn Val Pro Ser Tyr Phe 20 25 30 Ile Glu Asn Gly Val Ala Tyr Lys Gly Lys Asn Ile Glu Glu Val Val 35 40 45 Ile Gly Val Gly Pro Ala Phe Gly Lys His Ile Lys Lys Thr Ile Asn 50 55 60 Gly Leu Asp His Arg Asp Val Ile Lys Glu Ile Ile Ala Gly Ile Glu 65 70 75 80 Glu Glu Gly Met Val His Arg Ile Val Arg Val Leu Lys Thr Ser Asp 85 90 95 Val Ala Phe Ile Gly Lys Glu Ala Ala Leu Leu Ser Gly Ser Gly Ile 100 105 110 Gly Ile Gly Ile Gln Ser Lys Gly Thr Thr Val Ile His Gln Lys Asp 115 120 125 Leu Tyr Pro Leu Ser Asn Leu Glu Leu Phe Pro Gln Ala Pro Leu Leu 130 135 140 Asn Leu Glu Leu Tyr Arg Glu Ile Gly Lys Asn Ala Ala Arg Tyr Ala 145 150 155 160 Lys Gly Met Met Val Lys Pro Ile Leu Ile Gln Asn Asp Tyr Met Val 165 170 175 Arg Pro Lys Tyr Gln Val Lys Ala Ala Ile Met His Ile Lys Glu Thr 180 185 190 Glu Lys Ile Leu Lys Asn Ala Gln Ser Ile Gln Leu Thr Ile Asp Leu 195 200 205 69627DNAT. saccharolyticum 69atgtacgtag atgaagaact gttaaaagaa attactaaac gtgttataga agaattaaat 60aataagcata aaactgataa tgtgccttcg tattttattg aaaatggagt tgcctataag 120ggtaaaaata tagaggaagt cgtcattggt gttgggcctg catttggaaa gcatataaaa 180aagactataa atggccttga ccatagagat gtcataaaag aaataattgc aggcatcgaa 240gaagaaggta tggttcatag aattgtaaga gttctaaaga cttctgatgt ggcgttcata 300ggcaaagaag ctgctttatt aagcggatcg ggaataggca taggcataca atcaaaaggt 360actacagtga ttcatcaaaa agatttatat cctttaagca atttagaact gtttccacaa 420gctccactgc taaatttaga attatacagg gaaataggca aaaatgcggc gagatatgct 480aaaggcatga tggtaaagcc tattttgatt caaaatgatt acatggtgag acctaaatac 540caagtgaaag ctgctataat gcatataaaa gagacggaaa agatattgaa aaatgctcaa 600tcaatccaat tgacgataga cttgtaa 62770132PRTT. saccharolyticum 70Met Glu Glu Tyr Pro Leu Ser Lys Ser Ala Phe Asp Lys Leu Val Thr 1 5 10 15 Lys Thr Gly Lys His Leu Asn Glu Ile Asn Ile Glu Asn Val Met Lys 20 25 30 Gly Asn Val Lys Pro Asp Asp Ile Lys Ile Ser Lys Glu Val Leu Leu 35 40 45 Met Gln Gly Gln Ile Ala Glu Arg Tyr Gly Arg His Gln Met Lys Glu 50 55 60 Asn Phe Thr Arg Ala Ser Glu Leu Thr Asp Val Pro Asp Glu Lys Ile 65 70 75 80 Leu Glu Ile Tyr Glu Ser Leu Arg Pro Phe Arg Ser Thr Lys Glu Glu 85 90 95 Leu Ile Asn Leu Ala Tyr Glu Leu Arg Asp Lys Tyr Asn Ala Ile Asn 100 105 110 Cys Ala Asn Leu Ile Leu Glu Ala Ala Glu Val Tyr Glu Lys Arg Asn 115 120 125 Ile Leu Lys Thr 130 71399DNAT. saccharolyticum 71atggaagaat atccgctatc aaaaagtgct tttgataaat tggtgacaaa aacaggcaaa 60catttgaatg aaataaatat tgaaaatgta atgaagggaa acgtaaaacc cgatgatatc 120aagatatcca aagaagtgct tttaatgcaa gggcaaattg cagaaagata cggcaggcat 180cagatgaagg agaatttcac aagagcatcg gagcttacag atgttccaga tgaaaagatt 240ttggaaatat atgagagctt aaggccgttt agatctacaa aggaagagct tataaatctt 300gcctatgaat taagagataa gtacaatgcc attaactgtg caaacttgat acttgaggct 360gctgaagtat atgaaaaaag aaatattttg aaaacttaa 39972605PRTT. saccharolyticum 72Met Lys Leu Ile Ala Gly Val Asp Ile Gly Asn Ser Thr Thr Glu Val 1 5 10 15 Cys Ile Ala Ala Ile Lys Asp Asp Asn Thr Leu Glu Phe Leu Ser Ser 20 25 30 Ser Leu Thr Ala Thr Thr Gly Val Lys Gly Thr Val Asp Asn Val Thr 35 40 45 Gly Val Ile Asn Gly Leu Thr Glu Ala Leu Lys Lys Ile Gly Lys Asn 50 55 60 Ile Arg Asp Leu Ser Leu Ile Arg Ile Asn Glu Ala Ala Pro Val Val 65 70 75 80 Cys Gly Ala Ala Met Glu Thr Ile Thr Glu Thr Val Ile Thr Gly Ser 85 90 95 Thr Met Ile Gly His Asn Pro Ser Thr Pro Gly Gly Val Gly Leu Gly 100 105 110 Val Gly Glu Ile Ile His Ile Asn Asp Leu Ala Asp Ala Thr Lys Gly 115 120 125 Lys Asn Tyr Ile Val Val Ile Pro Lys Glu Ile Gly Tyr Glu Glu Ala 130 135 140 Ser Ile Met Ile Asn Lys Ser Phe Glu Asn Asp Ile Asp Val Lys Ala 145 150 155 160 Ala Ile Val Gln Ser Asp Glu Ala Val Leu Ile Asn Asn Arg Leu Lys 165 170 175 Lys Ile Ile Pro Ile Val Asp Glu Val Arg Gln Ile Glu Lys Ile Pro 180 185 190 Ser Gly Val Val Ala Ala Val Glu Val Ala Pro Glu Gly Lys Ser Ile 195 200 205 Ser Thr Leu Ser Asn Pro Tyr Gly Ile Ala Thr Ile Phe Asp Leu Thr 210 215 220 Pro Glu Glu Thr Lys Tyr Val Ile Pro Ile Ser Lys Ser Leu Met Gly 225 230 235 240 Lys Lys Ser Ala Val Val Ile Lys Thr Pro Arg Gly Gln Val Lys Glu 245 250 255 Arg Ile Ile Pro Ala Gly Asn Leu Leu Ile Met Gly Pro Thr Met Ser 260 265 270 Ser Lys Val Ser Val Asp Ser Gly Ala Glu Ala Ile Met Glu Ser Val 275 280 285 Glu Glu Val Gly Thr Ile Asp Asp Val Glu Gly Glu Glu Asn Thr Asn 290 295 300 Val Gly Asn Met Ile Lys Asn Leu Lys Asn Lys Met Ala Asn Ile Thr 305 310 315 320 Gly Gln Lys Val Asp Lys Ile Lys Ile Lys Asp Ile Phe Ala Val Asp 325 330 335 Thr Thr Val Pro Val Lys Val Glu Gly Gly Leu Ala Gly Glu Thr Ser 340 345 350 Met Glu Lys Ala Val Val Leu Ala Ala Met Val Lys Thr Asp Thr Leu 355 360 365 Pro Met Ile Glu Ile Ala Glu Lys Leu Gln Arg Lys Leu Gly Val Phe 370 375 380 Val Lys Ile Ala Gly Val Glu Ala Val Met Ala Thr Leu Gly Ala Leu 385 390 395 400 Thr Thr Pro Gly Thr Lys Leu Pro Leu Ala Ile Leu Asp Ile Gly Gly 405 410 415 Gly Ser Thr Asp Ala Ala Leu Ile Asp Glu Lys Gly Ile Val Lys Ser 420 425 430 Ile His Met Ala Gly Ala Gly Glu Leu Val Thr Met Leu Ile Asp Ser 435 440 445 Glu Leu Gly Leu Asn Asp Arg Tyr Leu Ser Glu Glu Ile Lys Arg Asn 450 455 460 Pro Ile Gly Lys Val Glu Ser Leu Phe His Ile Arg Met Glu Asn Arg 465 470 475 480 Glu Ile Lys Phe Phe Asp Lys Pro Leu Asn Pro Arg Tyr Tyr Gly Arg 485 490 495 Ile Val Ile Leu Lys Glu Asn Asp Met Ile Pro Val Phe Lys Glu Asp 500 505 510 Leu Thr Met Glu Lys Ile Ile Tyr Val Arg Arg Gln Ala Lys Asp Lys 515 520 525 Val Phe Val Lys Asn Ala Ile Arg Ala Leu Lys Lys Ile Ala Pro Glu 530 535 540 Asn Asn Leu Arg Arg Ile Pro Asn Val Val Leu Val Gly Gly Ser Ala 545 550 555 560 Leu Asp Phe Glu Ile Pro Glu Met Ile Leu Ser Glu Leu Ser Lys Tyr 565 570 575 Lys Ile Ile Ala Gly Arg Gly Asn Ile Arg Lys Ile Glu Gly Pro Arg 580 585 590 Asn Ala Val Ala Thr Gly Leu Val Met Ser Tyr Leu Gly 595 600 605 731817DNAT. saccharolyticum 73atgaaactca tagcaggtgt tgatattggc aattctacaa cagaagtgtg tatagccgct 60attaaagatg acaatacatt agaattttta agcagttcct tgacagctac gacaggtgta 120aaaggcactg tggataatgt gacaggggtt attaatggat tgactgaggc actaaaaaaa 180attggcaaga atattaggga tttaagcctc attagaatca atgaagccgc cccagttgtc 240tgtggtgctg ctatggagac aataacggaa actgttatca ctggttcgac tatgataggt 300cataatccat ccacgccggg tggtgtcgga cttggagtag gcgagataat acatataaat 360gatttagctg atgctactaa aggcaaaaat tacattgtgg ttatacctaa ggagattggc 420tatgaagaag cttcaataat gataaacaaa tcttttgaaa acgatattga tgtaaaagct 480gctatagttc aaagcgatga agcagtttta atcaacaaca ggcttaaaaa gattatacca 540attgttgacg aagtaaggca gatagaaaag attccatcgg gtgttgtagc ggctgtagag 600gtggcaccag aaggcaagtc cataagcacg ttatcaaatc cttatggtat cgcaacaata 660tttgacttaa ctccagaaga gacaaagtat gtcataccga tttcgaaaag tttgatgggg 720aaaaagtcag cagttgtcat aaaaacaccg aggggacaag tgaaagaaag aataattccg 780gctggtaatc tcttaatcat ggggcctact atgtcatcaa aagtaagtgt tgattctggt 840gctgaagcta taatggaatc agttgaagaa gtcggcacaa ttgatgacgt agaaggtgaa 900gaaaatacaa atgttgggaa tatgataaaa aatctaaaaa acaagatggc aaatataact 960gggcaaaaag tagataagat aaagattaaa gatatcttcg ctgttgatac gacagtccct 1020gttaaagtag agggcggact tgctggtgag acttcaatgg aaaaagcagt cgtgttggcg 1080gctatggtaa agacagatac gcttcgatga tagaaattgc agaaaagctt caaagaaagt 1140tgggtgtatt tgtaaaaata gctggagtag aagctgtgat ggctacatta ggtgcgctta 1200caactccagg cacaaagttg ccacttgcaa tactggatat cggtgggggt tctacagatg 1260cagctttgat tgatgaaaaa ggcattgtaa aatctataca catggcaggt gctggagaat 1320tagtcacaat gcttattgat tcagaattag ggttaaatga tagatatttg tctgaagaaa 1380taaagagaaa tccgattgga aaagttgaaa gcctatttca cataagaatg gaaaataggg 1440agataaagtt ttttgacaaa cctttaaatc ctcgatatta cggtaggatc gtaattttaa 1500aagaaaatga catgatccct gtatttaaag aagatttgac aatggaaaag attatttacg 1560tgcgaagaca agcgaaggat aaagttttcg ttaaaaatgc tattagagct ttgaaaaaaa 1620ttgctccgga aaataattta aggcgaatac caaatgtagt cttggttggc ggttctgctt 1680tggactttga aattccagag atgattttat cagagctatc aaaatacaaa atcatagcag 1740gcagagggaa tataagaaaa atcgaagggc caagaaatgc tgtagcgaca ggtcttgtga 1800tgtcttattt agggtga 181774112PRTT. saccharolyticum 74Met Glu Phe Ile Lys Pro Gln Ile Val Ile Phe Ala Asn Thr Glu Asn 1 5 10 15 Lys Tyr Ile Ile Asn Glu Val Ile Ala Gly Ile Glu Glu Glu Gly Ala 20 25 30 Leu Tyr Arg Leu Ser Tyr Asn Glu Cys Ala Asp Val Met Lys Met Ala 35 40 45 Tyr Asp Ala Ala Lys Ala Ser Val Leu Gly Ile Gly Ile Gly Ile Ser 50 55 60 Gly Asp Leu Val Cys Leu His Ser Lys Asn Leu Glu Ile Asn Thr Pro 65 70 75 80 Leu Ile Leu Ser Lys Thr Ser Glu Asn Phe Asp Pro Arg Leu Val Gly 85 90 95 Cys Asn Ala Ala Lys Tyr Val Lys Gly Leu Pro Leu Lys Tyr Leu Asp 100 105 110 75339DNAT. saccharolyticum 75atggaattta taaagcctca aatagtgatt tttgcaaata cagaaaacaa atatataata 60aacgaggtta tagctggcat tgaagaagaa ggtgcattat atagattatc ttacaatgaa 120tgtgctgatg ttatgaaaat ggcttatgat gcagcaaaag catctgtatt aggtatcgga 180ataggcatat ctggagattt agtgtgtttg cactctaaaa acttggaaat caatacacct 240ttgattcttt caaagacaag tgaaaacttt gatccacgac tcgttggatg caatgctgca 300aaatatgtaa agggtttgcc acttaaatac ttagattag 3397656PRTT. saccharolyticum 76Met Ser Val Tyr Thr Lys Thr Gly Asp Asp Gly Tyr Thr Leu Leu Leu 1 5 10 15 Asn Gly Glu Arg Ile Pro Lys Asp Asp Leu Arg Ile Glu Thr Leu Gly 20 25 30 Asn Leu Asp Glu Leu Thr Ser Tyr Leu Gly Phe Ala Lys Ala Gln Ile 35 40 45 Asn Asp Asp Ser Ile Lys Lys Arg 50 55 77171DNAT. saccharolyticum 77atgagtgttt atactaaaac tggtgatgat ggttacacgt tgctattaaa tggagaaaga 60attccaaagg acgatttgag aatagagaca ttgggaaatt tggatgaatt gacaagctat 120ttaggatttg caaaagctca aataaatgat gattccataa aaaagagata g 17178225PRTT. saccharolyticum 78Met Val Lys Ile Lys Asn Gly Phe Val Ile Pro Gly Lys Asn Gln Ile 1 5 10 15 Ser Ala Leu Leu Asp Ile Val Arg Thr Ile Thr Arg Lys Thr Glu Arg 20 25 30 Ser Leu Ile Lys Val Asp Lys Lys Tyr Pro Val Asn Ile Asn Ser Lys 35 40 45 Val Tyr Ile Asn Arg Leu Ser Asp Tyr Leu Phe Val Leu Ala Arg Tyr 50 55 60 Met Glu Ile Arg Thr Glu Ile Glu Glu Lys Val Lys Asp Val Ile Arg 65 70 75 80 Lys His Tyr Gly Lys Asn Lys Gly Glu Ile Lys Leu Asn Leu Asp Ile 85 90 95 Ala Lys Asn Leu Met Ala Lys Val Glu Lys Lys Ala Glu Ser Ile Asn 100 105 110 Leu Pro Val Ala Ile Ala Ile Val Asp Met His Gly Asn Leu Ile Ala 115 120 125 Ala His Phe Met Asp Gly Thr Leu Leu Glu Ser Met Asn Leu Ala Ile 130 135 140 Asn Lys Ala Tyr Thr Ser Val Val Leu Lys Met Ser Thr Gln Glu Leu 145 150 155 160 Ser Lys Leu Ala Gln Pro Gly Gln Pro Leu Tyr Gly Ile Asn Thr Thr 165 170 175 Asp Asn Arg Ile Val Val Phe Gly Gly Gly Cys Pro Ile Lys His Gln 180 185 190 Gly Glu Ile Val Gly Gly Ile Gly Val Ser Gly Gly Thr Val Glu Gln 195 200 205 Asp Ile Glu Leu Ser Ile Tyr Gly Ala Asp Val Phe Glu Glu Val Ile 210 215 220 Ser 225 79678DNAT. saccharolyticum 79atggtaaaga ttaaaaatgg ttttgtaata cctggtaaaa accaaatctc agcattatta 60gatattgtaa ggactataac gagaaaaact gagagaagct taatcaaagt tgacaagaaa 120tatcctgtaa atattaattc gaaagtttac atcaatagat tgtctgatta tttgtttgtt 180ttagcaaggt atatggaaat aagaacggaa atagaagaaa aagtaaaaga cgtgataaga 240aagcattatg gaaagaacaa aggcgaaata aagctaaatt tagatatagc aaaaaattta 300atggctaagg tagaaaagaa ggcagaaagc attaatctac cggttgctat tgcaatagtt 360gacatgcatg gcaatttgat agcggctcat tttatggatg gtacacttct tgaaagcatg 420aatctagcta taaataaagc ttatacatca gtggtgctta aaatgtcgac gcaagagtta 480tcaaaacttg cacaaccagg gcagcctctt tacgggataa atacaactga taatagaatc 540gtagtgtttg gaggtgggtg ccctataaaa catcaaggtg aaatagttgg tggaattgga 600gttagcggtg gtacagtaga acaagatata gaactttcta tttatggtgc agatgtattt 660gaggaggtta tatcatga 67880467PRTT. saccharolyticum 80Met Lys Val Lys Glu Glu Asp Ile Glu Ala Ile Val Lys Lys Val Leu 1 5 10 15 Ser Glu Phe Asn Phe Glu Lys Asn Thr Lys Ser Phe Arg Asp Phe Gly 20 25 30 Val Phe Gln Asp Met Asn Asp Ala Ile Arg Ala Ala Lys Asp Ala Gln 35 40 45 Lys Lys Leu Arg Asn Met Ser Met Glu Ser Arg Glu Lys Ile Ile Gln 50 55 60 Asn Ile Arg Lys Lys Ile Met Glu Asn Lys Lys Ile Leu Ala Glu Met 65 70 75 80 Gly Val Ser Glu Thr Gly Met Gly Lys Val Glu His Lys Ile Ile Lys 85 90 95 His Glu Leu Val Ala Leu Lys Thr Pro Gly Thr Glu Asp Ile Val Thr 100 105 110 Thr Ala Trp Ser Gly Asp Lys Gly Leu Thr Leu Val Glu Met Gly Pro 115 120 125 Phe Gly Val Ile Gly Thr Ile Thr Pro Ser Thr Asn Pro Ser Glu Thr 130 135 140 Val Leu Cys Asn Ser Ile Gly Met Ile Ala Ala Gly Asn Ser Val Val 145 150 155 160 Phe Asn Pro His Pro Gly Ala Val Asn Val Ser Asn Tyr Ala Val Lys 165 170 175 Leu Val Asn Glu Ala Val Met Glu Ala Gly Gly Pro Glu Asn Leu Val 180 185 190 Ala Ser Val Glu Lys Pro Thr Leu Glu Thr Gly Asn Ile Met Phe Lys 195 200 205 Ser Pro Asp Val Ser Leu Leu Val Ala Thr Gly Gly Pro Gly Val Val 210 215 220 Thr Ser Val Leu Ser Ser Gly Lys Arg Ala Ile Gly Ala Gly Ala Gly 225

230 235 240 Asn Pro Pro Val Val Val Asp Glu Thr Ala Asp Ile Lys Lys Ala Ala 245 250 255 Lys Asp Ile Val Asp Gly Ala Thr Phe Asp Asn Asn Leu Pro Cys Ile 260 265 270 Ala Glu Lys Glu Val Val Ser Val Asp Lys Ile Thr Asp Glu Leu Ile 275 280 285 Tyr Tyr Met Gln Gln Asn Gly Cys Tyr Lys Ile Glu Gly Arg Glu Ile 290 295 300 Glu Lys Leu Ile Glu Leu Val Leu Asp His Lys Gly Gly Lys Ile Thr 305 310 315 320 Leu Asn Arg Lys Trp Val Gly Lys Asp Ala His Leu Ile Leu Lys Ala 325 330 335 Ile Gly Ile Asp Ala Asp Glu Ser Val Arg Cys Ile Ile Phe Glu Ala 340 345 350 Glu Lys Asp Asn Pro Leu Val Val Glu Glu Leu Met Met Pro Ile Leu 355 360 365 Gly Ile Val Arg Ala Lys Asn Val Asp Glu Ala Ile Met Ile Ala Thr 370 375 380 Glu Leu Glu His Gly Asn Arg His Ser Ala His Met His Ser Lys Asn 385 390 395 400 Val Asp Asn Leu Thr Lys Phe Gly Lys Ile Ile Asp Thr Ala Ile Phe 405 410 415 Val Lys Asn Ala Pro Ser Tyr Ala Ala Leu Gly Tyr Gly Gly Glu Gly 420 425 430 Tyr Cys Thr Phe Thr Ile Ala Ser Arg Thr Gly Glu Gly Leu Thr Ser 435 440 445 Ala Arg Thr Phe Thr Lys Ser Arg Arg Cys Val Leu Ala Asp Gly Leu 450 455 460 Ser Ile Arg 465 811404DNAT. saccharolyticum 81atgaaagtta aagaggaaga tattgaagcg atcgtcaaaa aagtcttatc ggaatttaat 60tttgaaaaaa atactaaaag tttcagagat tttggcgtat ttcaagatat gaatgatgct 120attcgtgctg caaaagatgc ccagaaaaaa ttgagaaata tgtccatgga gtcgagagaa 180aagattatac agaatataag aaaaaagatt atggagaata aaaaaatact tgcagagatg 240ggcgtcagtg aaactggcat ggggaaagta gagcacaaaa taataaaaca tgagcttgta 300gcacttaaga cacctggtac cgaagatata gtgacaacag catggtctgg cgataaggga 360ctgacattgg ttgaaatggg gccatttggt gtaataggta cgattactcc ttcgacaaat 420ccaagtgaaa ccgtcctttg caatagcata ggtatgatag ccgcaggtaa ttcagtcgta 480tttaatccac atccaggtgc ggtaaatgta tctaattacg ctgtcaagtt agtaaatgaa 540gcggtgatgg aagctggcgg ccctgagaat ttagtcgcat ctgttgaaaa acctacactt 600gaaactggaa atattatgtt caagagtcct gatgtttcgc tattagtagc gacaggcgga 660cctggtgtag taacatcggt tctctcatct ggcaaaaggg caataggagc aggagcagga 720aatccaccag ttgtagttga tgaaacggca gatataaaaa aagctgcgaa agatatagtc 780gatggtgcta catttgacaa caatttgcct tgtattgctg aaaaggaagt agtttctgta 840gataaaataa cagatgaact gatttactac atgcaacaga atggctgcta caagattgag 900gggcgagaaa ttgaaaagct cattgaactt gtattggatc acaaaggtgg caagataaca 960ttaaacagga aatgggttgg caaagatgct catttaatac taaaagctat aggcatagat 1020gctgatgaaa gcgtaaggtg cataattttt gaggcggaaa aagacaatcc gttagtggta 1080gaagagctga tgatgcctat tttaggaata gtaagagcca aaaatgtaga tgaagcgata 1140atgattgcga cagagttaga acatggcaat aggcattcag cacatatgca ttctaaaaac 1200gttgataatt taacaaagtt tggaaaaata attgacactg ctatatttgt aaaaaatgct 1260ccatcgtatg ccgcgttagg atatggtggt gaaggttatt gcacatttac gattgcaagc 1320agaacaggtg aaggattgac atctgcaagg acttttacta aaagtcgtag atgtgtcttg 1380gcagatggat tatcaataag atag 140482403PRTT. saccharolyticum 82Met Glu Val Asn Gln Ile Asp Ile Glu Glu Ile Val Lys Lys Ile Leu 1 5 10 15 Asn Asp Leu Arg Asn Glu Pro Lys Glu Asn Ile Lys Glu Ser Asn Ser 20 25 30 Lys Ile Pro Ser Ile Cys Arg Ala Ala Val Leu Thr Asp Val Lys Lys 35 40 45 Ile Glu Val Lys Glu Phe Asn Ile Pro Glu Ile Asn Asp Asp Glu Met 50 55 60 Leu Val Lys Val Glu Gly Cys Gly Val Cys Gly Thr Asp Val His Glu 65 70 75 80 Tyr Lys Gly Asp Pro Phe Gly Leu Ile Pro Leu Val Leu Gly His Glu 85 90 95 Gly Thr Gly Glu Ile Val Lys Leu Gly Lys Asn Val Arg Arg Asp Ser 100 105 110 Ala Gly Lys Glu Ile Lys Glu Gly Asp Lys Ile Val Thr Ser Val Val 115 120 125 Pro Cys Gly Glu Cys Asp Ile Cys Leu Asn His Pro Asp Lys Thr Asn 130 135 140 Leu Cys Glu Asn Ser Lys Ile Tyr Gly Leu Ile Ser Asp Asp Asn Tyr 145 150 155 160 His Leu Asn Gly Trp Phe Ser Glu Tyr Ile Val Ile Arg Lys Gly Ser 165 170 175 Thr Phe Tyr Lys Val Asn Asp Ile Asn Leu Asn Leu Arg Leu Leu Val 180 185 190 Glu Pro Ala Ala Val Val Val His Ala Val Glu Arg Ala Lys Ser Thr 195 200 205 Gly Leu Met Lys Phe Asn Ser Lys Val Leu Val Gln Gly Cys Gly Pro 210 215 220 Ile Gly Leu Leu Leu Leu Ser Val Val Lys Thr Leu Gly Val Glu Asn 225 230 235 240 Ile Ile Ala Val Asp Gly Asp Glu Asn Arg Leu Asn Met Ala Lys Arg 245 250 255 Leu Gly Ala Thr Ala Leu Ile Asn Phe Thr Lys Tyr Ser Asn Ile Asp 260 265 270 Glu Leu Val Asp Ala Val Lys Lys Ala Ser Asp Gly Ile Gly Ala Asp 275 280 285 Phe Ala Phe Gln Cys Thr Gly Val Pro Ser Ala Ala Ser Asn Ile Trp 290 295 300 Lys Phe Val Arg Arg Gly Gly Gly Leu Cys Glu Val Gly Phe Phe Val 305 310 315 320 Asn Asn Gly Asp Cys Lys Ile Asn Pro His Tyr Asp Ile Cys Asn Lys 325 330 335 Glu Ile Thr Ala Val Gly Ser Trp Thr Tyr Thr Pro Gln Asp Tyr Leu 340 345 350 Thr Thr Phe Asp Phe Leu Lys Arg Ala Lys Glu Ile Gly Leu Pro Ile 355 360 365 Glu Glu Leu Ile Thr His Arg Phe Ser Leu Asp Lys Met Asn Glu Ala 370 375 380 Met Glu Val Asn Met Lys Gln Glu Gly Ile Lys Val Val Tyr Ile Asn 385 390 395 400 Asp Arg Phe 831212DNAT. saccharolyticum 83atggaagtca atcagataga cattgaggag atagttaaga aaatattaaa tgatttaaga 60aatgagccta aagaaaacat taaagagagc aattcaaaaa taccatctat ctgcagagct 120gctgtactta cagatgttaa aaaaatagaa gtaaaagaat ttaatattcc agaaataaat 180gatgatgaaa tgcttgtcaa ggtggaaggc tgtggcgttt gcggtactga tgttcatgaa 240tacaaaggag atccttttgg acttatacca ttggttttag gacacgaagg tacaggtgag 300atagtcaagc tggggaaaaa cgtgagacga gattctgctg gtaaagaaat caaagaaggc 360gataagattg ttacatctgt cgttccgtgc ggtgaatgcg atatatgttt gaatcatcca 420gacaagacaa atttgtgtga aaactcaaag atttacggct taatatccga tgataattac 480catttaaatg gttggttctc agagtacatc gtcataagga aaggctcaac attttataag 540gtcaatgata taaaccttaa tttgaggctt ttggtagaac cggctgcagt agtcgtacat 600gcagtagagc gcgcaaaatc cacaggtctt atgaaattca acagtaaagt tctcgtacaa 660ggctgtggcc ctataggatt actgctattg tcggttgtaa agacgcttgg agtagaaaat 720atcatagccg tcgacggcga tgagaataga ctcaacatgg ctaaaagatt aggtgctaca 780gcactcatta attttactaa atacagcaat attgatgagc ttgttgatgc tgttaaaaaa 840gcaagcgatg gaattggcgc agattttgca tttcaatgta caggcgttcc ttctgcagcg 900tctaatattt ggaagtttgt aaggcgggga ggtggtttat gcgaagttgg attttttgta 960aataatggtg attgtaagat aaacccccat tatgatattt gcaataagga gataacagca 1020gttggctcat ggacttacac tcctcaagac tatttgacaa cttttgattt tctcaaaaga 1080gctaaagaaa taggacttcc aattgaagag ctgataacac atagattttc acttgataaa 1140atgaatgaag ctatggaagt taatatgaag caggaaggga taaaagtagt gtatataaat 1200gacagatttt ag 121284101PRTT. saccharolyticum 84Met Gln Ala Val Gly Leu Ile Glu Val Tyr Gly Leu Val Ala Ala Phe 1 5 10 15 Val Ala Ala Asp Ala Ala Cys Lys Lys Ala Asn Val Val Ile Glu Ser 20 25 30 Phe Asp Asn Asn Lys Pro Leu Asn Ala Glu Ala Leu Pro Val Pro Leu 35 40 45 Ile Ile Val Val Lys Leu Arg Gly Asp Leu Glu Asp Val Lys Ile Ala 50 55 60 Val Asp Ala Ala Val Asp Ala Ala Asn Lys Ile Ser Gly Val Val Ala 65 70 75 80 Thr Asn Ile Ile Ala Lys Pro Glu Glu Asp Thr Glu Lys Leu Leu Lys 85 90 95 Leu Asn Cys Leu Lys 100 85306DNAT. saccharolyticum 85atgcaggctg ttggattgat tgaagtttat ggattagtag cggcatttgt ggcagcagat 60gctgcatgca aaaaagcgaa tgtcgtaata gagtcttttg acaacaataa gccattaaat 120gctgaagcat tgccagttcc attgataata gtcgttaagc tcagaggaga tcttgaggat 180gtaaaaatag cggtagatgc tgcagttgat gcagctaata aaatatctgg tgtagttgct 240acaaatataa tagcaaaacc agaagaagat actgaaaagc tattaaagct aaattgtctt 300aaataa 3068692PRTT. saccharolyticum 86Met Val Gln Glu Ala Leu Gly Met Val Glu Thr Arg Gly Leu Val Ala 1 5 10 15 Ala Ile Glu Ala Ala Asp Ala Met Val Lys Ala Ala Asp Val Thr Leu 20 25 30 Ile Gly Thr Glu Lys Ile Gly Ser Gly Leu Val Thr Val Met Val Arg 35 40 45 Gly Asp Val Gly Ala Val Lys Ala Ala Thr Glu Val Gly Ala Ser Ala 50 55 60 Ala Ser Lys Leu Gly Glu Leu Val Ala Val His Val Ile Pro Arg Pro 65 70 75 80 His Thr Asp Val Glu Lys Ile Leu Pro Thr Ile Lys 85 90 87279DNAT. saccharolyticum 87atggtacaag aagcattggg aatggtagaa acgagaggat tggtagcagc aatagaagca 60gcagatgcta tggtaaaggc tgcggatgtc actttgatag gaactgaaaa aataggttca 120ggacttgtaa cagtcatggt aagaggagat gtcggtgcag taaaagcagc gacagaagtt 180ggcgcaagtg cagcttcaaa attgggagag ttagtggctg ttcacgtaat accaaggcct 240catactgatg ttgaaaagat actgccgaca attaaataa 27988105PRTT. saccharolyticum 88Met Tyr Ala Ile Gly Leu Ile Glu Val Asn Gly Phe Val Thr Ala Val 1 5 10 15 Glu Thr Leu Asp Ala Met Leu Lys Thr Ala Asn Val Glu Phe Val Thr 20 25 30 Trp Glu Lys Lys Leu Gly Gly Arg Leu Val Thr Ile Ile Ile Lys Gly 35 40 45 Asp Val Ser Ala Val Glu Glu Ala Ile Leu Thr Gly Lys Ile Glu Ala 50 55 60 Asp Lys Ile Thr Arg Thr Val Ala Tyr Ala Val Ile Pro Asn Pro His 65 70 75 80 Pro Glu Thr Ile Lys Met Val Asn Ile Ser Ala Gly Lys Leu Phe Lys 85 90 95 Ala Asp Gly Gly Glu Ile Asn Glu Phe 100 105 89318DNAT. saccharolyticum 89atgtatgcaa ttggacttat tgaagtaaat gggtttgtca cagcggttga aacactggat 60gcaatgttga aaacagccaa tgtagagttt gtaacatggg agaaaaaact tggaggcaga 120cttgtgacaa tcattattaa aggagatgtt tcagcagttg aagaagcaat tttaactgga 180aagattgaag ctgacaagat tacacggaca gtagcatacg cagttattcc aaatccacat 240ccagaaacta taaagatggt aaatattagt gcaggaaagc tatttaaagc agatggtggt 300gaaataaatg agttctga 31890127PRTT. saccharolyticum 90Met Ser Ser Glu Glu Lys Asp Thr Asn Ala Lys Asp Val Lys Val Glu 1 5 10 15 Lys Gln Lys Asn Asn Leu Thr Lys Thr Ser Asn Lys Glu Phe Lys Glu 20 25 30 Glu Leu Ile Met Glu Gln Gln Ala Leu Gly Met Val Glu Thr Arg Gly 35 40 45 Leu Val Ala Ala Ile Glu Ala Ala Asp Ala Met Val Lys Ala Ala Asn 50 55 60 Val Thr Leu Ile Gly Thr Glu Lys Ile Gly Ser Gly Leu Val Thr Val 65 70 75 80 Met Val Arg Gly Asp Val Gly Ala Val Lys Ala Ala Thr Glu Thr Gly 85 90 95 Ala Asn Ala Ala Lys Lys Leu Gly Glu Leu Val Ala Val His Val Ile 100 105 110 Pro Arg Pro His Ala Asp Val Glu Lys Ile Leu Pro Thr Ile Lys 115 120 125 91384DNAT. saccharolyticum 91atgagttctg aagaaaagga tacgaatgca aaagatgtta aagtcgaaaa gcagaaaaat 60aatttaacga aaacatcaaa taaagaattt aaggaggaat tgattatgga acaacaagca 120ttaggaatgg tagagacgag aggattggta gcagcgatag aagctgctga tgcaatggta 180aaggctgcta atgtcacgtt aataggaact gaaaaaatag gttcaggact tgtaacagtc 240atggtaagag gagatgttgg tgcagtaaaa gcagcgacag agactggagc aaatgcagct 300aaaaagttag gggagttagt agctgttcac gtaataccaa gacctcatgc agatgtagag 360aaaatactgc ctacgataaa gtag 38492217PRTT. saccharolyticum 92Val Ile Thr Val Asn Glu Lys Leu Ile Glu Ile Ile Ser Lys Thr Ile 1 5 10 15 Ala Asp Thr Ile Ser Glu Arg Asn Ser Leu Lys Ile Pro Val Gly Val 20 25 30 Ser Ala Arg His Val His Leu Thr Lys Glu His Leu Asp Ile Leu Phe 35 40 45 Gly Lys Asp Tyr Ile Leu Lys Lys Lys Lys Glu Leu Met Gly Gly Gln 50 55 60 Phe Ala Ala Glu Glu Cys Val Thr Ile Ile Gly Phe Lys Leu Asn Ala 65 70 75 80 Ile Glu Lys Val Arg Val Leu Gly Pro Leu Arg Asp Lys Thr Gln Val 85 90 95 Glu Ile Ser Lys Thr Asp Ala Ile Ser Leu Gly Leu Asn Pro Pro Ile 100 105 110 Arg Glu Ser Gly Asp Ile Lys Gly Ser Ser Pro Ile Thr Ile Val Gly 115 120 125 Pro Arg Gly Ala Ile Ser Leu Lys Glu Gly Cys Ile Ile Ala Lys Arg 130 135 140 His Ile His Met Ser Pro Glu Asp Ser Lys Arg Phe Asn Val Lys Asp 145 150 155 160 Asp Asp Ile Ile Ser Val Lys Ile Asn Gly Gln Arg Gly Gly Ile Leu 165 170 175 Glu Asn Val Gln Ile Arg Val Asp Glu Lys Tyr Thr Leu Glu Met His 180 185 190 Ile Asp Thr Asp Glu Ala Asn Cys Met Gly Leu Lys Ser Gly Asp Phe 195 200 205 Val Glu Ile Val Arg Asp Asn Arg Ser 210 215 93654DNAT. saccharolyticum 93gtgataacag tgaacgaaaa attgatagag attatatcaa aaactatagc ggatacgatt 60agtgaaagga attcgcttaa gataccagta ggcgtatcag cccgacatgt acatctgact 120aaagaacatt tggatatatt atttggaaaa gattatatcc ttaaaaagaa aaaggaattg 180atgggtggac agttcgcagc agaggaatgt gtgacaatta tcggatttaa attaaatgct 240attgagaaag tgagagtttt gggtccttta agagataaaa cgcaggtaga aatatcgaag 300accgatgcaa taagtttagg gttaaaccct cctatacggg aatcaggtga tataaaaggt 360tcatcgccaa ttacaattgt agggccgaga ggagcaatat cattaaaaga aggatgtata 420atagcaaaac gacatattca catgtcaccg gaagattcca aaagattcaa tgttaaagac 480gacgatataa tatcagtaaa aataaatggt cagcgaggcg gaattttaga aaatgtacag 540attagagttg acgaaaagta tacacttgag atgcatattg acacagatga agctaattgc 600atgggactaa aaagcggcga ttttgttgaa atagtaagag ataataggag ttga 6549492PRTT. saccharolyticum 94Leu Ile Ile Ala Lys Val Val Gly Thr Val Ile Ser Thr Arg Lys Asn 1 5 10 15 Gln Asn Leu Ile Gly Asn Lys Phe Leu Ile Val Glu Pro Val Ser Glu 20 25 30 Met Asn Tyr Asp Ser Lys Asn Arg Val Val Ala Ile Asp Asn Val Gly 35 40 45 Ala Gly Val Gly Glu Ile Val Leu Val Thr Phe Gly Ser Ser Ala Arg 50 55 60 Ile Gly Cys Gly Met Pro Asp Ser Pro Val Asp Ala Ala Ile Val Gly 65 70 75 80 Ile Val Asp Ser Ile Lys Asp Ile Ile Ile Asp Asp 85 90 95279DNAT. saccharolyticum 95ttgataatag ctaaagttgt tggtactgtt atttctaccc gcaagaatca aaatttaata 60ggcaataaat ttttaatagt agaaccagta agtgaaatga attatgacag taaaaatagg 120gttgttgcaa tagataatgt aggtgcaggt gtaggagaga tagtattagt tacctttgga 180agttcagcaa gaatcggttg tggtatgcca gattcgcctg tagatgcggc aattgtcgga 240attgttgata gcataaaaga tattatcatt gatgattag 27996456PRTT. saccharolyticum 96Met Met Asn Ile Asp Glu Leu Lys Asn Ile Val Phe Glu Asn Gly Ile 1 5 10 15 Val Gly Ala Gly Gly Ala Gly Phe Pro Thr His Ala Lys Leu Thr Thr 20 25 30 Gly Ile Asp Thr Ile Ile Leu Asn Gly Ala Glu Cys Glu Pro Leu Leu 35 40 45 Arg Val Asp Arg Gln Leu Leu Ala Ile Tyr Thr Asp Glu Ile Leu Met 50 55 60 Thr Leu Ser Phe Ile Val Asp Thr Leu Gly Ala Lys Arg Gly Ile Val 65 70 75 80 Ala Ile Lys Ser Ala Tyr Lys Thr Ala Ile Ser

Ser Val Lys Asn Leu 85 90 95 Ile Gly Asn Tyr Lys Asn Leu Glu Leu Lys Val Leu Pro Asp Val Tyr 100 105 110 Pro Ala Gly Asp Glu Val Val Leu Ile Tyr Glu Thr Thr Gly Arg Ile 115 120 125 Val Pro Glu Gly Ser Ile Pro Ile Ser Val Gly Thr Leu Val Met Asn 130 135 140 Val Glu Thr Val Leu Asn Val Tyr Asn Ala Ile Tyr Leu Lys His Pro 145 150 155 160 Val Thr Glu Lys Tyr Val Thr Val Thr Gly Asp Val Lys Tyr Pro Ser 165 170 175 Thr Phe Lys Ala Lys Val Gly Thr Ser Val Ala Arg Leu Ile Glu Lys 180 185 190 Ala Gly Gly Cys Leu Glu Lys Asp Cys Glu Val Ile Met Gly Gly Pro 195 200 205 Met Thr Gly Lys Ile Val Asp Val Lys Thr Pro Ile Thr Lys Thr Thr 210 215 220 Lys Ala Ile Ile Val Leu Pro Lys Asp His Pro Val Ile Thr Lys Arg 225 230 235 240 Lys Thr Asn Ile Arg Ile Gly Leu Lys Arg Ala Met Ser Val Cys Ser 245 250 255 Gln Cys Gln Met Cys Thr Asp Leu Cys Pro Arg Asn Leu Leu Gly His 260 265 270 Ser Ile Lys Pro His Lys Val Met Asn Ala Val Ala Asn Ser Ile Ile 275 280 285 Asp Asp Thr Ala Ala Tyr Thr Met Thr Met Leu Cys Ser Glu Cys Gly 290 295 300 Leu Cys Glu Met Tyr Ser Cys His Gln Ser Leu Ser Pro Arg Lys Ile 305 310 315 320 Ile Ser Gln Ile Lys Ile Lys Leu Arg Gln Asn Gly Val Lys Asn Pro 325 330 335 His Asn Lys Arg Pro Glu Thr Ala Asn Val Met Arg Asp Glu Arg Leu 340 345 350 Val Pro Met Glu Arg Leu Ile Ser Arg Leu Ser Leu Lys Lys Tyr Asp 355 360 365 Val Asp Ala Pro Met Asn Phe Asp Thr Val Ile Pro Ser His His Val 370 375 380 Val Met Gln Leu Ser Gln His Val Gly Ala Lys Ala Ile Pro Val Val 385 390 395 400 Lys Val Gly Asp Ile Val Lys Glu Gly Asp Leu Ile Gly Asp Val Pro 405 410 415 Asn Asn Lys Leu Gly Ala Lys Leu His Ala Ser Ile Asp Gly Ile Ile 420 425 430 Ile Asp Val Thr Asp Asp Ser Ile Val Ile Lys Pro Arg Gly Asp Phe 435 440 445 Asp Gly Gln Ser Asp Arg Ile Gly 450 455 971371DNAT. saccharolyticum 97atgatgaata ttgatgaact taaaaatatc gtatttgaaa atggaatagt cggtgcaggc 60ggagctggat ttcctacaca tgcaaaactt actacaggta tagatacaat catattaaat 120ggcgctgaat gtgaaccgct tttaagagta gataggcagc tacttgcaat atatactgat 180gaaatattga tgactttatc attcatagtt gatactttag gagccaaacg tggcattgta 240gcaataaaat cagcatacaa aactgccatc agctcagtta agaatttgat tggtaattat 300aaaaacttgg agttaaaggt attgccagac gtttatcctg ctggtgatga agttgtatta 360atatatgaaa cgactggaag aattgtgcca gaaggttcta tacctatttc tgttggcacg 420ttggtaatga atgtggaaac tgtgcttaat gtttataatg ctatttattt aaaacatcca 480gtcacagaaa agtatgtaac agtaacggga gatgtcaaat atcccagcac atttaaagca 540aaagtaggaa catctgtagc tcgtcttatt gaaaaagcag gaggatgctt agaaaaagat 600tgtgaagtga taatgggtgg tcctatgact gggaaaatag ttgatgtaaa gactccaata 660acaaaaacta caaaagctat tatcgttctc ccaaaagacc accctgtgat aacaaagaga 720aagacaaaca taaggatagg gttaaaacga gcaatgtctg tttgctctca atgccaaatg 780tgcacagatc tatgtcctag aaatttatta ggtcattcca tcaaacctca taaagtcatg 840aatgcagttg caaatagtat tattgatgat accgctgcat atacgatgac aatgttatgt 900tctgaatgtg gattgtgcga gatgtattca tgtcatcaaa gtttgtcgcc gagaaagata 960ataagccaga taaagataaa attaaggcaa aatggtgtaa aaaatccaca caacaaaaga 1020ccagaaacag caaatgtcat gcgagatgag agattagtgc cgatggaaag gcttatttca 1080agactttcgc tcaaaaaata cgatgtagat gctccgatga attttgatac tgttattcct 1140tcacatcacg ttgtcatgca actaagtcag catgttggtg ccaaagcgat acctgtagta 1200aaggtaggag atattgtgaa agaaggagat ctgataggcg atgtgcctaa taataagctg 1260ggtgctaaat tgcatgccag tattgacggc attataatag atgtaactga tgacagtatt 1320gttatcaaac caagaggtga ttttgatgga caaagcgata ggattggttg a 137198182PRTT. saccharolyticum 98Met Asp Lys Ala Ile Gly Leu Val Glu Tyr Lys Ser Val Ala Thr Gly 1 5 10 15 Ile Thr Ala Ala Asp Asp Met Ala Lys Thr Ala Asp Val Glu Ile Ile 20 25 30 Glu Ala Tyr Thr Val Cys Pro Gly Lys Tyr Ile Val Leu Leu Ala Gly 35 40 45 Lys Leu Ser Ala Val Asn Ser Ala Ile Glu Lys Gly Ile Asn Gln Tyr 50 55 60 Ser Glu Asn Val Ile Asp Ser Phe Ile Leu Gly Asn Pro His Glu Thr 65 70 75 80 Ile Tyr Lys Ala Met Ser Gly Thr Ser Val Ile Glu Asp Val Glu Ala 85 90 95 Leu Gly Ile Ile Glu Thr Phe Ser Ala Ala Ser Ile Ile Leu Ala Ala 100 105 110 Asp Thr Ala Ala Lys Ala Ala Lys Val Asn Leu Val Glu Ile Arg Ile 115 120 125 Ala Arg Gly Met Cys Gly Lys Ser Tyr Leu Leu Leu Thr Gly Glu Leu 130 135 140 Ala Ala Val Glu Ala Ser Ile Asn Ala Gly Cys Lys Ala Leu Glu Arg 145 150 155 160 Thr Gly Met Leu Leu Asn Lys Ser Ile Ile Pro Asn Pro Asp Arg Ala 165 170 175 Ile Trp Asp Lys Ile Ile 180 99549DNAT. saccharolyticum 99atggacaaag cgataggatt ggttgaatac aaatcagttg ctacaggtat aactgctgct 60gatgacatgg ctaaaactgc tgatgtggaa ataatagaag catatacagt atgtccgggg 120aaatacattg ttctgttagc tgggaaatta agtgcagtta attcggcgat agaaaagggc 180ataaatcagt attcggaaaa tgtcattgat agctttatat tgggaaatcc gcatgaaaca 240atatataaag ctatgagtgg cacgtctgta attgaagatg tagaagcact tggtatcata 300gagacatttt ctgcagcatc aataatactt gcagcagata cggctgcaaa agctgcaaaa 360gtgaatctgg tagagataag aatagccaga ggtatgtgcg gcaagtcata tctactgctt 420acaggagaac ttgctgctgt tgaagcatct ataaatgcag gatgcaaagc tttggagaga 480acgggtatgc ttttaaataa gtctataata cccaatccag atagagctat ttgggataag 540ataatttaa 549100425PRTT. saccharolyticum 100Met Tyr Glu Ala Glu Lys Asp Lys Ile Leu Asn Asp Tyr Tyr Asn Ala 1 5 10 15 Lys Glu Ile Tyr Ala Lys Phe Asp Ile Asp Ile Asp Lys Val Leu Asp 20 25 30 Lys Met Lys Lys Ile Arg Ile Ser Leu His Cys Trp Gln Gly Asp Asp 35 40 45 Val Thr Gly Phe Glu Lys Ser Ala Asn Gly Leu Ser Gly Gly Gly Ile 50 55 60 Leu Ala Thr Gly Asn Trp Pro Gly Arg Ala Arg Asn Gly Glu Glu Leu 65 70 75 80 Arg Gln Asp Ile Glu Lys Ala Leu Ser Leu Ile Pro Gly Lys His Lys 85 90 95 Ile Asn Leu His Ala Ile Tyr Ala Glu Thr Asp Gly Glu Phe Val Asp 100 105 110 Arg Asp Glu Ile Asn Val Glu His Phe Arg Lys Trp Ile Tyr Trp Ala 115 120 125 Lys Glu Asn Gly Leu Gly Leu Asp Phe Asn Pro Thr Phe Phe Ser His 130 135 140 Pro Lys Ala Asn Asp Gly Tyr Thr Leu Ser Ser Lys Asp Glu Asn Ile 145 150 155 160 Arg Lys Phe Trp Ile Gln His Gly Lys Arg Cys Arg Glu Ile Ala Asn 165 170 175 Glu Ile Gly Arg Glu Leu Lys Thr Gln Cys Val Asn Asn Val Trp Ile 180 185 190 Pro Asp Gly Ser Lys Asp Leu Pro Ala Asn Arg Ile Glu His Arg Lys 195 200 205 Ile Leu Lys Glu Ser Leu Asp Glu Ile Phe Ser Val Lys Tyr Asp Lys 210 215 220 Ser Asn Ile Val Asp Ser Val Glu Ser Lys Leu Phe Gly Ile Gly Ser 225 230 235 240 Glu Ser Tyr Val Val Gly Ser His Glu Phe Tyr Met Asn Tyr Ala Ser 245 250 255 Arg Asn Asp Val Met Leu Cys Leu Asp Met Gly His Phe His Pro Thr 260 265 270 Glu Asn Ile Ala Asp Lys Ile Ser Ser Ile Leu Thr Phe Asn Asp Asn 275 280 285 Leu Leu Ile His Val Ser Arg Gly Val Arg Trp Asp Ser Asp His Val 290 295 300 Val Ile Leu Asn Glu Asp Leu Leu Ser Leu Ala Lys Glu Ile Arg Arg 305 310 315 320 Cys Asp Ala Tyr Asp Lys Val Tyr Ile Ala Leu Asp Phe Phe Asp Ala 325 330 335 Ser Ile Asn Arg Ile Met Ala Trp Val Ile Gly Ala Arg Ala Thr Leu 340 345 350 Lys Ala Ile Leu Ile Ser Leu Leu Glu Pro Val His Leu Leu Met Glu 355 360 365 Glu Glu Asn Lys Gly Asn Phe Gly Ala Arg Leu Ala Leu Met Glu Glu 370 375 380 Phe Lys Thr Leu Pro Phe Tyr Ser Val Trp Asn Lys Tyr Cys Met Asp 385 390 395 400 Glu Asn Val Pro Ile Gly Thr Ser Trp Ile Asp Asp Val Lys Glu Tyr 405 410 415 Glu Lys Glu Ile Val Lys Asn Arg Ala 420 425 1011278DNAT. saccharolyticum 101atgtatgaag cagaaaaaga taaaatttta aatgattatt ataatgcaaa agagatttat 60gcaaagtttg acatagatat tgataaagta ttagataaaa tgaagaagat tcgtatttca 120cttcactgct ggcaaggcga tgatgtaact ggattcgaaa aaagtgccaa tggattaagc 180ggtggaggta ttttggcgac aggaaactgg cctggtagag caagaaatgg tgaagaatta 240aggcaagaca ttgaaaaagc cttaagcctt ataccaggca aacacaaaat caatttacat 300gccatttacg cagaaacgga tggtgaattt gtagacagag atgaaataaa cgtggagcat 360ttcaggaaat ggatttactg ggcaaaagaa aatggccttg gccttgactt caatcctacg 420tttttttcgc atcctaaagc aaatgatggc tatacgcttt caagcaaaga tgaaaacata 480agaaaatttt ggatccaaca tggtaaaaga tgccgtgaaa tcgcaaatga aataggaaga 540gagctaaaaa ctcaatgtgt gaataatgtt tggattcctg atggttcaaa agatttgcct 600gctaatagga ttgaacacag aaaaatactt aaagaatctt tagatgagat attttcagta 660aaatatgaca aatcaaatat cgttgattct gttgaaagca aattatttgg cattggatct 720gaaagctatg tggttggttc acatgagttt tatatgaact atgcgtcgag aaatgatgta 780atgctgtgcc ttgatatggg acattttcat cctactgaga atattgctga taagatatca 840tcaatactta cattcaatga caatttgttg attcatgtaa gccgtggtgt ccggtgggat 900agcgaccatg tagtcatttt aaatgaagat ttgctttcat tagcaaaaga aataagaaga 960tgtgatgctt atgacaaagt gtatattgca ttagatttct ttgatgcaag cataaatagg 1020ataatggcat gggtaatagg tgcaagagcg acgctaaaag ccatattaat atcactatta 1080gagcctgtgc atctacttat ggaagaggag aataaaggaa attttggtgc aagacttgct 1140ttgatggagg aattcaaaac attgccattt tactctgttt ggaacaaata ctgcatggac 1200gaaaatgtgc ctattggtac atcgtggatt gatgatgtta aagaatatga aaaagaaatt 1260gtaaaaaata gggcttaa 1278102475PRTT. saccharolyticum 102Met Lys Asp Ile Val Tyr Asn Leu Ala Phe Asp Phe Gly Ala Ser Ser 1 5 10 15 Gly Arg Leu Met Leu Ser Ala Phe Asp Gly Glu Lys Ile Thr Ile Glu 20 25 30 Glu Ile Tyr Arg Phe Pro Asn Glu Pro Val Lys Leu Gly Gln Ser Phe 35 40 45 Tyr Trp Asp Phe Leu Arg Leu Phe His Glu Leu Lys Asn Gly Leu Lys 50 55 60 Ile Ala Ser Lys Arg Lys Ile Lys Ile Ser Gly Ile Gly Ile Asp Thr 65 70 75 80 Trp Gly Val Asp Tyr Gly Leu Leu Asp Lys Asn Asp Gln Leu Ile Ser 85 90 95 Asn Pro Phe His Tyr Arg Asp Lys Arg Thr Asp Gly Ile Ile Lys Asp 100 105 110 Phe Glu Asn Met Ala Leu Leu Glu Glu Ile Tyr Asn Val Thr Gly Ile 115 120 125 Gln Phe Met Glu Phe Asn Thr Ile Phe Gln Leu Tyr Cys Asp Tyr Lys 130 135 140 Lys Arg Pro Glu Leu Leu Asp Asn Ala Lys Thr Leu Leu Phe Ile Pro 145 150 155 160 Asp Leu Phe Asn Phe Tyr Leu Thr Asn Glu Lys Tyr Asn Glu Tyr Thr 165 170 175 Val Ala Ser Thr Ser Gln Met Leu Asp Ala Asn Lys Lys Asp Trp Ala 180 185 190 Asn Asp Leu Ile Glu Lys Leu Asn Leu Pro Glu Gly Ile Phe Gln Lys 195 200 205 Ile Leu Met Pro Gly Asn Thr Ile Gly Tyr Leu Thr Lys Glu Ile Gln 210 215 220 Glu Glu Thr Gly Leu Ser Glu Val Pro Val Ile Ser Val Gly Ser His 225 230 235 240 Asp Thr Ala Ser Ala Val Ala Gly Thr Pro Ile Glu Asn Gly Ser Ser 245 250 255 Ala Tyr Leu Ile Cys Gly Thr Trp Ser Leu Leu Gly Val Glu Ser Glu 260 265 270 Lys Pro Ile Ile Asn Glu Asn Thr Lys Lys Tyr Asn Phe Thr Asn Glu 275 280 285 Gly Gly Val Glu Gly Leu Ile Arg Leu Leu Lys Asn Ile Asn Gly Leu 290 295 300 Trp Ile Ile Gln Gln Leu Lys Gln Ser Trp Asn Ser Asn Gly Ile Lys 305 310 315 320 Ile Gly Phe Pro Glu Ile Ser Gln Met Ala Ser Lys Ala Glu His Glu 325 330 335 Glu Phe Ile Ile Asn Pro Asp Asp Lys Leu Phe Ile Ala Pro Asp Asp 340 345 350 Met Ala Glu Ala Ile Arg Gln Tyr Cys Thr Lys Thr Gly Gln Gly Leu 355 360 365 Pro Gln Asn Ile Gly Asp Ile Ala Arg Ala Ala Tyr Asn Gly Ile Val 370 375 380 Glu Gln Tyr Lys Asn Cys Leu Asn Asn Leu Glu Asp Ile Val Gly Gln 385 390 395 400 Glu Ile Asp Asn Ile His Met Val Gly Gly Gly Ile Gln Asp Lys Phe 405 410 415 Leu Cys Lys Leu Thr Ala Asp Val Thr Gly Lys Lys Val Ile Thr Gly 420 425 430 Pro Val Glu Ala Ser Ile Tyr Gly Asn Val Ile Val Gln Leu Met Ala 435 440 445 Leu Gly Tyr Ile Lys Asp Leu Arg Glu Gly Arg Lys Ile Ile Lys Asn 450 455 460 Ser Ile Glu Asn Asp Glu Glu Met Phe Ala Lys 465 470 475 1031428DNAT. saccharolyticum 103atgaaagata ttgtgtataa tctggctttt gattttggag cttcaagtgg ccgtcttatg 60ctatccgcgt ttgatggcga aaaaatcaca attgaagaga tttatagatt tccaaatgag 120ccagtcaagc tgggacaatc attttattgg gattttttaa ggctttttca cgaattaaaa 180aacggattaa aaatagcatc aaagaggaaa atcaaaatat ccggcattgg tatagacact 240tggggtgtcg attatggatt gcttgataaa aatgatcaat tgatttcaaa tccttttcat 300tacagagata aaagaacgga tggcataata aaagattttg aaaatatggc gttactggag 360gaaatctaca acgtaactgg tatacagttt atggaattta atacaatatt ccaattgtat 420tgcgattata aaaagcgtcc agaattattg gataatgcaa agacattgtt gtttattcca 480gatttattta acttttattt gacaaatgag aaatacaatg aatatactgt tgcatccaca 540tcgcaaatgt tggatgctaa caagaaagat tgggcaaatg atcttataga aaagttaaat 600ttgccagaag gtatttttca aaagatactg atgccaggaa atacaattgg ttatctaaca 660aaagaaattc aagaagaaac aggattgtct gaagttcccg tgatttctgt tggcagccat 720gatacggcat cagcagttgc aggtacacct attgaaaacg gttcaagtgc ttatttgatt 780tgtggtactt ggtcattatt aggtgttgaa agtgaaaaac ctataataaa tgaaaataca 840aagaagtaca attttacaaa tgaaggcggt gtcgaaggcc ttataaggct acttaaaaat 900attaatggtc tgtggataat tcagcaatta aaacaaagtt ggaattcaaa tggcattaaa 960ataggatttc cagaaatcag ccagatggca tctaaagcag agcacgaaga atttatcata 1020aatcctgatg acaaattgtt tatagctcca gatgatatgg ctgaggcgat aaggcaatat 1080tgtacaaaaa caggacaggg tttgccgcag aatattggcg acatagcaag agccgcttac 1140aatggtatag ttgaacaata caaaaattgc ttaaacaatt tagaagatat tgtagggcaa 1200gaaatagata atattcacat ggttggtggt gggatacagg ataagttcct gtgcaagctg 1260actgcagatg ttacagggaa aaaagtcata acaggccctg tagaagcttc aatctatggc 1320aatgtgatag tccagcttat ggcattggga tatataaaag acttgagaga aggaagaaag 1380ataataaaga attctataga gaatgatgaa gagatgtttg ctaaatag 1428104254PRTT. saccharolyticum 104Val Ser Asn Ile Tyr Thr Leu Val Val Val Glu Asp Glu Tyr Glu Ile 1 5 10 15 Arg Thr Gly Leu Val Asn Cys Phe Pro Trp Asn Lys Met Gly Phe Val 20 25 30 Val Ala Glu Glu Phe Glu Asn Gly Gly Glu Cys Phe Glu Tyr Leu Cys 35 40 45 Lys Asn Lys Val Asp Thr Ile Leu Cys Asp Ile Lys Met Pro Val Met 50 55 60 Ser Gly Ile Glu Leu Ala Lys Lys Ile Phe Glu Ser Asn Ile Ser Thr 65 70 75 80 Lys Ile Val Ile Ile Ser Gly Tyr Thr Asp Phe Glu Tyr Ala Arg Gln 85

90 95 Ala Leu Arg Tyr Gly Val Lys Asp Tyr Ile Val Lys Pro Thr Lys Tyr 100 105 110 Asn Glu Ile Ile Asp Val Phe Ser Arg Ile Lys Lys Glu Leu Asp Asn 115 120 125 Glu Asn Thr Lys Glu Ile Leu Asn Asn Ser Cys Asn Asn Glu Ile Asp 130 135 140 Gln Tyr Ser Ser Ile Ile Ser Ile Ile Glu Lys Tyr Val Asp Glu His 145 150 155 160 Tyr Arg Asp Val Thr Leu Glu Asp Val Ala Lys Val Val Tyr Met Asn 165 170 175 Pro Tyr Tyr Leu Ser Lys Tyr Phe Lys Gln Lys Thr Gly Met Asn Phe 180 185 190 Ser Asp Tyr Ile Thr Glu Val Arg Met Lys Lys Ala Val Glu Phe Leu 195 200 205 Lys Asn Pro Leu Tyr Lys Thr Tyr Glu Ile Ser Tyr Met Ile Gly Tyr 210 215 220 Lys Asn Pro Lys Asn Phe Thr Arg Ala Phe Lys Lys Tyr Tyr Lys Lys 225 230 235 240 Ser Pro Arg Glu Phe Val Asn Ser Ala Ile Asn Phe Lys Glu 245 250 105765DNAT. saccharolyticum 105gtgtctaata tttatacgct tgtagtagta gaagatgaat atgagataag aacaggatta 60gttaactgct ttccatggaa caaaatgggt tttgttgttg cagaagaatt tgaaaatgga 120ggagaatgtt ttgagtattt gtgtaaaaat aaggttgata caattttatg tgatataaaa 180atgccagtta tgtctggtat agagttggca aagaaaattt ttgaaagtaa tataagcact 240aaaatagtta taatcagtgg ttatactgat tttgaatatg ccagacaggc gttaagatat 300ggtgttaaag attatatagt aaaacctact aaatataatg aaataattga tgttttcagc 360agaataaaaa aagaattaga caatgaaaat acaaaggaaa tattgaataa ctcatgtaac 420aatgaaattg atcagtacag cagcataatt tcaatcatag aaaaatatgt tgatgaacat 480tacagagatg tgacattgga agatgtagct aaagtagttt atatgaatcc gtattattta 540agcaaatatt ttaaacaaaa aaccggtatg aatttttctg attatataac tgaggtcaga 600atgaaaaaag ctgtagagtt tctaaaaaat cctttgtata aaacttatga aataagttat 660atgattggat ataaaaatcc aaaaaatttt actagagcat ttaaaaaata ttataaaaaa 720tccccaagag aatttgtaaa ttcagcaata aattttaagg aatga 765106588PRTT. saccharolyticum 106Met Arg Glu Leu Asn Asn Lys Phe Phe Tyr Lys Asn Leu Phe Val Leu 1 5 10 15 Ala Leu Pro Leu Ile Leu Ile Val Ile Val Leu Gly Ser Phe Ser Ile 20 25 30 Leu Ile Thr Glu Arg Tyr Val Arg Asp Glu Ile Tyr Lys Asn Ser Arg 35 40 45 Glu Ile Leu Lys Gln Ser Ser Asn Asp Leu Ser Ile Leu Phe Asn Asp 50 55 60 Ile Asn Lys Ile Tyr Leu Thr Phe Gly Thr Asn Lys Asp Val Thr Leu 65 70 75 80 Tyr Leu Glu Arg Ile Leu Asn Thr Asn Lys Tyr Ser Leu Asp Asp Met 85 90 95 Trp His Leu Ser Met Ile Glu Ser Leu Phe Asp Ser Thr Ser Phe Ser 100 105 110 Glu Pro Tyr Ile Gln Ser Ile Tyr Leu Tyr Phe Asn Asn Pro Asn Lys 115 120 125 Asn Phe Leu Val Thr Gly Asn Gly Ile Asn Ser Val Thr Asn Tyr Ile 130 135 140 Asp Asn Lys Trp Tyr Asp Ser Phe Leu Asn Ala Pro Lys Asp Glu Ile 145 150 155 160 Ser Trp Ile Glu Val Arg Asn Leu Lys Met Tyr Ser Phe Asp Lys Lys 165 170 175 Gly Ile Lys Val Leu Ser Ile Tyr Lys Lys Ile Ala Asn Phe Asn Gly 180 185 190 Asp Lys Ile Asp Gly Val Leu Val Leu Asn Ile Tyr Leu Asp Tyr Ile 195 200 205 Glu Asn Leu Leu Asn Thr Ser Thr Ile Phe Pro Asp Gln Lys Ile Leu 210 215 220 Ile Leu Asp Ala His Asp Asn Leu Ile Cys Gln Asn Ile Asn Gly Asn 225 230 235 240 Phe Thr Gly Lys Ile Asp Leu Asp Asn Tyr Ser Lys Ala Asn Ile Ile 245 250 255 Thr Lys Leu Glu Ser Pro Asn Tyr Asn Ile Lys Tyr Val Ser Ile Val 260 265 270 Pro Lys Lys Tyr Leu Tyr Glu Val Pro Ile Lys Leu Leu Lys Met Thr 275 280 285 Leu Val Leu Leu Leu Thr Ser Ile Phe Phe Val Ile Leu Ile Thr Phe 290 295 300 Arg Ile Thr Lys Arg Asn Tyr Glu Asn Val Asn Lys Ile Leu Lys Ile 305 310 315 320 Ile Glu Ala Glu Lys Thr Asn Glu Ile Phe Pro Glu Ile Pro Val Glu 325 330 335 Ser Arg Asp Glu Tyr Ser Tyr Ile Ile Tyr Asn Ile Ile Asn Ser Tyr 340 345 350 Ile Glu Lys Ser Gln Leu Lys Met Glu Leu Ala Glu Lys Lys Tyr Lys 355 360 365 Met Lys Ala Met Glu Leu Leu Ala Leu Gln Ser Gln Ile Ser Pro His 370 375 380 Phe Leu Ser Asn Ala Leu Glu Ile Ile Tyr Leu Arg Ala Leu Ser Tyr 385 390 395 400 Thr Asn Gly Pro Asn Asp Val Thr Lys Met Ile Glu Asn Leu Ser Gln 405 410 415 Ile Leu Lys Tyr Leu Leu Ser Asn Pro Asn Glu Thr Val Thr Val Lys 420 425 430 Glu Glu Ile Glu Asn Thr Lys Ala Tyr Ile Gln Ile Leu Lys Val Arg 435 440 445 Tyr Arg Asp Lys Phe Lys Val Asn Leu Ile Tyr Asp Glu Ser Ile Leu 450 455 460 Ser Cys Leu Met Met Lys Leu Met Leu Gln His Leu Ile Glu Asn Ser 465 470 475 480 Ile Lys His Gly Leu Lys Lys Lys Asn Tyr Glu Gly Ser Ile Lys Ile 485 490 495 Lys Ile Lys Ala Val Asp Lys Lys Lys Ile Lys Ile Ser Val Ile Asp 500 505 510 Asn Gly Ile Gly Met Ser Lys Glu Arg Leu Asn Tyr Val Lys Arg Ile 515 520 525 Leu Asp Ser Asp Phe Asp Phe Tyr Glu His Ile Gly Leu Met Asn Thr 530 535 540 Asn Glu Arg Leu Lys Leu Leu Tyr Gly Lys Asp Cys Glu Ile Leu Ile 545 550 555 560 Arg Ser Lys Leu Asn Ile Gly Thr Ala Val Tyr Ile Ile Phe Pro Tyr 565 570 575 Gln Leu Lys Asn Gln Asn Asn Asp Asp Tyr Asn Lys 580 585 1071767DNAT. saccharolyticum 107atgagagaat taaacaataa atttttttat aaaaatcttt ttgttttggc attgccatta 60attttaattg ttattgtatt aggttcattt tcaatattaa taacagaaag atatgttaga 120gatgaaatat acaaaaatag tagagaaata ttaaagcaaa gcagtaatga tttgtcaatt 180ttatttaatg atataaataa aatttattta acatttggaa caaacaaaga tgtgacattg 240tatttggaaa ggatcttaaa tacaaataaa tattctttag atgatatgtg gcatcttagc 300atgatagaaa gtttatttga ttctacgtcg ttttcagaac cttatataca atcaatttat 360ttgtatttta acaatcctaa taaaaatttt ttagtgacag gaaatggtat taattctgta 420acaaattata ttgataataa atggtatgac agctttttaa atgcaccaaa agatgagatt 480tcttggatag aggttagaaa tttaaaaatg tatagtttcg ataaaaaggg gataaaagtc 540ctaagtatat acaaaaaaat tgcaaacttt aacggggata aaattgatgg tgtgcttgta 600ctaaatatat atttggacta tattgaaaat ttgctaaata cttcaacaat atttcctgac 660caaaaaattc ttatattaga tgcccacgac aatttaatat gtcaaaatat taatgggaat 720ttcactggga agatagactt agataattat agcaaagcaa acatcataac aaaattagaa 780tctccaaatt ataatataaa atatgtatct attgttccta aaaaatacct ttatgaagtt 840cctataaagc ttttaaagat gactttagtt ttacttttga cgtcaatttt ttttgtgata 900ttgataacat ttagaatcac taaacgaaat tacgaaaatg taaataaaat attaaagatt 960atagaggcag aaaagacaaa tgagatattt ccagaaattc cagtagaaag tagagatgag 1020tacagctata taatttacaa cattattaat agttatattg aaaaaagtca attgaaaatg 1080gaattagcag aaaagaagta taaaatgaaa gcaatggagt tattagcact gcaatcgcaa 1140attagtcctc attttttgtc taatgcgttg gagattattt atcttagggc attgtcatac 1200acaaacggtc ctaatgatgt cacaaaaatg attgaaaatt tgtcacagat tttaaagtat 1260ttgttaagta atccaaatga aacagtaact gtaaaagaag aaattgaaaa tacaaaggca 1320tatatacaaa tattgaaggt caggtataga gataaattta aagtaaatct aatttatgat 1380gaaagtattt tatcatgtct catgatgaaa ctgatgctgc aacatttaat agaaaattct 1440ataaaacatg ggcttaagaa gaaaaattat gaaggatcaa taaaaatcaa aataaaagca 1500gttgataaaa agaaaataaa aatttcagta atcgataatg gcataggaat gtccaaagag 1560aggctaaatt atgtaaaaag aattcttgac tctgacttcg atttttatga acatattgga 1620ctaatgaata caaatgaacg gttaaaactt ctctatggga aagattgtga aatattaata 1680agaagtaaat tgaatattgg tactgccgta tatataattt ttccatatca attaaaaaat 1740cagaataatg atgattataa taagtga 1767108687PRTT. saccharolyticum 108Met Gly Ile Asn Arg Tyr Asp Leu Val Lys Arg His Asn Val Ile Leu 1 5 10 15 Glu Lys Ala Asp Ile Glu Asn Pro Leu Ser Val Gly Asn Gly Glu Ile 20 25 30 Ala Phe Thr Ala Asp Ile Thr Gly Met Gln Thr Phe Ile Asp Asp Tyr 35 40 45 Lys Ser Ile Pro Leu Cys Thr Met Ser Gln Trp Gly Phe His Thr Thr 50 55 60 Pro Ala Gln Asn Asp Lys Gly Tyr Tyr Thr Leu Glu Asp Leu Asn Leu 65 70 75 80 Lys Tyr Tyr Asp Ala Phe Asp Arg Lys Val Gly Tyr Val Thr Ser Ala 85 90 95 Glu Asn Gln Glu Asn Val Phe Asn Trp Leu Arg Ser Asn Pro His Arg 100 105 110 Ile Asn Leu Gly Asn Ile Gly Leu Asn Ile Ile Leu Asp Asp Gly Thr 115 120 125 Lys Ala Glu Leu Lys Asp Ile Phe Glu Ile His Gln Val Leu Asp Leu 130 135 140 Trp Asn Gly Ile Leu Ile Ser Asp Phe Lys Val Glu Lys Val Pro Val 145 150 155 160 His Val Glu Thr Phe Cys His Pro Tyr Glu Asp Met Ile Asn Phe Ser 165 170 175 Val Glu Ser Glu Leu Leu Lys Gln Asn Lys Ile Tyr Ile Glu Val Lys 180 185 190 Phe Pro Tyr Gly Ala Ala Asn Ile Ser Gly Ser Asp Trp Asp Arg Asn 195 200 205 Asp Arg His Asp Thr Asn Val Val Asp Tyr Gly Arg Asp Phe Val Glu 210 215 220 Leu Leu Arg Thr Val Asp Glu Asp Val Tyr Phe Val Lys Ile Glu Tyr 225 230 235 240 Ser Lys Gly Val Tyr Leu Asn Arg Ile Gly Glu Asn His Phe Ala Leu 245 250 255 Lys Gln Lys Glu Tyr Asn Gly Arg Ile Glu Phe Ser Cys Leu Phe Ser 260 265 270 Lys Gln Lys Pro Leu Lys Cys Leu His Ser Phe Ser Glu Ser Lys Arg 275 280 285 Met Cys Lys Glu Tyr Trp Asn Ser Phe Trp Arg Gly Gly Gly Ala Ile 290 295 300 Asp Phe Ser Lys Cys Glu Asp Lys Arg Ala Phe Glu Leu Glu Arg Arg 305 310 315 320 Val Ile Leu Ser Gln Tyr Leu Thr Ala Ile Gln Cys Ser Gly Ser Met 325 330 335 Pro Pro Gln Glu Thr Gly Leu Thr Cys Asn Ser Trp Tyr Gly Lys Phe 340 345 350 His Leu Glu Met His Trp Trp His Ala Val His Phe Ala Leu Trp Gly 355 360 365 Arg Met Pro Leu Leu Ser Arg Ser Ile Trp Trp Tyr Arg Ser Ile Phe 370 375 380 Asn Val Ser Arg Asp Ile Ala Arg Lys Gln Gly Tyr Lys Gly Val Arg 385 390 395 400 Trp Pro Lys Met Val Gly Pro Asp Gly Arg Asp Ser Pro Ser Pro Ile 405 410 415 Gly Pro Leu Leu Val Trp Gln Gln Pro His Leu Ile Tyr Tyr Ser Glu 420 425 430 Leu Phe Phe Arg Glu Asn Pro Thr Glu Glu Thr Leu Asp Met Phe Lys 435 440 445 Asp Ile Val Ile Asn Thr Ala Asp Phe Ile Ala Ser Phe Val Ala Tyr 450 455 460 Asp Arg Lys Asn Asp Arg Tyr Ile Leu Ala Pro Pro Leu Ile Pro Ala 465 470 475 480 Gln Glu Asn His Asp Pro Asn Val Thr Leu Asn Pro Val Phe Glu Leu 485 490 495 Glu Tyr Phe Ser Phe Ala Leu Glu Ile Ala Val Lys Trp Ile Glu Arg 500 505 510 Leu Gly Leu Asn Val Asn Gln Glu Trp Asn Glu Ile Arg Phe Lys Leu 515 520 525 Ala Asn Leu Pro Ser Lys Asp Gly Val Tyr Ile Ser His Glu Lys Cys 530 535 540 Ile Asn Thr Tyr Glu Lys Phe Asn Phe Asp His Pro Ser Met Leu Ala 545 550 555 560 Ala Leu Gly Met Leu Pro Gly Arg Lys Val Asp Lys Glu Thr Met Arg 565 570 575 Arg Thr Leu His Arg Val Leu Lys Glu Trp Lys Phe Glu Glu Met Trp 580 585 590 Gly Trp Asp Phe Pro Met Met Ala Met Thr Ala Thr Arg Leu Gly Glu 595 600 605 Pro Glu Thr Ala Ile Asn Ile Leu Leu Met Asp Ser Pro Lys Asn Thr 610 615 620 Tyr Met Val Asn Gly His Asn Asn Gln Ile Pro Asn Lys Glu Leu Pro 625 630 635 640 Val Tyr Leu Pro Gly Asn Gly Gly Leu Leu Ala Ala Met Ala Leu Met 645 650 655 Thr Ala Gly Trp Asp Gly Asn Ser Gln Ser Thr Pro Gly Phe Pro Lys 660 665 670 Asn Gly Met Trp Asn Val Glu Trp Glu Gly Leu Lys Ala Met Ile 675 680 685 1092064DNAT. saccharolyticum 109atgggaatta acagatatga tcttgtaaaa aggcataatg taattttgga aaaagcagat 60atcgaaaatc cattgtcagt aggtaatgga gaaattgctt ttacagctga tataacggga 120atgcaaactt ttattgatga ctataagagc attcctttat gtaccatgtc acagtggggg 180tttcatacta cgccggcaca gaatgataag ggctattata ctttggaaga tttgaacctc 240aagtattacg atgcatttga ccgaaaggtt ggatatgtaa catcagcaga aaatcaagag 300aatgtattta attggttgag gagtaatcct catagaatta atttaggtaa tataggatta 360aatataattc ttgatgatgg cacaaaagca gaattgaaag atattttcga aatacaccaa 420gtattagatt tgtggaacgg aatattgata agtgacttta aagtcgaaaa agtccctgtt 480cacgttgaga ctttttgcca tccatatgaa gatatgataa atttttctgt tgaatcagaa 540ctgctaaaac aaaataaaat ttatattgaa gtaaaatttc catatggtgc ggccaatata 600tcaggctccg attgggatag aaatgataga catgatacaa atgtggttga ttatggcaga 660gattttgtcg aattattgag aactgtcgat gaagatgttt attttgtaaa aatagagtac 720tcaaaaggcg tttatttaaa tagaatcggg gaaaatcatt ttgcattaaa gcaaaaagag 780tataatggga gaatagaatt ttcgtgcttg ttttcgaagc aaaaacctct taagtgcttg 840cattcattta gtgaaagcaa aaggatgtgt aaagaatatt ggaatagctt ttggagagga 900ggtggtgcaa tagatttttc aaagtgtgag gataaaagag cttttgaatt ggagagaagg 960gtaatacttt cgcaatatct tacagctatt caatgttcgg gttctatgcc gccgcaagaa 1020acagggctca cctgtaatag ctggtatggt aaatttcatt tggaaatgca ttggtggcat 1080gctgtacatt ttgctttatg gggtagaatg cctttgctga gtagaagtat atggtggtac 1140aggagcattt tcaatgtatc acgtgacatt gcgagaaagc aaggatacaa aggtgtacgc 1200tggcctaaaa tggttggacc agatggaagg gatagccctt ctccgatagg accattgctt 1260gtttggcagc agcctcatct tatatattac agtgaactgt tttttagaga aaatcctacg 1320gaagaaacat tagatatgtt taaagacata gtaattaata ctgctgattt tattgcatca 1380tttgttgcat atgatagaaa aaatgataga tatatacttg cgccaccttt gattccagca 1440caagaaaatc atgatcctaa cgttacatta aatccggtat ttgaattgga gtatttttcg 1500tttgcgctgg aaatagcagt taaatggatt gaaaggttag gactaaatgt gaaccaagag 1560tggaatgaaa tacgttttaa attagctaat ttaccttcaa aagacggtgt atatatatcg 1620catgaaaaat gtattaacac ttatgagaaa tttaattttg accatccatc tatgcttgca 1680gcattgggga tgctaccagg ccgcaaggtt gataaagaaa ctatgagaag gactttacat 1740agagtattaa aagagtggaa atttgaggaa atgtggggtt gggattttcc gatgatggct 1800atgactgcaa caagattagg cgaaccggag acagcaataa atattctttt gatggattca 1860ccaaaaaata cttatatggt aaatggccat aataaccaaa taccgaataa agaactacca 1920gtatatttgc ctggaaatgg tggactattg gcggcaatgg ccctcatgac agctggttgg 1980gatgggaata gccaaagcac acctggattt cctaaaaatg ggatgtggaa tgttgaatgg 2040gaagggttaa aagcgatgat atga 2064110293PRTT. saccharolyticum 110Met Ile Lys Arg Lys Asp Leu Tyr Ile Arg Asp Pro Phe Val Val Pro 1 5 10 15 Val Pro Asn Glu Lys Ile Tyr Tyr Met Phe Gly Thr Thr Asp Ile Asn 20 25 30 Cys Trp Asn Asp Glu Lys Ala Thr Gly Phe Asp Tyr Tyr Lys Ser Ser 35 40 45 Asp Leu Glu Asn Phe Glu Gly Pro Phe Ile Ala Phe Arg Pro Asp Lys 50 55 60 Asn Phe Ile Trp Asp Lys Asn Phe Trp Ala Pro Glu Val His Lys Tyr 65 70 75 80 Asn Asp Met Tyr Tyr Met Phe Ala Thr Phe Phe Ala Asp Gly Arg Asn 85 90 95 Arg Gly Thr Gln Ile Leu Val Ser Glu Lys Ile Ser Gly Pro Tyr Arg 100 105 110 Pro Trp Ser Ile Glu Pro Val Thr Pro Lys Asp Trp Met Cys Leu Asp 115 120

125 Gly Thr Phe Tyr Val Asp Glu Asn Gly Glu Pro Trp Met Ile Phe Cys 130 135 140 His Glu Trp Val Gln Ile Tyr Asp Gly Glu Ile Cys Ala Val Arg Leu 145 150 155 160 Ser Lys Asp Leu Lys Thr Thr Ile Gly Asn Pro Ile Thr Leu Phe Lys 165 170 175 Ala Ser Ser Ala Asn Trp Thr Arg Ser Ile Lys Lys Ile Lys Asp His 180 185 190 Glu Cys Tyr Val Thr Asp Gly Pro Phe Ile Tyr Arg Ser Glu Glu Gly 195 200 205 Lys Leu Tyr Met Leu Trp Ser Ser Phe Ile Glu Asn Asn Ile Tyr Ala 210 215 220 Val Gly Ile Ser Leu Ser Arg Thr Gly Lys Ile Thr Gly Pro Trp Val 225 230 235 240 His Ser Glu Asn Pro Ile Phe Ala Gly Asp Gly Gly His Gly Met Ile 245 250 255 Phe Lys Thr Phe Glu Gly Asn Leu Thr Leu Ala Val His Thr Pro Asn 260 265 270 Lys Arg Lys Glu Glu Arg Pro Leu Phe Ile Thr Leu Glu Lys Ser Val 275 280 285 Leu Asn Asp Thr Leu 290 111882DNAT. saccharolyticum 111atgataaaac gaaaggatct ttatatacgt gatccatttg tagttccagt accgaatgaa 60aaaatatatt atatgtttgg aactactgat ataaattgct ggaatgatga gaaagcaact 120ggatttgatt actataaatc atctgattta gaaaattttg aaggaccttt tattgcattt 180agaccagata aaaactttat ttgggataaa aatttttggg ctccagaagt gcacaaatac 240aatgacatgt attatatgtt tgctacattt ttcgctgatg gcagaaatag aggaacgcaa 300attttagtat ctgaaaaaat aagtgggcca tatagaccat ggagtattga accggtgacg 360ccgaaggatt ggatgtgttt agatgggact ttttatgtag atgagaatgg ggaaccctgg 420atgatatttt gccatgaatg ggtacaaata tatgatgggg aaatttgtgc tgtaagattg 480tcgaaagatt taaaaacaac gataggaaat cctattacac tttttaaagc ttccagtgct 540aattggacaa gaagtattaa aaagattaaa gatcatgaat gctacgttac ggatggccct 600tttatttata ggtctgaaga gggaaagctt tatatgttgt ggtccagttt tattgaaaac 660aatatatacg ctgttggtat atcattatcg agaacaggca aaataaccgg cccgtgggta 720cacagtgaaa atccaatttt cgcaggtgat ggtgggcatg gtatgatatt taagaccttt 780gaagggaatc taacattggc agtacacaca cctaataaaa ggaaagaaga acggcccctt 840tttataactt tagaaaaatc tgtgcttaat gataccttat aa 882112442PRTT. saccharolyticum 112Met Phe Lys Lys Ile Thr Ser Leu Leu Ile Ser Leu Leu Leu Ile Ile 1 5 10 15 Ser Leu Val Thr Gly Cys Ser Ser Ser Ser Asn Ser Ser Ser Ser Ser 20 25 30 Lys Asn Ser Ser Glu Asn Asn Thr Ser Pro Lys Thr Val Thr Leu Arg 35 40 45 Phe Met Trp Trp Gly Gly Asp Ala Arg His Lys Ala Thr Leu Asp Ala 50 55 60 Ile Ser Leu Tyr Glu Lys Glu His Pro Asn Val Lys Ile Asn Ala Glu 65 70 75 80 Tyr Gly Gly Val Thr Asp Tyr Leu Gln Lys Leu Ile Thr Gln Leu Ser 85 90 95 Ser Gly Thr Ala Pro Asp Leu Ile Gln Ile Asp Val Thr Trp Leu Gln 100 105 110 Gln Leu Phe Ser Gln Gly Asp Phe Phe Ala Asp Leu Ser Lys Leu Lys 115 120 125 Asp Ile Asn Val Asn Ala Phe Asp Gln Asn Phe Leu Lys Asn Tyr Cys 130 135 140 Tyr Val Asn Asn Lys Leu Ile Gly Leu Pro Thr Gly Ile Asn Asn Ser 145 150 155 160 Ala Met Tyr Ile Asn Lys Asp Phe Phe Asn Lys Phe Gly Ile Asp Asp 165 170 175 Lys Thr Val Trp Thr Trp Asp Asn Leu Leu Gln Thr Ala Lys Met Val 180 185 190 His Glu Lys Asp Lys Asn Ala Tyr Leu Leu Asp Ala Asp Ser Thr Ile 195 200 205 Cys Asp Tyr Ile Leu Val Thr Tyr Val Gly Gln Lys Thr Gly Asn Gln 210 215 220 Trp Val Lys Asp Asp Tyr Thr Leu Gly Phe Asp Lys Gln Thr Leu Thr 225 230 235 240 Glu Ala Phe Lys Tyr Leu Asn Asp Leu Phe Glu Val Gly Ala Ile Glu 245 250 255 Pro Phe Ser Gln Ser Ala Pro Tyr Glu Gly Lys Pro Asp Gln Asn Pro 260 265 270 Met Trp Leu Asn Gly Gln Thr Gly Met Leu Trp Asn Trp Ser Ser Ile 275 280 285 Tyr Ala Gly Val Lys Ala Asn Ile Lys Asn Leu Ser Leu Ala Leu Pro 290 295 300 Pro Ile Asp Pro Asn Ala Lys Gln Thr Gly Ile Val Val Arg Pro Ser 305 310 315 320 Gln Leu Ile Ala Ile Asn Lys Asp Ser Lys Asn Ile Asp Glu Ala Ala 325 330 335 Lys Phe Leu Asn Trp Phe Phe Thr Asn Thr Asp Ala Ile Lys Thr Leu 340 345 350 Lys Asp Val Arg Gly Val Pro Ala Thr Ala Asp Ala Arg Lys Ile Leu 355 360 365 Ser Glu Asn Asn Leu Leu Asp Ser Thr Leu Thr Asp Asn Ala Asn Gln 370 375 380 Ala Met Glu Lys Met Ala Pro Pro Glu Asn Gly Ile Ser Gly Asn Gln 385 390 395 400 Glu Leu Glu Lys Ile Asn Thr Asp Ile Ile Gln Glu Leu Ala Tyr Lys 405 410 415 Lys Ile Thr Pro Glu Gln Ala Ala Asp Glu Leu Ile Asn Thr Tyr Lys 420 425 430 Gln Lys Leu Pro Glu Leu Lys Ser Gln Gln 435 440 1131329DNAT. saccharolyticum 113atgtttaaaa aaattacatc tctgttaata tcgcttcttt tgataatttc attagttaca 60ggatgtagca gttcttcgaa ttcttcgagt tcatcgaaaa atagttctga aaataatacc 120agcccaaaaa ccgtaacatt aagatttatg tggtggggtg gagatgccag acataaagca 180acacttgatg ccataagtct ttatgaaaaa gaacatccca atgtaaagat taatgctgaa 240tatggcggcg ttactgacta tctccaaaag ctgataactc aattaagcag tggtacagca 300cctgatctta tacaaataga tgtaacatgg ttgcagcaac tttttagcca aggtgatttt 360tttgcagatt taagtaagtt aaaagatatc aatgtgaatg catttgatca aaattttctt 420aaaaattatt gctatgtcaa caataagttg ataggtttgc ctacaggaat aaacaattcg 480gcaatgtata ttaacaaaga cttttttaat aaatttggca tagacgataa gacggtttgg 540acatgggata atctcttgca aaccgctaag atggtgcatg aaaaggataa aaatgcttat 600cttttagatg ctgattctac tatttgtgat tatatattgg tcacatacgt ggggcaaaaa 660actggaaatc agtgggtgaa agatgattac actttaggtt ttgataaaca aacattgaca 720gaggcattca aatatttaaa cgatttgttc gaagtaggcg ctatagagcc attttctcaa 780agtgctccat acgaaggaaa acctgatcaa aatcctatgt ggcttaatgg tcaaacgggt 840atgctttgga actggtcatc tatatatgct ggtgtaaaag caaacataaa gaacctgtca 900ttggcattgc cacctattga ccctaatgca aaacagacag gcatagttgt aagaccatca 960cagcttattg ctattaacaa ggattctaaa aatatcgatg aagcagcaaa atttttaaat 1020tggttcttta cgaatacaga tgctataaaa acacttaaag atgtcagagg agttccagct 1080accgcagatg cacgcaaaat tttatcagaa aataatttgt tggattcgac tttaactgat 1140aatgcaaatc aagctatgga aaagatggca cctcctgaaa acggtataag tggtaatcaa 1200gagttagaaa agataaatac tgatatcata caagaactgg cttataaaaa gataacgcca 1260gagcaggctg ctgatgaatt gataaatact tataaacaga aacttccaga attaaaaagc 1320cagcaataa 1329114295PRTT. saccharolyticum 114Met Ser Tyr Asn Lys Lys Arg Asn Leu Met Gly Tyr Leu Tyr Ile Ser 1 5 10 15 Pro Trp Ile Ile Gly Phe Leu Ile Phe Thr Leu Tyr Pro Phe Ala Met 20 25 30 Thr Phe Ile Tyr Ser Phe Cys Asn Tyr Ser Ile Thr Lys Ser Pro Val 35 40 45 Phe Ile Gly Leu Gly Asn Tyr Ile Thr Met Phe Thr Lys Asp Met Tyr 50 55 60 Phe Trp Pro Ser Leu Ile Asn Thr Ile Lys Tyr Val Leu Met Thr Val 65 70 75 80 Pro Leu Lys Leu Cys Phe Ala Leu Phe Val Ala Met Ile Leu Asn Ile 85 90 95 Asp Ile Lys Gly Val Asn Val Phe Arg Thr Thr Tyr Tyr Leu Pro Ser 100 105 110 Ile Phe Gly Gly Ser Val Ala Leu Ser Val Ile Trp Lys Phe Leu Phe 115 120 125 Met Asp Asn Gly Ile Met Asn Lys Phe Leu Ser Tyr Phe His Ile His 130 135 140 Gly Pro Ser Trp Leu Gly Asn Pro His Ile Ser Leu Phe Thr Ile Ser 145 150 155 160 Leu Leu Ser Val Trp Glu Phe Gly Ser Ser Met Val Ile Phe Leu Ala 165 170 175 Ala Leu Lys Gln Val Pro Asn Glu Leu Tyr Glu Ala Ser Met Leu Asp 180 185 190 Gly Ala Ser Lys Ile Arg Arg Phe Phe Ser Ile Thr Leu Pro Met Ile 195 200 205 Ser Pro Val Leu Leu Phe Asn Leu Val Met Gln Thr Ile Asn Ala Phe 210 215 220 Gln Glu Phe Thr Gly Pro Tyr Val Ile Thr Gly Gly Gly Pro Met Asn 225 230 235 240 Ser Thr Tyr Val Tyr Ser Met Leu Ile Tyr Asp Asn Ala Phe Arg Tyr 245 250 255 Phe Arg Met Gly Tyr Ser Ser Ala Leu Ser Trp Ile Leu Phe Leu Leu 260 265 270 Ile Leu Ile Val Thr Val Ile Ile Phe Lys Ser Ser Asn Thr Trp Val 275 280 285 Tyr Tyr Glu Asn Gly Gly Arg 290 295 115888DNAT. saccharolyticum 115atgagttata ataaaaagag aaatttgatg gggtatttat atattagtcc atggattata 60ggctttttaa tatttactct gtatccattt gctatgactt ttatctattc attttgtaac 120tacagtatta caaaatcacc tgtatttatt ggattaggca attatataac tatgtttact 180aaagatatgt atttttggcc atctttaatt aatactataa aatatgtatt aatgacagtt 240cctttaaaat tatgttttgc actttttgtt gcaatgatct taaatattga tattaaagga 300gttaatgtgt ttagaacaac ttattatctg ccttctattt ttggaggaag tgttgcttta 360tctgttatat ggaaattttt attcatggat aatggtatta tgaataaatt tctttcatac 420tttcatatac acgggccaag ttggcttgga aacccacaca tatcattatt tactataagt 480ttattgtcag tgtgggaatt tgggtcttct atggtaatat ttttggcagc cctaaaacag 540gtcccgaatg agttgtatga agcatctatg ttagatggtg caagcaaaat aagaaggttt 600ttctcaataa ctttacctat gatatcgcct gtgctattat ttaatttggt tatgcagact 660ataaatgctt ttcaggaatt tacaggtcca tacgtgataa ctggtggagg accgatgaac 720tctacttatg tgtacagtat gttgatttat gataatgcgt ttaggtattt taggatgggt 780tattcatctg ccttgtcttg gattttattt ttgttaatat tgattgttac agttataata 840tttaaatctt caaatacatg ggtgtattac gaaaatggag gtagatga 888116285PRTT. saccharolyticum 116Met Lys Ala Lys Asn Ser Gln Asn Asn Asp Ile Ile Arg Lys Val Phe 1 5 10 15 Ile Tyr Val Phe Leu Val Ala Phe Gly Ile Phe Met Ile Tyr Pro Leu 20 25 30 Leu Trp Val Phe Ala Ser Ser Phe Lys Ser Asn Asp Glu Ile Phe Lys 35 40 45 Ser Ile Ser Leu Ile Pro Lys His Ile Val Thr Asn Ser Tyr Phe Glu 50 55 60 Gly Trp Lys Gly Thr Gly Gln Tyr Ser Phe Gly Thr Phe Ile Leu Asn 65 70 75 80 Ser Ile Thr Leu Val Val Pro Val Val Val Phe Thr Ala Ile Ser Ser 85 90 95 Thr Ile Val Ala Tyr Gly Phe Ala Arg Phe Glu Phe Pro Leu Lys Thr 100 105 110 Ile Leu Phe Thr Leu Met Ile Ser Thr Met Met Leu Pro Gly Thr Ala 115 120 125 Val Leu Ile Pro Arg Tyr Ile Leu Phe Asn Trp Leu Gly Trp Ile Asn 130 135 140 Thr Tyr Lys Pro Phe Ile Val Pro Ala Leu Phe Gly Thr Thr Pro Phe 145 150 155 160 Phe Ile Phe Met Met Val Gln Phe Leu Arg Gly Leu Pro Lys Glu Leu 165 170 175 Glu Glu Ser Ala Thr Ile Asp Gly Cys Asn Ser Phe Gln Ile Leu Met 180 185 190 Lys Ile Leu Ile Pro Leu Cys Lys Pro Ala Ile Ile Ser Met Cys Ile 195 200 205 Phe Gln Phe Ile Trp Thr Trp Asn Asp Phe Phe Asn Pro Leu Ile Tyr 210 215 220 Ile Asn Ser Val Glu Lys Tyr Thr Val Ser Leu Gly Leu Asn Met Thr 225 230 235 240 Ile Asp Gly Thr Ser Val Val Asn Trp Asn Gln Ile Met Ala Met Thr 245 250 255 Ile Ile Ser Met Ile Pro Ser Ile Ile Ile Phe Phe Ser Ala Gln Lys 260 265 270 Tyr Phe Val Glu Gly Ile Ala Thr Thr Gly Leu Lys Asn 275 280 285 117858DNAT. saccharolyticum 117atgaaagcaa agaatagtca aaataacgat ataatcagaa aagtatttat atatgttttc 60ttggtggctt ttggtatttt catgatatat cctttacttt gggtttttgc atcatcattt 120aaatcaaatg atgaaatctt taaatcgata agccttatac caaaacacat tgtgacaaat 180tcatattttg aaggatggaa aggtacggga caatactctt ttggtacatt tattttaaac 240agcattacgc ttgttgtacc tgttgttgta tttactgcta tatcatcaac aattgtagcc 300tatggatttg caagatttga gtttccgctt aaaactattt tgtttacttt gatgatatct 360actatgatgt tgccgggcac tgcagttttg ataccaagat atatattgtt taattggtta 420ggctggataa acacttataa accatttatt gttcccgctt tgttcggaac aacgcctttt 480ttcattttta tgatggttca atttttgaga ggtcttccta aagaattaga agaatcggct 540acaattgatg gttgcaattc atttcaaata cttatgaaga ttttaatacc attgtgtaaa 600cctgcaatta tttctatgtg tatatttcag ttcatttgga cttggaatga cttttttaat 660ccattgatat atatcaacag tgtagaaaaa tatacagttt ctctcgggct taatatgaca 720attgatggga cttcagttgt aaattggaac caaataatgg caatgacaat tatttcaatg 780ataccgagca tcataatatt tttttcagcg caaaaatact tcgttgaagg tattgcaaca 840actggattaa agaactaa 858118465PRTT. saccharolyticum 118Met Arg Tyr Thr Asp Gly Lys Val His Asp Ile Thr Ile Ala Tyr Ile 1 5 10 15 Gly Gly Gly Ser Arg Gly Trp Ala Trp Asn Leu Met Thr Asp Leu Ala 20 25 30 Lys Glu Glu Ser Ile Ser Gly Thr Val Lys Leu Tyr Asp Ile Asp Tyr 35 40 45 Asp Ala Ala His Asp Asn Glu Ile Ile Gly Asn Ala Leu Ser Met Arg 50 55 60 Gln Asp Val Lys Gly Lys Trp Leu Tyr Lys Ala Cys Glu Thr Leu Glu 65 70 75 80 Glu Ser Leu Lys Gly Ala Asp Phe Val Ile Ile Ser Ile Leu Pro Gly 85 90 95 Thr Phe Asp Glu Met Glu Ser Asp Val His Ala Pro Glu Lys Tyr Gly 100 105 110 Ile Tyr Gln Ser Val Gly Asp Thr Val Gly Pro Gly Gly Ile Val Arg 115 120 125 Ala Leu Arg Thr Ile Pro Met Phe Val Asp Ile Ala Asn Ala Ile Lys 130 135 140 Glu His Cys Pro Asp Ala Trp Val Ile Asn Tyr Thr Asn Pro Met Thr 145 150 155 160 Leu Cys Val Arg Thr Leu Tyr Glu Ile Phe Pro Gln Ile Lys Ala Phe 165 170 175 Gly Cys Cys His Glu Val Phe Gly Thr Gln Lys Leu Leu Ser Arg Ala 180 185 190 Leu Gln Asp Ile Glu Gly Ile Glu Asn Val Pro Arg Glu Glu Ile Lys 195 200 205 Ile Asn Val Leu Gly Ile Asn His Phe Thr Trp Ile Asp Asn Ala Arg 210 215 220 Tyr Lys Asp Ile Asp Leu Met Tyr Val Tyr Lys Gln Phe Val Asn Lys 225 230 235 240 Tyr Tyr Glu Ser Gly Phe Val Ser Asp Ala Asn Asn Asn Trp Met Asn 245 250 255 Asn Ser Phe Val Ser Ala Glu Arg Val Lys Phe Asp Leu Phe Leu Arg 260 265 270 Tyr Gly Val Ile Ala Ala Ala Gly Asp Arg His Leu Ala Glu Phe Val 275 280 285 Pro Gly Tyr Trp Tyr Leu Lys Asp Pro Glu Thr Val Arg Glu Trp Met 290 295 300 Phe Gly Leu Thr Thr Val Ser Trp Arg Lys Glu Asp Leu Lys Arg Arg 305 310 315 320 Leu Glu Arg Ser Lys Arg Leu Lys Thr Gly Glu Glu Lys Phe Glu Leu 325 330 335 Lys Glu Thr Gly Glu Glu Gly Val Arg Gln Ile Lys Ala Leu Leu Gly 340 345 350 Leu Gly Asp Leu Val Thr Asn Val Asn Met Pro Asn His Gly Gln Ile 355 360 365 Glu Gly Ile Pro Tyr Gly Ala Val Val Glu Thr Asn Ala Leu Phe Ser 370 375 380 Gly Asn Lys Leu Lys Pro Val Leu Ser Gly Lys Leu Pro Asp Asn Val 385 390 395 400 Asn Ser Leu Val Leu Arg Gln Val Tyr Asn Gln Glu Thr Thr Leu Lys 405 410 415 Ala Ala Leu Lys Arg Asp Phe Asp Leu Ala Phe Ser Ala Phe Val Asn 420 425 430 Asp Pro Leu Val Thr Ile Ser Leu Lys Asp Ala Lys Lys Leu Phe Lys 435 440 445 Glu Met Leu Glu Asn Thr Lys Lys Tyr Leu Asp Gly Trp Lys Ile Lys 450

455 460 Ala 465 1191398DNAT. saccharolyticum 119atgagatata cagatggaaa ggttcatgac attactattg cttatatcgg tggtggttca 60agaggatggg cgtggaattt aatgactgac ttagcaaaag aggaaagtat ttctggtaca 120gtaaagttat acgacataga ttacgatgcg gcacatgaca atgagataat aggcaatgct 180ttatcaatga gacaggatgt taaaggcaaa tggctttata aagcttgtga gacgttagaa 240gagtcactaa aaggtgctga ttttgtcata atatctattt tgccaggtac gttcgacgag 300atggaatctg atgttcatgc accagaaaag tatggcattt atcagtcagt aggtgataca 360gtaggacctg gtggaatagt cagagcttta aggacgattc cgatgtttgt ggacattgcc 420aatgcgatta aagagcattg tccagatgca tgggtcataa attatacaaa tcctatgaca 480ctttgtgtaa ggacattgta tgaaattttc cctcaaatta aagcatttgg atgctgccat 540gaagtttttg gcacacagaa gctattatct cgtgctctgc aggatataga aggcattgaa 600aatgttccga gggaagagat aaagataaat gttttaggta taaatcattt tacgtggatc 660gacaatgcaa ggtacaaaga catagattta atgtatgttt ataaacaatt tgtgaataag 720tactatgaaa gcggatttgt cagcgatgct aacaataatt ggatgaacaa ttcatttgta 780tctgcagaga gagtaaagtt tgatctgttt ttgaggtatg gagtaatagc tgcagcggga 840gatagacatc tggcggaatt tgtgccggga tattggtatt taaaagatcc agagacagtc 900agagaatgga tgtttggctt aacgactgta agttggagaa aagaagactt aaaacgcagg 960cttgaaagaa gtaaaaggct taagacaggt gaggaaaaat ttgagttaaa ggaaacaggc 1020gaagaaggtg ttaggcaaat taaagcacta ttaggcttag gcgatttagt gactaatgtc 1080aacatgccga accatggaca gattgaagga ataccatacg gtgcggtagt tgaaacaaac 1140gctttatttt caggtaataa actaaagcct gtattatcag gaaaattgcc tgacaatgta 1200aacagcctcg tgttaaggca agtatacaac caagaaacga cgttgaaagc tgctttaaag 1260agagattttg atttggcttt tagtgctttt gtaaatgatc cacttgttac aatatcttta 1320aaagatgcaa aaaaattatt taaggaaatg cttgaaaata cgaagaaata tctagatgga 1380tggaaaataa aagcttga 1398120392PRTC. acetobutylicum 120Met Lys Glu Val Val Ile Ala Ser Ala Val Arg Thr Ala Ile Gly Ser 1 5 10 15 Tyr Gly Lys Ser Leu Lys Asp Val Pro Ala Val Asp Leu Gly Ala Thr 20 25 30 Ala Ile Lys Glu Ala Val Lys Lys Ala Gly Ile Lys Pro Glu Asp Val 35 40 45 Asn Glu Val Ile Leu Gly Asn Val Leu Gln Ala Gly Leu Gly Gln Asn 50 55 60 Pro Ala Arg Gln Ala Ser Phe Lys Ala Gly Leu Pro Val Glu Ile Pro 65 70 75 80 Ala Met Thr Ile Asn Lys Val Cys Gly Ser Gly Leu Arg Thr Val Ser 85 90 95 Leu Ala Ala Gln Ile Ile Lys Ala Gly Asp Ala Asp Val Ile Ile Ala 100 105 110 Gly Gly Met Glu Asn Met Ser Arg Ala Pro Tyr Leu Ala Asn Asn Ala 115 120 125 Arg Trp Gly Tyr Arg Met Gly Asn Ala Lys Phe Val Asp Glu Met Ile 130 135 140 Thr Asp Gly Leu Trp Asp Ala Phe Asn Asp Tyr His Met Gly Ile Thr 145 150 155 160 Ala Glu Asn Ile Ala Glu Arg Trp Asn Ile Ser Arg Glu Glu Gln Asp 165 170 175 Glu Phe Ala Leu Ala Ser Gln Lys Lys Ala Glu Glu Ala Ile Lys Ser 180 185 190 Gly Gln Phe Lys Asp Glu Ile Val Pro Val Val Ile Lys Gly Arg Lys 195 200 205 Gly Glu Thr Val Val Asp Thr Asp Glu His Pro Arg Phe Gly Ser Thr 210 215 220 Ile Glu Gly Leu Ala Lys Leu Lys Pro Ala Phe Lys Lys Asp Gly Thr 225 230 235 240 Val Thr Ala Gly Asn Ala Ser Gly Leu Asn Asp Cys Ala Ala Val Leu 245 250 255 Val Ile Met Ser Ala Glu Lys Ala Lys Glu Leu Gly Val Lys Pro Leu 260 265 270 Ala Lys Ile Val Ser Tyr Gly Ser Ala Gly Val Asp Pro Ala Ile Met 275 280 285 Gly Tyr Gly Pro Phe Tyr Ala Thr Lys Ala Ala Ile Glu Lys Ala Gly 290 295 300 Trp Thr Val Asp Glu Leu Asp Leu Ile Glu Ser Asn Glu Ala Phe Ala 305 310 315 320 Ala Gln Ser Leu Ala Val Ala Lys Asp Leu Lys Phe Asp Met Asn Lys 325 330 335 Val Asn Val Asn Gly Gly Ala Ile Ala Leu Gly His Pro Ile Gly Ala 340 345 350 Ser Gly Ala Arg Ile Leu Val Thr Leu Val His Ala Met Gln Lys Arg 355 360 365 Asp Ala Lys Lys Gly Leu Ala Thr Leu Cys Ile Gly Gly Gly Gln Gly 370 375 380 Thr Ala Ile Leu Leu Glu Lys Cys 385 390 121218PRTC. acetobutylicum 121Met Asn Ser Lys Ile Ile Arg Phe Glu Asn Leu Arg Ser Phe Phe Lys 1 5 10 15 Asp Gly Met Thr Ile Met Ile Gly Gly Phe Leu Asn Cys Gly Thr Pro 20 25 30 Thr Lys Leu Ile Asp Phe Leu Val Asn Leu Asn Ile Lys Asn Leu Thr 35 40 45 Ile Ile Ser Asn Asp Thr Cys Tyr Pro Asn Thr Gly Ile Gly Lys Leu 50 55 60 Ile Ser Asn Asn Gln Val Lys Lys Leu Ile Ala Ser Tyr Ile Gly Ser 65 70 75 80 Asn Pro Asp Thr Gly Lys Lys Leu Phe Asn Asn Glu Leu Glu Val Glu 85 90 95 Leu Ser Pro Gln Gly Thr Leu Val Glu Arg Ile Arg Ala Gly Gly Ser 100 105 110 Gly Leu Gly Gly Val Leu Thr Lys Thr Gly Leu Gly Thr Leu Ile Glu 115 120 125 Lys Gly Lys Lys Lys Ile Ser Ile Asn Gly Thr Glu Tyr Leu Leu Glu 130 135 140 Leu Pro Leu Thr Ala Asp Val Ala Leu Ile Lys Gly Ser Ile Val Asp 145 150 155 160 Glu Ala Gly Asn Thr Phe Tyr Lys Gly Thr Thr Lys Asn Phe Asn Pro 165 170 175 Tyr Met Ala Met Ala Ala Lys Thr Val Ile Val Glu Ala Glu Asn Leu 180 185 190 Val Ser Cys Glu Lys Leu Glu Lys Glu Lys Ala Met Thr Pro Gly Val 195 200 205 Leu Ile Asn Tyr Ile Val Lys Glu Pro Ala 210 215 122221PRTC. acetobutylicum 122Met Ile Asn Asp Lys Asn Leu Ala Lys Glu Ile Ile Ala Lys Arg Val 1 5 10 15 Ala Arg Glu Leu Lys Asn Gly Gln Leu Val Asn Leu Gly Val Gly Leu 20 25 30 Pro Thr Met Val Ala Asp Tyr Ile Pro Lys Asn Phe Lys Ile Thr Phe 35 40 45 Gln Ser Glu Asn Gly Ile Val Gly Met Gly Ala Ser Pro Lys Ile Asn 50 55 60 Glu Ala Asp Lys Asp Val Val Asn Ala Gly Gly Asp Tyr Thr Thr Val 65 70 75 80 Leu Pro Asp Gly Thr Phe Phe Asp Ser Ser Val Ser Phe Ser Leu Ile 85 90 95 Arg Gly Gly His Val Asp Val Thr Val Leu Gly Ala Leu Gln Val Asp 100 105 110 Glu Lys Gly Asn Ile Ala Asn Trp Ile Val Pro Gly Lys Met Leu Ser 115 120 125 Gly Met Gly Gly Ala Met Asp Leu Val Asn Gly Ala Lys Lys Val Ile 130 135 140 Ile Ala Met Arg His Thr Asn Lys Gly Gln Pro Lys Ile Leu Lys Lys 145 150 155 160 Cys Thr Leu Pro Leu Thr Ala Lys Ser Gln Ala Asn Leu Ile Val Thr 165 170 175 Glu Leu Gly Val Ile Glu Val Ile Asn Asp Gly Leu Leu Leu Thr Glu 180 185 190 Ile Asn Lys Asn Thr Thr Ile Asp Glu Ile Arg Ser Leu Thr Ala Ala 195 200 205 Asp Leu Leu Ile Ser Asn Glu Leu Arg Pro Met Ala Val 210 215 220 123244PRTC. acetobutylicum 123Met Leu Lys Asp Glu Val Ile Lys Gln Ile Ser Thr Pro Leu Thr Ser 1 5 10 15 Pro Ala Phe Pro Arg Gly Pro Tyr Lys Phe His Asn Arg Glu Tyr Phe 20 25 30 Asn Ile Val Tyr Arg Thr Asp Met Asp Ala Leu Arg Lys Val Val Pro 35 40 45 Glu Pro Leu Glu Ile Asp Glu Pro Leu Val Arg Phe Glu Ile Met Ala 50 55 60 Met His Asp Thr Ser Gly Leu Gly Cys Tyr Thr Glu Ser Gly Gln Ala 65 70 75 80 Ile Pro Val Ser Phe Asn Gly Val Lys Gly Asp Tyr Leu His Met Met 85 90 95 Tyr Leu Asp Asn Glu Pro Ala Ile Ala Val Gly Arg Glu Leu Ser Ala 100 105 110 Tyr Pro Lys Lys Leu Gly Tyr Pro Lys Leu Phe Val Asp Ser Asp Thr 115 120 125 Leu Val Gly Thr Leu Asp Tyr Gly Lys Leu Arg Val Ala Thr Ala Thr 130 135 140 Met Gly Tyr Lys His Lys Ala Leu Asp Ala Asn Glu Ala Lys Asp Gln 145 150 155 160 Ile Cys Arg Pro Asn Tyr Met Leu Lys Ile Ile Pro Asn Tyr Asp Gly 165 170 175 Ser Pro Arg Ile Cys Glu Leu Ile Asn Ala Lys Ile Thr Asp Val Thr 180 185 190 Val His Glu Ala Trp Thr Gly Pro Thr Arg Leu Gln Leu Phe Asp His 195 200 205 Ala Met Ala Pro Leu Asn Asp Leu Pro Val Lys Glu Ile Val Ser Ser 210 215 220 Ser His Ile Leu Ala Asp Ile Ile Leu Pro Arg Ala Glu Val Ile Tyr 225 230 235 240 Asp Tyr Leu Lys 124244PRTC. acetobutylicum 124Met Leu Lys Asp Glu Val Ile Lys Gln Ile Ser Thr Pro Leu Thr Ser 1 5 10 15 Pro Ala Phe Pro Arg Gly Pro Tyr Lys Phe His Asn Arg Glu Tyr Phe 20 25 30 Asn Ile Val Tyr Arg Thr Asp Met Asp Ala Leu Arg Lys Val Val Pro 35 40 45 Glu Pro Leu Glu Ile Asp Glu Pro Leu Val Arg Phe Glu Ile Met Ala 50 55 60 Met His Asp Thr Ser Gly Leu Gly Cys Tyr Thr Glu Ser Gly Gln Ala 65 70 75 80 Ile Pro Val Ser Phe Asn Gly Val Lys Gly Asp Tyr Leu His Met Met 85 90 95 Tyr Leu Asp Asn Glu Pro Ala Ile Ala Val Gly Arg Glu Leu Ser Ala 100 105 110 Tyr Pro Lys Lys Leu Gly Tyr Pro Lys Leu Phe Val Asp Ser Asp Thr 115 120 125 Leu Val Gly Thr Leu Asp Tyr Gly Lys Leu Arg Val Ala Thr Ala Thr 130 135 140 Met Gly Tyr Lys His Lys Ala Leu Asp Ala Asn Glu Ala Lys Asp Gln 145 150 155 160 Ile Cys Arg Pro Asn Tyr Met Leu Lys Ile Ile Pro Asn Tyr Asp Gly 165 170 175 Ser Pro Arg Ile Cys Glu Leu Ile Asn Ala Lys Ile Thr Asp Val Thr 180 185 190 Val His Glu Ala Trp Thr Gly Pro Thr Arg Leu Gln Leu Phe Asp His 195 200 205 Ala Met Ala Pro Leu Asn Asp Leu Pro Val Lys Glu Ile Val Ser Ser 210 215 220 Ser His Ile Leu Ala Asp Ile Ile Leu Pro Arg Ala Glu Val Ile Tyr 225 230 235 240 Asp Tyr Leu Lys 125899PRTM. thermoacetica 125Met Val Asn Leu Thr Ile Asp Gly Gln Arg Val Thr Ala Pro Glu Gly 1 5 10 15 Met Thr Ile Leu Glu Val Ala Arg Glu Asn Gly Ile His Ile Pro Thr 20 25 30 Leu Cys His His Pro Lys Leu Arg Pro Leu Gly Tyr Cys Arg Leu Cys 35 40 45 Leu Val Asp Ile Glu Gly Ala Ala Lys Pro Met Thr Ala Cys Asn Thr 50 55 60 Pro Val Ala Glu Gly Met Val Ile Arg Thr Ser Thr Pro Val Ile Glu 65 70 75 80 Glu Met Arg Lys Gly Ile Ile Glu Met Leu Leu Ser Leu His Pro Glu 85 90 95 Asp Cys Leu Thr Cys Glu Lys Ala Gly Asn Cys Gln Leu Gln Asp Cys 100 105 110 Ala Tyr Thr Tyr Gly Val Lys His Gly Glu Leu Pro Val Lys Arg Glu 115 120 125 Glu Leu Pro Val Leu Lys Glu Asn Pro Phe Ile Val Arg Asp Tyr Asn 130 135 140 Lys Cys Ile Val Cys Gly Arg Cys Val Arg Ala Cys Gln Glu Val Gln 145 150 155 160 Val Gln Arg Val Val Asp Leu Val Gly Lys Gly Ser Ala Ala Arg Val 165 170 175 Gly Ala Thr Lys Ala Gly Ala Glu Val Ser Leu Glu Glu Gly Gly Cys 180 185 190 Val Phe Cys Gly Asn Cys Val Gln Val Cys Pro Val Gly Ala Leu Thr 195 200 205 Glu Lys Ala Gly Leu Gly Gln Gly Arg Glu Trp Glu Phe Lys Lys Val 210 215 220 Arg Ser Ile Cys Ser Tyr Cys Gly Val Gly Cys Asn Leu Thr Leu Tyr 225 230 235 240 Val Lys Asp Gly Lys Val Val Lys Val Arg Gly Tyr Glu Asn Pro Glu 245 250 255 Val Asn Asn Gly Trp Leu Cys Val Lys Gly Arg Phe Gly Phe Asp Tyr 260 265 270 Ile His Asn Pro Asp Arg Ile Thr Arg Pro Leu Ile Arg Glu Gly Asp 275 280 285 Arg Glu Lys Gly Tyr Phe Arg Glu Ala Ser Trp Glu Glu Ala Leu Ala 290 295 300 Leu Val Ser Gln Lys Leu Thr Gln Ile Lys Gly Ser Tyr Gly Ser Glu 305 310 315 320 Ala Leu Gly Phe Leu Cys Ser Ala Lys Cys Thr Asn Glu Glu Asn Tyr 325 330 335 Leu Leu Gln Lys Leu Ala Arg Gly Val Leu Gly Thr Asn Asn Val Asp 340 345 350 His Cys Ala Arg Leu His Ser Ser Thr Val Ala Gly Leu Ala Thr Thr 355 360 365 Phe Gly Ser Gly Ala Met Thr Asn Ser Ile Ala Asp Ile Ala Ser Ala 370 375 380 Asp Cys Ile Phe Val Ile Gly Ser Asn Thr Thr Glu Asn His Pro Val 385 390 395 400 Ile Ala Leu Lys Val Lys Glu Ala Val Arg Arg Gly Ala Arg Leu Ile 405 410 415 Val Ala Asp Pro Arg Arg Ile Glu Leu Val Asn Phe Ser Tyr Leu Trp 420 425 430 Leu Arg Gln Lys Pro Gly Thr Asp Leu Ala Leu Leu Asn Gly Leu Leu 435 440 445 His Val Ile Ile Lys Glu Glu Leu Tyr Asp Lys Glu Phe Ile Ala Gln 450 455 460 Arg Thr Glu Gly Phe Glu Ala Leu Lys Leu Ala Val Glu Glu Tyr Thr 465 470 475 480 Pro Ala Lys Val Ser Glu Val Thr Gly Val Pro Ala Gly Asp Ile Ile 485 490 495 Glu Ala Ala Arg Thr Tyr Ala Arg Gly Pro Ser Ser Thr Ile Leu Tyr 500 505 510 Ala Met Gly Ile Thr Gln His Ile Thr Gly Thr Ala Asn Val Met Ala 515 520 525 Leu Ala Asn Leu Ala Met Ala Cys Gly Gln Val Gly Lys Glu Gly Ser 530 535 540 Gly Val Asn Pro Leu Arg Gly Gln Ser Asn Val Gln Gly Ala Cys Asp 545 550 555 560 Met Gly Gly Leu Pro Asn Val Leu Pro Gly Tyr Gln Pro Val Thr Asp 565 570 575 Pro Gly Val Arg His Lys Phe Ser Glu Ala Trp Gly Val Pro Asp Leu 580 585 590 Pro Gly Glu Pro Gly Leu Thr Leu Met Glu Met Met Ala Ala Ala Gln 595 600 605 Glu Gly Lys Leu Lys Gly Met Tyr Ile Leu Gly Glu Asn Pro Val Leu 610 615 620 Thr Asp Pro Asp Val Ser His Val Lys Glu Ala Leu Lys Asn Leu Glu 625 630 635 640 Phe Leu Val Val Gln Asp Ile Phe Leu Thr Glu Thr Ala Arg Met Ala 645 650 655 Asp Val Val Leu Pro Gly Ala Ser Phe Ala Glu Lys Glu Gly Thr Phe 660 665 670 Thr Ser Thr Glu Arg Arg Val Gln Leu Leu His Lys Ala Ile Glu Pro 675 680 685 Pro Gly Glu Ala Arg Pro Asp Trp Leu Ile Leu Asn Asp Leu Leu Leu 690 695 700 Leu Met Gly Tyr Pro Arg Lys Tyr Ser Ser Pro Gly Glu Ile Met Gln 705 710 715 720 Glu Ile Ala Gly Leu Thr Pro Ser Tyr Ala Gly Ile Thr Tyr Glu Arg 725 730

735 Leu Glu Asp Lys Gly Leu Gln Trp Pro Val Leu Ser Leu Glu His Pro 740 745 750 Gly Thr Pro Val Leu His Arg Glu Lys Phe Ser Arg Gly Tyr Gly Gln 755 760 765 Phe Gln Val Val His Tyr Arg Pro Pro Ala Glu Glu Pro Asp Glu Glu 770 775 780 Tyr Pro Phe Leu Phe Thr Thr Gly Arg Asn Leu Tyr His Tyr His Thr 785 790 795 800 Val Ile Ser Arg Lys Ser Arg Gly Leu Glu Glu Met Cys Pro Ala Pro 805 810 815 Val Val Glu Ile Asn Asp Asn Asp Ala Ala Arg Leu Gly Ile Arg Glu 820 825 830 Gly Glu Met Ile Glu Ile Val Ser Arg Arg Gly Lys Val Arg Val Lys 835 840 845 Ala Leu Val Thr Asp Arg Ile Pro Arg Gly Gln Val Phe Met Asn Phe 850 855 860 His Phe His Glu Ala Ala Ala Asn Leu Leu Thr Ile Ala Ala Leu Asp 865 870 875 880 Pro Val Ala Lys Ile Pro Glu Tyr Lys Thr Cys Ala Val Ala Ile Lys 885 890 895 Val Lys Lys 126152PRTOryza sativa 126Met Glu Leu Thr Thr Arg Thr Ile Ala Glu Arg Lys His Ile Ala Leu 1 5 10 15 Val Ala His Asp His Arg Lys Gln Ala Leu Leu Glu Trp Val Glu Ser 20 25 30 His Lys Thr Ile Leu Ala Gln His Gln Leu Tyr Ala Thr Gly Thr Thr 35 40 45 Gly Asn Leu Ile Gln Arg Ala Ser Gly Ile Pro Val Thr Ser Met Leu 50 55 60 Ser Gly Pro Met Gly Gly Asp Gln Gln Val Gly Ala Leu Ile Ala Glu 65 70 75 80 Gly Lys Ile Asp Met Leu Ile Phe Phe Trp Asp Pro Leu Asn Ala Val 85 90 95 Pro His Asp Pro Asp Val Lys Ala Leu Leu Arg Leu Ala Thr Val Trp 100 105 110 Asn Ile Pro Val Ala Thr Asn Arg Ser Thr Ala Asp Phe Leu Ile Asp 115 120 125 Ser Pro Leu Phe Lys Ser Glu Val Ala Ile Ala Ile Pro Asp Tyr Gln 130 135 140 Arg Tyr Leu Gln Asp Arg Leu Lys 145 150 127350PRTT. saccharolyticum 127Met Lys Thr Ser Glu Leu Leu Ala Met Val Val Glu Lys Gly Ala Ser 1 5 10 15 Asp Leu His Ile Thr Val Gly Val Pro Pro Val Leu Arg Ile Asn Gly 20 25 30 Gln Leu Ile Lys Leu Asn Leu Pro Gln Leu Thr Pro Gln Asp Thr Glu 35 40 45 Glu Ile Thr Lys Asp Leu Leu Ser Ser Asp Glu Leu Lys Lys Leu Glu 50 55 60 Asp Met Gly Asp Ile Asp Leu Ser Tyr Ser Val Lys Gly Leu Gly Arg 65 70 75 80 Phe Arg Ile Asn Ala Tyr Lys Gln Arg Gly Thr Tyr Ser Leu Ala Ile 85 90 95 Arg Ser Val Ala Leu Arg Ile Pro Thr Ile Asp Glu Leu Gly Leu Pro 100 105 110 Glu Val Ile Lys Glu Leu Ala Leu Lys Thr Arg Gly Leu Ile Ile Val 115 120 125 Thr Gly Pro Thr Gly Ser Gly Lys Ser Thr Thr Leu Ala Ser Met Ile 130 135 140 Asp Leu Ile Asn Glu Glu Arg Asn Cys His Ile Leu Thr Leu Glu Asp 145 150 155 160 Pro Ile Glu Tyr Leu His Lys His Lys Lys Ser Ile Val Asn Gln Arg 165 170 175 Glu Ile Gly His Asp Ala Ala Ser Tyr Ala Ser Ala Leu Arg Ala Ala 180 185 190 Leu Arg Glu Asp Pro Asp Val Ile Leu Val Gly Glu Met Arg Asp Leu 195 200 205 Glu Thr Ile Gln Ile Ala Ile Thr Ala Ala Glu Thr Gly His Leu Val 210 215 220 Leu Ser Thr Leu His Thr Ile Gly Ser Ala Lys Thr Ile Asp Arg Ile 225 230 235 240 Ile Asp Val Phe Pro Pro His Gln Gln Gln Gln Ile Lys Val Gln Leu 245 250 255 Ser Asn Val Leu Glu Gly Ile Val Ser Gln Gln Leu Leu Pro Lys Ile 260 265 270 Asp Asn Ser Gly Arg Val Val Ala Val Glu Val Met Ile Ala Thr Pro 275 280 285 Ala Ile Arg Asn Leu Ile Arg Glu Gly Lys Ser Phe Gln Ile Gln Ser 290 295 300 Met Val Gln Thr Gly Asn Lys Phe Gly Met Val Thr Met Asp Met Trp 305 310 315 320 Ile Ser Gln Leu Leu Lys Arg Asn Leu Ile Ser Met Asp Asp Ala Leu 325 330 335 Thr Tyr Cys Val Asp Arg Glu Asn Phe Ser Arg Leu Val Val 340 345 350 128365PRTPseudomonas putida 128Met Asp Arg Ala Ile Gln Ser Pro Gly Lys Tyr Val Gln Gly Ala Asp 1 5 10 15 Ala Leu Gln Arg Leu Gly Asp Tyr Leu Lys Pro Leu Ala Asp Ser Trp 20 25 30 Leu Val Ile Ala Asp Lys Phe Val Leu Gly Phe Ala Glu Asp Thr Ile 35 40 45 Arg Gln Ser Leu Ser Lys Ala Gly Leu Ala Met Asp Ile Val Ala Phe 50 55 60 Asn Gly Glu Cys Ser Gln Gly Glu Val Asp Arg Leu Cys Gln Leu Ala 65 70 75 80 Thr Gln Asn Gly Arg Ser Ala Ile Val Gly Ile Gly Gly Gly Lys Thr 85 90 95 Leu Asp Thr Ala Lys Ala Val Ala Phe Phe Gln Lys Val Pro Val Ala 100 105 110 Val Ala Pro Thr Ile Ala Ser Thr Asp Ala Pro Cys Ser Ala Leu Ser 115 120 125 Val Leu Tyr Thr Asp Glu Gly Glu Phe Asp Arg Tyr Leu Met Leu Pro 130 135 140 Thr Asn Pro Ala Leu Val Val Val Asp Thr Ala Ile Val Ala Arg Ala 145 150 155 160 Pro Ala Arg Leu Leu Ala Ala Gly Ile Gly Asp Ala Leu Ala Thr Trp 165 170 175 Phe Glu Ala Arg Ala Ala Ser Arg Ser Ser Ala Ala Thr Met Ala Gly 180 185 190 Gly Pro Ala Thr Gln Thr Ala Leu Asn Leu Ala Arg Phe Cys Tyr Asp 195 200 205 Thr Leu Leu Glu Glu Gly Glu Lys Ala Met Leu Ala Val Gln Ala Gln 210 215 220 Val Val Thr Pro Ala Leu Glu Arg Ile Val Glu Ala Asn Thr Tyr Leu 225 230 235 240 Ser Gly Val Gly Phe Glu Ser Gly Gly Val Ala Ala Ala His Ala Val 245 250 255 His Asn Gly Leu Thr Ala Val Ala Glu Thr His His Phe Tyr His Gly 260 265 270 Glu Lys Val Ala Phe Gly Val Leu Val Gln Leu Ala Leu Glu Asn Ala 275 280 285 Ser Asn Ala Glu Met Gln Glu Val Met Ser Leu Cys His Ala Val Gly 290 295 300 Leu Pro Ile Thr Leu Ala Gln Leu Asp Ile Thr Glu Asp Ile Pro Thr 305 310 315 320 Lys Met Arg Ala Val Ala Glu Leu Ala Cys Ala Pro Gly Glu Thr Ile 325 330 335 His Asn Met Pro Gly Gly Val Thr Val Glu Gln Val Tyr Gly Ala Leu 340 345 350 Leu Val Ala Asp Gln Leu Gly Gln His Phe Leu Glu Phe 355 360 365 129564PRTT. saccharolyticum 129Met Ile Lys Lys Lys Leu Gly Asp Leu Leu Val Glu Val Gly Leu Leu 1 5 10 15 Asp Glu Ser Gln Leu Asn Asn Ala Ile Lys Ile Gln Lys Lys Thr Gly 20 25 30 Glu Lys Leu Gly Lys Ile Leu Val Lys Glu Gly Tyr Leu Thr Glu Glu 35 40 45 Gln Ile Ile Glu Ala Leu Glu Phe Gln Leu Gly Ile Pro His Ile Asp 50 55 60 Met Lys Lys Val Phe Ile Asp Ala Asn Val Ala Lys Leu Ile Pro Glu 65 70 75 80 Ser Met Ala Lys Arg His Val Ala Ile Pro Ile Lys Lys Glu Asn Asn 85 90 95 Ser Ile Phe Val Ala Met Ala Asp Pro Leu Asn Ile Phe Ala Ile Asp 100 105 110 Asp Ile Lys Leu Val Thr Lys Leu Asp Val Lys Pro Leu Ile Ala Ser 115 120 125 Glu Asp Gly Ile Leu Lys Ala Ile Asp Arg Val Phe Gly Lys Glu Glu 130 135 140 Ala Glu Arg Ala Val Gln Asp Phe Lys Lys Glu Leu Ser His Asp Ser 145 150 155 160 Ala Glu Asp Asp Gly Asn Leu Leu Arg Asp Ile Ser Glu Asp Glu Ile 165 170 175 Asn Asn Ala Pro Ala Val Arg Leu Val Asn Ser Ile Ile Glu Gln Ala 180 185 190 Val Lys Asn Arg Ala Ser Asp Val His Ile Glu Pro Thr Glu Asn Asp 195 200 205 Leu Arg Ile Arg Phe Arg Ile Asp Gly Glu Leu His Glu Ala Met Arg 210 215 220 Val Phe Lys Ser Thr Gln Gly Pro Val Ile Thr Arg Ile Lys Ile Met 225 230 235 240 Ala Asn Met Asn Ile Ala Glu Arg Arg Ile Pro Gln Asp Gly Lys Ile 245 250 255 Glu Met Asn Ala Gly Gly Lys Asn Ile Asp Ile Arg Val Ser Ser Leu 260 265 270 Pro Thr Ile Tyr Gly Glu Lys Leu Val Leu Arg Ile Leu Asp Lys Ser 275 280 285 Gly Tyr Ile Ile Thr Lys Asp Lys Leu Gly Leu Gly Asn Asp Asp Leu 290 295 300 Lys Leu Phe Asp Asn Leu Leu Lys His Pro Asn Gly Ile Ile Leu Leu 305 310 315 320 Thr Gly Pro Thr Gly Ser Gly Lys Thr Thr Thr Leu Tyr Ala Met Leu 325 330 335 Asn Glu Leu Asn Lys Pro Asp Lys Asn Ile Ile Thr Val Glu Asp Pro 340 345 350 Val Glu Tyr Thr Leu Glu Gly Leu Asn Gln Val Gln Val Asn Glu Lys 355 360 365 Ala Gly Leu Thr Phe Ala Ser Ala Leu Arg Ser Ile Leu Arg Gln Asp 370 375 380 Pro Asp Ile Ile Met Ile Gly Glu Ile Arg Asp Arg Glu Thr Ala Glu 385 390 395 400 Ile Ala Ile Arg Ser Ser Ile Thr Gly His Leu Val Leu Ser Thr Leu 405 410 415 His Thr Asn Asp Ser Ala Gly Ala Ile Thr Arg Leu Ile Asp Met Gly 420 425 430 Ile Glu Pro Tyr Leu Val Ser Ser Ser Val Val Gly Val Ile Ala Gln 435 440 445 Arg Leu Ala Arg Lys Ile Cys Asp Asn Cys Lys Ile Glu Tyr Asp Ala 450 455 460 Ser Lys Arg Glu Lys Ile Ile Leu Gly Ile Asp Ala Asp Glu Ser Leu 465 470 475 480 Lys Leu Tyr Arg Ser Lys Gly Cys Ala Val Cys Asn Lys Thr Gly Tyr 485 490 495 Arg Gly Arg Val Pro Ile Tyr Glu Ile Met Met Met Thr Pro Lys Ile 500 505 510 Lys Glu Leu Thr Asn Glu Lys Ala Pro Ala Asp Val Ile Leu Asn Glu 515 520 525 Ala Val Ser Asn Gly Met Ser Thr Leu Lys Glu Ser Ala Lys Lys Leu 530 535 540 Val Leu Ser Gly Val Thr Thr Val Asp Glu Met Leu Arg Leu Thr Tyr 545 550 555 560 Asp Asp Ala Tyr 130218PRTC. acetobutylicum 130Met Asn Ser Lys Ile Ile Arg Phe Glu Asn Leu Arg Ser Phe Phe Lys 1 5 10 15 Asp Gly Met Thr Ile Met Ile Gly Gly Phe Leu Asn Cys Gly Thr Pro 20 25 30 Thr Lys Leu Ile Asp Phe Leu Val Asn Leu Asn Ile Lys Asn Leu Thr 35 40 45 Ile Ile Ser Asn Asp Thr Cys Tyr Pro Asn Thr Gly Ile Gly Lys Leu 50 55 60 Ile Ser Asn Asn Gln Val Lys Lys Leu Ile Ala Ser Tyr Ile Gly Ser 65 70 75 80 Asn Pro Asp Thr Gly Lys Lys Leu Phe Asn Asn Glu Leu Glu Val Glu 85 90 95 Leu Ser Pro Gln Gly Thr Leu Val Glu Arg Ile Arg Ala Gly Gly Ser 100 105 110 Gly Leu Gly Gly Val Leu Thr Lys Thr Gly Leu Gly Thr Leu Ile Glu 115 120 125 Lys Gly Lys Lys Lys Ile Ser Ile Asn Gly Thr Glu Tyr Leu Leu Glu 130 135 140 Leu Pro Leu Thr Ala Asp Val Ala Leu Ile Lys Gly Ser Ile Val Asp 145 150 155 160 Glu Ala Gly Asn Thr Phe Tyr Lys Gly Thr Thr Lys Asn Phe Asn Pro 165 170 175 Tyr Met Ala Met Ala Ala Lys Thr Val Ile Val Glu Ala Glu Asn Leu 180 185 190 Val Ser Cys Glu Lys Leu Glu Lys Glu Lys Ala Met Thr Pro Gly Val 195 200 205 Leu Ile Asn Tyr Ile Val Lys Glu Pro Ala 210 215 131221PRTC. acetobutylicum 131Met Ile Asn Asp Lys Asn Leu Ala Lys Glu Ile Ile Ala Lys Arg Val 1 5 10 15 Ala Arg Glu Leu Lys Asn Gly Gln Leu Val Asn Leu Gly Val Gly Leu 20 25 30 Pro Thr Met Val Ala Asp Tyr Ile Pro Lys Asn Phe Lys Ile Thr Phe 35 40 45 Gln Ser Glu Asn Gly Ile Val Gly Met Gly Ala Ser Pro Lys Ile Asn 50 55 60 Glu Ala Asp Lys Asp Val Val Asn Ala Gly Gly Asp Tyr Thr Thr Val 65 70 75 80 Leu Pro Asp Gly Thr Phe Phe Asp Ser Ser Val Ser Phe Ser Leu Ile 85 90 95 Arg Gly Gly His Val Asp Val Thr Val Leu Gly Ala Leu Gln Val Asp 100 105 110 Glu Lys Gly Asn Ile Ala Asn Trp Ile Val Pro Gly Lys Met Leu Ser 115 120 125 Gly Met Gly Gly Ala Met Asp Leu Val Asn Gly Ala Lys Lys Val Ile 130 135 140 Ile Ala Met Arg His Thr Asn Lys Gly Gln Pro Lys Ile Leu Lys Lys 145 150 155 160 Cys Thr Leu Pro Leu Thr Ala Lys Ser Gln Ala Asn Leu Ile Val Thr 165 170 175 Glu Leu Gly Val Ile Glu Val Ile Asn Asp Gly Leu Leu Leu Thr Glu 180 185 190 Ile Asn Lys Asn Thr Thr Ile Asp Glu Ile Arg Ser Leu Thr Ala Ala 195 200 205 Asp Leu Leu Ile Ser Asn Glu Leu Arg Pro Met Ala Val 210 215 220 132244PRTC. acetobutylicum 132Met Leu Lys Asp Glu Val Ile Lys Gln Ile Ser Thr Pro Leu Thr Ser 1 5 10 15 Pro Ala Phe Pro Arg Gly Pro Tyr Lys Phe His Asn Arg Glu Tyr Phe 20 25 30 Asn Ile Val Tyr Arg Thr Asp Met Asp Ala Leu Arg Lys Val Val Pro 35 40 45 Glu Pro Leu Glu Ile Asp Glu Pro Leu Val Arg Phe Glu Ile Met Ala 50 55 60 Met His Asp Thr Ser Gly Leu Gly Cys Tyr Thr Glu Ser Gly Gln Ala 65 70 75 80 Ile Pro Val Ser Phe Asn Gly Val Lys Gly Asp Tyr Leu His Met Met 85 90 95 Tyr Leu Asp Asn Glu Pro Ala Ile Ala Val Gly Arg Glu Leu Ser Ala 100 105 110 Tyr Pro Lys Lys Leu Gly Tyr Pro Lys Leu Phe Val Asp Ser Asp Thr 115 120 125 Leu Val Gly Thr Leu Asp Tyr Gly Lys Leu Arg Val Ala Thr Ala Thr 130 135 140 Met Gly Tyr Lys His Lys Ala Leu Asp Ala Asn Glu Ala Lys Asp Gln 145 150 155 160 Ile Cys Arg Pro Asn Tyr Met Leu Lys Ile Ile Pro Asn Tyr Asp Gly 165 170 175 Ser Pro Arg Ile Cys Glu Leu Ile Asn Ala Lys Ile Thr Asp Val Thr 180 185 190 Val His Glu Ala Trp Thr Gly Pro Thr Arg Leu Gln Leu Phe Asp His 195 200 205 Ala Met Ala Pro Leu Asn Asp Leu Pro Val Lys Glu Ile Val Ser Ser 210 215 220 Ser His Ile Leu Ala Asp Ile Ile Leu Pro Arg Ala Glu Val Ile Tyr 225 230 235 240 Asp Tyr Leu Lys 133246PRTEscherichia

coli 133Met Ser Val Ile Gly Arg Ile His Ser Phe Glu Ser Cys Gly Thr Val 1 5 10 15 Asp Gly Pro Gly Ile Arg Phe Ile Thr Phe Phe Gln Gly Cys Leu Met 20 25 30 Arg Cys Leu Tyr Cys His Asn Arg Asp Thr Trp Asp Thr His Gly Gly 35 40 45 Lys Glu Val Thr Val Glu Asp Leu Met Lys Glu Val Val Thr Tyr Arg 50 55 60 His Phe Met Asn Ala Ser Gly Gly Gly Val Thr Ala Ser Gly Gly Glu 65 70 75 80 Ala Ile Leu Gln Ala Glu Phe Val Arg Asp Trp Phe Arg Ala Cys Lys 85 90 95 Lys Glu Gly Ile His Thr Cys Leu Asp Thr Asn Gly Phe Val Arg Arg 100 105 110 Tyr Asp Pro Val Ile Asp Glu Leu Leu Glu Val Thr Asp Leu Val Met 115 120 125 Leu Asp Leu Lys Gln Met Asn Asp Glu Ile His Gln Asn Leu Val Gly 130 135 140 Val Ser Asn His Arg Thr Leu Glu Phe Ala Lys Tyr Leu Ala Asn Lys 145 150 155 160 Asn Val Lys Val Trp Ile Arg Tyr Val Val Val Pro Gly Trp Ser Asp 165 170 175 Asp Asp Asp Ser Ala His Arg Leu Gly Glu Phe Thr Arg Asp Met Gly 180 185 190 Asn Val Glu Lys Ile Glu Leu Leu Pro Tyr His Glu Leu Gly Lys His 195 200 205 Lys Trp Val Ala Met Gly Glu Glu Tyr Lys Leu Asp Gly Val Lys Pro 210 215 220 Pro Lys Lys Glu Thr Met Glu Arg Val Lys Gly Ile Leu Glu Gln Tyr 225 230 235 240 Gly His Lys Val Met Phe 245 134760PRTEscherichia coli 134Met Ser Glu Leu Asn Glu Lys Leu Ala Thr Ala Trp Glu Gly Phe Thr 1 5 10 15 Lys Gly Asp Trp Gln Asn Glu Val Asn Val Arg Asp Phe Ile Gln Lys 20 25 30 Asn Tyr Thr Pro Tyr Glu Gly Asp Glu Ser Phe Leu Ala Gly Ala Thr 35 40 45 Glu Ala Thr Thr Thr Leu Trp Asp Lys Val Met Glu Gly Val Lys Leu 50 55 60 Glu Asn Arg Thr His Ala Pro Val Asp Phe Asp Thr Ala Val Ala Ser 65 70 75 80 Thr Ile Thr Ser His Asp Ala Gly Tyr Ile Asn Lys Gln Leu Glu Lys 85 90 95 Ile Val Gly Leu Gln Thr Glu Ala Pro Leu Lys Arg Ala Leu Ile Pro 100 105 110 Phe Gly Gly Ile Lys Met Ile Glu Gly Ser Cys Lys Ala Tyr Asn Arg 115 120 125 Glu Leu Asp Pro Met Ile Lys Lys Ile Phe Thr Glu Tyr Arg Lys Thr 130 135 140 His Asn Gln Gly Val Phe Asp Val Tyr Thr Pro Asp Ile Leu Arg Cys 145 150 155 160 Arg Lys Ser Gly Val Leu Thr Gly Leu Pro Asp Ala Tyr Gly Arg Gly 165 170 175 Arg Ile Ile Gly Asp Tyr Arg Arg Val Ala Leu Tyr Gly Ile Asp Tyr 180 185 190 Leu Met Lys Asp Lys Leu Ala Gln Phe Thr Ser Leu Gln Ala Asp Leu 195 200 205 Glu Asn Gly Val Asn Leu Glu Gln Thr Ile Arg Leu Arg Glu Glu Ile 210 215 220 Ala Glu Gln His Arg Ala Leu Gly Gln Met Lys Glu Met Ala Ala Lys 225 230 235 240 Tyr Gly Tyr Asp Ile Ser Gly Pro Ala Thr Asn Ala Gln Glu Ala Ile 245 250 255 Gln Trp Thr Tyr Phe Gly Tyr Leu Ala Ala Val Lys Ser Gln Asn Gly 260 265 270 Ala Ala Met Ser Phe Gly Arg Thr Ser Thr Phe Leu Asp Val Tyr Ile 275 280 285 Glu Arg Asp Leu Lys Ala Gly Lys Ile Thr Glu Gln Glu Ala Gln Glu 290 295 300 Met Val Asp His Leu Val Met Lys Leu Arg Met Val Arg Phe Leu Arg 305 310 315 320 Thr Pro Glu Tyr Asp Glu Leu Phe Ser Gly Asp Pro Ile Trp Ala Thr 325 330 335 Glu Ser Ile Gly Gly Met Gly Leu Asp Gly Arg Thr Leu Val Thr Lys 340 345 350 Asn Ser Phe Arg Phe Leu Asn Thr Leu Tyr Thr Met Gly Pro Ser Pro 355 360 365 Glu Pro Asn Met Thr Ile Leu Trp Ser Glu Lys Leu Pro Leu Asn Phe 370 375 380 Lys Lys Phe Ala Ala Lys Val Ser Ile Asp Thr Ser Ser Leu Gln Tyr 385 390 395 400 Glu Asn Asp Asp Leu Met Arg Pro Asp Phe Asn Asn Asp Asp Tyr Ala 405 410 415 Ile Ala Cys Cys Val Ser Pro Met Ile Val Gly Lys Gln Met Gln Phe 420 425 430 Phe Gly Ala Arg Ala Asn Leu Ala Lys Thr Met Leu Tyr Ala Ile Asn 435 440 445 Gly Gly Val Asp Glu Lys Leu Lys Met Gln Val Gly Pro Lys Ser Glu 450 455 460 Pro Ile Lys Gly Asp Val Leu Asn Tyr Asp Glu Val Met Glu Arg Met 465 470 475 480 Asp His Phe Met Asp Trp Leu Ala Lys Gln Tyr Ile Thr Ala Leu Asn 485 490 495 Ile Ile His Tyr Met His Asp Lys Tyr Ser Tyr Glu Ala Ser Leu Met 500 505 510 Ala Leu His Asp Arg Asp Val Ile Arg Thr Met Ala Cys Gly Ile Ala 515 520 525 Gly Leu Ser Val Ala Ala Asp Ser Leu Ser Ala Ile Lys Tyr Ala Lys 530 535 540 Val Lys Pro Ile Arg Asp Glu Asp Gly Leu Ala Ile Asp Phe Glu Ile 545 550 555 560 Glu Gly Glu Tyr Pro Gln Phe Gly Asn Asn Asp Pro Arg Val Asp Asp 565 570 575 Leu Ala Val Asp Leu Val Glu Arg Phe Met Lys Lys Ile Gln Lys Leu 580 585 590 His Thr Tyr Arg Asp Ala Ile Pro Thr Gln Ser Val Leu Thr Ile Thr 595 600 605 Ser Asn Val Val Tyr Gly Lys Lys Thr Gly Asn Thr Pro Asp Gly Arg 610 615 620 Arg Ala Gly Ala Pro Phe Gly Pro Gly Ala Asn Pro Met His Gly Arg 625 630 635 640 Asp Gln Lys Gly Ala Val Ala Ser Leu Thr Ser Val Ala Lys Leu Pro 645 650 655 Phe Ala Tyr Ala Lys Asp Gly Ile Ser Tyr Thr Phe Ser Ile Val Pro 660 665 670 Asn Ala Leu Gly Lys Asp Asp Glu Val Arg Lys Thr Asn Leu Ala Gly 675 680 685 Leu Met Asp Gly Tyr Phe His His Glu Ala Ser Ile Glu Gly Gly Gln 690 695 700 His Leu Asn Val Asn Val Met Asn Arg Glu Met Leu Leu Asp Ala Met 705 710 715 720 Glu Asn Pro Glu Lys Tyr Pro Gln Leu Thr Ile Arg Val Ser Gly Tyr 725 730 735 Ala Val Arg Phe Asn Ser Leu Thr Lys Glu Gln Gln Gln Asp Val Ile 740 745 750 Thr Arg Thr Phe Thr Gln Ser Met 755 760 135398PRTSaccharomyces cerevisiae 135Met Ser Gln Asn Val Tyr Ile Val Ser Thr Ala Arg Thr Pro Ile Gly 1 5 10 15 Ser Phe Gln Gly Ser Leu Ser Ser Lys Thr Ala Val Glu Leu Gly Ala 20 25 30 Val Ala Leu Lys Gly Ala Leu Ala Lys Val Pro Glu Leu Asp Ala Ser 35 40 45 Lys Asp Phe Asp Glu Ile Ile Phe Gly Asn Val Leu Ser Ala Asn Leu 50 55 60 Gly Gln Ala Pro Ala Arg Gln Val Ala Leu Ala Ala Gly Leu Ser Asn 65 70 75 80 His Ile Val Ala Ser Thr Val Asn Lys Val Cys Ala Ser Ala Met Lys 85 90 95 Ala Ile Ile Leu Gly Ala Gln Ser Ile Lys Cys Gly Asn Ala Asp Val 100 105 110 Val Val Ala Gly Gly Cys Glu Ser Met Thr Asn Ala Pro Tyr Tyr Met 115 120 125 Pro Ala Ala Arg Ala Gly Ala Lys Phe Gly Gln Thr Val Leu Val Asp 130 135 140 Gly Val Glu Arg Asp Gly Leu Asn Asp Ala Tyr Asp Gly Leu Ala Met 145 150 155 160 Gly Val His Ala Glu Lys Cys Ala Arg Asp Trp Asp Ile Thr Arg Glu 165 170 175 Gln Gln Asp Asn Phe Ala Ile Glu Ser Tyr Gln Lys Ser Gln Lys Ser 180 185 190 Gln Lys Glu Gly Lys Phe Asp Asn Glu Ile Val Pro Val Thr Ile Lys 195 200 205 Gly Phe Arg Gly Lys Pro Asp Thr Gln Val Thr Lys Asp Glu Glu Pro 210 215 220 Ala Arg Leu His Val Glu Lys Leu Arg Ser Ala Arg Thr Val Phe Gln 225 230 235 240 Lys Glu Asn Gly Thr Val Thr Ala Ala Asn Ala Ser Pro Ile Asn Asp 245 250 255 Gly Ala Ala Ala Val Ile Leu Val Ser Glu Lys Val Leu Lys Glu Lys 260 265 270 Asn Leu Lys Pro Leu Ala Ile Ile Lys Gly Trp Gly Glu Ala Ala His 275 280 285 Gln Pro Ala Asp Phe Thr Trp Ala Pro Ser Leu Ala Val Pro Lys Ala 290 295 300 Leu Lys His Ala Gly Ile Glu Asp Ile Asn Ser Val Asp Tyr Phe Glu 305 310 315 320 Phe Asn Glu Ala Phe Ser Val Val Gly Leu Val Asn Thr Lys Ile Leu 325 330 335 Lys Leu Asp Pro Ser Lys Val Asn Val Tyr Gly Gly Ala Val Ala Leu 340 345 350 Gly His Pro Leu Gly Cys Ser Gly Ala Arg Val Val Val Thr Leu Leu 355 360 365 Ser Ile Leu Gln Gln Glu Gly Gly Lys Ile Gly Val Ala Ala Ile Cys 370 375 380 Asn Gly Gly Gly Gly Ala Ser Ser Ile Val Ile Glu Lys Ile 385 390 395 136348PRTSaccharomyces cerevisiae 136Met Ser Ile Pro Glu Thr Gln Lys Gly Val Ile Phe Tyr Glu Ser His 1 5 10 15 Gly Lys Leu Glu Tyr Lys Asp Ile Pro Val Pro Lys Pro Lys Ala Asn 20 25 30 Glu Leu Leu Ile Asn Val Lys Tyr Ser Gly Val Cys His Thr Asp Leu 35 40 45 His Ala Trp His Gly Asp Trp Pro Leu Pro Val Lys Leu Pro Leu Val 50 55 60 Gly Gly His Glu Gly Ala Gly Val Val Val Gly Met Gly Glu Asn Val 65 70 75 80 Lys Gly Trp Lys Ile Gly Asp Tyr Ala Gly Ile Lys Trp Leu Asn Gly 85 90 95 Ser Cys Met Ala Cys Glu Tyr Cys Glu Leu Gly Asn Glu Ser Asn Cys 100 105 110 Pro His Ala Asp Leu Ser Gly Tyr Thr His Asp Gly Ser Phe Gln Gln 115 120 125 Tyr Ala Thr Ala Asp Ala Val Gln Ala Ala His Ile Pro Gln Gly Thr 130 135 140 Asp Leu Ala Gln Val Ala Pro Ile Leu Cys Ala Gly Ile Thr Val Tyr 145 150 155 160 Lys Ala Leu Lys Ser Ala Asn Leu Met Ala Gly His Trp Val Ala Ile 165 170 175 Ser Gly Ala Ala Gly Gly Leu Gly Ser Leu Ala Val Gln Tyr Ala Lys 180 185 190 Ala Met Gly Tyr Arg Val Leu Gly Ile Asp Gly Gly Glu Gly Lys Glu 195 200 205 Glu Leu Phe Arg Ser Ile Gly Gly Glu Val Phe Ile Asp Phe Thr Lys 210 215 220 Glu Lys Asp Ile Val Gly Ala Val Leu Lys Ala Thr Asp Gly Gly Ala 225 230 235 240 His Gly Val Ile Asn Val Ser Val Ser Glu Ala Ala Ile Glu Ala Ser 245 250 255 Thr Arg Tyr Val Arg Ala Asn Gly Thr Thr Val Leu Val Gly Met Pro 260 265 270 Ala Gly Ala Lys Cys Cys Ser Asp Val Phe Asn Gln Val Val Lys Ser 275 280 285 Ile Ser Ile Val Gly Ser Tyr Val Gly Asn Arg Ala Asp Thr Arg Glu 290 295 300 Ala Leu Asp Phe Phe Ala Arg Gly Leu Val Lys Ser Pro Ile Lys Val 305 310 315 320 Val Gly Leu Ser Thr Leu Pro Glu Ile Tyr Glu Lys Met Glu Lys Gly 325 330 335 Gln Ile Val Gly Arg Tyr Val Val Asp Thr Ser Lys 340 345 137348PRTSaccharomyces cerevisiae 137Met Ser Ile Pro Glu Thr Gln Lys Ala Ile Ile Phe Tyr Glu Ser Asn 1 5 10 15 Gly Lys Leu Glu His Lys Asp Ile Pro Val Pro Lys Pro Lys Pro Asn 20 25 30 Glu Leu Leu Ile Asn Val Lys Tyr Ser Gly Val Cys His Thr Asp Leu 35 40 45 His Ala Trp His Gly Asp Trp Pro Leu Pro Thr Lys Leu Pro Leu Val 50 55 60 Gly Gly His Glu Gly Ala Gly Val Val Val Gly Met Gly Glu Asn Val 65 70 75 80 Lys Gly Trp Lys Ile Gly Asp Tyr Ala Gly Ile Lys Trp Leu Asn Gly 85 90 95 Ser Cys Met Ala Cys Glu Tyr Cys Glu Leu Gly Asn Glu Ser Asn Cys 100 105 110 Pro His Ala Asp Leu Ser Gly Tyr Thr His Asp Gly Ser Phe Gln Glu 115 120 125 Tyr Ala Thr Ala Asp Ala Val Gln Ala Ala His Ile Pro Gln Gly Thr 130 135 140 Asp Leu Ala Glu Val Ala Pro Ile Leu Cys Ala Gly Ile Thr Val Tyr 145 150 155 160 Lys Ala Leu Lys Ser Ala Asn Leu Arg Ala Gly His Trp Ala Ala Ile 165 170 175 Ser Gly Ala Ala Gly Gly Leu Gly Ser Leu Ala Val Gln Tyr Ala Lys 180 185 190 Ala Met Gly Tyr Arg Val Leu Gly Ile Asp Gly Gly Pro Gly Lys Glu 195 200 205 Glu Leu Phe Thr Ser Leu Gly Gly Glu Val Phe Ile Asp Phe Thr Lys 210 215 220 Glu Lys Asp Ile Val Ser Ala Val Val Lys Ala Thr Asn Gly Gly Ala 225 230 235 240 His Gly Ile Ile Asn Val Ser Val Ser Glu Ala Ala Ile Glu Ala Ser 245 250 255 Thr Arg Tyr Cys Arg Ala Asn Gly Thr Val Val Leu Val Gly Leu Pro 260 265 270 Ala Gly Ala Lys Cys Ser Ser Asp Val Phe Asn His Val Val Lys Ser 275 280 285 Ile Ser Ile Val Gly Ser Tyr Val Gly Asn Arg Ala Asp Thr Arg Glu 290 295 300 Ala Leu Asp Phe Phe Ala Arg Gly Leu Val Lys Ser Pro Ile Lys Val 305 310 315 320 Val Gly Leu Ser Ser Leu Pro Glu Ile Tyr Glu Lys Met Glu Lys Gly 325 330 335 Gln Ile Ala Gly Arg Tyr Val Val Asp Thr Ser Lys 340 345 138375PRTSaccharomyces cerevisiae 138Met Leu Arg Thr Ser Thr Leu Phe Thr Arg Arg Val Gln Pro Ser Leu 1 5 10 15 Phe Ser Arg Asn Ile Leu Arg Leu Gln Ser Thr Ala Ala Ile Pro Lys 20 25 30 Thr Gln Lys Gly Val Ile Phe Tyr Glu Asn Lys Gly Lys Leu His Tyr 35 40 45 Lys Asp Ile Pro Val Pro Glu Pro Lys Pro Asn Glu Ile Leu Ile Asn 50 55 60 Val Lys Tyr Ser Gly Val Cys His Thr Asp Leu His Ala Trp His Gly 65 70 75 80 Asp Trp Pro Leu Pro Val Lys Leu Pro Leu Val Gly Gly His Glu Gly 85 90 95 Ala Gly Val Val Val Lys Leu Gly Ser Asn Val Lys Gly Trp Lys Val 100 105 110 Gly Asp Leu Ala Gly Ile Lys Trp Leu Asn Gly Ser Cys Met Thr Cys 115 120 125 Glu Phe Cys Glu Ser Gly His Glu Ser Asn Cys Pro Asp Ala Asp Leu 130 135 140 Ser Gly Tyr Thr His Asp Gly Ser Phe Gln Gln Phe Ala Thr Ala Asp 145 150 155 160 Ala Ile Gln Ala Ala Lys Ile Gln Gln Gly Thr Asp Leu Ala Glu Val 165 170 175 Ala Pro Ile Leu

Cys Ala Gly Val Thr Val Tyr Lys Ala Leu Lys Glu 180 185 190 Ala Asp Leu Lys Ala Gly Asp Trp Val Ala Ile Ser Gly Ala Ala Gly 195 200 205 Gly Leu Gly Ser Leu Ala Val Gln Tyr Ala Thr Ala Met Gly Tyr Arg 210 215 220 Val Leu Gly Ile Asp Ala Gly Glu Glu Lys Glu Lys Leu Phe Lys Lys 225 230 235 240 Leu Gly Gly Glu Val Phe Ile Asp Phe Thr Lys Thr Lys Asn Met Val 245 250 255 Ser Asp Ile Gln Glu Ala Thr Lys Gly Gly Pro His Gly Val Ile Asn 260 265 270 Val Ser Val Ser Glu Ala Ala Ile Ser Leu Ser Thr Glu Tyr Val Arg 275 280 285 Pro Cys Gly Thr Val Val Leu Val Gly Leu Pro Ala Asn Ala Tyr Val 290 295 300 Lys Ser Glu Val Phe Ser His Val Val Lys Ser Ile Asn Ile Lys Gly 305 310 315 320 Ser Tyr Val Gly Asn Arg Ala Asp Thr Arg Glu Ala Leu Asp Phe Phe 325 330 335 Ser Arg Gly Leu Ile Lys Ser Pro Ile Lys Ile Val Gly Leu Ser Glu 340 345 350 Leu Pro Lys Val Tyr Asp Leu Met Glu Lys Gly Lys Ile Leu Gly Arg 355 360 365 Tyr Val Val Asp Thr Ser Lys 370 375 139382PRTSaccharomyces cerevisiae 139Met Ser Ser Val Thr Gly Phe Tyr Ile Pro Pro Ile Ser Phe Phe Gly 1 5 10 15 Glu Gly Ala Leu Glu Glu Thr Ala Asp Tyr Ile Lys Asn Lys Asp Tyr 20 25 30 Lys Lys Ala Leu Ile Val Thr Asp Pro Gly Ile Ala Ala Ile Gly Leu 35 40 45 Ser Gly Arg Val Gln Lys Met Leu Glu Glu Arg Asp Leu Asn Val Ala 50 55 60 Ile Tyr Asp Lys Thr Gln Pro Asn Pro Asn Ile Ala Asn Val Thr Ala 65 70 75 80 Gly Leu Lys Val Leu Lys Glu Gln Asn Ser Glu Ile Val Val Ser Ile 85 90 95 Gly Gly Gly Ser Ala His Asp Asn Ala Lys Ala Ile Ala Leu Leu Ala 100 105 110 Thr Asn Gly Gly Glu Ile Gly Asp Tyr Glu Gly Val Asn Gln Ser Lys 115 120 125 Lys Ala Ala Leu Pro Leu Phe Ala Ile Asn Thr Thr Ala Gly Thr Ala 130 135 140 Ser Glu Met Thr Arg Phe Thr Ile Ile Ser Asn Glu Glu Lys Lys Ile 145 150 155 160 Lys Met Ala Ile Ile Asp Asn Asn Val Thr Pro Ala Val Ala Val Asn 165 170 175 Asp Pro Ser Thr Met Phe Gly Leu Pro Pro Ala Leu Thr Ala Ala Thr 180 185 190 Gly Leu Asp Ala Leu Thr His Cys Ile Glu Ala Tyr Val Ser Thr Ala 195 200 205 Ser Asn Pro Ile Thr Asp Ala Cys Ala Leu Lys Gly Ile Asp Leu Ile 210 215 220 Asn Glu Ser Leu Val Ala Ala Tyr Lys Asp Gly Lys Asp Lys Lys Ala 225 230 235 240 Arg Thr Asp Met Cys Tyr Ala Glu Tyr Leu Ala Gly Met Ala Phe Asn 245 250 255 Asn Ala Ser Leu Gly Tyr Val His Ala Leu Ala His Gln Leu Gly Gly 260 265 270 Phe Tyr His Leu Pro His Gly Val Cys Asn Ala Val Leu Leu Pro His 275 280 285 Val Gln Glu Ala Asn Met Gln Cys Pro Lys Ala Lys Lys Arg Leu Gly 290 295 300 Glu Ile Ala Leu His Phe Gly Ala Ser Gln Glu Asp Pro Glu Glu Thr 305 310 315 320 Ile Lys Ala Leu His Val Leu Asn Arg Thr Met Asn Ile Pro Arg Asn 325 330 335 Leu Lys Glu Leu Gly Val Lys Thr Glu Asp Phe Glu Ile Leu Ala Glu 340 345 350 His Ala Met His Asp Ala Cys His Leu Thr Asn Pro Val Gln Phe Thr 355 360 365 Lys Glu Gln Val Val Ala Ile Ile Lys Lys Ala Tyr Glu Tyr 370 375 380 140351PRTSaccharomyces cerevisiae 140Met Pro Ser Gln Val Ile Pro Glu Lys Gln Lys Ala Ile Val Phe Tyr 1 5 10 15 Glu Thr Asp Gly Lys Leu Glu Tyr Lys Asp Val Thr Val Pro Glu Pro 20 25 30 Lys Pro Asn Glu Ile Leu Val His Val Lys Tyr Ser Gly Val Cys His 35 40 45 Ser Asp Leu His Ala Trp His Gly Asp Trp Pro Phe Gln Leu Lys Phe 50 55 60 Pro Leu Ile Gly Gly His Glu Gly Ala Gly Val Val Val Lys Leu Gly 65 70 75 80 Ser Asn Val Lys Gly Trp Lys Val Gly Asp Phe Ala Gly Ile Lys Trp 85 90 95 Leu Asn Gly Thr Cys Met Ser Cys Glu Tyr Cys Glu Val Gly Asn Glu 100 105 110 Ser Gln Cys Pro Tyr Leu Asp Gly Thr Gly Phe Thr His Asp Gly Thr 115 120 125 Phe Gln Glu Tyr Ala Thr Ala Asp Ala Val Gln Ala Ala His Ile Pro 130 135 140 Pro Asn Val Asn Leu Ala Glu Val Ala Pro Ile Leu Cys Ala Gly Ile 145 150 155 160 Thr Val Tyr Lys Ala Leu Lys Arg Ala Asn Val Ile Pro Gly Gln Trp 165 170 175 Val Thr Ile Ser Gly Ala Cys Gly Gly Leu Gly Ser Leu Ala Ile Gln 180 185 190 Tyr Ala Leu Ala Met Gly Tyr Arg Val Ile Gly Ile Asp Gly Gly Asn 195 200 205 Ala Lys Arg Lys Leu Phe Glu Gln Leu Gly Gly Glu Ile Phe Ile Asp 210 215 220 Phe Thr Glu Glu Lys Asp Ile Val Gly Ala Ile Ile Lys Ala Thr Asn 225 230 235 240 Gly Gly Ser His Gly Val Ile Asn Val Ser Val Ser Glu Ala Ala Ile 245 250 255 Glu Ala Ser Thr Arg Tyr Cys Arg Pro Asn Gly Thr Val Val Leu Val 260 265 270 Gly Met Pro Ala His Ala Tyr Cys Asn Ser Asp Val Phe Asn Gln Val 275 280 285 Val Lys Ser Ile Ser Ile Val Gly Ser Cys Val Gly Asn Arg Ala Asp 290 295 300 Thr Arg Glu Ala Leu Asp Phe Phe Ala Arg Gly Leu Ile Lys Ser Pro 305 310 315 320 Ile His Leu Ala Gly Leu Ser Asp Val Pro Glu Ile Phe Ala Lys Met 325 330 335 Glu Lys Gly Glu Ile Val Gly Arg Tyr Val Val Glu Thr Ser Lys 340 345 350 141360PRTSaccharomyces cerevisiae 141Met Ser Tyr Pro Glu Lys Phe Glu Gly Ile Ala Ile Gln Ser His Glu 1 5 10 15 Asp Trp Lys Asn Pro Lys Lys Thr Lys Tyr Asp Pro Lys Pro Phe Tyr 20 25 30 Asp His Asp Ile Asp Ile Lys Ile Glu Ala Cys Gly Val Cys Gly Ser 35 40 45 Asp Ile His Cys Ala Ala Gly His Trp Gly Asn Met Lys Met Pro Leu 50 55 60 Val Val Gly His Glu Ile Val Gly Lys Val Val Lys Leu Gly Pro Lys 65 70 75 80 Ser Asn Ser Gly Leu Lys Val Gly Gln Arg Val Gly Val Gly Ala Gln 85 90 95 Val Phe Ser Cys Leu Glu Cys Asp Arg Cys Lys Asn Asp Asn Glu Pro 100 105 110 Tyr Cys Thr Lys Phe Val Thr Thr Tyr Ser Gln Pro Tyr Glu Asp Gly 115 120 125 Tyr Val Ser Gln Gly Gly Tyr Ala Asn Tyr Val Arg Val His Glu His 130 135 140 Phe Val Val Pro Ile Pro Glu Asn Ile Pro Ser His Leu Ala Ala Pro 145 150 155 160 Leu Leu Cys Gly Gly Leu Thr Val Tyr Ser Pro Leu Val Arg Asn Gly 165 170 175 Cys Gly Pro Gly Lys Lys Val Gly Ile Val Gly Leu Gly Gly Ile Gly 180 185 190 Ser Met Gly Thr Leu Ile Ser Lys Ala Met Gly Ala Glu Thr Tyr Val 195 200 205 Ile Ser Arg Ser Ser Arg Lys Arg Glu Asp Ala Met Lys Met Gly Ala 210 215 220 Asp His Tyr Ile Ala Thr Leu Glu Glu Gly Asp Trp Gly Glu Lys Tyr 225 230 235 240 Phe Asp Thr Phe Asp Leu Ile Val Val Cys Ala Ser Ser Leu Thr Asp 245 250 255 Ile Asp Phe Asn Ile Met Pro Lys Ala Met Lys Val Gly Gly Arg Ile 260 265 270 Val Ser Ile Ser Ile Pro Glu Gln His Glu Met Leu Ser Leu Lys Pro 275 280 285 Tyr Gly Leu Lys Ala Val Ser Ile Ser Tyr Ser Ala Leu Gly Ser Ile 290 295 300 Lys Glu Leu Asn Gln Leu Leu Lys Leu Val Ser Glu Lys Asp Ile Lys 305 310 315 320 Ile Trp Val Glu Thr Leu Pro Val Gly Glu Ala Gly Val His Glu Ala 325 330 335 Phe Glu Arg Met Glu Lys Gly Asp Val Arg Tyr Arg Phe Thr Leu Val 340 345 350 Gly Tyr Asp Lys Glu Phe Ser Asp 355 360 142361PRTSaccharomyces cerevisiae 142Met Leu Tyr Pro Glu Lys Phe Gln Gly Ile Gly Ile Ser Asn Ala Lys 1 5 10 15 Asp Trp Lys His Pro Lys Leu Val Ser Phe Asp Pro Lys Pro Phe Gly 20 25 30 Asp His Asp Val Asp Val Glu Ile Glu Ala Cys Gly Ile Cys Gly Ser 35 40 45 Asp Phe His Ile Ala Val Gly Asn Trp Gly Pro Val Pro Glu Asn Gln 50 55 60 Ile Leu Gly His Glu Ile Ile Gly Arg Val Val Lys Val Gly Ser Lys 65 70 75 80 Cys His Thr Gly Val Lys Ile Gly Asp Arg Val Gly Val Gly Ala Gln 85 90 95 Ala Leu Ala Cys Phe Glu Cys Glu Arg Cys Lys Ser Asp Asn Glu Gln 100 105 110 Tyr Cys Thr Asn Asp His Val Leu Thr Met Trp Thr Pro Tyr Lys Asp 115 120 125 Gly Tyr Ile Ser Gln Gly Gly Phe Ala Ser His Val Arg Leu His Glu 130 135 140 His Phe Ala Ile Gln Ile Pro Glu Asn Ile Pro Ser Pro Leu Ala Ala 145 150 155 160 Pro Leu Leu Cys Gly Gly Ile Thr Val Phe Ser Pro Leu Leu Arg Asn 165 170 175 Gly Cys Gly Pro Gly Lys Arg Val Gly Ile Val Gly Ile Gly Gly Ile 180 185 190 Gly His Met Gly Ile Leu Leu Ala Lys Ala Met Gly Ala Glu Val Tyr 195 200 205 Ala Phe Ser Arg Gly His Ser Lys Arg Glu Asp Ser Met Lys Leu Gly 210 215 220 Ala Asp His Tyr Ile Ala Met Leu Glu Asp Lys Gly Trp Thr Glu Gln 225 230 235 240 Tyr Ser Asn Ala Leu Asp Leu Leu Val Val Cys Ser Ser Ser Leu Ser 245 250 255 Lys Val Asn Phe Asp Ser Ile Val Lys Ile Met Lys Ile Gly Gly Ser 260 265 270 Ile Val Ser Ile Ala Ala Pro Glu Val Asn Glu Lys Leu Val Leu Lys 275 280 285 Pro Leu Gly Leu Met Gly Val Ser Ile Ser Ser Ser Ala Ile Gly Ser 290 295 300 Arg Lys Glu Ile Glu Gln Leu Leu Lys Leu Val Ser Glu Lys Asn Val 305 310 315 320 Lys Ile Trp Val Glu Lys Leu Pro Ile Ser Glu Glu Gly Val Ser His 325 330 335 Ala Phe Thr Arg Met Glu Ser Gly Asp Val Lys Tyr Arg Phe Thr Leu 340 345 350 Val Asp Tyr Asp Lys Lys Phe His Lys 355 360 143417PRTSaccharomyces cerevisiae 143Met Arg Ala Leu Ala Tyr Phe Gly Lys Gly Asn Ile Arg Phe Thr Asn 1 5 10 15 His Leu Lys Glu Pro His Ile Val Ala Pro Asp Glu Leu Val Ile Asp 20 25 30 Ile Glu Trp Cys Gly Ile Cys Gly Thr Asp Leu His Glu Tyr Thr Asp 35 40 45 Gly Pro Ile Phe Phe Pro Glu Asp Gly His Thr His Glu Ile Ser His 50 55 60 Asn Pro Leu Pro Gln Ala Met Gly His Glu Met Ala Gly Thr Val Leu 65 70 75 80 Glu Val Gly Pro Gly Val Lys Asn Leu Lys Val Gly Asp Lys Val Val 85 90 95 Val Glu Pro Thr Gly Thr Cys Arg Asp Arg Tyr Arg Trp Pro Leu Ser 100 105 110 Pro Asn Val Asp Lys Glu Trp Cys Ala Ala Cys Lys Lys Gly Tyr Tyr 115 120 125 Asn Ile Cys Ser Tyr Leu Gly Leu Cys Gly Ala Gly Val Gln Ser Gly 130 135 140 Gly Phe Ala Glu Arg Val Val Met Asn Glu Ser His Cys Tyr Lys Val 145 150 155 160 Pro Asp Phe Val Pro Leu Asp Val Ala Ala Leu Ile Gln Pro Leu Ala 165 170 175 Val Cys Trp His Ala Ile Arg Val Cys Glu Phe Lys Ala Gly Ser Thr 180 185 190 Ala Leu Ile Ile Gly Ala Gly Pro Ile Gly Leu Gly Thr Ile Leu Ala 195 200 205 Leu Asn Ala Ala Gly Cys Lys Asp Ile Val Val Ser Glu Pro Ala Lys 210 215 220 Val Arg Arg Glu Leu Ala Glu Lys Met Gly Ala Arg Val Tyr Asp Pro 225 230 235 240 Thr Ala His Ala Ala Lys Glu Ser Ile Asp Tyr Leu Arg Ser Ile Ala 245 250 255 Asp Gly Gly Asp Gly Phe Asp Tyr Thr Phe Asp Cys Ser Gly Leu Glu 260 265 270 Val Thr Leu Asn Ala Ala Ile Gln Cys Leu Thr Phe Arg Gly Thr Ala 275 280 285 Val Asn Leu Ala Met Trp Gly His His Lys Ile Gln Phe Ser Pro Met 290 295 300 Asp Ile Thr Leu His Glu Arg Lys Tyr Thr Gly Ser Met Cys Tyr Thr 305 310 315 320 His His Asp Phe Glu Ala Val Ile Glu Ala Leu Glu Glu Gly Arg Ile 325 330 335 Asp Ile Asp Arg Ala Arg His Met Ile Thr Gly Arg Val Asn Ile Glu 340 345 350 Asp Gly Leu Asp Gly Ala Ile Met Lys Leu Ile Asn Glu Lys Glu Ser 355 360 365 Thr Ile Lys Ile Ile Leu Thr Pro Asn Asn His Gly Glu Leu Asn Arg 370 375 380 Glu Ala Asp Asn Glu Lys Lys Glu Ile Ser Glu Leu Ser Ser Arg Lys 385 390 395 400 Asp Gln Glu Arg Leu Arg Glu Ser Ile Asn Glu Ala Lys Leu Arg His 405 410 415 Thr 144386PRTSaccharomyces cerevisiae 144Met Ser Ala Ala Thr Val Gly Lys Pro Ile Lys Cys Ile Ala Ala Val 1 5 10 15 Ala Tyr Asp Ala Lys Lys Pro Leu Ser Val Glu Glu Ile Thr Val Asp 20 25 30 Ala Pro Lys Ala His Glu Val Arg Ile Lys Ile Glu Tyr Thr Ala Val 35 40 45 Cys His Thr Asp Ala Tyr Thr Leu Ser Gly Ser Asp Pro Glu Gly Leu 50 55 60 Phe Pro Cys Val Leu Gly His Glu Gly Ala Gly Ile Val Glu Ser Val 65 70 75 80 Gly Asp Asp Val Ile Thr Val Lys Pro Gly Asp His Val Ile Ala Leu 85 90 95 Tyr Thr Ala Glu Cys Gly Lys Cys Lys Phe Cys Thr Ser Gly Lys Thr 100 105 110 Asn Leu Cys Gly Ala Val Arg Ala Thr Gln Gly Lys Gly Val Met Pro 115 120 125 Asp Gly Thr Thr Arg Phe His Asn Ala Lys Gly Glu Asp Ile Tyr His 130 135 140 Phe Met Gly Cys Ser Thr Phe Ser Glu Tyr Thr Val Val Ala Asp Val 145 150 155 160 Ser Val Val Ala Ile Asp Pro Lys Ala Pro Leu Asp Ala Ala Cys Leu 165 170 175 Leu Gly Cys Gly Val Thr Thr Gly Phe Gly Ala Ala Leu Lys Thr Ala 180 185 190 Asn Val Gln Lys Gly Asp Thr Val Ala Val Phe Gly Cys Gly Thr Val 195 200

205 Gly Leu Ser Val Ile Gln Gly Ala Lys Leu Arg Gly Ala Ser Lys Ile 210 215 220 Ile Ala Ile Asp Ile Asn Asn Lys Lys Lys Gln Tyr Cys Ser Gln Phe 225 230 235 240 Gly Ala Thr Asp Phe Val Asn Pro Lys Glu Asp Leu Ala Lys Asp Gln 245 250 255 Thr Ile Val Glu Lys Leu Ile Glu Met Thr Asp Gly Gly Leu Asp Phe 260 265 270 Thr Phe Asp Cys Thr Gly Asn Thr Lys Ile Met Arg Asp Ala Leu Glu 275 280 285 Ala Cys His Lys Gly Trp Gly Gln Ser Ile Ile Ile Gly Val Ala Ala 290 295 300 Ala Gly Glu Glu Ile Ser Thr Arg Pro Phe Gln Leu Val Thr Gly Arg 305 310 315 320 Val Trp Lys Gly Ser Ala Phe Gly Gly Ile Lys Gly Arg Ser Glu Met 325 330 335 Gly Gly Leu Ile Lys Asp Tyr Gln Lys Gly Ala Leu Lys Val Glu Glu 340 345 350 Phe Ile Thr His Arg Arg Pro Phe Lys Glu Ile Asn Gln Ala Phe Glu 355 360 365 Asp Leu His Asn Gly Asp Cys Leu Arg Thr Val Leu Lys Ser Asp Glu 370 375 380 Ile Lys 385 145342PRTSaccharomyces cerevisiae 145Met Val Leu Val Lys Gln Val Arg Leu Gly Asn Ser Gly Leu Lys Ile 1 5 10 15 Ser Pro Ile Val Ile Gly Cys Met Ser Tyr Gly Ser Lys Lys Trp Ala 20 25 30 Asp Trp Val Ile Glu Asp Lys Thr Gln Ile Phe Lys Ile Met Lys His 35 40 45 Cys Tyr Asp Lys Gly Leu Arg Thr Phe Asp Thr Ala Asp Phe Tyr Ser 50 55 60 Asn Gly Leu Ser Glu Arg Ile Ile Lys Glu Phe Leu Glu Tyr Tyr Ser 65 70 75 80 Ile Lys Arg Glu Thr Val Val Ile Met Thr Lys Ile Tyr Phe Pro Val 85 90 95 Asp Glu Thr Leu Asp Leu His His Asn Phe Thr Leu Asn Glu Phe Glu 100 105 110 Glu Leu Asp Leu Ser Asn Gln Arg Gly Leu Ser Arg Lys His Ile Ile 115 120 125 Ala Gly Val Glu Asn Ser Val Lys Arg Leu Gly Thr Tyr Ile Asp Leu 130 135 140 Leu Gln Ile His Arg Leu Asp His Glu Thr Pro Met Lys Glu Ile Met 145 150 155 160 Lys Ala Leu Asn Asp Val Val Glu Ala Gly His Val Arg Tyr Ile Gly 165 170 175 Ala Ser Ser Met Leu Ala Thr Glu Phe Ala Glu Leu Gln Phe Thr Ala 180 185 190 Asp Lys Tyr Gly Trp Phe Gln Phe Ile Ser Ser Gln Ser Tyr Tyr Asn 195 200 205 Leu Leu Tyr Arg Glu Asp Glu Arg Glu Leu Ile Pro Phe Ala Lys Arg 210 215 220 His Asn Ile Gly Leu Leu Pro Trp Ser Pro Asn Ala Arg Gly Met Leu 225 230 235 240 Thr Arg Pro Leu Asn Gln Ser Thr Asp Arg Ile Lys Ser Asp Pro Thr 245 250 255 Phe Lys Ser Leu His Leu Asp Asn Leu Glu Glu Glu Gln Lys Glu Ile 260 265 270 Ile Asn Arg Val Glu Lys Val Ser Lys Asp Lys Lys Val Ser Met Ala 275 280 285 Met Leu Ser Ile Ala Trp Val Leu His Lys Gly Cys His Pro Ile Val 290 295 300 Gly Leu Asn Thr Thr Ala Arg Val Asp Glu Ala Ile Ala Ala Leu Gln 305 310 315 320 Val Thr Leu Thr Glu Glu Glu Ile Lys Tyr Leu Glu Glu Pro Tyr Lys 325 330 335 Pro Gln Arg Gln Arg Cys 340 146359PRTSaccharomyces cerevisiae 146Met Gly Val Glu Gln Ile Leu Lys Arg Lys Thr Gly Val Ile Val Gly 1 5 10 15 Glu Asp Val His Asn Leu Phe Thr Tyr Ala Lys Glu His Lys Phe Ala 20 25 30 Ile Pro Ala Ile Asn Val Thr Ser Ser Ser Thr Ala Val Ala Ala Leu 35 40 45 Glu Ala Ala Arg Asp Ser Lys Ser Pro Ile Ile Leu Gln Thr Ser Asn 50 55 60 Gly Gly Ala Ala Tyr Phe Ala Gly Lys Gly Ile Ser Asn Glu Gly Gln 65 70 75 80 Asn Ala Ser Ile Lys Gly Ala Ile Ala Ala Ala His Tyr Ile Arg Ser 85 90 95 Ile Ala Pro Ala Tyr Gly Ile Pro Val Val Leu His Ser Asp His Cys 100 105 110 Ala Lys Lys Leu Leu Pro Trp Phe Asp Gly Met Leu Glu Ala Asp Glu 115 120 125 Ala Tyr Phe Lys Glu His Gly Glu Pro Leu Phe Ser Ser His Met Leu 130 135 140 Asp Leu Ser Glu Glu Thr Asp Glu Glu Asn Ile Ser Thr Cys Val Lys 145 150 155 160 Tyr Phe Lys Arg Met Ala Ala Met Asp Gln Trp Leu Glu Met Glu Ile 165 170 175 Gly Ile Thr Gly Gly Glu Glu Asp Gly Val Asn Asn Glu Asn Ala Asp 180 185 190 Lys Glu Asp Leu Tyr Thr Lys Pro Glu Gln Val Tyr Asn Val Tyr Lys 195 200 205 Ala Leu His Pro Ile Ser Pro Asn Phe Ser Ile Ala Ala Ala Phe Gly 210 215 220 Asn Cys His Gly Leu Tyr Ala Gly Asp Ile Ala Leu Arg Pro Glu Ile 225 230 235 240 Leu Ala Glu His Gln Lys Tyr Thr Arg Glu Gln Val Gly Cys Lys Glu 245 250 255 Glu Lys Pro Leu Phe Leu Val Phe His Gly Gly Ser Gly Ser Thr Val 260 265 270 Gln Glu Phe His Thr Gly Ile Asp Asn Gly Val Val Lys Val Asn Leu 275 280 285 Asp Thr Asp Cys Gln Tyr Ala Tyr Leu Thr Gly Ile Arg Asp Tyr Val 290 295 300 Leu Asn Lys Lys Asp Tyr Ile Met Ser Pro Val Gly Asn Pro Glu Gly 305 310 315 320 Pro Glu Lys Pro Asn Lys Lys Phe Phe Asp Pro Arg Val Trp Val Arg 325 330 335 Glu Gly Glu Lys Thr Met Gly Ala Lys Ile Thr Lys Ser Leu Glu Thr 340 345 350 Phe Arg Thr Thr Asn Thr Leu 355 147248PRTSaccharomyces cerevisiae 147Met Ala Arg Thr Phe Phe Val Gly Gly Asn Phe Lys Leu Asn Gly Ser 1 5 10 15 Lys Gln Ser Ile Lys Glu Ile Val Glu Arg Leu Asn Thr Ala Ser Ile 20 25 30 Pro Glu Asn Val Glu Val Val Ile Cys Pro Pro Ala Thr Tyr Leu Asp 35 40 45 Tyr Ser Val Ser Leu Val Lys Lys Pro Gln Val Thr Val Gly Ala Gln 50 55 60 Asn Ala Tyr Leu Lys Ala Ser Gly Ala Phe Thr Gly Glu Asn Ser Val 65 70 75 80 Asp Gln Ile Lys Asp Val Gly Ala Lys Trp Val Ile Leu Gly His Ser 85 90 95 Glu Arg Arg Ser Tyr Phe His Glu Asp Asp Lys Phe Ile Ala Asp Lys 100 105 110 Thr Lys Phe Ala Leu Gly Gln Gly Val Gly Val Ile Leu Cys Ile Gly 115 120 125 Glu Thr Leu Glu Glu Lys Lys Ala Gly Lys Thr Leu Asp Val Val Glu 130 135 140 Arg Gln Leu Asn Ala Val Leu Glu Glu Val Lys Asp Trp Thr Asn Val 145 150 155 160 Val Val Ala Tyr Glu Pro Val Trp Ala Ile Gly Thr Gly Leu Ala Ala 165 170 175 Thr Pro Glu Asp Ala Gln Asp Ile His Ala Ser Ile Arg Lys Phe Leu 180 185 190 Ala Ser Lys Leu Gly Asp Lys Ala Ala Ser Glu Leu Arg Ile Leu Tyr 195 200 205 Gly Gly Ser Ala Asn Gly Ser Asn Ala Val Thr Phe Lys Asp Lys Ala 210 215 220 Asp Val Asp Gly Phe Leu Val Gly Gly Ala Ser Leu Lys Pro Glu Phe 225 230 235 240 Val Asp Ile Ile Asn Ser Arg Asn 245 148376PRTSaccharomyces cerevisiae 148Met Ser Lys Gly Lys Val Leu Leu Val Leu Tyr Glu Gly Gly Lys His 1 5 10 15 Ala Glu Glu Gln Glu Lys Leu Leu Gly Cys Ile Glu Asn Glu Leu Gly 20 25 30 Ile Arg Asn Phe Ile Glu Glu Gln Gly Tyr Glu Leu Val Thr Thr Ile 35 40 45 Asp Lys Asp Pro Glu Pro Thr Ser Thr Val Asp Arg Glu Leu Lys Asp 50 55 60 Ala Glu Ile Val Ile Thr Thr Pro Phe Phe Pro Ala Tyr Ile Ser Arg 65 70 75 80 Asn Arg Ile Ala Glu Ala Pro Asn Leu Lys Leu Cys Val Thr Ala Gly 85 90 95 Val Gly Ser Asp His Val Asp Leu Glu Ala Ala Asn Glu Arg Lys Ile 100 105 110 Thr Val Thr Glu Val Thr Gly Ser Asn Val Val Ser Val Ala Glu His 115 120 125 Val Met Ala Thr Ile Leu Val Leu Ile Arg Asn Tyr Asn Gly Gly His 130 135 140 Gln Gln Ala Ile Asn Gly Glu Trp Asp Ile Ala Gly Val Ala Lys Asn 145 150 155 160 Glu Tyr Asp Leu Glu Asp Lys Ile Ile Ser Thr Val Gly Ala Gly Arg 165 170 175 Ile Gly Tyr Arg Val Leu Glu Arg Leu Val Ala Phe Asn Pro Lys Lys 180 185 190 Leu Leu Tyr Tyr Asp Tyr Gln Glu Leu Pro Ala Glu Ala Ile Asn Arg 195 200 205 Leu Asn Glu Ala Ser Lys Leu Phe Asn Gly Arg Gly Asp Ile Val Gln 210 215 220 Arg Val Glu Lys Leu Glu Asp Met Val Ala Gln Ser Asp Val Val Thr 225 230 235 240 Ile Asn Cys Pro Leu His Lys Asp Ser Arg Gly Leu Phe Asn Lys Lys 245 250 255 Leu Ile Ser His Met Lys Asp Gly Ala Tyr Leu Val Asn Thr Ala Arg 260 265 270 Gly Ala Ile Cys Val Ala Glu Asp Val Ala Glu Ala Val Lys Ser Gly 275 280 285 Lys Leu Ala Gly Tyr Gly Gly Asp Val Trp Asp Lys Gln Pro Ala Pro 290 295 300 Lys Asp His Pro Trp Arg Thr Met Asp Asn Lys Asp His Val Gly Asn 305 310 315 320 Ala Met Thr Val His Ile Ser Gly Thr Ser Leu Asp Ala Gln Lys Arg 325 330 335 Tyr Ala Gln Gly Val Lys Asn Ile Leu Asn Ser Tyr Phe Ser Lys Lys 340 345 350 Phe Asp Tyr Arg Pro Gln Asp Ile Ile Val Gln Asn Gly Ser Tyr Ala 355 360 365 Thr Arg Ala Tyr Gly Gln Lys Lys 370 375 149327PRTSaccharomyces cerevisiae 149Met Ser Ser Leu Val Thr Leu Asn Asn Gly Leu Lys Met Pro Leu Val 1 5 10 15 Gly Leu Gly Cys Trp Lys Ile Asp Lys Lys Val Cys Ala Asn Gln Ile 20 25 30 Tyr Glu Ala Ile Lys Leu Gly Tyr Arg Leu Phe Asp Gly Ala Cys Asp 35 40 45 Tyr Gly Asn Glu Lys Glu Val Gly Glu Gly Ile Arg Lys Ala Ile Ser 50 55 60 Glu Gly Leu Val Ser Arg Lys Asp Ile Phe Val Val Ser Lys Leu Trp 65 70 75 80 Asn Asn Phe His His Pro Asp His Val Lys Leu Ala Leu Lys Lys Thr 85 90 95 Leu Ser Asp Met Gly Leu Asp Tyr Leu Asp Leu Tyr Tyr Ile His Phe 100 105 110 Pro Ile Ala Phe Lys Tyr Val Pro Phe Glu Glu Lys Tyr Pro Pro Gly 115 120 125 Phe Tyr Thr Gly Ala Asp Asp Glu Lys Lys Gly His Ile Thr Glu Ala 130 135 140 His Val Pro Ile Ile Asp Thr Tyr Arg Ala Leu Glu Glu Cys Val Asp 145 150 155 160 Glu Gly Leu Ile Lys Ser Ile Gly Val Ser Asn Phe Gln Gly Ser Leu 165 170 175 Ile Gln Asp Leu Leu Arg Gly Cys Arg Ile Lys Pro Val Ala Leu Gln 180 185 190 Ile Glu His His Pro Tyr Leu Thr Gln Glu His Leu Val Glu Phe Cys 195 200 205 Lys Leu His Asp Ile Gln Val Val Ala Tyr Ser Ser Phe Gly Pro Gln 210 215 220 Ser Phe Ile Glu Met Asp Leu Gln Leu Ala Lys Thr Thr Pro Thr Leu 225 230 235 240 Phe Glu Asn Asp Val Ile Lys Lys Val Ser Gln Asn His Pro Gly Ser 245 250 255 Thr Thr Ser Gln Val Leu Leu Arg Trp Ala Thr Gln Arg Gly Ile Ala 260 265 270 Val Ile Pro Lys Ser Ser Lys Lys Glu Arg Leu Leu Gly Asn Leu Glu 275 280 285 Ile Glu Lys Lys Phe Thr Leu Thr Glu Gln Glu Leu Lys Asp Ile Ser 290 295 300 Ala Leu Asn Ala Asn Ile Arg Phe Asn Asp Pro Trp Thr Trp Leu Asp 305 310 315 320 Gly Lys Phe Pro Thr Phe Ala 325 150350PRTSaccharomyces cerevisiae 150Met Ser Lys Lys Pro Ile Val Leu Lys Leu Gly Lys Asp Ala Phe Gly 1 5 10 15 Asp Gln Ala Trp Gly Glu Leu Glu Lys Ile Ala Asp Val Ile Thr Ile 20 25 30 Pro Glu Ser Thr Thr Arg Glu Gln Phe Leu Arg Glu Val Lys Asp Pro 35 40 45 Gln Asn Lys Leu Ser Gln Val Gln Val Ile Thr Arg Thr Ala Arg Ser 50 55 60 Val Lys Asn Thr Gly Arg Phe Asp Glu Glu Leu Ala Leu Ala Leu Pro 65 70 75 80 Ser Ser Val Val Ala Val Cys His Thr Gly Ala Gly Tyr Asp Gln Ile 85 90 95 Asp Val Glu Pro Phe Lys Lys Arg His Ile Gln Val Ala Asn Val Pro 100 105 110 Asp Leu Val Ser Asn Ala Thr Ala Asp Thr His Val Phe Leu Leu Leu 115 120 125 Gly Ala Leu Arg Asn Phe Gly Ile Gly Asn Arg Arg Leu Ile Glu Gly 130 135 140 Asn Trp Pro Glu Ala Gly Pro Ala Cys Gly Ser Pro Phe Gly Tyr Asp 145 150 155 160 Pro Glu Gly Lys Thr Val Gly Ile Leu Gly Leu Gly Arg Ile Gly Arg 165 170 175 Cys Ile Leu Glu Arg Leu Lys Pro Phe Gly Phe Glu Asn Phe Ile Tyr 180 185 190 His Asn Arg His Gln Leu Pro Ser Glu Glu Glu His Gly Cys Glu Tyr 195 200 205 Val Gly Phe Glu Glu Phe Leu Lys Arg Ser Asp Ile Val Ser Val Asn 210 215 220 Val Pro Leu Asn His Asn Thr His His Leu Ile Asn Ala Glu Thr Ile 225 230 235 240 Glu Lys Met Lys Asp Gly Val Val Ile Val Asn Thr Ala Arg Gly Ala 245 250 255 Val Ile Asp Glu Gln Ala Met Thr Asp Ala Leu Arg Ser Gly Lys Ile 260 265 270 Arg Ser Ala Gly Leu Asp Val Phe Glu Tyr Glu Pro Lys Ile Ser Lys 275 280 285 Glu Leu Leu Ser Met Ser Gln Val Leu Gly Leu Pro His Met Gly Thr 290 295 300 His Ser Val Glu Thr Arg Lys Lys Met Glu Glu Leu Val Val Glu Asn 305 310 315 320 Ala Lys Asn Val Ile Leu Thr Gly Lys Val Leu Thr Ile Val Pro Glu 325 330 335 Leu Gln Asn Glu Asp Trp Pro Asn Glu Ser Lys Pro Leu Val 340 345 350 151396PRTSaccharomyces cerevisiae 151Met Ile Thr Ser Ile Asp Ile Ala Asp Val Thr Tyr Ser Ala Lys Pro 1 5 10 15 Arg Ile Leu Val Pro Tyr Lys Thr Gln Trp Glu Val Ala Ser His Leu 20 25 30 Pro Glu Tyr Arg Lys Leu Ala Glu Arg Val Glu Phe Tyr Lys Tyr Glu 35 40 45 Met Ser Thr Lys Asp Asp Phe Val Lys Phe Leu Glu Thr His Arg Ile 50 55 60 Asn Gly Phe Trp Leu Thr Glu Glu Phe Phe Thr Val Leu Gly Asn Pro 65 70 75 80 Ser Ser Tyr Ile Glu Phe Phe Pro Ala Ser Leu Lys

Val Ile Leu Val 85 90 95 Pro Trp Val Gly Cys Asp Phe Ile Asp Gly Lys Leu Leu Arg Ser Lys 100 105 110 Gly Ile Thr Leu Cys Asn Ile Gly Pro His Ala Ala Asp His Val Thr 115 120 125 Glu Leu Ala Ile Phe Leu Ala Ile Ser Cys Phe Arg Met Thr Ser Phe 130 135 140 Trp Glu Tyr Cys Phe Lys Tyr Val Glu Asn Gly Asn Val Glu Gln Cys 145 150 155 160 Lys Lys Tyr Ile Ser Ser Asp Ser Tyr Glu Ile Val Thr Asp Ser Tyr 165 170 175 His Gly Gln Glu Met Lys Phe Pro Ser Arg Thr Asp Lys Cys Lys Pro 180 185 190 Asn Lys Asp Arg Lys Val Val His Leu Ala Glu Lys Tyr Thr Val Gly 195 200 205 Gly Lys Lys Met Glu Ser Pro Met Asn Lys Lys Val Leu Ile Leu Gly 210 215 220 Phe Gly Ser Ile Gly Gln Asn Ile Gly Ser Asn Leu His Lys Val Phe 225 230 235 240 Asn Met Ser Ile Glu Tyr Tyr Lys Arg Thr Gly Pro Val Gln Lys Ser 245 250 255 Leu Leu Asp Tyr Asn Ala Lys Tyr His Ser Asp Leu Asp Asp Pro Asn 260 265 270 Thr Trp Lys Asn Ala Asp Leu Ile Ile Leu Ala Leu Pro Ser Thr Ala 275 280 285 Ser Thr Asn Asn Ile Ile Asn Arg Lys Ser Leu Ala Trp Cys Lys Asp 290 295 300 Gly Val Arg Ile Val Asn Val Gly Arg Gly Thr Cys Ile Asp Glu Asp 305 310 315 320 Val Leu Leu Asp Ala Leu Glu Ser Gly Lys Val Ala Ser Cys Gly Leu 325 330 335 Asp Val Phe Lys Asn Glu Glu Thr Arg Val Lys Gln Glu Leu Leu Arg 340 345 350 Arg Trp Asp Val Thr Ala Leu Pro His Ile Gly Ser Thr Val Ala Asp 355 360 365 Met Val Ile Lys Gln Thr Leu Ile Thr Leu Glu Asn Val Gln Asp Ile 370 375 380 Phe Val Glu Gly Gly Asp Gly Lys Tyr Val Leu Asn 385 390 395 152312PRTSaccharomyces cerevisiae 152Met Pro Ala Thr Leu His Asp Ser Thr Lys Ile Leu Ser Leu Asn Thr 1 5 10 15 Gly Ala Gln Ile Pro Gln Ile Gly Leu Gly Thr Trp Gln Ser Lys Glu 20 25 30 Asn Asp Ala Tyr Lys Ala Val Leu Thr Ala Leu Lys Asp Gly Tyr Arg 35 40 45 His Ile Asp Thr Ala Ala Ile Tyr Arg Asn Glu Asp Gln Val Gly Gln 50 55 60 Ala Ile Lys Asp Ser Gly Val Pro Arg Glu Glu Ile Phe Val Thr Thr 65 70 75 80 Lys Leu Trp Cys Thr Gln His His Glu Pro Glu Val Ala Leu Asp Gln 85 90 95 Ser Leu Lys Arg Leu Gly Leu Asp Tyr Val Asp Leu Tyr Leu Met His 100 105 110 Trp Pro Ala Arg Leu Asp Pro Ala Tyr Ile Lys Asn Glu Asp Ile Leu 115 120 125 Ser Val Pro Thr Lys Lys Asp Gly Ser Arg Ala Val Asp Ile Thr Asn 130 135 140 Trp Asn Phe Ile Lys Thr Trp Glu Leu Met Gln Glu Leu Pro Lys Thr 145 150 155 160 Gly Lys Thr Lys Ala Val Gly Val Ser Asn Phe Ser Ile Asn Asn Leu 165 170 175 Lys Asp Leu Leu Ala Ser Gln Gly Asn Lys Leu Thr Pro Ala Ala Asn 180 185 190 Gln Val Glu Ile His Pro Leu Leu Pro Gln Asp Glu Leu Ile Asn Phe 195 200 205 Cys Lys Ser Lys Gly Ile Val Val Glu Ala Tyr Ser Pro Leu Gly Ser 210 215 220 Thr Asp Ala Pro Leu Leu Lys Glu Pro Val Ile Leu Glu Ile Ala Lys 225 230 235 240 Lys Asn Asn Val Gln Pro Gly His Val Val Ile Ser Trp His Val Gln 245 250 255 Arg Gly Tyr Val Val Leu Pro Lys Ser Val Asn Pro Asp Arg Ile Lys 260 265 270 Thr Asn Arg Lys Ile Phe Thr Leu Ser Thr Glu Asp Phe Glu Ala Ile 275 280 285 Asn Asn Ile Ser Lys Glu Lys Gly Glu Lys Arg Val Val His Pro Asn 290 295 300 Trp Ser Pro Phe Glu Val Phe Lys 305 310 153505PRTSaccharomyces cerevisiae 153Met Pro Thr Leu Tyr Thr Asp Ile Glu Ile Pro Gln Leu Lys Ile Ser 1 5 10 15 Leu Lys Gln Pro Leu Gly Leu Phe Ile Asn Asn Glu Phe Cys Pro Ser 20 25 30 Ser Asp Gly Lys Thr Ile Glu Thr Val Asn Pro Ala Thr Gly Glu Pro 35 40 45 Ile Thr Ser Phe Gln Ala Ala Asn Glu Lys Asp Val Asp Lys Ala Val 50 55 60 Lys Ala Ala Arg Ala Ala Phe Asp Asn Val Trp Ser Lys Thr Ser Ser 65 70 75 80 Glu Gln Arg Gly Ile Tyr Leu Ser Asn Leu Leu Lys Leu Ile Glu Glu 85 90 95 Glu Gln Asp Thr Leu Ala Ala Leu Glu Thr Leu Asp Ala Gly Lys Pro 100 105 110 Tyr Ser Asn Ala Lys Gly Asp Leu Ala Gln Ile Leu Gln Leu Thr Arg 115 120 125 Tyr Phe Ala Gly Ser Ala Asp Lys Phe Asp Lys Gly Ala Thr Ile Pro 130 135 140 Leu Thr Phe Asn Lys Phe Ala Tyr Thr Leu Lys Val Pro Phe Gly Val 145 150 155 160 Val Ala Gln Ile Val Pro Trp Asn Tyr Pro Leu Ala Met Ala Cys Trp 165 170 175 Lys Leu Gln Gly Ala Leu Ala Ala Gly Asn Thr Val Ile Ile Lys Pro 180 185 190 Ala Glu Asn Thr Ser Leu Ser Leu Leu Tyr Phe Ala Thr Leu Ile Lys 195 200 205 Lys Ala Gly Phe Pro Pro Gly Val Val Asn Ile Val Pro Gly Tyr Gly 210 215 220 Ser Leu Val Gly Gln Ala Leu Ala Ser His Met Asp Ile Asp Lys Ile 225 230 235 240 Ser Phe Thr Gly Ser Thr Lys Val Gly Gly Phe Val Leu Glu Ala Ser 245 250 255 Gly Gln Ser Asn Leu Lys Asp Val Thr Leu Glu Cys Gly Gly Lys Ser 260 265 270 Pro Ala Leu Val Phe Glu Asp Ala Asp Leu Asp Lys Ala Ile Asp Trp 275 280 285 Ile Ala Ala Gly Ile Phe Tyr Asn Ser Gly Gln Asn Cys Thr Ala Asn 290 295 300 Ser Arg Val Tyr Val Gln Ser Ser Ile Tyr Asp Lys Phe Val Glu Lys 305 310 315 320 Phe Lys Glu Thr Ala Lys Lys Glu Trp Asp Val Ala Gly Lys Phe Asp 325 330 335 Pro Phe Asp Glu Lys Cys Ile Val Gly Pro Val Ile Ser Ser Thr Gln 340 345 350 Tyr Asp Arg Ile Lys Ser Tyr Ile Glu Arg Gly Lys Arg Glu Glu Lys 355 360 365 Leu Asp Met Phe Gln Thr Ser Glu Phe Pro Ile Gly Gly Ala Lys Gly 370 375 380 Tyr Phe Ile Pro Pro Thr Ile Phe Thr Asp Val Pro Gln Thr Ser Lys 385 390 395 400 Leu Leu Gln Asp Glu Ile Phe Gly Pro Val Val Val Val Ser Lys Phe 405 410 415 Thr Asn Tyr Asp Asp Ala Leu Lys Leu Ala Asn Asp Thr Cys Tyr Gly 420 425 430 Leu Ala Ser Ala Val Phe Thr Lys Asp Val Lys Lys Ala His Met Phe 435 440 445 Ala Arg Asp Ile Lys Ala Gly Thr Val Trp Ile Asn Ser Ser Asn Asp 450 455 460 Glu Asp Val Thr Val Pro Phe Gly Gly Phe Lys Met Ser Gly Ile Gly 465 470 475 480 Arg Glu Leu Gly Gln Ser Gly Val Asp Thr Tyr Leu Gln Thr Lys Ala 485 490 495 Val His Ile Asn Leu Ser Leu Asp Asn 500 505 154506PRTSaccharomyces cerevisiae 154Met Pro Thr Leu Tyr Thr Asp Ile Glu Ile Pro Gln Leu Lys Ile Ser 1 5 10 15 Leu Lys Gln Pro Leu Gly Leu Phe Ile Asn Asn Glu Phe Cys Pro Ser 20 25 30 Ser Asp Gly Lys Thr Ile Glu Thr Val Asn Pro Ala Thr Gly Glu Pro 35 40 45 Ile Thr Ser Phe Gln Ala Ala Asn Glu Lys Asp Val Asp Lys Ala Val 50 55 60 Lys Ala Ala Arg Ala Ala Phe Asp Asn Val Trp Ser Lys Thr Ser Ser 65 70 75 80 Glu Gln Arg Gly Ile Tyr Leu Ser Asn Leu Leu Lys Leu Ile Glu Glu 85 90 95 Glu Gln Asp Thr Leu Ala Ala Leu Glu Thr Leu Asp Ala Gly Lys Pro 100 105 110 Phe His Ser Asn Ala Lys Gln Asp Leu Ala Gln Ile Ile Glu Leu Thr 115 120 125 Arg Tyr Tyr Ala Gly Ala Val Asp Lys Phe Asn Met Gly Glu Thr Ile 130 135 140 Pro Leu Thr Phe Asn Lys Phe Ala Tyr Thr Leu Lys Val Pro Phe Gly 145 150 155 160 Val Val Ala Gln Ile Val Pro Trp Asn Tyr Pro Leu Ala Met Ala Cys 165 170 175 Arg Lys Met Gln Gly Ala Leu Ala Ala Gly Asn Thr Val Ile Ile Lys 180 185 190 Pro Ala Glu Asn Thr Ser Leu Ser Leu Leu Tyr Phe Ala Thr Leu Ile 195 200 205 Lys Lys Ala Gly Phe Pro Pro Gly Val Val Asn Val Ile Pro Gly Tyr 210 215 220 Gly Ser Val Val Gly Lys Ala Leu Gly Thr His Met Asp Ile Asp Lys 225 230 235 240 Ile Ser Phe Thr Gly Ser Thr Lys Val Gly Gly Ser Val Leu Glu Ala 245 250 255 Ser Gly Gln Ser Asn Leu Lys Asp Ile Thr Leu Glu Cys Gly Gly Lys 260 265 270 Ser Pro Ala Leu Val Phe Glu Asp Ala Asp Leu Asp Lys Ala Ile Glu 275 280 285 Trp Val Ala Asn Gly Ile Phe Phe Asn Ser Gly Gln Ile Cys Thr Ala 290 295 300 Asn Ser Arg Val Tyr Val Gln Ser Ser Ile Tyr Asp Lys Phe Val Glu 305 310 315 320 Lys Phe Lys Glu Thr Ala Lys Lys Glu Trp Asp Val Ala Gly Lys Phe 325 330 335 Asp Pro Phe Asp Glu Lys Cys Ile Val Gly Pro Val Ile Ser Ser Thr 340 345 350 Gln Tyr Asp Arg Ile Lys Ser Tyr Ile Glu Arg Gly Lys Lys Glu Glu 355 360 365 Lys Leu Asp Met Phe Gln Thr Ser Glu Phe Pro Ile Gly Gly Ala Lys 370 375 380 Gly Tyr Phe Ile Pro Pro Thr Ile Phe Thr Asp Val Pro Glu Thr Ser 385 390 395 400 Lys Leu Leu Arg Asp Glu Ile Phe Gly Pro Val Val Val Val Ser Lys 405 410 415 Phe Thr Asn Tyr Asp Asp Ala Leu Lys Leu Ala Asn Asp Thr Cys Tyr 420 425 430 Gly Leu Ala Ser Ala Val Phe Thr Lys Asp Val Lys Lys Ala His Met 435 440 445 Phe Ala Arg Asp Ile Lys Ala Gly Thr Val Trp Ile Asn Gln Thr Asn 450 455 460 Gln Glu Glu Ala Lys Val Pro Phe Gly Gly Phe Lys Met Ser Gly Ile 465 470 475 480 Gly Arg Glu Ser Gly Asp Thr Gly Val Asp Asn Tyr Leu Gln Ile Lys 485 490 495 Ser Val His Val Asp Leu Ser Leu Asp Lys 500 505 155519PRTSaccharomyces cerevisiae 155Met Phe Ser Arg Ser Thr Leu Cys Leu Lys Thr Ser Ala Ser Ser Ile 1 5 10 15 Gly Arg Leu Gln Leu Arg Tyr Phe Ser His Leu Pro Met Thr Val Pro 20 25 30 Ile Lys Leu Pro Asn Gly Leu Glu Tyr Glu Gln Pro Thr Gly Leu Phe 35 40 45 Ile Asn Asn Lys Phe Val Pro Ser Lys Gln Asn Lys Thr Phe Glu Val 50 55 60 Ile Asn Pro Ser Thr Glu Glu Glu Ile Cys His Ile Tyr Glu Gly Arg 65 70 75 80 Glu Asp Asp Val Glu Glu Ala Val Gln Ala Ala Asp Arg Ala Phe Ser 85 90 95 Asn Gly Ser Trp Asn Gly Ile Asp Pro Ile Asp Arg Gly Lys Ala Leu 100 105 110 Tyr Arg Leu Ala Glu Leu Ile Glu Gln Asp Lys Asp Val Ile Ala Ser 115 120 125 Ile Glu Thr Leu Asp Asn Gly Lys Ala Ile Ser Ser Ser Arg Gly Asp 130 135 140 Val Asp Leu Val Ile Asn Tyr Leu Lys Ser Ser Ala Gly Phe Ala Asp 145 150 155 160 Lys Ile Asp Gly Arg Met Ile Asp Thr Gly Arg Thr His Phe Ser Tyr 165 170 175 Thr Lys Arg Gln Pro Leu Gly Val Cys Gly Gln Ile Ile Pro Trp Asn 180 185 190 Phe Pro Leu Leu Met Trp Ala Trp Lys Ile Ala Pro Ala Leu Val Thr 195 200 205 Gly Asn Thr Val Val Leu Lys Thr Ala Glu Ser Thr Pro Leu Ser Ala 210 215 220 Leu Tyr Val Ser Lys Tyr Ile Pro Gln Ala Gly Ile Pro Pro Gly Val 225 230 235 240 Ile Asn Ile Val Ser Gly Phe Gly Lys Ile Val Gly Glu Ala Ile Thr 245 250 255 Asn His Pro Lys Ile Lys Lys Val Ala Phe Thr Gly Ser Thr Ala Thr 260 265 270 Gly Arg His Ile Tyr Gln Ser Ala Ala Ala Gly Leu Lys Lys Val Thr 275 280 285 Leu Glu Leu Gly Gly Lys Ser Pro Asn Ile Val Phe Ala Asp Ala Glu 290 295 300 Leu Lys Lys Ala Val Gln Asn Ile Ile Leu Gly Ile Tyr Tyr Asn Ser 305 310 315 320 Gly Glu Val Cys Cys Ala Gly Ser Arg Val Tyr Val Glu Glu Ser Ile 325 330 335 Tyr Asp Lys Phe Ile Glu Glu Phe Lys Ala Ala Ser Glu Ser Ile Lys 340 345 350 Val Gly Asp Pro Phe Asp Glu Ser Thr Phe Gln Gly Ala Gln Thr Ser 355 360 365 Gln Met Gln Leu Asn Lys Ile Leu Lys Tyr Val Asp Ile Gly Lys Asn 370 375 380 Glu Gly Ala Thr Leu Ile Thr Gly Gly Glu Arg Leu Gly Ser Lys Gly 385 390 395 400 Tyr Phe Ile Lys Pro Thr Val Phe Gly Asp Val Lys Glu Asp Met Arg 405 410 415 Ile Val Lys Glu Glu Ile Phe Gly Pro Val Val Thr Val Thr Lys Phe 420 425 430 Lys Ser Ala Asp Glu Val Ile Asn Met Ala Asn Asp Ser Glu Tyr Gly 435 440 445 Leu Ala Ala Gly Ile His Thr Ser Asn Ile Asn Thr Ala Leu Lys Val 450 455 460 Ala Asp Arg Val Asn Ala Gly Thr Val Trp Ile Asn Thr Tyr Asn Asp 465 470 475 480 Phe His His Ala Val Pro Phe Gly Gly Phe Asn Ala Ser Gly Leu Gly 485 490 495 Arg Glu Met Ser Val Asp Ala Leu Gln Asn Tyr Leu Gln Val Lys Ala 500 505 510 Val Arg Ala Lys Leu Asp Glu 515 156520PRTSaccharomyces cerevisiae 156Met Leu Ser Arg Thr Arg Ala Ala Ala Pro Asn Ser Arg Ile Phe Thr 1 5 10 15 Arg Ser Leu Leu Arg Leu Tyr Ser Gln Ala Pro Leu Arg Val Pro Ile 20 25 30 Thr Leu Pro Asn Gly Phe Thr Tyr Glu Gln Pro Thr Gly Leu Phe Ile 35 40 45 Asn Gly Glu Phe Val Ala Ser Lys Gln Lys Lys Thr Phe Asp Val Ile 50 55 60 Asn Pro Ser Asn Glu Glu Lys Ile Thr Thr Val Tyr Lys Ala Met Glu 65 70 75 80 Asp Asp Val Asp Glu Ala Val Ala Ala Ala Lys Lys Ala Phe Glu Thr 85 90 95 Lys Trp Ser Ile Val Glu Pro Glu Val Arg Ala Lys Ala Leu Phe Asn 100 105 110 Leu Ala Asp Leu Val Glu Lys His Gln Glu Thr Leu Ala Ala Ile Glu 115 120 125 Ser Met Asp Asn Gly Lys Ser Leu

Phe Cys Ala Arg Gly Asp Val Ala 130 135 140 Leu Val Ser Lys Tyr Leu Arg Ser Cys Gly Gly Trp Ala Asp Lys Ile 145 150 155 160 Tyr Gly Asn Val Ile Asp Thr Gly Lys Asn His Phe Thr Tyr Ser Ile 165 170 175 Lys Glu Pro Leu Gly Val Cys Gly Gln Ile Ile Pro Trp Asn Phe Pro 180 185 190 Leu Leu Met Trp Ser Trp Lys Ile Gly Pro Ala Leu Ala Thr Gly Asn 195 200 205 Thr Val Val Leu Lys Pro Ala Glu Thr Thr Pro Leu Ser Ala Leu Phe 210 215 220 Ala Ser Gln Leu Cys Gln Glu Ala Gly Ile Pro Ala Gly Val Val Asn 225 230 235 240 Ile Leu Pro Gly Ser Gly Arg Val Val Gly Glu Arg Leu Ser Ala His 245 250 255 Pro Asp Val Lys Lys Ile Ala Phe Thr Gly Ser Thr Ala Thr Gly Arg 260 265 270 His Ile Met Lys Val Ala Ala Asp Thr Val Lys Lys Val Thr Leu Glu 275 280 285 Leu Gly Gly Lys Ser Pro Asn Ile Val Phe Ala Asp Ala Asp Leu Asp 290 295 300 Lys Ala Val Lys Asn Ile Ala Phe Gly Ile Phe Tyr Asn Ser Gly Glu 305 310 315 320 Val Cys Cys Ala Gly Ser Arg Ile Tyr Ile Gln Asp Thr Val Tyr Glu 325 330 335 Glu Val Leu Gln Lys Leu Lys Asp Tyr Thr Glu Ser Leu Lys Val Gly 340 345 350 Asp Pro Phe Asp Glu Glu Val Phe Gln Gly Ala Gln Thr Ser Asp Lys 355 360 365 Gln Leu His Lys Ile Leu Asp Tyr Val Asp Val Ala Lys Ser Glu Gly 370 375 380 Ala Arg Leu Val Thr Gly Gly Ala Arg His Gly Ser Lys Gly Tyr Phe 385 390 395 400 Val Lys Pro Thr Val Phe Ala Asp Val Lys Gly Asp Met Arg Ile Val 405 410 415 Lys Glu Glu Val Phe Gly Pro Ile Val Thr Val Ser Lys Phe Ser Thr 420 425 430 Val Asp Glu Val Ile Ala Met Ala Asn Asp Ser Gln Tyr Gly Leu Ala 435 440 445 Ala Gly Ile His Thr Asn Asp Ile Asn Lys Ala Val Asp Val Ser Lys 450 455 460 Arg Val Lys Ala Gly Thr Val Trp Ile Asn Thr Tyr Asn Asn Phe His 465 470 475 480 Gln Asn Val Pro Phe Gly Gly Phe Gly Gln Ser Gly Ile Gly Arg Glu 485 490 495 Met Gly Glu Ala Ala Leu Ser Asn Tyr Thr Gln Thr Lys Ser Val Arg 500 505 510 Ile Ala Ile Asp Lys Pro Ile Arg 515 520 157500PRTSaccharomyces cerevisiae 157Met Thr Lys Leu His Phe Asp Thr Ala Glu Pro Val Lys Ile Thr Leu 1 5 10 15 Pro Asn Gly Leu Thr Tyr Glu Gln Pro Thr Gly Leu Phe Ile Asn Asn 20 25 30 Lys Phe Met Lys Ala Gln Asp Gly Lys Thr Tyr Pro Val Glu Asp Pro 35 40 45 Ser Thr Glu Asn Thr Val Cys Glu Val Ser Ser Ala Thr Thr Glu Asp 50 55 60 Val Glu Tyr Ala Ile Glu Cys Ala Asp Arg Ala Phe His Asp Thr Glu 65 70 75 80 Trp Ala Thr Gln Asp Pro Arg Glu Arg Gly Arg Leu Leu Ser Lys Leu 85 90 95 Ala Asp Glu Leu Glu Ser Gln Ile Asp Leu Val Ser Ser Ile Glu Ala 100 105 110 Leu Asp Asn Gly Lys Thr Leu Ala Leu Ala Arg Gly Asp Val Thr Ile 115 120 125 Ala Ile Asn Cys Leu Arg Asp Ala Ala Ala Tyr Ala Asp Lys Val Asn 130 135 140 Gly Arg Thr Ile Asn Thr Gly Asp Gly Tyr Met Asn Phe Thr Thr Leu 145 150 155 160 Glu Pro Ile Gly Val Cys Gly Gln Ile Ile Pro Trp Asn Phe Pro Ile 165 170 175 Met Met Leu Ala Trp Lys Ile Ala Pro Ala Leu Ala Met Gly Asn Val 180 185 190 Cys Ile Leu Lys Pro Ala Ala Val Thr Pro Leu Asn Ala Leu Tyr Phe 195 200 205 Ala Ser Leu Cys Lys Lys Val Gly Ile Pro Ala Gly Val Val Asn Ile 210 215 220 Val Pro Gly Pro Gly Arg Thr Val Gly Ala Ala Leu Thr Asn Asp Pro 225 230 235 240 Arg Ile Arg Lys Leu Ala Phe Thr Gly Ser Thr Glu Val Gly Lys Ser 245 250 255 Val Ala Val Asp Ser Ser Glu Ser Asn Leu Lys Lys Ile Thr Leu Glu 260 265 270 Leu Gly Gly Lys Ser Ala His Leu Val Phe Asp Asp Ala Asn Ile Lys 275 280 285 Lys Thr Leu Pro Asn Leu Val Asn Gly Ile Phe Lys Asn Ala Gly Gln 290 295 300 Ile Cys Ser Ser Gly Ser Arg Ile Tyr Val Gln Glu Gly Ile Tyr Asp 305 310 315 320 Glu Leu Leu Ala Ala Phe Lys Ala Tyr Leu Glu Thr Glu Ile Lys Val 325 330 335 Gly Asn Pro Phe Asp Lys Ala Asn Phe Gln Gly Ala Ile Thr Asn Arg 340 345 350 Gln Gln Phe Asp Thr Ile Met Asn Tyr Ile Asp Ile Gly Lys Lys Glu 355 360 365 Gly Ala Lys Ile Leu Thr Gly Gly Glu Lys Val Gly Asp Lys Gly Tyr 370 375 380 Phe Ile Arg Pro Thr Val Phe Tyr Asp Val Asn Glu Asp Met Arg Ile 385 390 395 400 Val Lys Glu Glu Ile Phe Gly Pro Val Val Thr Val Ala Lys Phe Lys 405 410 415 Thr Leu Glu Glu Gly Val Glu Met Ala Asn Ser Ser Glu Phe Gly Leu 420 425 430 Gly Ser Gly Ile Glu Thr Glu Ser Leu Ser Thr Gly Leu Lys Val Ala 435 440 445 Lys Met Leu Lys Ala Gly Thr Val Trp Ile Asn Thr Tyr Asn Asp Phe 450 455 460 Asp Ser Arg Val Pro Phe Gly Gly Val Lys Gln Ser Gly Tyr Gly Arg 465 470 475 480 Glu Met Gly Glu Glu Val Tyr His Ala Tyr Thr Glu Val Lys Ala Val 485 490 495 Arg Ile Lys Leu 500 158532PRTSaccharomyces cerevisiae 158Met Ser Asn Asp Gly Ser Lys Ile Leu Asn Tyr Thr Pro Val Ser Lys 1 5 10 15 Ile Asp Glu Ile Val Glu Ile Ser Arg Asn Phe Phe Phe Glu Lys Gln 20 25 30 Leu Lys Leu Ser His Glu Asn Asn Pro Arg Lys Lys Asp Leu Glu Phe 35 40 45 Arg Gln Leu Gln Leu Lys Lys Leu Tyr Tyr Ala Val Lys Asp His Glu 50 55 60 Glu Glu Leu Ile Asp Ala Met Tyr Lys Asp Phe His Arg Asn Lys Ile 65 70 75 80 Glu Ser Val Leu Asn Glu Thr Thr Lys Leu Met Asn Asp Ile Leu His 85 90 95 Leu Ile Glu Ile Leu Pro Lys Leu Ile Lys Pro Arg Arg Val Ser Asp 100 105 110 Ser Ser Pro Pro Phe Met Phe Gly Lys Thr Ile Val Glu Lys Ile Ser 115 120 125 Arg Gly Ser Val Leu Ile Ile Ala Pro Phe Asn Phe Pro Leu Leu Leu 130 135 140 Ala Phe Ala Pro Leu Ala Ala Ala Leu Ala Ala Gly Asn Thr Ile Val 145 150 155 160 Leu Lys Pro Ser Glu Leu Thr Pro His Thr Ala Val Val Met Glu Asn 165 170 175 Leu Leu Thr Thr Ala Gly Phe Pro Asp Gly Leu Ile Gln Val Val Gln 180 185 190 Gly Ala Ile Asp Glu Thr Thr Arg Leu Leu Asp Cys Gly Lys Phe Asp 195 200 205 Leu Ile Phe Tyr Thr Gly Ser Pro Arg Val Gly Ser Ile Val Ala Glu 210 215 220 Lys Ala Ala Lys Ser Leu Thr Pro Cys Val Leu Glu Leu Gly Gly Lys 225 230 235 240 Ser Pro Thr Phe Ile Thr Glu Asn Phe Lys Ala Ser Asn Ile Lys Ile 245 250 255 Ala Leu Lys Arg Ile Phe Phe Gly Ala Phe Gly Asn Ser Gly Gln Ile 260 265 270 Cys Val Ser Pro Asp Tyr Leu Leu Val His Lys Ser Ile Tyr Pro Lys 275 280 285 Val Ile Lys Glu Cys Glu Ser Val Leu Asn Glu Phe Tyr Pro Ser Phe 290 295 300 Asp Glu Gln Thr Asp Phe Thr Arg Met Ile His Glu Pro Ala Tyr Lys 305 310 315 320 Lys Ala Val Ala Ser Ile Asn Ser Thr Asn Gly Ser Lys Ile Val Pro 325 330 335 Ser Lys Ile Ser Ile Asn Ser Asp Thr Glu Asp Leu Cys Leu Val Pro 340 345 350 Pro Thr Ile Val Tyr Asn Ile Gly Trp Asp Asp Pro Leu Met Lys Gln 355 360 365 Glu Asn Phe Ala Pro Val Leu Pro Ile Ile Glu Tyr Glu Asp Leu Asp 370 375 380 Glu Thr Ile Asn Lys Ile Ile Glu Glu His Asp Thr Pro Leu Val Gln 385 390 395 400 Tyr Ile Phe Ser Asp Ser Gln Thr Glu Ile Asn Arg Ile Leu Thr Arg 405 410 415 Leu Arg Ser Gly Asp Cys Val Val Gly Asp Thr Val Ile His Val Gly 420 425 430 Ile Thr Asp Ala Pro Phe Gly Gly Ile Gly Thr Ser Gly Tyr Gly Asn 435 440 445 Tyr Gly Gly Tyr Tyr Gly Phe Asn Thr Phe Ser His Glu Arg Thr Ile 450 455 460 Phe Lys Gln Pro Tyr Trp Asn Asp Phe Thr Leu Phe Met Arg Tyr Pro 465 470 475 480 Pro Asn Ser Ala Gln Lys Glu Lys Leu Val Arg Phe Ala Met Glu Arg 485 490 495 Lys Pro Trp Phe Asp Arg Asn Gly Asn Asn Lys Trp Gly Leu Arg Gln 500 505 510 Tyr Phe Ser Leu Ser Ala Ala Val Ile Leu Ile Ser Thr Ile Tyr Ala 515 520 525 His Cys Ser Ser 530 159500PRTSaccharomyces cerevisiae 159Met Ser Phe Asp Asp Leu His Lys Ala Thr Glu Arg Ala Val Ile Gln 1 5 10 15 Ala Val Asp Gln Ile Cys Asp Asp Phe Glu Val Thr Pro Glu Lys Leu 20 25 30 Asp Glu Leu Thr Ala Tyr Phe Ile Glu Gln Met Glu Lys Gly Leu Ala 35 40 45 Pro Pro Lys Glu Gly His Thr Leu Ala Ser Asp Lys Gly Leu Pro Met 50 55 60 Ile Pro Ala Phe Val Thr Gly Ser Pro Asn Gly Thr Glu Arg Gly Val 65 70 75 80 Leu Leu Ala Ala Asp Leu Gly Gly Thr Asn Phe Arg Ile Cys Ser Val 85 90 95 Asn Leu His Gly Asp His Thr Phe Ser Met Glu Gln Met Lys Ser Lys 100 105 110 Ile Pro Asp Asp Leu Leu Asp Asp Glu Asn Val Thr Ser Asp Asp Leu 115 120 125 Phe Gly Phe Leu Ala Arg Arg Thr Leu Ala Phe Met Lys Lys Tyr His 130 135 140 Pro Asp Glu Leu Ala Lys Gly Lys Asp Ala Lys Pro Met Lys Leu Gly 145 150 155 160 Phe Thr Phe Ser Tyr Pro Val Asp Gln Thr Ser Leu Asn Ser Gly Thr 165 170 175 Leu Ile Arg Trp Thr Lys Gly Phe Arg Ile Ala Asp Thr Val Gly Lys 180 185 190 Asp Val Val Gln Leu Tyr Gln Glu Gln Leu Ser Ala Gln Gly Met Pro 195 200 205 Met Ile Lys Val Val Ala Leu Thr Asn Asp Thr Val Gly Thr Tyr Leu 210 215 220 Ser His Cys Tyr Thr Ser Asp Asn Thr Asp Ser Met Thr Ser Gly Glu 225 230 235 240 Ile Ser Glu Pro Val Ile Gly Cys Ile Phe Gly Thr Gly Thr Asn Gly 245 250 255 Cys Tyr Met Glu Glu Ile Asn Lys Ile Thr Lys Leu Pro Gln Glu Leu 260 265 270 Arg Asp Lys Leu Ile Lys Glu Gly Lys Thr His Met Ile Ile Asn Val 275 280 285 Glu Trp Gly Ser Phe Asp Asn Glu Leu Lys His Leu Pro Thr Thr Lys 290 295 300 Tyr Asp Val Val Ile Asp Gln Lys Leu Ser Thr Asn Pro Gly Phe His 305 310 315 320 Leu Phe Glu Lys Arg Val Ser Gly Met Phe Leu Gly Glu Val Leu Arg 325 330 335 Asn Ile Leu Val Asp Leu His Ser Gln Gly Leu Leu Leu Gln Gln Tyr 340 345 350 Arg Ser Lys Glu Gln Leu Pro Arg His Leu Thr Thr Pro Phe Gln Leu 355 360 365 Ser Ser Glu Val Leu Ser His Ile Glu Ile Asp Asp Ser Thr Gly Leu 370 375 380 Arg Glu Thr Glu Leu Ser Leu Leu Gln Ser Leu Arg Leu Pro Thr Thr 385 390 395 400 Pro Thr Glu Arg Val Gln Ile Gln Lys Leu Val Arg Ala Ile Ser Arg 405 410 415 Arg Ser Ala Tyr Leu Ala Ala Val Pro Leu Ala Ala Ile Leu Ile Lys 420 425 430 Thr Asn Ala Leu Asn Lys Arg Tyr His Gly Glu Val Glu Ile Gly Cys 435 440 445 Asp Gly Ser Val Val Glu Tyr Tyr Pro Gly Phe Arg Ser Met Leu Arg 450 455 460 His Ala Leu Ala Leu Ser Pro Leu Gly Ala Glu Gly Glu Arg Lys Val 465 470 475 480 His Leu Lys Ile Ala Lys Asp Gly Ser Gly Val Gly Ala Ala Leu Cys 485 490 495 Ala Leu Val Ala 500 160554PRTSaccharomyces cerevisiae 160Met Ser Asn Asn Ser Phe Thr Asn Phe Lys Leu Ala Thr Glu Leu Pro 1 5 10 15 Ala Trp Ser Lys Leu Gln Lys Ile Tyr Glu Ser Gln Gly Lys Thr Leu 20 25 30 Ser Val Lys Gln Glu Phe Gln Lys Asp Ala Lys Arg Phe Glu Lys Leu 35 40 45 Asn Lys Thr Phe Thr Asn Tyr Asp Gly Ser Lys Ile Leu Phe Asp Tyr 50 55 60 Ser Lys Asn Leu Val Asn Asp Glu Ile Ile Ala Ala Leu Ile Glu Leu 65 70 75 80 Ala Lys Glu Ala Asn Val Thr Gly Leu Arg Asp Ala Met Phe Lys Gly 85 90 95 Glu His Ile Asn Ser Thr Glu Asp Arg Ala Val Tyr His Val Ala Leu 100 105 110 Arg Asn Arg Ala Asn Lys Pro Met Tyr Val Asp Gly Val Asn Val Ala 115 120 125 Pro Glu Val Asp Ser Val Leu Lys His Met Lys Glu Phe Ser Glu Gln 130 135 140 Val Arg Ser Gly Glu Trp Lys Gly Tyr Thr Gly Lys Lys Ile Thr Asp 145 150 155 160 Val Val Asn Ile Gly Ile Gly Gly Ser Asp Leu Gly Pro Val Met Val 165 170 175 Thr Glu Ala Leu Lys His Tyr Ala Gly Val Leu Asp Val His Phe Val 180 185 190 Ser Asn Ile Asp Gly Thr His Ile Ala Glu Thr Leu Lys Val Val Asp 195 200 205 Pro Glu Thr Thr Leu Phe Leu Ile Ala Ser Lys Thr Phe Thr Thr Ala 210 215 220 Glu Thr Ile Thr Asn Ala Asn Thr Ala Lys Asn Trp Phe Leu Ser Lys 225 230 235 240 Thr Gly Asn Asp Pro Ser His Ile Ala Lys His Phe Ala Ala Leu Ser 245 250 255 Thr Asn Glu Thr Glu Val Ala Lys Phe Gly Ile Asp Thr Lys Asn Met 260 265 270 Phe Gly Phe Glu Ser Trp Val Gly Gly Arg Tyr Ser Val Trp Ser Ala 275 280 285 Ile Gly Leu Ser Val Ala Leu Tyr Ile Gly Tyr Asp Asn Phe Glu Ala 290 295 300 Phe Leu Lys Gly Ala Glu Ala Val Asp Asn His Phe Thr Gln Thr Pro 305 310 315 320 Leu Glu Asp Asn Ile Pro Leu Leu Gly Gly Leu Leu Ser Val Trp Tyr 325 330 335 Asn Asn Phe Phe Gly Ala Gln Thr His Leu Val Ala Pro Phe Asp Gln 340 345 350 Tyr Leu His Arg Phe Pro Ala Tyr Leu Gln Gln Leu Ser Met Glu Ser 355 360 365 Asn

Gly Lys Ser Val Thr Arg Gly Asn Val Phe Thr Asp Tyr Ser Thr 370 375 380 Gly Ser Ile Leu Phe Gly Glu Pro Ala Thr Asn Ala Gln His Ser Phe 385 390 395 400 Phe Gln Leu Val His Gln Gly Thr Lys Leu Ile Pro Ser Asp Phe Ile 405 410 415 Leu Ala Ala Gln Ser His Asn Pro Ile Glu Asn Lys Leu His Gln Lys 420 425 430 Met Leu Ala Ser Asn Phe Phe Ala Gln Ala Glu Ala Leu Met Val Gly 435 440 445 Lys Asp Glu Glu Gln Val Lys Ala Glu Gly Ala Thr Gly Gly Leu Val 450 455 460 Pro His Lys Val Phe Ser Gly Asn Arg Pro Thr Thr Ser Ile Leu Ala 465 470 475 480 Gln Lys Ile Thr Pro Ala Thr Leu Gly Ala Leu Ile Ala Tyr Tyr Glu 485 490 495 His Val Thr Phe Thr Glu Gly Ala Ile Trp Asn Ile Asn Ser Phe Asp 500 505 510 Gln Trp Gly Val Glu Leu Gly Lys Val Leu Ala Lys Val Ile Gly Lys 515 520 525 Glu Leu Asp Asn Ser Ser Thr Ile Ser Thr His Asp Ala Ser Thr Asn 530 535 540 Gly Leu Ile Asn Gln Phe Lys Glu Trp Met 545 550 161987PRTSaccharomyces cerevisiae 161Met Gln Ser Gln Asp Ser Cys Tyr Gly Val Ala Phe Arg Ser Ile Ile 1 5 10 15 Thr Asn Asp Glu Ala Leu Phe Lys Lys Thr Ile His Phe Tyr His Thr 20 25 30 Leu Gly Phe Ala Thr Val Lys Asp Phe Asn Lys Phe Lys His Gly Glu 35 40 45 Asn Ser Leu Leu Ser Ser Gly Thr Ser Gln Asp Ser Leu Arg Glu Val 50 55 60 Trp Leu Glu Ser Phe Lys Leu Ser Glu Val Asp Ala Ser Gly Phe Arg 65 70 75 80 Ile Pro Gln Gln Glu Ala Thr Asn Lys Ala Gln Ser Gln Gly Ala Leu 85 90 95 Leu Lys Ile Arg Leu Val Met Ser Ala Pro Ile Asp Glu Thr Phe Asp 100 105 110 Thr Asn Glu Thr Ala Thr Ile Thr Tyr Phe Ser Thr Asp Leu Asn Lys 115 120 125 Ile Val Glu Lys Phe Pro Lys Gln Ala Glu Lys Leu Ser Asp Thr Leu 130 135 140 Val Phe Leu Lys Asp Pro Met Gly Asn Asn Ile Thr Phe Ser Gly Leu 145 150 155 160 Ala Asn Ala Thr Asp Ser Ala Pro Thr Ser Lys Asp Ala Phe Leu Glu 165 170 175 Ala Thr Ser Glu Asp Glu Ile Ile Ser Arg Ala Ser Ser Asp Ala Ser 180 185 190 Asp Leu Leu Arg Gln Thr Leu Gly Ser Ser Gln Lys Lys Lys Lys Ile 195 200 205 Ala Val Met Thr Ser Gly Gly Asp Ser Pro Gly Met Asn Ala Ala Val 210 215 220 Arg Ala Val Val Arg Thr Gly Ile His Phe Gly Cys Asp Val Phe Ala 225 230 235 240 Val Tyr Glu Gly Tyr Glu Gly Leu Leu Arg Gly Gly Lys Tyr Leu Lys 245 250 255 Lys Met Ala Trp Glu Asp Val Arg Gly Trp Leu Ser Glu Gly Gly Thr 260 265 270 Leu Ile Gly Thr Ala Arg Ser Met Glu Phe Arg Lys Arg Glu Gly Arg 275 280 285 Arg Gln Ala Ala Gly Asn Leu Ile Ser Gln Gly Ile Asp Ala Leu Val 290 295 300 Val Cys Gly Gly Asp Gly Ser Leu Thr Gly Ala Asp Leu Phe Arg His 305 310 315 320 Glu Trp Pro Ser Leu Val Asp Glu Leu Val Ala Glu Gly Arg Phe Thr 325 330 335 Lys Glu Glu Val Ala Pro Tyr Lys Asn Leu Ser Ile Val Gly Leu Val 340 345 350 Gly Ser Ile Asp Asn Asp Met Ser Gly Thr Asp Ser Thr Ile Gly Ala 355 360 365 Tyr Ser Ala Leu Glu Arg Ile Cys Glu Met Val Asp Tyr Ile Asp Ala 370 375 380 Thr Ala Lys Ser His Ser Arg Ala Phe Val Val Glu Val Met Gly Arg 385 390 395 400 His Cys Gly Trp Leu Ala Leu Met Ala Gly Ile Ala Thr Gly Ala Asp 405 410 415 Tyr Ile Phe Ile Pro Glu Arg Ala Val Pro His Gly Lys Trp Gln Asp 420 425 430 Glu Leu Lys Glu Val Cys Gln Arg His Arg Ser Lys Gly Arg Arg Asn 435 440 445 Asn Thr Ile Ile Val Ala Glu Gly Ala Leu Asp Asp Gln Leu Asn Pro 450 455 460 Val Thr Ala Asn Asp Val Lys Asp Ala Leu Ile Glu Leu Gly Leu Asp 465 470 475 480 Thr Lys Val Thr Ile Leu Gly His Val Gln Arg Gly Gly Thr Ala Val 485 490 495 Ala His Asp Arg Trp Leu Ala Thr Leu Gln Gly Val Asp Ala Val Lys 500 505 510 Ala Val Leu Glu Phe Thr Pro Glu Thr Pro Ser Pro Leu Ile Gly Ile 515 520 525 Leu Glu Asn Lys Ile Ile Arg Met Pro Leu Val Glu Ser Val Lys Leu 530 535 540 Thr Lys Ser Val Ala Thr Ala Ile Glu Asn Lys Asp Phe Asp Lys Ala 545 550 555 560 Ile Ser Leu Arg Asp Thr Glu Phe Ile Glu Leu Tyr Glu Asn Phe Leu 565 570 575 Ser Thr Thr Val Lys Asp Asp Gly Ser Glu Leu Leu Pro Val Ser Asp 580 585 590 Arg Leu Asn Ile Gly Ile Val His Val Gly Ala Pro Ser Ala Ala Leu 595 600 605 Asn Ala Ala Thr Arg Ala Ala Thr Leu Tyr Cys Leu Ser His Gly His 610 615 620 Lys Pro Tyr Ala Ile Met Asn Gly Phe Ser Gly Leu Ile Gln Thr Gly 625 630 635 640 Glu Val Lys Glu Leu Ser Trp Ile Asp Val Glu Asn Trp His Asn Leu 645 650 655 Gly Gly Ser Glu Ile Gly Thr Asn Arg Ser Val Ala Ser Glu Asp Leu 660 665 670 Gly Thr Ile Ala Tyr Tyr Phe Gln Lys Asn Lys Leu Asp Gly Leu Ile 675 680 685 Ile Leu Gly Gly Phe Glu Gly Phe Arg Ser Leu Lys Gln Leu Arg Asp 690 695 700 Gly Arg Thr Gln His Pro Ile Phe Asn Ile Pro Met Cys Leu Ile Pro 705 710 715 720 Ala Thr Val Ser Asn Asn Val Pro Gly Thr Glu Tyr Ser Leu Gly Val 725 730 735 Asp Thr Cys Leu Asn Ala Leu Val Asn Tyr Thr Asp Asp Ile Lys Gln 740 745 750 Ser Ala Ser Ala Thr Arg Arg Arg Val Phe Val Cys Glu Val Gln Gly 755 760 765 Gly His Ser Gly Tyr Ile Ala Ser Phe Thr Gly Leu Ile Thr Gly Ala 770 775 780 Val Ser Val Tyr Thr Pro Glu Lys Lys Ile Asp Leu Ala Ser Ile Arg 785 790 795 800 Glu Asp Ile Thr Leu Leu Lys Glu Asn Phe Arg His Asp Lys Gly Glu 805 810 815 Asn Arg Asn Gly Lys Leu Leu Val Arg Asn Glu Gln Ala Ser Ser Val 820 825 830 Tyr Ser Thr Gln Leu Leu Ala Asp Ile Ile Ser Glu Ala Ser Lys Gly 835 840 845 Lys Phe Gly Val Arg Thr Ala Ile Pro Gly His Val Gln Gln Gly Gly 850 855 860 Val Pro Ser Ser Lys Asp Arg Val Thr Ala Ser Arg Phe Ala Val Lys 865 870 875 880 Cys Ile Lys Phe Ile Glu Gln Trp Asn Lys Lys Asn Glu Ala Ser Pro 885 890 895 Asn Thr Asp Ala Lys Val Leu Arg Phe Lys Phe Asp Thr His Gly Glu 900 905 910 Lys Val Pro Thr Val Glu His Glu Asp Asp Ser Ala Ala Val Ile Cys 915 920 925 Val Asn Gly Ser His Val Ser Phe Lys Pro Ile Ala Asn Leu Trp Glu 930 935 940 Asn Glu Thr Asn Val Glu Leu Arg Lys Gly Phe Glu Val His Trp Ala 945 950 955 960 Glu Tyr Asn Lys Ile Gly Asp Ile Leu Ser Gly Arg Leu Lys Leu Arg 965 970 975 Ala Glu Val Ala Ala Leu Ala Ala Glu Asn Lys 980 985 162959PRTSaccharomyces cerevisiae 162Met Thr Val Thr Thr Pro Phe Val Asn Gly Thr Ser Tyr Cys Thr Val 1 5 10 15 Thr Ala Tyr Ser Val Gln Ser Tyr Lys Ala Ala Ile Asp Phe Tyr Thr 20 25 30 Lys Phe Leu Ser Leu Glu Asn Arg Ser Ser Pro Asp Glu Asn Ser Thr 35 40 45 Leu Leu Ser Asn Asp Ser Ile Ser Leu Lys Ile Leu Leu Arg Pro Asp 50 55 60 Glu Lys Ile Asn Lys Asn Val Glu Ala His Leu Lys Glu Leu Asn Ser 65 70 75 80 Ile Thr Lys Thr Gln Asp Trp Arg Ser His Ala Thr Gln Ser Leu Val 85 90 95 Phe Asn Thr Ser Asp Ile Leu Ala Val Lys Asp Thr Leu Asn Ala Met 100 105 110 Asn Ala Pro Leu Gln Gly Tyr Pro Thr Glu Leu Phe Pro Met Gln Leu 115 120 125 Tyr Thr Leu Asp Pro Leu Gly Asn Val Val Gly Val Thr Ser Thr Lys 130 135 140 Asn Ala Val Ser Thr Lys Pro Thr Pro Pro Pro Ala Pro Glu Ala Ser 145 150 155 160 Ala Glu Ser Gly Leu Ser Ser Lys Val His Ser Tyr Thr Asp Leu Ala 165 170 175 Tyr Arg Met Lys Thr Thr Asp Thr Tyr Pro Ser Leu Pro Lys Pro Leu 180 185 190 Asn Arg Pro Gln Lys Ala Ile Ala Val Met Thr Ser Gly Gly Asp Ala 195 200 205 Pro Gly Met Asn Ser Asn Val Arg Ala Ile Val Arg Ser Ala Ile Phe 210 215 220 Lys Gly Cys Arg Ala Phe Val Val Met Glu Gly Tyr Glu Gly Leu Val 225 230 235 240 Arg Gly Gly Pro Glu Tyr Ile Lys Glu Phe His Trp Glu Asp Val Arg 245 250 255 Gly Trp Ser Ala Glu Gly Gly Thr Asn Ile Gly Thr Ala Arg Cys Met 260 265 270 Glu Phe Lys Lys Arg Glu Gly Arg Leu Leu Gly Ala Gln His Leu Ile 275 280 285 Glu Ala Gly Val Asp Ala Leu Ile Val Cys Gly Gly Asp Gly Ser Leu 290 295 300 Thr Gly Ala Asp Leu Phe Arg Ser Glu Trp Pro Ser Leu Ile Glu Glu 305 310 315 320 Leu Leu Lys Thr Asn Arg Ile Ser Asn Glu Gln Tyr Glu Arg Met Lys 325 330 335 His Leu Asn Ile Cys Gly Thr Val Gly Ser Ile Asp Asn Asp Met Ser 340 345 350 Thr Thr Asp Ala Thr Ile Gly Ala Tyr Ser Ala Leu Asp Arg Ile Cys 355 360 365 Lys Ala Ile Asp Tyr Val Glu Ala Thr Ala Asn Ser His Ser Arg Ala 370 375 380 Phe Val Val Glu Val Met Gly Arg Asn Cys Gly Trp Leu Ala Leu Leu 385 390 395 400 Ala Gly Ile Ala Thr Ser Ala Asp Tyr Ile Phe Ile Pro Glu Lys Pro 405 410 415 Ala Thr Ser Ser Glu Trp Gln Asp Gln Met Cys Asp Ile Val Ser Lys 420 425 430 His Arg Ser Arg Gly Lys Arg Thr Thr Ile Val Val Val Ala Glu Gly 435 440 445 Ala Ile Ala Ala Asp Leu Thr Pro Ile Ser Pro Ser Asp Val His Lys 450 455 460 Val Leu Val Asp Arg Leu Gly Leu Asp Thr Arg Ile Thr Thr Leu Gly 465 470 475 480 His Val Gln Arg Gly Gly Thr Ala Val Ala Tyr Asp Arg Ile Leu Ala 485 490 495 Thr Leu Gln Gly Leu Glu Ala Val Asn Ala Val Leu Glu Ser Thr Pro 500 505 510 Asp Thr Pro Ser Pro Leu Ile Ala Val Asn Glu Asn Lys Ile Val Arg 515 520 525 Lys Pro Leu Met Glu Ser Val Lys Leu Thr Lys Ala Val Ala Glu Ala 530 535 540 Ile Gln Ala Lys Asp Phe Lys Arg Ala Met Ser Leu Arg Asp Thr Glu 545 550 555 560 Phe Ile Glu His Leu Asn Asn Phe Met Ala Ile Asn Ser Ala Asp His 565 570 575 Asn Glu Pro Lys Leu Pro Lys Asp Lys Arg Leu Lys Ile Ala Ile Val 580 585 590 Asn Val Gly Ala Pro Ala Gly Gly Ile Asn Ser Ala Val Tyr Ser Met 595 600 605 Ala Thr Tyr Cys Met Ser Gln Gly His Arg Pro Tyr Ala Ile Tyr Asn 610 615 620 Gly Trp Ser Gly Leu Ala Arg His Glu Ser Val Arg Ser Leu Asn Trp 625 630 635 640 Lys Asp Met Leu Gly Trp Gln Ser Arg Gly Gly Ser Glu Ile Gly Thr 645 650 655 Asn Arg Val Thr Pro Glu Glu Ala Asp Leu Gly Met Ile Ala Tyr Tyr 660 665 670 Phe Gln Lys Tyr Glu Phe Asp Gly Leu Ile Ile Val Gly Gly Phe Glu 675 680 685 Ala Phe Glu Ser Leu His Gln Leu Glu Arg Ala Arg Glu Ser Tyr Pro 690 695 700 Ala Phe Arg Ile Pro Met Val Leu Ile Pro Ala Thr Leu Ser Asn Asn 705 710 715 720 Val Pro Gly Thr Glu Tyr Ser Leu Gly Ser Asp Thr Ala Leu Asn Ala 725 730 735 Leu Met Glu Tyr Cys Asp Val Val Lys Gln Ser Ala Ser Ser Thr Arg 740 745 750 Gly Arg Ala Phe Val Val Asp Cys Gln Gly Gly Asn Ser Gly Tyr Leu 755 760 765 Ala Thr Tyr Ala Ser Leu Ala Val Gly Ala Gln Val Ser Tyr Val Pro 770 775 780 Glu Glu Gly Ile Ser Leu Glu Gln Leu Ser Glu Asp Ile Glu Tyr Leu 785 790 795 800 Ala Gln Ser Phe Glu Lys Ala Glu Gly Arg Gly Arg Phe Gly Lys Leu 805 810 815 Ile Leu Lys Ser Thr Asn Ala Ser Lys Ala Leu Ser Ala Thr Lys Leu 820 825 830 Ala Glu Val Ile Thr Ala Glu Ala Asp Gly Arg Phe Asp Ala Lys Pro 835 840 845 Ala Tyr Pro Gly His Val Gln Gln Gly Gly Leu Pro Ser Pro Ile Asp 850 855 860 Arg Thr Arg Ala Thr Arg Met Ala Ile Lys Ala Val Gly Phe Ile Lys 865 870 875 880 Asp Asn Gln Ala Ala Ile Ala Glu Ala Arg Ala Ala Glu Glu Asn Phe 885 890 895 Asn Ala Asp Asp Lys Thr Ile Ser Asp Thr Ala Ala Val Val Gly Val 900 905 910 Lys Gly Ser His Val Val Tyr Asn Ser Ile Arg Gln Leu Tyr Asp Tyr 915 920 925 Glu Thr Glu Val Ser Met Arg Met Pro Lys Val Ile His Trp Gln Ala 930 935 940 Thr Arg Leu Ile Ala Asp His Leu Val Gly Arg Lys Arg Val Asp 945 950 955 163563PRTSaccharomyces cerevisiae 163Met Ser Glu Ile Thr Leu Gly Lys Tyr Leu Phe Glu Arg Leu Lys Gln 1 5 10 15 Val Asn Val Asn Thr Val Phe Gly Leu Pro Gly Asp Phe Asn Leu Ser 20 25 30 Leu Leu Asp Lys Ile Tyr Glu Val Glu Gly Met Arg Trp Ala Gly Asn 35 40 45 Ala Asn Glu Leu Asn Ala Ala Tyr Ala Ala Asp Gly Tyr Ala Arg Ile 50 55 60 Lys Gly Met Ser Cys Ile Ile Thr Thr Phe Gly Val Gly Glu Leu Ser 65 70 75 80 Ala Leu Asn Gly Ile Ala Gly Ser Tyr Ala Glu His Val Gly Val Leu 85 90 95 His Val Val Gly Val Pro Ser Ile Ser Ala Gln Ala Lys Gln Leu Leu 100 105 110 Leu His His Thr Leu Gly Asn Gly Asp Phe Thr Val Phe His Arg Met 115 120 125 Ser Ala Asn Ile Ser Glu Thr Thr Ala Met Ile Thr Asp Ile Ala Thr 130 135 140 Ala Pro Ala Glu Ile Asp Arg Cys Ile Arg Thr Thr Tyr Val Thr Gln 145 150 155

160 Arg Pro Val Tyr Leu Gly Leu Pro Ala Asn Leu Val Asp Leu Asn Val 165 170 175 Pro Ala Lys Leu Leu Gln Thr Pro Ile Asp Met Ser Leu Lys Pro Asn 180 185 190 Asp Ala Glu Ser Glu Lys Glu Val Ile Asp Thr Ile Leu Ala Leu Val 195 200 205 Lys Asp Ala Lys Asn Pro Val Ile Leu Ala Asp Ala Cys Cys Ser Arg 210 215 220 His Asp Val Lys Ala Glu Thr Lys Lys Leu Ile Asp Leu Thr Gln Phe 225 230 235 240 Pro Ala Phe Val Thr Pro Met Gly Lys Gly Ser Ile Asp Glu Gln His 245 250 255 Pro Arg Tyr Gly Gly Val Tyr Val Gly Thr Leu Ser Lys Pro Glu Val 260 265 270 Lys Glu Ala Val Glu Ser Ala Asp Leu Ile Leu Ser Val Gly Ala Leu 275 280 285 Leu Ser Asp Phe Asn Thr Gly Ser Phe Ser Tyr Ser Tyr Lys Thr Lys 290 295 300 Asn Ile Val Glu Phe His Ser Asp His Met Lys Ile Arg Asn Ala Thr 305 310 315 320 Phe Pro Gly Val Gln Met Lys Phe Val Leu Gln Lys Leu Leu Thr Thr 325 330 335 Ile Ala Asp Ala Ala Lys Gly Tyr Lys Pro Val Ala Val Pro Ala Arg 340 345 350 Thr Pro Ala Asn Ala Ala Val Pro Ala Ser Thr Pro Leu Lys Gln Glu 355 360 365 Trp Met Trp Asn Gln Leu Gly Asn Phe Leu Gln Glu Gly Asp Val Val 370 375 380 Ile Ala Glu Thr Gly Thr Ser Ala Phe Gly Ile Asn Gln Thr Thr Phe 385 390 395 400 Pro Asn Asn Thr Tyr Gly Ile Ser Gln Val Leu Trp Gly Ser Ile Gly 405 410 415 Phe Thr Thr Gly Ala Thr Leu Gly Ala Ala Phe Ala Ala Glu Glu Ile 420 425 430 Asp Pro Lys Lys Arg Val Ile Leu Phe Ile Gly Asp Gly Ser Leu Gln 435 440 445 Leu Thr Val Gln Glu Ile Ser Thr Met Ile Arg Trp Gly Leu Lys Pro 450 455 460 Tyr Leu Phe Val Leu Asn Asn Asp Gly Tyr Thr Ile Glu Lys Leu Ile 465 470 475 480 His Gly Pro Lys Ala Gln Tyr Asn Glu Ile Gln Gly Trp Asp His Leu 485 490 495 Ser Leu Leu Pro Thr Phe Gly Ala Lys Asp Tyr Glu Thr His Arg Val 500 505 510 Ala Thr Thr Gly Glu Trp Asp Lys Leu Thr Gln Asp Lys Ser Phe Asn 515 520 525 Asp Asn Ser Lys Ile Arg Met Ile Glu Ile Met Leu Pro Val Phe Asp 530 535 540 Ala Pro Gln Asn Leu Val Glu Gln Ala Lys Leu Thr Ala Ala Thr Asn 545 550 555 560 Ala Lys Gln 164563PRTSaccharomyces cerevisiae 164Met Ser Glu Ile Thr Leu Gly Lys Tyr Leu Phe Glu Arg Leu Ser Gln 1 5 10 15 Val Asn Cys Asn Thr Val Phe Gly Leu Pro Gly Asp Phe Asn Leu Ser 20 25 30 Leu Leu Asp Lys Leu Tyr Glu Val Lys Gly Met Arg Trp Ala Gly Asn 35 40 45 Ala Asn Glu Leu Asn Ala Ala Tyr Ala Ala Asp Gly Tyr Ala Arg Ile 50 55 60 Lys Gly Met Ser Cys Ile Ile Thr Thr Phe Gly Val Gly Glu Leu Ser 65 70 75 80 Ala Leu Asn Gly Ile Ala Gly Ser Tyr Ala Glu His Val Gly Val Leu 85 90 95 His Val Val Gly Val Pro Ser Ile Ser Ser Gln Ala Lys Gln Leu Leu 100 105 110 Leu His His Thr Leu Gly Asn Gly Asp Phe Thr Val Phe His Arg Met 115 120 125 Ser Ala Asn Ile Ser Glu Thr Thr Ala Met Ile Thr Asp Ile Ala Asn 130 135 140 Ala Pro Ala Glu Ile Asp Arg Cys Ile Arg Thr Thr Tyr Thr Thr Gln 145 150 155 160 Arg Pro Val Tyr Leu Gly Leu Pro Ala Asn Leu Val Asp Leu Asn Val 165 170 175 Pro Ala Lys Leu Leu Glu Thr Pro Ile Asp Leu Ser Leu Lys Pro Asn 180 185 190 Asp Ala Glu Ala Glu Ala Glu Val Val Arg Thr Val Val Glu Leu Ile 195 200 205 Lys Asp Ala Lys Asn Pro Val Ile Leu Ala Asp Ala Cys Ala Ser Arg 210 215 220 His Asp Val Lys Ala Glu Thr Lys Lys Leu Met Asp Leu Thr Gln Phe 225 230 235 240 Pro Val Tyr Val Thr Pro Met Gly Lys Gly Ala Ile Asp Glu Gln His 245 250 255 Pro Arg Tyr Gly Gly Val Tyr Val Gly Thr Leu Ser Arg Pro Glu Val 260 265 270 Lys Lys Ala Val Glu Ser Ala Asp Leu Ile Leu Ser Ile Gly Ala Leu 275 280 285 Leu Ser Asp Phe Asn Thr Gly Ser Phe Ser Tyr Ser Tyr Lys Thr Lys 290 295 300 Asn Ile Val Glu Phe His Ser Asp His Ile Lys Ile Arg Asn Ala Thr 305 310 315 320 Phe Pro Gly Val Gln Met Lys Phe Ala Leu Gln Lys Leu Leu Asp Ala 325 330 335 Ile Pro Glu Val Val Lys Asp Tyr Lys Pro Val Ala Val Pro Ala Arg 340 345 350 Val Pro Ile Thr Lys Ser Thr Pro Ala Asn Thr Pro Met Lys Gln Glu 355 360 365 Trp Met Trp Asn His Leu Gly Asn Phe Leu Arg Glu Gly Asp Ile Val 370 375 380 Ile Ala Glu Thr Gly Thr Ser Ala Phe Gly Ile Asn Gln Thr Thr Phe 385 390 395 400 Pro Thr Asp Val Tyr Ala Ile Val Gln Val Leu Trp Gly Ser Ile Gly 405 410 415 Phe Thr Val Gly Ala Leu Leu Gly Ala Thr Met Ala Ala Glu Glu Leu 420 425 430 Asp Pro Lys Lys Arg Val Ile Leu Phe Ile Gly Asp Gly Ser Leu Gln 435 440 445 Leu Thr Val Gln Glu Ile Ser Thr Met Ile Arg Trp Gly Leu Lys Pro 450 455 460 Tyr Ile Phe Val Leu Asn Asn Asn Gly Tyr Thr Ile Glu Lys Leu Ile 465 470 475 480 His Gly Pro His Ala Glu Tyr Asn Glu Ile Gln Gly Trp Asp His Leu 485 490 495 Ala Leu Leu Pro Thr Phe Gly Ala Arg Asn Tyr Glu Thr His Arg Val 500 505 510 Ala Thr Thr Gly Glu Trp Glu Lys Leu Thr Gln Asp Lys Asp Phe Gln 515 520 525 Asp Asn Ser Lys Ile Arg Met Ile Glu Val Met Leu Pro Val Phe Asp 530 535 540 Ala Pro Gln Asn Leu Val Lys Gln Ala Gln Leu Thr Ala Ala Thr Asn 545 550 555 560 Ala Lys Gln 165563PRTSaccharomyces cerevisiae 165Met Ser Glu Ile Thr Leu Gly Lys Tyr Leu Phe Glu Arg Leu Lys Gln 1 5 10 15 Val Asn Val Asn Thr Ile Phe Gly Leu Pro Gly Asp Phe Asn Leu Ser 20 25 30 Leu Leu Asp Lys Ile Tyr Glu Val Asp Gly Leu Arg Trp Ala Gly Asn 35 40 45 Ala Asn Glu Leu Asn Ala Ala Tyr Ala Ala Asp Gly Tyr Ala Arg Ile 50 55 60 Lys Gly Leu Ser Val Leu Val Thr Thr Phe Gly Val Gly Glu Leu Ser 65 70 75 80 Ala Leu Asn Gly Ile Ala Gly Ser Tyr Ala Glu His Val Gly Val Leu 85 90 95 His Val Val Gly Val Pro Ser Ile Ser Ala Gln Ala Lys Gln Leu Leu 100 105 110 Leu His His Thr Leu Gly Asn Gly Asp Phe Thr Val Phe His Arg Met 115 120 125 Ser Ala Asn Ile Ser Glu Thr Thr Ser Met Ile Thr Asp Ile Ala Thr 130 135 140 Ala Pro Ser Glu Ile Asp Arg Leu Ile Arg Thr Thr Phe Ile Thr Gln 145 150 155 160 Arg Pro Ser Tyr Leu Gly Leu Pro Ala Asn Leu Val Asp Leu Lys Val 165 170 175 Pro Gly Ser Leu Leu Glu Lys Pro Ile Asp Leu Ser Leu Lys Pro Asn 180 185 190 Asp Pro Glu Ala Glu Lys Glu Val Ile Asp Thr Val Leu Glu Leu Ile 195 200 205 Gln Asn Ser Lys Asn Pro Val Ile Leu Ser Asp Ala Cys Ala Ser Arg 210 215 220 His Asn Val Lys Lys Glu Thr Gln Lys Leu Ile Asp Leu Thr Gln Phe 225 230 235 240 Pro Ala Phe Val Thr Pro Leu Gly Lys Gly Ser Ile Asp Glu Gln His 245 250 255 Pro Arg Tyr Gly Gly Val Tyr Val Gly Thr Leu Ser Lys Gln Asp Val 260 265 270 Lys Gln Ala Val Glu Ser Ala Asp Leu Ile Leu Ser Val Gly Ala Leu 275 280 285 Leu Ser Asp Phe Asn Thr Gly Ser Phe Ser Tyr Ser Tyr Lys Thr Lys 290 295 300 Asn Val Val Glu Phe His Ser Asp Tyr Val Lys Val Lys Asn Ala Thr 305 310 315 320 Phe Leu Gly Val Gln Met Lys Phe Ala Leu Gln Asn Leu Leu Lys Val 325 330 335 Ile Pro Asp Val Val Lys Gly Tyr Lys Ser Val Pro Val Pro Thr Lys 340 345 350 Thr Pro Ala Asn Lys Gly Val Pro Ala Ser Thr Pro Leu Lys Gln Glu 355 360 365 Trp Leu Trp Asn Glu Leu Ser Lys Phe Leu Gln Glu Gly Asp Val Ile 370 375 380 Ile Ser Glu Thr Gly Thr Ser Ala Phe Gly Ile Asn Gln Thr Ile Phe 385 390 395 400 Pro Lys Asp Ala Tyr Gly Ile Ser Gln Val Leu Trp Gly Ser Ile Gly 405 410 415 Phe Thr Thr Gly Ala Thr Leu Gly Ala Ala Phe Ala Ala Glu Glu Ile 420 425 430 Asp Pro Asn Lys Arg Val Ile Leu Phe Ile Gly Asp Gly Ser Leu Gln 435 440 445 Leu Thr Val Gln Glu Ile Ser Thr Met Ile Arg Trp Gly Leu Lys Pro 450 455 460 Tyr Leu Phe Val Leu Asn Asn Asp Gly Tyr Thr Ile Glu Lys Leu Ile 465 470 475 480 His Gly Pro His Ala Glu Tyr Asn Glu Ile Gln Thr Trp Asp His Leu 485 490 495 Ala Leu Leu Pro Ala Phe Gly Ala Lys Lys Tyr Glu Asn His Lys Ile 500 505 510 Ala Thr Thr Gly Glu Trp Asp Ala Leu Thr Thr Asp Ser Glu Phe Gln 515 520 525 Lys Asn Ser Val Ile Arg Leu Ile Glu Leu Lys Leu Pro Val Phe Asp 530 535 540 Ala Pro Glu Ser Leu Ile Lys Gln Ala Gln Leu Thr Ala Ala Thr Asn 545 550 555 560 Ala Lys Gln 166440PRTSaccharomyces cerevisiae 166Met Leu Ala Val Arg Arg Leu Thr Arg Tyr Thr Phe Leu Lys Arg Thr 1 5 10 15 His Pro Val Leu Tyr Thr Arg Arg Ala Tyr Lys Ile Leu Pro Ser Arg 20 25 30 Ser Thr Phe Leu Arg Arg Ser Leu Leu Gln Thr Gln Leu His Ser Lys 35 40 45 Met Thr Ala His Thr Asn Ile Lys Gln His Lys His Cys His Glu Asp 50 55 60 His Pro Ile Arg Arg Ser Asp Ser Ala Val Ser Ile Val His Leu Lys 65 70 75 80 Arg Ala Pro Phe Lys Val Thr Val Ile Gly Ser Gly Asn Trp Gly Thr 85 90 95 Thr Ile Ala Lys Val Ile Ala Glu Asn Thr Glu Leu His Ser His Ile 100 105 110 Phe Glu Pro Glu Val Arg Met Trp Val Phe Asp Glu Lys Ile Gly Asp 115 120 125 Glu Asn Leu Thr Asp Ile Ile Asn Thr Arg His Gln Asn Val Lys Tyr 130 135 140 Leu Pro Asn Ile Asp Leu Pro His Asn Leu Val Ala Asp Pro Asp Leu 145 150 155 160 Leu His Ser Ile Lys Gly Ala Asp Ile Leu Val Phe Asn Ile Pro His 165 170 175 Gln Phe Leu Pro Asn Ile Val Lys Gln Leu Gln Gly His Val Ala Pro 180 185 190 His Val Arg Ala Ile Ser Cys Leu Lys Gly Phe Glu Leu Gly Ser Lys 195 200 205 Gly Val Gln Leu Leu Ser Ser Tyr Val Thr Asp Glu Leu Gly Ile Gln 210 215 220 Cys Gly Ala Leu Ser Gly Ala Asn Leu Ala Pro Glu Val Ala Lys Glu 225 230 235 240 His Trp Ser Glu Thr Thr Val Ala Tyr Gln Leu Pro Lys Asp Tyr Gln 245 250 255 Gly Asp Gly Lys Asp Val Asp His Lys Ile Leu Lys Leu Leu Phe His 260 265 270 Arg Pro Tyr Phe His Val Asn Val Ile Asp Asp Val Ala Gly Ile Ser 275 280 285 Ile Ala Gly Ala Leu Lys Asn Val Val Ala Leu Ala Cys Gly Phe Val 290 295 300 Glu Gly Met Gly Trp Gly Asn Asn Ala Ser Ala Ala Ile Gln Arg Leu 305 310 315 320 Gly Leu Gly Glu Ile Ile Lys Phe Gly Arg Met Phe Phe Pro Glu Ser 325 330 335 Lys Val Glu Thr Tyr Tyr Gln Glu Ser Ala Gly Val Ala Asp Leu Ile 340 345 350 Thr Thr Cys Ser Gly Gly Arg Asn Val Lys Val Ala Thr Tyr Met Ala 355 360 365 Lys Thr Gly Lys Ser Ala Leu Glu Ala Glu Lys Glu Leu Leu Asn Gly 370 375 380 Gln Ser Ala Gln Gly Ile Ile Thr Cys Arg Glu Val His Glu Trp Leu 385 390 395 400 Gln Thr Cys Glu Leu Thr Gln Glu Phe Pro Leu Phe Glu Ala Val Tyr 405 410 415 Gln Ile Val Tyr Asn Asn Val Arg Met Glu Asp Leu Pro Glu Met Ile 420 425 430 Glu Glu Leu Asp Ile Asp Asp Glu 435 440 167250PRTSaccharomyces cerevisiae 167Met Pro Leu Thr Thr Lys Pro Leu Ser Leu Lys Ile Asn Ala Ala Leu 1 5 10 15 Phe Asp Val Asp Gly Thr Ile Ile Ile Ser Gln Pro Ala Ile Ala Ala 20 25 30 Phe Trp Arg Asp Phe Gly Lys Asp Lys Pro Tyr Phe Asp Ala Glu His 35 40 45 Val Ile His Ile Ser His Gly Trp Arg Thr Tyr Asp Ala Ile Ala Lys 50 55 60 Phe Ala Pro Asp Phe Ala Asp Glu Glu Tyr Val Asn Lys Leu Glu Gly 65 70 75 80 Glu Ile Pro Glu Lys Tyr Gly Glu His Ser Ile Glu Val Pro Gly Ala 85 90 95 Val Lys Leu Cys Asn Ala Leu Asn Ala Leu Pro Lys Glu Lys Trp Ala 100 105 110 Val Ala Thr Ser Gly Thr Arg Asp Met Ala Lys Lys Trp Phe Asp Ile 115 120 125 Leu Lys Ile Lys Arg Pro Glu Tyr Phe Ile Thr Ala Asn Asp Val Lys 130 135 140 Gln Gly Lys Pro His Pro Glu Pro Tyr Leu Lys Gly Arg Asn Gly Leu 145 150 155 160 Gly Phe Pro Ile Asn Glu Gln Asp Pro Ser Lys Ser Lys Val Val Val 165 170 175 Phe Glu Asp Ala Pro Ala Gly Ile Ala Ala Gly Lys Ala Ala Gly Cys 180 185 190 Lys Ile Val Gly Ile Ala Thr Thr Phe Asp Leu Asp Phe Leu Lys Glu 195 200 205 Lys Gly Cys Asp Ile Ile Val Lys Asn His Glu Ser Ile Arg Val Gly 210 215 220 Glu Tyr Asn Ala Glu Thr Asp Glu Val Glu Leu Ile Phe Asp Asp Tyr 225 230 235 240 Leu Tyr Ala Lys Asp Asp Leu Leu Lys Trp 245 250

Claims

1. A recombinant microorganism: wherein said microorganism expresses one or more native and/or heterologous enzymes; wherein said one or more enzymes function in one or more engineered metabolic pathways to convert a carbohydrate source to 1,2-propanediol, isopropropanol and ethanol or to 1,2-propanediol, isopropropanol and glycerol; and wherein said one or more native and/or heterologous enzymes is activated, upregulated or downregulated.

2. The recombinant microorganism of claim 1, wherein said carbohydrate source is lignocellulose.

3. The recombinant microorganism of claim 1, wherein said carbohydrate source is converted to dihydroxyacetone phosphate and glyceraldehyde phosphate.

4. The recombinant microorganism of claim 1, wherein one of said engineered metabolic pathways comprises the conversion of dihydroxyacetone phosphate to 1-propanediol.

5. The recombinant microorganism of claim 1, wherein one of said engineered metabolic pathways comprises conversion of pyruvate to isopropanol.

6. The recombinant microorganism of claim 1, wherein one of said engineered metabolic pathway comprises conversion of pyruvate to ethanol.

7. The recombinant microorganism of claim 1, wherein one of said engineered metabolic pathway comprises conversion of dihydroxyacetone phosphate to glycerol.

8. The recombinant microorganism of claim 1, wherein the engineered metabolic pathway that produces 1,2 propanediol comprises the following steps: (a) conversion of dihydroxyacetone phosphate to methyglyoxal; (b) conversion of methylglyoxal to acetol or lactaldehyde; and (c) reduction of acetol or lactaldehyde to propanediol.

9. The recombinant microorganism of claim 1, wherein the engineered metabolic pathway that produces isopropanol comprises: (a) conversion of acetyl-CoA to acetate; (b) conversion of acetyl-CoA to acetoacetyl-CoA; (c) conversion of acetoacetyl-CoA to acetoacetate; (d) conversion of acetoacetate to acetone; and (e) reduction of acetone to isopropanol.

10. The recombinant microorganism of claim 1, wherein the engineered metabolic pathway that produces ethanol comprises: (a) conversion of acetyl-CoA to ethanol.

11. The recombinant microorganism of claim 1, wherein the engineered metabolic pathway that produces glycerol comprises: (a) conversion of dihydroxyacetone phosphate to glycerol.

12. The recombinant microorganism of claim 1, wherein said microorganism produces 1,2-propanediol, isopropropanol and ethanol.

13. The recombinant microorganism of claim 12, wherein the combined production of 1,2-propanediol, isopropropanol and ethanol is governed by the stoichiometric equation 2C.sub.6H.sub.12O.sub.6.fwdarw.C.sub.3H.sub.8O+C.sub.3H.sub.8O.sub.2+C.su- b.2H.sub.6O+4CO.sub.2+H.sub.2+3ATP.

14. The recombinant microorganism of claim 1, wherein said microorganism is a thermophilic or mesophilic bacterium.

15. The recombinant microorganism of claim 14, wherein said thermophilic or mesophilic bacterium is a species of the genera Thermoanaerobacterium, Thermoanaerobacter, Clostridium, Geobacilliis, Saccharococcus, Paenibacillus, Bacillus, Caldicelhilosiruptor, Anaerocellum, or Anoxybacillus,

16. The recombinant microorganism of claim 15, wherein said microorganism is a bacterium selected from the group consisting of: Thermoanaerobacterium thermosulfurigenes, Thermoanaerobacterium aotearoense, Thermoanaerobacterium polysaccharolyticum, Thermoanaerobacterium zeae, Thermoanaerobacterium xylanolyticiim, Thermoanaerobacterium saccharolyticiim, Thermoanaerobium brockii, Thermoanaerobacterium thermosaccharolyticum, Thermoanaerobacter thermohydrosulfuricus, Thermoanaerobacter ethanolicus, Thermoanaerobacter brocki, Clostridium thermocellum, Clostridium cellulolyticiim, Clostridium phytofermentans, Clostridium straminosolvens, Geobacillus thermoglucosidasiiis, Geobacillus stearothermophilus, Saccharococcus caldoxylosilyticus, Saccharocciis thermophilus, Paenibacillus campinasensis, Bacillus flavothermus, Anoxybacillus kamchatkensis, Anoxybacillus gonensis, Caldicellulosiruptor acetigenus, Caldicellulosiruptor saccharolyticus, Caldicellulosiruptor kristjanssonii, Caldicellulosiruptor owensensis, Caldicellulosiruptor lactoaceticus, and Anaerocellum thermophilum.

17. The recombinant microorganism of claim 16, wherein said microorganism is selected from the group consisting of Clostridium thermocellum, and Thermoanaerobacterium saccharolyticum.

18. The recombinant microorganism of claim 1, wherein said microorganism produces 1,2-propanediol, isopropropanol and glycerol.

19. The recombinant microorganism of claim 18, wherein the combined production of 1,2-propanediol, isopropropanol and ethanol is governed by the stoichiometric equation 2C.sub.6H.sub.12O.sub.6.fwdarw.C.sub.3H.sub.8O+C.sub.3H.sub.8O.sub.2+C.su- b.3H.sub.8O.sub.3+3CO.sub.2+ATP.

20. The recombinant microorganism of claim 1, wherein said microorganism is selected from the group consisting of Saccharomyces cerevisiae, Kluyveromyces lactis, Kluyveromyces marxianus, Pichia pastoris, Yarrowia lipolytica, Hansemila polymorphs Phaffta rhodozyma, Candida ittliis, Arxula adeninivorans, Pichia stipitis, Debaryomyces hansenii, Debaryomyces polymorphic, Schizosaccharomyces pombe, Candida albicans, and Schwanniomyces occidentalis.

21-105. (canceled)