Electron Consuming Ethanol Production Pathway To Displace Glycerol Formation In S. Cerevisiae

Abstract

The present invention provides for a mechanism to completely replace the electron accepting function of glycerol formation with an alternative pathway to ethanol formation, thereby reducing glycerol production and increasing ethanol production. In some embodiments, the invention provides for a recombinant microorganism comprising a down-regulation in one or more native enzymes in the glycerol-production pathway. In some embodiments, the invention provides for a recombinant microorganism comprising an up-regulation in one or more enzymes in the ethanol-production pathway.

Description

CROSS REFERENCE

[0001] This is a continuation of U.S. Pat. No. 15,828,729 filed Dec. 1, 2017, which is a continuation of U.S. Ser. No. 14/442,408 filed May 13, 2015, now U.S. Pat. No. 9,988,650, which is a .sctn. 371 of PCT/US13/70964 filed Nov. 20, 2013, which claims priority to U.S. provisional applications Nos. 61/728,450 filed Nov. 20, 2012 and 61,792,731 filed Mar. 15, 2013, each application of which is hereby incorporated by reference in its entirety.

REFERENCE TO SEQUENCE LISTING

[0002] The contents of the sequence listing attached herewith (entitled "115235-266_seq_listing.txt"; of size 349 kb, created on Sep. 24, 2019) are incorporated herein by reference in their entirety.

BACKGROUND OF THE INVENTION

[0003] Energy conversion, utilization and access underlie many of the great challenges of our time, including those associated with sustainability, environmental quality, security, and quality of life. New applications of emerging technologies are required to respond to these challenges. Biotechnology, one of the most powerful of the emerging technologies, can give rise to important new energy conversion processes. Plant biomass and derivatives thereof are a resource for the biological conversion of energy to forms useful to humanity.

[0004] Among forms of plant biomass, both grain-based biomass and lignocellulosic biomass (collectively "biomass") are well-suited for energy applications. Each feedstock has advantages and disadvantages. For example, because of its large-scale availability, low cost, and environmentally benign production lignocellulosic biomass has gained attention as a viable feed source for biofuel production. In particular, many energy production and utilization cycles based on cellulosic biomass have near-zero greenhouse gas emissions on a life-cycle basis.

[0005] However, grain-based feed stocks are more readily converted to fuels by existing microorganisms, although grain-based feed stock is more expensive than lignocellulosic feed stock and conversion to fuel competes with alternative uses for the grain.

[0006] 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 can occur in a single step in a process configuration called consolidated bioprocessing ("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.

[0007] 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 cellulase 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 cellulase systems. Moreover, cellulose-adherent cellulolytic microorganisms are likely to compete successfully for products of cellulose hydrolysis with non-adhered microbes, e.g., contaminants. Successful competition of desirable microbes increases 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.

[0008] One way to meet the demand for ethanol production is to convert sugars found in biomass, i.e., materials such as agricultural wastes, corn hulls, corncobs, cellulosic materials, and the like to produce ethanol. Efficient biomass conversion in large-scale industrial applications requires a microorganism that is able to tolerate high concentrations of sugar and ethanol, and which is able to ferment more than one sugar simultaneously.

[0009] Bakers' yeast (Saccharomyces cerevisiae) is the preferred microorganism for the production of ethanol (Hahn-Hagerdal, B., et al., Adv. Biochem. Eng. Biotechnol. 73: 53-84 (2001)). Pathways for ethanol production in S. cerevisiae can be seen in FIGS. 1-3. Attributes in favor of this microbe are (i) high productivity at close to theoretical yields (0.51 g ethanol produced/g glucose used), (ii) high osmo- and ethanol tolerance, (iii) natural robustness in industrial processes, and also (iv) being generally regarded as safe (GRAS) due to its long association with wine and bread making, and beer brewing. Furthermore, S. cerevisiae exhibits tolerance to inhibitors commonly found in hydrolysates resulting from biomass pretreatment. Exemplary metabolic pathways for the production of ethanol are depicted in FIGS. 1-4.

[0010] However, glycerol is a required metabolic end-product of native yeast ethanolic fermentation (FIG. 5A). During anaerobic growth on carbohydrates, production of ethanol and carbon dioxide is redox neutral, while the reactions that create cell biomass and associated carbon dioxide are more oxidized relative to carbohydrates. The production of glycerol, which is more reduced relative to carbohydrates, functions as an electron sink to off-set cell biomass formation, so that overall redox neutrality is conserved. This is essential from a theoretical consideration of conservation of mass, and in practice strains unable to produce glycerol are unable (or only very poorly able) to grow under anaerobic conditions.

[0011] There is a strong commercial incentive to not produce glycerol, as it represents lost ethanol yield. In industrial corn ethanol fermentations, this yield loss can be up to 6% of theoretical, for a market of .about.14 billion gallons/yr. At selling price of $2.50/gal, this is a total market value of $2 B/yr.

[0012] Strategies from the literature to address this problem include decreasing glycerol formation by engineering ammonia fixation to function with NADH instead of NADPH via up-regulation of GLN1, encoding glutamine synthetase, or GLT1, encoding glutamate synthase with deletion of GDH1, encoding the NADPH-dependent glutamate dehydrogenase. (Nissen, T. L., et al., Metabolic Engineering 2: 69-77 (2000)). Another strategy engineering cells to produce excess NADPH during glycolysis via expression of a NADPH linked glyceraldehyde-3-phosphate dehydrogenase. (Bro, C., et al., Metabolic Engineering 8: 102-111 (2006)).

[0013] However, most glycerol reduction strategies either only partially reduce the requirement for glycerol formation, or create a by-product other than ethanol. The present invention overcomes the shortcomings of these other strategies by redirecting electrons typically placed on glycerol into ethanol formation, thereby reducing glycerol production and increasing ethanol production (FIG. 5B, FIG. 6).

BRIEF SUMMARY OF THE INVENTION

[0014] Aspects of the invention are directed to a recombinant microorganism comprising a heterologous nucleic acid encoding a phosphoketolase; at least one heterologous nucleic acid encoding an enzyme in an acetyl-CoA production pathway; a heterologous nucleic acid encoding a bifunctional acetaldehyde-alcohol dehydrogenase; and, at least one genetic modification that leads to the down-regulation of an enzyme in a glycerol-production pathway. In some embodiments, the phosphoketolase is a single-specificity phosphoketolase with the Enzyme Commission Number 4.1.2.9. In some embodiments, the phosphoketolase is dual-specificity phosphoketolase with the Enzyme Commission Number 4.1.2.22. Aspects of the invention are directed to a recombinant microorganism comprising at least one heterologous nucleic acid encoding an enzyme in an acetyl-CoA production pathway; a heterologous nucleic acid encoding a bifunctional acetaldehyde-alcohol dehydrogenase; and, optionally, at least one genetic modification that leads to the down-regulation of an enzyme in a glycerol-production pathway. In some embodiments, the enzyme in the acetyl-CoA production pathway is phosphotransacetylase with the Enzyme Commission Number 2.3.1.8. In some embodiments, the enzyme in the acetyl-CoA production pathway is acetate kinase with the Enzyme Commission Number 2.7.2.12. In some embodiments, the bifunctional acetaldehyde-alcohol dehydrogenase is selected from a group of enzymes having both of the following Enzyme Commission Numbers: EC 1.2.1.10 and 1.1.1.1. In some embodiments, the bifunctional acetaldehyde-alcohol dehydrogenase is an NADPH dependent bifunctional acetaldehyde-alcohol dehydrogenase selected from a group of enzymes having the following Enzyme Commission Numbers: EC 1.2.1.10 and 1.1.1.2. In some embodiments, the enzyme in the glycerol-production pathway is glycerol-3-phosphate dehydrogenase with the Enzyme Commission Number 1.1.1.8. In some embodiments, the enzyme in the glycerol-production pathway is glycerol-3-phosphate phosphatase with the Enzyme Commission Number 3.1.3.21.

[0015] In some embodiments, the microorganism further comprises at least one additional up-regulated enzyme. In some embodiments, transaldolase with the Enzyme Commission Number 2.2.1.2 is up-regulated. In some embodiments, transketolase with the Enzyme Commission Number 2.2.1.1 is up-regulated. In some embodiments, ribose-5-P isomerase with the Enzyme Commission Number 5.3.1.6 is up-regulated. In some embodiments, ribulose-5-P 3-epimerase with the Enzyme Commission Number 5.1.3.1 is up-regulated.

[0016] In some embodiments, at least one enzyme in a glycolysis pathway is up-regulated in the microorganism. In some embodiments, the enzyme in the glycolysis pathway is pyruvate decarboxylase with the Enzyme Commission Number 4.1.1.1. In some embodiments, the enzyme in the glycolysis pathway is alcohol dehydrogenase selected from a group of enzymes having the following Enzyme Commission Numbers: 1.1.1.1 and 1.1.1.2.

[0017] In some embodiments, the microorganism additionally comprises at least one genetic modification that leads to the down-regulation of aldehyde dehydrogenase selected from a group of enzymes having the following Enzyme Commission Numbers: 1.2.1.3, 1.2.1.4 and 1.2.1.10. In some embodiments, the microorganism additionally comprises at least one genetic modification that leads to the up-regulation of aldehyde dehydrogenase selected from a group of enzymes having the following Enzyme Commission Numbers: 1.2.1.3, 1.2.1.4 and 1.2.1.10. In some embodiments, the aldehyde dehydrogenase is acetaldehyde dehydrogenase.

[0018] In some embodiments, the microorganism additionally comprises at least one genetic modification that leads to the down-regulation of formate dehydrogenase selected from a group of enzymes having the following Enzyme Commission Numbers: 1.2.1.43 and 1.2.1.2.

[0019] In some embodiments, the microorganism additionally comprises at least one genetic modification that leads to the up-regulation of pyruvate formate lyase with the Enzyme Commission Number 2.3.1.54. In some embodiments, the microorganism additionally comprises at least one genetic modification that leads to the up-regulation of pyruvate formate lyase activating enzyme with the Enzyme Commission Number 1.97.1.4.

[0020] In some embodiments, the microorganism is yeast. In some embodiments, the microorganism is from the genus Saccharomyces. In some embodiments, the microorganism is Saccharomyces cerevisiae.

[0021] In some embodiments, the recombinant microorganisms described herein produce ethanol at a higher yield than an otherwise identical microorganism lacking said genetic modifications. In some embodiments, the microorganism produces an ethanol titer 1%-10% more than an otherwise identical microorganism lacking said genetic modifications.

[0022] In some embodiments, the recombinant microorganisms described herein produce glycerol at a lower yield than an otherwise identical microorganism lacking said genetic modifications. In some embodiments, the microorganism produces a glycerol titer of 10-100% less than an otherwise identical microorganism lacking said genetic modifications.

[0023] In some embodiments, the invention is directed to the host cells described herein and a carbon-containing feedstock. In some embodiments, the feedstock is selected from the group consisting of woody biomass, grasses, sugar-processing residues, municipal waste, agricultural wastes or any combination thereof. In some embodiments, the feedstock comprises recycled wood pulp fiber, sawdust, hardwood, softwood, rice straw, rice hulls, barley straw, corn cobs, cereal straw, wheat straw, canola straw, oat straw, oat hulls, corn fiber, stover, succulents, agave, cane bagasse, switchgrass, miscanthus, paper sludge, municipal waste or any combination thereof.

[0024] In some embodiments, the invention is directed to a method of producing a fermentation product using a composition described herein, wherein the host cell of the composition is capable of fermenting the carbon containing feedstock to yield the fermentation product.

[0025] In some embodiments, the invention is directed to a method of producing ethanol comprising providing any host cell described herein; culturing the host cell in the presence of a carbon containing feedstock for sufficient time to produce ethanol; and, optionally, extracting the ethanol.

[0026] In some embodiments, the invention is directed to a method of reducing glycerol production comprising providing any host cell described herein, wherein said glycerol production is 10-100% less than an otherwise identical microorganism lacking said genetic modifications. In some embodiments, the glycerol production of the host cell is reduced as compared to an otherwise identical microorganism lacking said genetic modifications, and wherein the ethanol titer increased by at least 1-10% when the host cell is cultured in the presence of a carbon containing feedstock for a sufficient time to produce ethanol.

[0027] In some embodiments, the invention is directed to a co-culture comprising at least two host cells wherein one of the host cells comprises a host cell described herein and another host cell that is genetically distinct from the host cell of the invention. In some embodiments, the genetically distinct host cell of the co-culture is a yeast or bacterium. In some embodiments, the genetically distinct host cell is any organism from the genus Saccharomyces, Issatchenkia, Pichia, Clavispora, Candida, Hansenula, Kluyveromyces, Trichoderma, Thermoascus, Escherichia, Clostridium, Caldicellulosiruptor, Zymomonas, Thermoanaerobacter and Thermoanaerobacterium.

BRIEF DESCRIPTION OF THE DRAWINGS/FIGURES

[0028] FIG. 1 depicts a simplified metabolic pathway from glucose to ethanol utilizing single-specificity phosphoketolase. The names of enzymes catalyzing steps applicable to the invention are shown in italics.

[0029] FIG. 2 depicts a simplified metabolic pathway from glucose to ethanol utilizing dual-specificity phosphoketolase. The names of enzymes catalyzing steps applicable to the invention are shown in italics.

[0030] FIG. 3 depicts a simplified metabolic pathway from glucose to ethanol that does not utilize phosphoketolase. The names of enzymes catalyzing steps applicable to the invention are shown in italics.

[0031] FIG. 4 depicts alternative conversions of acetyl-phosphate ("Acetyl-P") to ethanol.

[0032] FIG. 5 depicts (A) simplified carbon and redox pathways utilized by wildtype S. cerevisiae during anaerobic growth. Ethanol formation is redox neutral while cell biomass formation generates net NADH, which is balanced by glycerol formation; and (B) S. cerevisiae engineered with the PHK-PTA-ADHE pathway balances NADH generated during cell biomass formation with production of ethanol.

[0033] FIG. 6 depicts mechanisms by which glycerol formation could be deleted or down-regulated in the context of the overall metabolic pathway.

[0034] FIG. 7 depicts equivalent metabolic interconversions using single specificity and dual-specificity phosphoketolase.

[0035] FIG. 8 depicts a schematic diagram for PCR construction and integration of KT-MX and NT-MX integration cassettes into both copies of each chromosome at a given target locus. (A) The transformed PCR assembly contains four PCR products, a 5' flank (p1) which is homologous to sequences upstream of the target site, KT-MX cassette (p2), NT-MX cassette (p3), and a 3' flank (p4) homologous to sequences downstream of the target site. Each component is amplified individually using primers which create homologous overlapping extensions of each PCR product. The bent dashed lines represent homology between the KT/NT-MX cassettes and the 5' flank and the bent solid lines represent homology with the 3' flank. (B) Schematic of the chromosome after replacement of the target site with KT-MX and NT-MX.

[0036] FIG. 9 depicts a schematic diagram of the strategy used to replace integrated KT-MX and NT-MX selection cassettes with a "Mascoma Assembly" on both chromosomes at a target locus. A Mascoma Assembly ("MA") is used to identify a series of overlapping PCR products that form the desired construction once they are recombined in the organism. In this document a MA number is given to represent a combination of molecular components to be assembled via recombination. (A) The transformed Mascoma Assembly contains a quantity of PCR products which is dependent on the desired engineering event (pX), a 5' flank (p1) which is homologous to sequences upstream of the target site, and a 3' flank (p4) homologous to sequences downstream of the target site. Each component is amplified individually using primers which create homologous overlapping extensions. The overlapping bent lines represent homology at the end of those PCR products. (B) Schematic diagram of chromosome following selection on FUDR and replacement of genetic markers with the Mascoma Assembly.

[0037] FIG. 10 depicts the integration schemes for B. adolescentis PTA and (A) B. adolescentis PHK, (B) L. plantarum PHK1, and (C) A. niger PHK at the S. cerevisiae FCY1 site.

[0038] FIG. 11 depicts the improved anaerobic growth rate of S. cerevisiae strains containing integrated PTA and PHK, M4008, M4409, and M4410 compared to a strain that does not contain integrated PTA and PHK, M3293.

[0039] FIG. 12 depicts the aerobic growth rate of S. cerevisiae strains containing integrated PTA and PHK, M4008, M4409, and M4410 compared to a strain that does not contain integrated PTA and PHK, M3293.

[0040] FIG. 13 depicts the ethanol production of S. cerevisiae strains containing integrated PTA and PHK, M4008, M4409, and M4410 compared to a strain that does not contain integrated PTA and PHK, M3293, for fermentation using 31% solids corn mash. M2390 is a wildtype control.

[0041] FIG. 14 depicts the production of acetic acid in S. cerevisiae strains containing integrated PTA and PHK, M4008, M4409, and M4410 compared to a strain that does not contain integrated PTA and PHK, M3293, during fermentation using 31% solids corn mash. M2390 is a wildtype control.

[0042] FIG. 15 depicts the improved growth of in S. cerevisiae strains containing integrated PTA and PHK, M4008, M4409, and M4410 compared to a strain that does not contain integrated PTA and PHK, M3293, during fermentation in defined media. M2390 is a wildtype control.

[0043] FIG. 16 depicts the ethanol production in defined media of S. cerevisiae strains containing integrated PTA and PHK, M4008, M4409, and M4410 compared to a strain that does not contain integrated PTA and PHK, M3293, during fermentation in defined media. M2390 is a wildtype control.

[0044] FIG. 17 depicts the glycerol production in defined media of S. cerevisiae strains containing integrated PTA and PHK, M4008, M4409, and M4410 compared to a strain that does not contain integrated PTA and PHK, M3293, during fermentation in defined media. M2390 is a wildtype control.

[0045] FIG. 18 depicts an L. plantarum PHK1 expression plasmid.

[0046] FIG. 19 depicts an L. plantarum PHK2 expression plasmid.

[0047] FIG. 20 depicts an A. niger PHK expression plasmid.

[0048] FIG. 21 depicts an L. casei PHK expression plasmid.

[0049] FIG. 22 depicts an N. crassa PHK expression plasmid.

[0050] FIG. 23 depicts a B. adolescentis PHK expression plasmid.

[0051] FIG. 24 depicts a B. adolescentis PTA expression plasmid.

[0052] FIG. 25 depicts the ethanol production of S. cerevisiae strains M4408, M4579, M4581, M4582, M4584, M4788, M4789, M4790, M4791, M4792, M4793, M4794, and M4795 that contain pathway components in a M3293 background.

[0053] FIG. 26 depicts a schematic diagram of the MA0281 insertion cassette.

[0054] FIG. 27 depicts a schematic diagram of the MA0449.2 insertion cassette.

[0055] FIG. 28 depicts a schematic diagram of the MA0449.3 insertion cassette.

[0056] FIG. 29 depicts a schematic diagram of the MA0449.1 insertion cassette.

[0057] FIG. 30 depicts a schematic diagram of the MA0467.6 insertion cassette.

[0058] FIG. 31 depicts a schematic diagram of the MA0435.1 insertion cassette.

[0059] FIG. 32 depicts a schematic diagram of the MA0415.4 insertion cassette.

[0060] FIG. 33 depicts a schematic diagram of the MA0415.3 insertion cassette.

[0061] FIG. 34 depicts a schematic diagram of the MA0434.6 insertion cassette.

DETAILED DESCRIPTION OF THE INVENTION

Definitions

[0062] Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood to one of ordinary skill in the art of microbial metabolic engineering. Although methods and materials similar or equivalent to those described herein can be used in the practice of the disclosed methods and compositions, exemplary methods, devices and materials are described herein.

[0063] The embodiments described, and references in the specification to "one embodiment", "an embodiment", "an example embodiment", etc., indicate that the embodiments described can include a particular feature, structure, or characteristic, but every embodiment does not necessarily include the particular feature, structure, or characteristic. Moreover, such phrases are not necessarily referring to the same embodiment. Further, when a particular feature, structure, or characteristic is described in connection with an embodiment, it is understood that it is within the knowledge of one skilled in the art to effect such feature, structure, or characteristic in connection with other embodiments whether or not explicitly described.

[0064] The description of "a" or "an" item herein may refer to a single item or multiple items. It is understood that wherever embodiments are described herein with the language "comprising," otherwise analogous embodiments described in terms of "consisting of" and/or "consisting essentially of" are also provided. Thus, for example, reference to "a polynucleotide" includes a plurality of such polynucleotides and reference to "the microorganism" includes reference to one or more microorganisms, and so forth.

[0065] The term "heterologous" is used in reference to a polynucleotide or a gene not normally found in the host organism. "Heterologous" includes up-regulated or down-regulated endogenous genes. "Heterologous" also includes a native coding region, or portion thereof, that is reintroduced into the source organism in a form that is different from the corresponding native gene, e.g., not in its natural location in the organism's genome. "Heterologous" also includes any gene that has been modified and placed into an organism. A heterologous gene may include a native coding region that is a portion of a chimeric gene including a non-native regulatory region that is reintroduced into the native host or modifications to the native regulatory sequences that affect the expression level of the gene. Foreign genes can comprise native genes inserted into a non-native organism, or chimeric genes. A heterologous polynucleotide, gene, polypeptide, or an enzyme may be derived or isolated from any source, e.g., eukaryotes, prokaryotes, viruses, or synthetic polynucleotide fragments, and includes up-regulated endogenous genes.

[0066] The terms "gene(s)" or "polynucleotide" or "nucleic acid" 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. Also, the terms are intended to include a specific gene for a selected purpose. The gene may be endogenous to the host cell or may 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 may, for example, be in the form of linear DNA or RNA. The term "gene" is also intended to cover multiple copies of a particular gene, e.g., all of the DNA sequences in a cell encoding a particular gene product.

[0067] The term "expression" is intended to include the expression of a gene at least at the level of mRNA production, generally subsequently translated into a protein product.

[0068] As used herein, an "expression vector" is a vector capable of directing the expression of genes to which it is operably linked.

[0069] In some embodiments, the microorganisms contain enzymes involved in cellulose digestion, metabolism and/or hydrolysis. A "cellulolytic enzyme" can be any enzyme involved in cellulose digestion, metabolism, and/or hydrolysis. The term "cellulase" refers to a class of enzymes produced chiefly by fungi, bacteria, and protozoans that catalyze cellulolysis (i.e. the hydrolysis) of cellulose. However, there are also cellulases produced by other types of organisms such as plants and animals. Several different kinds of cellulases are known, which differ structurally and mechanistically. There are general types of cellulases based on the type of reaction catalyzed: endocellulase breaks internal bonds to disrupt the crystalline structure of cellulose and expose individual cellulose polysaccharide chains; exocellulase cleaves 2-4 units from the ends of the exposed chains produced by endocellulase, resulting in the tetrasaccharides or disaccharide such as cellobiose. There are two main types of exocellulases (or cellobiohydrolases, abbreviated CBH)--one type working processively from the reducing end, and one type working processively from the non-reducing end of cellulose; cellobiase or beta-glucosidase hydrolyses the exocellulase product into individual monosaccharides; oxidative cellulases that depolymerize cellulose by radical reactions, as for instance cellobiose dehydrogenase (acceptor); cellulose phosphorylases that depolymerize cellulose using phosphates instead of water. In the most familiar case of cellulase activity, the enzyme complex breaks down cellulose to beta-glucose. A "cellulase" can be any enzyme involved in cellulose digestion, metabolism and/or hydrolysis, including, for example, an endoglucanase, glucosidase, cellobiohydrolase, xylanase, glucanase, xylosidase, xylan esterase, arabinofuranosidase, galactosidase, cellobiose phosphorylase, cellodextrin phosphorylase, mannanase, mannosidase, xyloglucanase, endoxylanase, glucuronidase, acetylxylanesterase, arabinofuranohydrolase, swollenin, glucuronyl esterase, expansin, pectinase, and feruoyl esterase protein.

[0070] A "plasmid" or "vector" refers to an extrachromosomal element often carrying one or more genes, and is usually in the form of a circular double-stranded DNA molecule. Plasmids and vectors may also contain additional genetic elements such as autonomously replicating sequences, genome integrating sequences, phage or nucleotide sequences. They may also be linear, circular, or supercoiled, of a single- or double-stranded DNA or RNA, derived from any source. Plasmids and vectors may be constructed by known techniques in which a number of nucleotide sequences have been joined or recombined into a unique construction. Plasmids and vectors generally also include a promoter fragment and DNA sequence for a selected gene product along with appropriate 3' untranslated sequence. Generally, the plasmids of the present invention are stable and self-replicating.

[0071] As used herein, the term "anaerobic" refers to an organism, biochemical reaction or process that is active or occurs under conditions of an absence of gaseous O.sub.2.

[0072] "Anaerobic conditions" are defined as conditions under which the oxygen concentration in the fermentation medium is too low for the microorganism to use as a terminal electron acceptor. Anaerobic conditions may be achieved by sparging a fermentation medium with an inert gas such as nitrogen until oxygen is no longer available to the microorganism as a terminal electron acceptor. Alternatively, anaerobic conditions may be achieved by the microorganism consuming the available oxygen of fermentation until oxygen is unavailable to the microorganism as a terminal electron acceptor.

[0073] "Aerobic metabolism" refers to a biochemical process in which oxygen is used as a terminal electron acceptor to convert energy, typically in the form of ATP, from carbohydrates. Aerobic metabolism typically occurs, for example, via the electron transport chain in mitochondria in eukaryotes, wherein a single glucose molecule is metabolized completely into carbon dioxide in the presence of oxygen.

[0074] In contrast, "anaerobic metabolism" refers to a biochemical process in which oxygen is not the final acceptor of electrons generated. Anaerobic metabolism can be divided into anaerobic respiration, in which compounds other than oxygen serve as the terminal electron acceptor, and substrate level phosphorylation, in which no exogenous electron acceptor is used and products of an intermediate oxidation state are generated via a "fermentative pathway."

[0075] In "fermentative pathways", the amount of NAD(P)H generated by glycolysis is balanced by the consumption of the same amount of NAD(P)H in subsequent steps. For example, in one of the fermentative pathways of certain yeast strains, NAD(P)H generated through glycolysis donates its electrons to acetaldehyde, yielding ethanol. Fermentative pathways are usually active under anaerobic conditions but may also occur under aerobic conditions, under conditions where NADH is not fully oxidized via the respiratory chain.

[0076] As used herein, the term "end-product" refers to a chemical compound that is not or cannot be used by a cell, and so is excreted or allowed to diffuse into the extracellular environment. Common examples of end-products from anaerobic fermentation include, but are not limited to, ethanol, acetic acid, formic acid, lactic acid, hydrogen and carbon dioxide.

[0077] As used herein, "cofactors" are compounds involved in biochemical reactions that are recycled within the cells and remain at approximately steady state levels. Common examples of cofactors involved in anaerobic fermentation include, but are not limited to, NAD.sup.+ and NADP.sup.+. In metabolism, a cofactor can act in oxidation-reduction reactions to accept or donate electrons. When organic compounds are broken down by oxidation in metabolism, their energy can be transferred to NAD.sup.+ by its reduction to NADH, to NADP.sup.- by its reduction to NADPH, or to another cofactor, FAD.sup.+, by its reduction to FADH.sub.2. The reduced cofactors can then be used as a substrate for a reductase.

[0078] As used herein, a "pathway" is a group of biochemical reactions that together can convert one compound into another compound in a step-wise process. A product of the first step in a pathway may be a substrate for the second step, and a product of the second step may be a substrate for the third, and so on. Pathways of the present invention include, but are not limited to, the pyruvate metabolism pathway the lactate production pathway, the ethanol production pathway, and the glycerol-production pathway.

[0079] The term "recombination" or "recombinant" refers to the physical exchange of DNA between two identical (homologous), or nearly identical, DNA molecules. Recombination can be used for targeted gene deletion or to modify the sequence of a gene. The term "recombinant microorganism" and "recombinant host cell" are used interchangeably herein and refer to microorganisms that have been genetically modified to express or over-express endogenous polynucleotides, or to express heterologous polynucleotides, such as those included in a vector, or which have a modification in expression of an endogenous gene.

[0080] By "expression modification" it is meant that the expression of the gene, or level of a RNA molecule or equivalent RNA molecules encoding one or more polypeptides or polypeptide subunits, or activity of one or more polypeptides or polypeptide subunits is up regulated or down-regulated, such that expression, level, or activity, is greater than or less than that observed in the absence of the modification.

[0081] In one aspect of the invention, genes or particular polynucleotide sequences are partially, substantially, or completely deleted, silenced, inactivated, or down-regulated in order to inactivate the enzymatic activity they encode. Complete 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, deletion, removal or substitution of nucleic acid sequences that disrupt the function and/or expression of the gene.

[0082] As used herein, the term "down-regulate" includes the deletion or mutation of a genetic sequence, or insertion of a disrupting genetic element, coding or non-coding, such that the production of a gene product is lessened by the deletion, mutation, or insertion. It includes a decrease in the expression level (i.e., molecular quantity) of a mRNA or protein. "Delete" or "deletion" as used herein refers to a removal of a genetic element such that a corresponding gene is completely prevented from being expressed. In some embodiments, deletion refers to a complete gene deletion. Down-regulation can also occur by causing the repression of genetic elements by chemical or other environmental means, for example by engineering a chemically-responsive promoter element (or other type of conditional promoter) to control the expression of a desired gene product. Down-regulation can also occur through use of a weak promoter.

[0083] As used herein, the term "up-regulate" includes the insertion, reintroduction, mutation, or increased expression of a genetic sequence, such that the production of a gene product is increased by the insertion, reintroduction, or mutation. "Insert" or "insertion" as used herein refers to an introduction of a genetic element such that a corresponding gene is expressed. Up-regulation can also occur by causing the increased expression of genetic elements through an alteration of the associated regulatory sequence.

[0084] As used herein, the term "glycerol-production pathway" refers to the collection of biochemical pathways that produce glycerol from DHAP. Components of the pathway consist of all substrates, cofactors, byproducts, intermediates, end-products, and enzymes in the pathway.

[0085] As used herein, the term "ethanol production pathway" refers the collection of biochemical pathways that produce ethanol from pyruvate. Components of the pathway consist of all substrates, cofactors, byproducts, intermediates, end-products, and enzymes in the pathway.

[0086] As used herein, the term "acetyl-CoA production pathway" refers to the collection of biochemical pathways that produce acetyl-CoA from acetyl-phosphate. Components of the pathway consist of all substrates, cofactors, byproducts, intermediates, end-products, and enzymes in the pathway.

[0087] As used herein, the term "pyruvate metabolism pathway" refers to the collection of biochemical pathways that convert pyruvate into any product, including, but not limited to, ethanol, lactic acid, acetic acid and formate. It also includes the collection of pathways that result in the production of pyruvate, such as glycolysis. Components of the pathway consist of all substrates, cofactors, byproducts, intermediates, end-products, and enzymes in the pathway.

[0088] As used herein, the term "glycolysis" or "glycolytic pathway" refers to the canonical pathway of basic metabolism in which a sugar such as glucose is broken down into more oxidized products, converting energy and compounds required for cell growth. Components of the pathway consist of all substrates, cofactors, byproducts, intermediates end-products, and enzymes in the pathway.

[0089] As used herein, the term "phosphoketolase", "single-specificity phosphoketolase" or "dual-specificity phosphoketolase" is intended to include the enzymes that catalyze the conversion of D-xylulose 5-phosphate to D-glyceraldehyde 3-phosphate. Dual specificity phosphoketolase additionally includes the enzymes that catalyze the conversion of D-fructose 6-phosphate to D-erythrose 4-phosphate. Phosphoketolase, single-specificity phosphoketolase and dual-specificity phosphoketolase are referred to collectively as "PHKs" or "phosphoketolase" (FIG. 7). PHKs include those enzymes that correspond to Enzyme Commission Number (EC) 4.1.2.9 and 4.1.2.22. In some embodiments, PHK is from A. niger (SEQ ID NOs: 3 and 13), N. crassa (SEQ ID NOs: 4 and 14), L. casei PHK (SEQ ID NOs: 5 and 11), L. plantarum PHK1 (SEQ ID NOs: 7 and 9), L. plantarum PHK2 (SEQ ID NOs: 6 and 15), B. adolescentis (SEQ ID NOs: 8 and 12), B. bifidum (SEQ ID NOs: 61 and 62), B. gallicum (SEQ ID NOs: 63 and 64), B. animalis (SEQ ID NOs: 65 and 66), L. pentosum (SEQ ID NOs: 67 and 68), L. acidophilus (SEQ ID NOs: 69 and 70), P. chrysogenum (SEQ ID NOs: 71 and 72), A. nidulans (SEQ ID NOs: 73 and 74), A. clavatus (SEQ ID NOs: 77 and 78), L. mesenteroides (SEQ ID NOs: 93 and 94), or O. oenii (SEQ ID NOs: 101 and 102).

[0090] As used herein, the term "alcohol dehydrogenase" or "ADH" is intended to include the enzymes that catalyze the conversion of ethanol into acetylaldehyde. Very commonly, the same enzyme catalyzes the reverse reaction from acetaldehyde to ethanol, which is the direction more relevant to fermentation. Alcohol dehydrogenase includes those enzymes that correspond to EC 1.1.1.1 and 1.1.1.2 and exemplified by the enzymes disclosed in GenBank Accession No. U49975.

[0091] As used herein, the term "aldehyde dehydrogenase", "ALD" or "ALDH" is intended to include the enzymes that catalyze the oxidation of aldehydes. Aldehyde dehydrogenase enzymes include "acetaldehyde dehydrogenase", which catalyzes the conversion of acetaldehyde into acetyl-CoA. Very commonly, the same enzyme catalyzes the reverse reaction from acetyl-CoA to acetaldehyde, which is the direction more relevant to fermentation. Aldehyde dehydrogenase includes those enzymes that correspond to EC 1.2.1.3, 1.2.1.4 and 1.2.1.10. In some embodiments, acetaldehyde dehydrogenase is from S. cerevisiae (ALD2: SEQ ID NO: 17 and 18; ALD3: SEQ ID NO: 19 and 20; ALD4: SEQ ID NO: 21 and 22; ALD5: SEQ ID NO: 23 and 24; or ALD6 SEQ ID NO: 25 and 26).

[0092] As used herein, the term "phosphotransacetylase" or "PTA" is intended to include the enzymes capable of converting acetyl-CoA to acetylphosphate. PTA includes those enzymes that correspond to EC 2.3.1.8. In some embodiments, PTA is from B. adolescentis (SEQ ID NOs: 1 and 10), C. cellulolyticum (SEQ ID NOs: 79 and 80), C. phytofermentans (SEQ ID NOs: 81 and 82), B. bifidum (SEQ ID NOs: 83 and 84), B. animalis (SEQ ID NOs: 85 and 86), L. mesenteroides (SEQ ID NOs: 95 and 96), or O. oenii (SEQ ID NOs: 103 and 34).

[0093] As used herein, the term "acetate kinase" or "ACK" is intended to include the enzymes capable of converting acetylphosphate to acetate. ACK includes those enzymes that correspond to EC 2.7.2.12. In some embodiments, ACK is from B. adolescentis (SEQ ID NOs: 2 and 16), C. cellulolyticum (SEQ ID NOs: 87 and 88), C. phytofermentans (SEQ ID NOs: 75 and 76), L. mesenteroides (SEQ ID NOs: 91 and 92), or O. oenii (SEQ ID NOs: 99 and 100).

[0094] As used herein, the term "glycerol-3-phosphate dehydrogenase" or "GPD" is intended to include those enzymes capable of converting dihydroxyacetone phosphate to glycerol-3-phosphate. GPD includes those enzymes that correspond to EC 1.1.1.8. In some embodiments, the GPD is GPD1 and/or GPD2 from S. cerevisiae (GDP1: SEQ ID NO: 27 and 28, GDP2: SEQ ID NO: 29 and 30).

[0095] As used herein, the term "glycerol-3-phosphate phosphatase" is intended to include those enzymes capable of converting glycerol-1-phosphate to glycerol. Glycerol-3-phosphate is intended to include those enzymes that correspond to EC 3.1.3.21.

[0096] As used herein, the term "formate dehydrogenase" or "FDH" is intended to include those enzymes capable of converting formate to bicarbonate (carbon dioxide). Formate dehydrogenase includes those enzymes that correspond to EC 1.2.1.43 and EC 1.2.1.2. In some embodiments, the FDH is from S. cerevisiae (FDH1: SEQ ID NO: 31, and 32, FDH2: SEQ ID NO: 33).

[0097] As used herein, the term "transaldolase" is intended to include those enzymes capable of converting glyceraldehyde-3-phosphate to fructose-6-phosphate. Transaldolase is intended to include those enzymes that correspond to EC 2.2.1.2. In some embodiments, the transaldolase is from S. cerevisiae (TAL1: SEQ ID NO: 35 and 36).

[0098] As used herein, the term "transketolase" is intended to include those enzymes capable of converting sedoheptulose-7-P and glyceraldehyde-3-P to D-ribose-5-P and D-xylulose-5-P and those enzymes capable of converting fructose 6-phosphate and glyceraldehyde-3-P to D-xylulose-5-P and aldose erythrose-4-phosphate. Transketolase is intended to include those enzymes that correspond to EC 2.2.1.1. In some embodiments, the transketolase is from S. cerevisiae (TKL1: SEQ ID NO: 37 and 38).

[0099] As used herein, the term "ribose-5-P isomerase" or "ribose-5-phosphate isomerase" is intended to include those enzymes capable of converting ribose-5-phosphate to ribulose-5-phosphate. Ribose-5-P isomerase is intended to include those enzymes that correspond to EC 5.3.1.6. In some embodiments, the ribose-5-P isomerase is from S. cerevisiae (RKI1: SEQ ID NO: 39 and 40).

[0100] As used herein, the term "ribulose-S-P 3 epimerase", "ribulose-5-phosphate 3 epimerase" or "ribulose-phosphate 3-epimerase" is intended to include those enzymes capable of converting D-ribulose 5-phosphate to D-xylulose 5-phosphate. Ribulose-5-P 3 epimerase is intended to include those enzymes that correspond to EC 5.1.3.1. In some embodiments, the ribulose-5-P 3 epimerase is from S. cerevisiae (RPE1: SEQ ID NO: 41 and 42).

[0101] As used herein, the term "pyruvate decarboxylase" or "PDC" is intended to include those enzymes capable of converting pyruvic acid to acetaldehyde. PDC is intended to include those enzymes that correspond to EC 4.1.1.1.

[0102] As used herein, the term "bifunctional" is intended to include enzymes that catalyze more than one biochemical reaction step. A specific example of a bifunctional enzyme used herein is an enzyme (adhE) that catalyzes both the alcohol dehydrogenase and acetaldehyde dehydrogenase reactions, and includes those enzymes that correspond to EC 1.2.1.10 and 1.1.1.1. In some embodiments, the bifunctional acetaldehyde-alcohol dehydrogenase is from B. adolescentis (adhE: SEQ ID NO: 43 and 44). In some embodiments, the bifunctional enzyme is a NADPH specific bifunctional acetaldehyde-alcohol dehydrogenase, and includes those enzymes that correspond to EC 1.2.1.10 and 1.1.1.2. In some embodiments, the NADPH specific bifunctional acetaldehyde-alcohol dehydrogenase is from L. mesenteroides (SEQ ID NO: 89 and 90) or Oenococcus oenii (SEQ ID NO: 97 and 98).

[0103] As used herein, the term "pyruvate formate lyase" or "PFL" is intended to include the enzymes capable of converting pyruvate to formate and acetyl-CoA. PFL includes those enzymes that correspond to EC 2.3.1.54 and exemplified by SEQ ID NO: 47 and SEQ ID NO: 48.

[0104] As used herein, the term "PFL-activating enzymes" is intended to include those enzymes capable of aiding in the activation of PFL. PFL-activating enzymes include those enzymes that correspond to EC 1.97.1.4 and exemplified by SEQ ID NO: 45 and SEQ ID NO: 46.

[0105] The term "feedstock" is defined as a raw material or mixture of raw materials supplied to a microorganism or fermentation process from which other products can be made. For example, a carbon source, such as biomass or the carbon compounds derived from biomass are a feedstock for a microorganism that produces a product in a fermentation process. A feedstock can contain nutrients other than a carbon source.

[0106] Biomass can include any type of biomass known in the art or described herein. The terms "lignocellulosic material," "lignocellulosic substrate" and "cellulosic biomass" mean any type of carbon containing feed stock including woody biomass, such as recycled wood pulp fiber, sawdust, hardwood, softwood, grasses, sugar-processing residues, 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, corn fiber, stover, succulents, agave, or any combination thereof.

[0107] The term "yield" is defined as the amount of product obtained per unit weight of raw material and may be expressed as gram product per gram substrate (g/g). Yield may be expressed as a percentage of the theoretical yield. "Theoretical yield" is defined as the maximum amount of product that can be generated per a given amount of substrate as dictated by the stoichiometry of the metabolic pathway used to make the product. For example, the theoretical yield for one typical conversion of glucose to ethanol is 0.51 g EtOH per 1 g glucose. As such, a yield of 4.8 g ethanol from 10 g of glucose would be expressed as 94% of theoretical or 94% theoretical yield.

[0108] The term "titer" is defined as the strength of a solution or the concentration of a substance in solution. For example, the titer of a product in a fermentation broth is described as gram of product in solution per liter of fermentation broth (g/L) or as g/kg broth.

[0109] As used herein, the term "flux" is the rate of flow of molecules through a metabolic pathway, akin to the flow of material in a process.

[0110] "Bacteria", or "eubacteria", refers to a domain of prokaryotic organisms. Bacteria include gram-positive (gram+) bacteria and gram-negative (gram-) bacteria.

[0111] "Yeast" refers to a domain of eukaryotic organisms that are unicellular fungi.

[0112] The terms "derivative" and "analog" refer to a polypeptide differing from the enzymes of the invention, but retaining essential properties thereof. Generally, derivatives and analogs are overall closely similar, and, in many regions, identical to the enzymes of the invention. The terms "derived from", "derivative" and "analog" when referring to enzymes of the invention include any polypeptides which retain at least some of the activity of the corresponding native polypeptide or the activity of its catalytic domain.

[0113] Derivatives of enzymes disclosed herein are polypeptides which may have been altered so as to exhibit features not found on the native polypeptide. Derivatives can be covalently modified by substitution (e.g. amino acid substitution), chemical, enzymatic, or other appropriate means with a moiety other than a naturally occurring amino acid (e.g., a detectable moiety such as an enzyme or radioisotope). Examples of derivatives include fusion proteins, or proteins which are based on a naturally occurring protein sequence, but which have been altered. For example, proteins can be designed by knowledge of a particular amino acid sequence, and/or a particular secondary, tertiary, and quaternary structure. Derivatives include proteins that are modified based on the knowledge of a previous sequence, natural or synthetic, which is then optionally modified, often, but not necessarily to confer some improved function. These sequences, or proteins, are then said to be derived from a particular protein or amino acid sequence. In some embodiments of the invention, a derivative must retain at least about 50% identity, at least about 60% identity, at least about 70% identity, at least about 80% identity, at least about 90% identity, at least about 95% identity, at least about 97% identity, or at least about 99% identity to the sequence the derivative is "derived from. " In some embodiments of the invention, an enzyme is said to be derived from an enzyme naturally found in a particular species if, using molecular genetic techniques, the DNA sequence for part or all of the enzyme is amplified and placed into a new host cell.

[0114] The term "percent identity", as known in the art, is a relationship between two or more polypeptide sequences or two or more polynucleotide sequences, as determined by comparing the sequences. In the art, "identity" also means the degree of sequence relatedness between polypeptide or polynucleotide sequences, as the case may be, as determined by the match between strings of such sequences.

[0115] As known in the art, "similarity" between two polypeptides is determined by comparing the amino acid sequence and conserved amino acid substitutes thereto of the polypeptide to the sequence of a second polypeptide.

[0116] "Identity" and "similarity" can be readily calculated by known methods, including but not limited to those described in: Computational Molecular Biology (Lesk, A. M., ed.) Oxford University Press, NY (1988); Biocomputing: Informatics and Genome Projects (Smith, D. W., ed.) Academic Press, NY (1993); Computer Analysis of Sequence Data, Part I (Griffin, A. M., and Griffin, H. G., eds.) Humana Press, NJ (1994); Sequence Analysis in Molecular Biology (von Heinje, G., ed.) Academic Press (1987); and Sequence Analysis Primer (Gribskov, M. and Devereux, J., eds.) Stockton Press, NY (1991). Preferred methods to determine identity are designed to give the best match between the sequences tested. Methods to determine identity and similarity are codified in publicly available computer programs. Sequence alignments and percent identity calculations may be performed using the Megalign program of the LASERGENE bioinformatics computing suite (DNASTAR Inc., Madison, Wis.). Multiple alignments of the sequences disclosed herein were performed using the Clustal method of alignment (Higgins and Sharp (1989) CABIOS. 5:151-153) with the default parameters (GAP PENALTY=10, GAP LENGTH PENALTY=10). Default parameters for pairwise alignments using the Clustal method were KTUPLE 1, GAP PENALTY=3, WINDOW=5 and DIAGONALS SAVED=5.

[0117] Suitable nucleic acid sequences or fragments thereof (isolated polynucleotides of the present invention) encode polypeptides that are at least about 70% to 75% identical to the amino acid sequences disclosed herein, at least about 80%, at least about 85%, or at least about 90% identical to the amino acid sequences disclosed herein, or at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or at least about 100% identical to the amino acid sequences disclosed herein. Suitable nucleic acid fragments are at least about 70%, at least about 75%, or at least about 80% identical to the nucleic acid sequences disclosed herein, at least about 80%, at least about 85%, or at least about 90% identical to the nucleic acid sequences disclosed herein, or at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or at least about 100% identical to the nucleic acid sequences disclosed herein. Suitable nucleic acid fragments not only have the above identities/similarities but typically encode a polypeptide having at least about 50 amino acids, at least about 100 amino acids, at least about 150 amino acids, at least about 200 amino acids, or at least about 250 amino acids.

Codon Optimization

[0118] In some embodiments of the present invention, exogenous genes may be codon-optimized in order to express the polypeptide they encode most efficiently in the host cell. Methods of codon optimization are well known in the art. (See, e.g. Welch et al. "Designing genes for successful protein expression." Methods Enzymol. 2011. 498:43-66.)

[0119] In general, highly expressed genes in an organism are biased towards codons that are recognized by the most abundant tRNA species in that organism. One measure of this bias is the "codon adaptation index" or "CAI," which measures the extent to which the codons used to encode each amino acid in a particular gene are those which occur most frequently in a reference set of highly expressed genes from an organism. The Codon Adaptation Index is described in more detail in Sharp et al., "The Codon Adaptation Index: a Measure of Directional Synonymous Codon Usage Bias, and Its Potential Applications." Nucleic Acids Research 1987. 15: 1281-1295, which is incorporated by reference herein in its entirety.

[0120] A codon optimized sequence may be further modified for expression in a particular organism, depending on that organism's biological constraints. For example, large runs of "As" or "Ts" (e.g., runs greater than 3, 4, 5, 6, 7, 8, 9, or 10 consecutive bases) can effect transcription negatively. Therefore, it can be useful to remove a run by, for example, replacing at least one nucleotide in the run with another nucleotide. Furthermore, specific restriction enzyme sites may be removed for molecular cloning purposes by replacing at least one nucleotide in the restriction site with another nucleotide. Examples of such restriction enzyme sites include PacI, Ascl, BamHI, BglII, EcoRI and XhoI. Additionally, the DNA sequence can be checked for direct repeats, inverted repeats and mirror repeats with lengths of about 5, 6, 7, 8, 9 or 10 bases or longer. Runs of "As" or "Ts", restriction sites and/or repeats can be modified by replacing at least one codon within the sequence with the "second best" codons, i.e., the codon that occurs at the second highest frequency for a particular amino acid within the particular organism for which the sequence is being optimized.

[0121] Deviations in the nucleotide sequence that comprise the codons encoding the amino acids of any polypeptide chain allow for variations in the sequence coding for the gene. Since each codon consists of three nucleotides, and the nucleotides comprising DNA are restricted to four specific bases, there are 64 possible combinations of nucleotides, 61 of which encode amino acids (the remaining three codons encode signals ending translation). The "genetic code" which shows which codons encode which amino acids is reproduced herein as Table 1. As a result, many amino acids are designated by more than one codon. For example, the amino acids alanine and proline are coded for by four triplets, serine and arginine by six triplets each, whereas tryptophan and methionine are coded for by just one triplet. This degeneracy allows for DNA base composition to vary over a wide range without altering the amino acid sequence of the proteins encoded by the DNA.

TABLE-US-00001 TABLE 1 The Standard Genetic Code T C A G T TTT Phe (F) TCT Ser (S) TAT Tyr (Y) TGT Cys (C) TTC'' TCC'' TAC'' TGC TTA Leu (L) TCA'' TAA Ter TGA Ter TTG'' TCG'' TAG Ter TGG Trp (W) C CTT Leu (L) CCT Pro (P) CAT His (H) CGT Arg (R) CTC'' CCC'' CAC'' CGC'' CTA'' CCA'' CAA Gln (Q) CGA'' CTG'' CCG'' CAG'' CGG'' A ATT Ile (I) ACT Thr (T) AAT Asn (N) AGT Ser (S) ATC'' ACC'' AAC'' AGC'' ATA'' ACA'' AAA Lys (K) AGA Arg (R) ATG Met (M) ACG'' AAG'' AGG'' G GTT Val (V) GCT Ala (A) GAT Asp (D) GGT Gly (G) GTC'' GCC'' GAC'' GGC'' GTA'' GCA'' GAA Glu (E) GGA'' GTG'' GCG'' GAG'' GGG''

[0122] Many organisms display a bias for use of particular codons to code for insertion of a particular amino acid in a growing peptide chain. Codon preference or codon bias, differences in codon usage between organisms, is afforded by degeneracy of the genetic code, and is well documented among many organisms. Codon bias often correlates with the efficiency of translation of messenger RNA (mRNA), which is in turn believed to be dependent on, inter alia, the properties of the codons being translated and the availability of particular transfer RNA (tRNA) molecules. The predominance of selected tRNAs in a cell is generally a reflection of the codons used most frequently in peptide synthesis. Accordingly, genes can be tailored for optimal gene expression in a given organism based on codon optimization.

Host Cells

[0123] In some embodiments of the invention, the host cell is a eukaryotic microorganism. In some embodiments, the host cell is a yeast. In some embodiments, the host cell is able to digest and ferment cellulose. In some embodiments, the host cell is from the genus Saccharomyces. In some embodiments, the host cell is Saccharomyces cerevisiae.

[0124] In some embodiments, the host cells of the invention are cultured at a temperature above about 20.degree. C., above about 25.degree. C., above about 27.degree. C., above about 30.degree. C., above about 33.degree. C., above about 35.degree. C., above about 37.degree. C., above about 40.degree. C., above about 43.degree. C., above about 45.degree. C., or above about 47.degree. C.

[0125] In some embodiments, the host cells of the invention contain genetic constructs that lead to the down-regulation of one or more genes encoding a polypeptide at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical to one or more of the polypeptides encompassed by EC 1.1.1.8, 3.1.3.21, 1.2.1.43, 1.2.1.10 and/or EC 1.2.1.2.

[0126] In some embodiments, the host cells of the invention contain genetic constructs that lead to the expression or up-regulation of one or more genes encoding a polypeptide at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical to one or more of the polypeptides encoded by SEQ ID NOs: 9, 10, 11, 12, 13, 14, 15, 16, 18, 20, 22, 24, 26, 34, 36, 38, 40, 42, 44, 46, 48, 62, 64, 66, 68, 70, 72, 74, 76, 78, 80, 82, 84, 86, 88, 90, 92, 94, 96, 98, 100, or 102. In some embodiments, the host cells of the invention contain genetic constructs that leads to the expression or up-regulation of a polypeptide encoding the activity associated with EC Nos.: 4.1.2.9, 4.1.2.22, 2.3.1.8, 1.2.1.10, 1.2.1.3, 1.2.1.4, 1.1.1.1, 1.1.1.2, 2.7.2.12, 2.2.1.2, 2.2.1.1, 5.3.1.6, 5.1.3.1, 2.3.1.54, 1.97.1.4, 1.2.1.3, 1.2.1.4, or 4.1.1.1.

[0127] In some embodiments, PHK is up-regulated. In some embodiments, single-specificity phosphoketolase is up-regulated. In some embodiments, dual-specificity phosphoketolase is up-regulated. In some embodiments, the up-regulated PHK is from an enzyme that corresponds to an EC number selected from the group consisting of: EC 4.1.2.9 and 4.1.2.22. In some embodiments, the PHK is from A. niger. In some embodiments, the PHK is from N. crassa. In some embodiments, the PHK is from L. casei. In some embodiments, the PHK is from L. plantarum. In some embodiments, the PHK is from B. adolescentis. In some embodiments, the PHK is derived from a genus selected from the group consisting of Aspergillus, Neurospora, Lactobacillus, Bifidobacterium, and Penicillium. In some embodiments, the phosphoketolase corresponds to a polypeptide selected from a group consisting of SEQ ID NOs: 9, 11, 12, 13, 14, 15, 62, 64, 66, 68, 70, 72, 74, 78, 94, or 102.

[0128] In some embodiments, phosphotransacetylase is up-regulated. In some embodiments, the up-regulated phosphotransacetylase is from an enzyme that corresponds to EC 2.3.1.8. In some embodiments, the phosphotransacetylase is from B. adolescentis. In some embodiments, the phosphotransacetylase is derived from a genus selected from the group consisting of Bifidobacterium, Lactobacillus, or Clostridium. In some embodiments, the phosphotransacetylase corresponds to a polypeptide selected from a group consisting of SEQ ID NOs: 10, 34, 80, 82, 84, 86, or 94.

[0129] In some embodiments of the invention, acetate kinase is up-regulated. In some embodiments, the up-regulated acetate kinase is from an enzyme that corresponds to EC 2.7.2.12. In some embodiments, the acetate kinase is from B. adolescentis. In some embodiments, the acetate kinase is derived from a genus selected from the group consisting of Bifidobacterium, Lactobacillus, or Clostridium. In some embodiments, the acetate kinase corresponds to a polypeptide selected from a group consisting of SEQ ID NOs: 16, 88, 76, 92, or 100.

[0130] In some embodiments, bifunctional acetaldehyde-alcohol dehydrogenase is up-regulated. In some embodiments, the up-regulated bifunctional acetaldehyde-alcohol dehydrogenase is from an enzyme that corresponds to an EC number selected from the group consisting of: EC 1.2.1.0 and 1.1.1.1. In some embodiments, the bifunctional acetaldehyde-alcohol dehydrogenase is a NADPH dependent bifunctional acetaldehyde-alcohol dehydrogenase selected from a group of enzymes having the following Enzyme Commission Numbers: EC 1.2.1.10 and 1.1.1.2. In some embodiments, the bifunctional acetaldehyde-alcohol dehydrogenase corresponds to a polypeptide selected from the group consisting of SEQ ID NOs: 44, 90, and 98

[0131] In some embodiments, transaldolase is up-regulated. In some embodiments, the up-regulated transaldolase is from an enzyme that corresponds to EC 2.2.1.2. In some embodiments, the transaldolase is from S. cerevisiae and corresponds to a polypeptide encoded by SEQ ID NO: 36. In some embodiments, transketolase is up-regulated. In some embodiments, the up-regulated transketolase is from an enzyme that corresponds to EC 2.2.1.1. In some embodiments, the transketolase is from S. cerevisiae and corresponds to a polypeptide encoded by SEQ ID NO: 38. In some embodiments, ribose-5-P isomerase is up-regulated. In some embodiments, the up-regulated ribose-5-P isomerase is from an enzyme that corresponds to EC 5.3.1.6. In some embodiments, the ribose-5-P isomerase is from S. cerevisiae and corresponds to a polypeptide encoded by SEQ ID NO: 40. In some embodiments, ribulose-5-P epimerase is up-regulated. In some embodiments, the up-regulated ribulose-5-P epimerase is from an enzyme that corresponds to EC 5.1.3.1. In some embodiments, the ribulose-5-P 3-epimerase is from S. cerevisiae and corresponds to a polypeptide encoded by SEQ ID NO: 42. In some embodiments, pyruvate decarboxylase is up-regulated. In some embodiments, the up-regulated pyruvate decarboxylase is from an enzyme that corresponds to EC 4.1.1.1. In some embodiments, alcohol dehydrogenase is up-regulated. In some embodiments, the up-regulated alcohol dehydrogenase is from an enzyme that corresponds to EC 1.1.1.1 and 1.1.1.2. In some embodiments, pyruvate formate lyase is up-regulated. In some embodiments, the up-regulated pyruvate formate lyase is from an enzyme that corresponds to EC 2.3.1.54. In some embodiments, the pyruvate formate lyase corresponds to a polypeptide encoded by SEQ ID NO: 48. In some embodiments, pyruvate formate lyase activating enzyme is up-regulated. In some embodiments, the up-regulated pyruvate formate lyase activating enzyme is from an enzyme that corresponds to EC 1.97.1.4. In some embodiments, the pyruvate formate lyase activating enzyme corresponds to a polypeptide encoded by SEQ ID NO: 46.

[0132] In some embodiments, the host cells of the invention contain genetic constructs that lead to the expression or down-regulation of one or more genes encoding a polypeptide at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical to one or more of the polypeptides encoded by SEQ ID NOs: 18, 20, 22, 24, 26, 28, 30, 32 or the polynucleotide encoded by SEQ ID NO: 33. In some embodiments, the host cells of the invention contain genetic constructs that leads to the expression or down-regulation of a polypeptide encoding the activity associated with EC Nos.: 1.2.1.3, 1.2.1.4, 1.2.1.10, 1.1.1.8 and 3.1.3.21.

[0133] In some embodiments, GPD is down-regulated. In some embodiments, the down-regulated GPD is from an enzyme that corresponds to EC 1.1.1.8. In some embodiments gpd1 and gpd2 from S. cerevisiae are down-regulated. In some embodiments, the glycerol-3-phosphate dehydrogenase is selected from the group consisting of glycerol-3-phosphate dehydrogenase 1, glycerol-3-phosphate dehydrogenase 2, and combinations thereof. In some embodiments, the glycerol-3-phosphate dehydrogenase 1 is from S. cerevisiae and corresponds to a polypeptide encoded by SEQ ID NO: 28. In some embodiments, the glycerol-3-phosphate dehydrogenase 2 is from S. cerevisiae and corresponds to a polypeptide encoded by SEQ ID NO: 30. In some embodiments, formate dehydrogenase is down-regulated. In some embodiments, the down-regulated formate dehydrogenase corresponds to an EC number selected from the group consisting of: EC 1.2.1.43 and EC 1.2.1.2. In some embodiments, formate dehydrogenase from S. cerevisiae is down-regulated. In some embodiments, the formate dehydrogenase from S. cerevisiae corresponds to a polypeptide corresponding to SEQ ID NO: 32 or a polynucleotide corresponding to SEQ ID NO: 33. In some embodiments, glycerol-3-phosphate phosphatase is down-regulated. In some embodiments, the down-regulated glycerol-3-phosphate phosphatase corresponds to EC 3.1.3.21.

[0134] In some embodiments, aldehyde dehydrogenase is down-regulated. In some embodiments, aldehyde dehydrogenase is up-regulated. In some embodiments, the aldehyde dehydrogenase is an enzyme that corresponds to EC 1.2.1.10, EC 1.2.1.3, or EC 1.2.1.4. In some embodiments, the aldehyde dehydrogenase is acetaldehyde dehydrogenase. In some embodiments, the acetaldehyde dehydrogenase enzyme is selected from Ald2, Ald3, Ald4, Ald5, and Ald6 in S. cerevisiae. In some embodiments, the acetaldehyde dehydrogenase from S. cerevisiae corresponds to a polypeptide sequence selected from the group consisting of: SEQ ID NO: 18, SEQ ID NO: 20, SEQ ID NO: 22, SEQ ID NO: 24, and SEQ ID NO: 26.

[0135] In some embodiments, phosphoketolase, phosphotransacetylase and bifunctional acetaldehyde-alcohol dehydrogenase are up-regulated, and at least one glycerol-3-phosphate dehydrogenase enzyme is down-regulated.

[0136] In some embodiments, phosphotransacetylase, bifunctional acetaldehyde-alcohol dehydrogenase, and at least one aldehyde dehydrogenase are up-regulated. In some embodiments, phosphotransacetylase, bifunctional acetaldehyde-alcohol dehydrogenase, at least one aldehyde dehydrogenase are up-regulated, and at least one glycerol-3-phosphate dehydrogenase enzyme is down-regulated. In some embodiments, phosphotransacetylase, acetate kinase, bifunctional acetaldehyde-alcohol dehydrogenase, and at least one aldehyde dehydrogenase are up-regulated. In some embodiments, phosphotransacetylase, acetate kinase and bifunctional acetaldehyde-alcohol dehydrogenase are up-regulated, and at least one glycerol-3-phosphate dehydrogenase enzyme is down-regulated. In some embodiments, at least one aldehyde dehydrogenase enzyme is also down-regulated. In some embodiments, the aldehyde dehydrogenase is acetaldehyde dehydrogenase.

[0137] In some embodiments, phosphoketolase, phosphotransacetylase, bifunctional acetaldehyde-alcohol dehydrogenase, pyruvate formate lyase, and pyruvate formate lyase activating enzyme are up-regulated, and at least one glycerol-3-phosphate dehydrogenase enzyme is down-regulated.

[0138] In some embodiments, phosphoketolase, acetate kinase and bifunctional acetaldehyde-alcohol dehydrogenase are up-regulated, and at least one glycerol-3-phosphate dehydrogenase enzyme is down-regulated.

[0139] In some embodiments, phosphoketolase, acetate kinase, bifunctional acetaldehyde-alcohol dehydrogenase, pyruvate formate lyase, and pyruvate formate lyase activating enzyme are up-regulated, and at least one glycerol-3-phosphate dehydrogenase enzyme is down-regulated.

[0140] In some embodiments, gpd1, gpd2, fdh1, and/or fdh2 are down-regulated. In some embodiments, gpd1, gpd2, fdh1 and fdh2 are down-regulated, and PHK, bifunctional acetaldehyde-alcohol dehydrogenase (AdhE), and PTA are up-regulated. In some embodiments, gpd1, gpd2, fdh1 and fdh2 are down-regulated, and B. adolescentis PHK, AdhE, and B. adolescentis PTA are up-regulated in S. cerevisiae cells. In some embodiments, gpd1, gpd2, fdh1 and fdh2 are down-regulated, and L. plantarum PHK1, AdhE, and B. adolescentis PTA are up-regulated in S. cerevisiae cells. In some embodiments, gpd1, gpd2, fdh1 and fdh2 are down-regulated, and L. plantarum PHK2, AdhE, B. adolescentis PTA are up-regulated in S. cerevisiae cells. In some embodiments, gpd1, gpd2, fdh1 and fdh2 are down-regulated, and A. niger PHK, AdhE, and B. adolescentis PTA are up-regulated in S. cerevisiae cells. In some embodiments, gpd1, gpd2, fdh1 and fdh2 are down-regulated, and L. casei PHK1, AdhE, and B. adolescentis PTA are up-regulated in S. cerevisiae cells. In some embodiments, gpd1, gpd2, fdh1 and fdh2 are down-regulated, and L. casei PHK2, AdhE, and B. adolescentis PTA are up-regulated in S. cerevisiae cells. In some embodiments, gpd1, gpd2, fdh1 and/or fdh2 are down-regulated, PHK, B. adolescentis AdhE, and B. adolescentis PTA are up-regulated in S. cerevisiae cells. In some embodiments, the up-regulated enzymes are integrated. In some embodiments, the up-regulated enzymes are expressed using an expression vector. In some embodiments, gpd1, gpd2, fdh1 and/or fdh2 are down-regulated, and B. adolescentis PHK, B adolescentis bifunctional acetaldehyde-alcohol dehydrogenase, B adolescentis PTA, B. adolescentis pyruvate formate lyase, and B adolescentis pyruvate formate lyase activating enzyme are up-regulated. In some embodiments, fcy is down-regulated.

[0141] In some embodiments, gpd1, gpd2, fdh1 and/or fdh2 are down-regulated, and PHK, bifunctional acetaldehyde-alcohol dehydrogenase (AdhE), ACK and PTA are up-regulated. In some embodiments, gpd1, gpd2, fdh1 and/or fdh2 are down-regulated, and B. adolescentis PHK, AdhE, B. adolescentis ACK, and B. adolescentis PTA are up-regulated in S. cerevisiae cells. In some embodiments, gpd1, gpd2, fdh1 and/or fdh2 are down-regulated, and L. plantarum PHK1, AdhE, B. adolescentis ACK and B. adolescentis PTA are up-regulated in S. cerevisiae cells. In some embodiments, gpd1, gpd2, fdh1 and/or fdh2 are down-regulated, and L. plantarum PHK2, AdhE, B. adolescentis ACK, and B. adolescentis PTA are up-regulated in S. cerevisiae cells. In some embodiments, gpd1, gpd2, fdh1 and/or fdh2 are down-regulated, and A. niger PHK, AdhE, B. adolescentis ACK and B. adolescentis PTA are up-regulated in S. cerevisiae cells. In some embodiments, gpd1, gpd2, fdh1 and/or fdh2 are down-regulated, and L. casei PHK1, AdhE, B. adolescentis ACK and B. adolescentis PTA are up-regulated in S. cerevisiae cells. In some embodiments, gpd1, gpd2, fdh1 and/or fdh2 are down-regulated, and L. casei PHK2, AdhE, B. adolescentis ACK, and B. adolescentis PTA are up-regulated in S. cerevisiae cells. In some embodiments, gpd1, gpd2, fdh1 and/or fdh2 are down-regulated, PHK, B. adolescentis AdhE, B. adolescentis PTA and B. adolescentis ACK are up-regulated in S. cerevisiae cells. In some embodiments, the up-regulated enzymes are integrated. In some embodiments, the up-regulated enzymes are expressed using an expression vector. In some embodiments, gpd1, gpd2, fdh1 and/or fdh2 are down-regulated, and B. adolescentis PHK, B adolescentis bifunctional acetaldehyde-alcohol dehydrogenase, B. adolescentis PTA, B. adolescentis ACK B. adolescentis pyruvate formate lyase, and B adolescentis pyruvate formate lyase activating enzyme are up-regulated.

[0142] In some embodiments, gpd1, gpd2, fdh1 and/or fdh2 are down-regulated, and bifunctional acetaldehyde-alcohol dehydrogenase (AdhE), ACK and PTA are up-regulated. In some embodiments, gpd1, gpd2, fdh1 and/or fdh2 are down-regulated, and AdhE, B. adolescentis ACK, and B. adolescentis PTA are up-regulated in S. cerevisiae cells. In some embodiments, the up-regulated enzymes are integrated. In some embodiments, the up-regulated enzymes are expressed using an expression vector. In some embodiments, gpd1, gpd2, fdh1 and/or fdh2 are down-regulated, B. adolescentis AdhE, B. adolescentis PTA and B. adolescentis ACK are up-regulated in S. cerevisiae cells. In some embodiments, gpd1, gpd2, fdh1 and/or fdh2 are down-regulated, and B adolescentis bifunctional acetaldehyde-alcohol dehydrogenase, B. adolescentis PTA, B. adolescentis ACK B. adolescentis pyruvate formate lyase, and B adolescentis pyruvate formate lyase activating enzyme are up-regulated. In some embodiments, S. cerevisiae ALD2, ALD3, ALD4, or combinations thereof are down-regulated and S. cerevisiae ALD6, ALD5, or combinations thereof are up-regulated.

[0143] In some embodiments, gpd1, gpd2, fdh1 and/or fdh2 are down-regulated, B. adolescentis AdhE, B. adolescentis PTA and B. adolescentis ACK are up-regulated in S. cerevisiae cells.

Ethanol Production

[0144] For a microorganism to produce ethanol most economically, it is desired to produce a high yield. In one embodiment, the only product produced is ethanol. Extra products lead to a reduction in product yield and an increase in capital and operating costs, particularly if the extra products have little or no value. Extra products also require additional capital and operating costs to separate these products from ethanol.

[0145] Ethanol production can be measured using any method known in the art. For example, the quantity of ethanol in fermentation samples can be assessed using HPLC analysis. Additionally, many ethanol assay kits are commercially available, for example, alcohol oxidase enzyme based assays. Methods of determining ethanol production are within the scope of those skilled in the art from the teachings herein.

[0146] In some embodiments of the invention where redirected carbon flux generates increased ethanol production, the ethanol output can be improved by growth-coupled selection. For example, continuous culture or serial dilution cultures can be performed to select for cells that grow faster and/or produce ethanol (or any desired product) more efficiently on a desired feedstock.

[0147] One embodiment of the present invention relates to a method of producing ethanol using a microorganism described herein wherein said microorganism is cultured in the presence of a carbon containing feedstock for sufficient time to produce ethanol and, optionally, extracting the ethanol.

[0148] Ethanol may be extracted by methods known in the art. (See, e.g., U.S. Appl. Pub. No. 2011/0171709, which is incorporated herein by reference.)

[0149] Another embodiment of the present invention relates to a method of producing ethanol using a co-culture composed of at least two microorganisms in which at least one of the organisms is an organism described herein, and at least one of the organisms is a genetically distinct microorganism. In some embodiments, the genetically distinct microorganism is a yeast or bacterium. In some embodiments the genetically distinct microorganism is any organism from the genus Issatchenkia, Pichia, Clavispora, Candida, Hansenula, Kluyveromyces, Saccharomyces, Trichoderma, Thermoascus, Escherichia, Clostridium, Caldicellulosiruptor, Thermoanaerobacter and Thermoanaerobacterium.

[0150] In some embodiments, the recombinant microorganism produces about 2 to about 3% higher ethanol titer than a wildtype, non-recombinant organism; at least about 1 to at least about 2% higher ethanol titer than a wildtype, non-recombinant organism; at least about 1 to at least about 5% higher ethanol titer than a wildtype, non-recombinant organism; at least about 1 to at least about 7% higher ethanol titer than a wildtype, non-recombinant organism; at least about 1 to at least about 10% higher ethanol titer than a wildtype, non-recombinant organism; at least about 1 to at least about 15% higher ethanol titer than a wildtype, non-recombinant organism; at least about 1 to at least about 20% higher ethanol titer than a wildtype, non-recombinant organism; at least about 1 to at least about 30% higher ethanol titer than a wildtype, non-recombinant organism; at least about 1 to at least about 50% higher ethanol titer than a wildtype, non-recombinant organism; at least about 1 to at least about 75% higher ethanol titer than a wildtype, non-recombinant organism; at least about 1 to at least about 100% higher ethanol titer than a wildtype, non-recombinant organism.

[0151] In some embodiments, the recombinant microorganism produces at least about 0.5 g/L ethanol to at least about 2 g/L ethanol, at least about 0.5 g/L ethanol to at least about 3 g/L ethanol, at least about 0.5 g/L ethanol to at least about 5 g/L ethanol, at least about 0.5 g/L ethanol to at least about 7 g/L ethanol, at least about 0.5 g/L ethanol to at least about 10 g/L ethanol, at least about 0.5 g/L ethanol to at least about 15 g/L ethanol, at least about 0.5 g/L ethanol to at least about 20 g/L ethanol, at least about 0.5 g/L ethanol to at least about 30 g/L ethanol, at least about 0.5 g/L ethanol to at least about 40 g/L ethanol, at least about 0.5 g/L ethanol to at least about 50 g/L ethanol, at least about 0.5 g/L ethanol to at least about 75 g/L ethanol, or at least about 0.5 g/L ethanol to at least about 99 g/L ethanol per 24 hour incubation on a carbon-containing feed stock.

[0152] In some embodiments, the recombinant microorganism produces ethanol at least about 55% to at least about 75% of theoretical yield, at least about 50% to at least about 80% of theoretical yield, at least about 45% to at least about 85% of theoretical yield, at least about 40% to at least about 90% of theoretical yield, at least about 35% to at least about 95% of theoretical yield, at least about 30% to at least about 99% of theoretical yield, or at least about 25% to at least about 99% of theoretical yield.

[0153] In some embodiments, methods of producing ethanol can comprise contacting a biomass feedstock with a host cell or co-culture of the invention and additionally contacting the biomass feedstock with externally produced saccharolytic enzymes. In some embodiments, the host cells are genetically engineered (e.g., transduced, transformed, or transfected) with the polynucleotides encoding saccharolytic enzymes.

[0154] A "saccharolytic enzyme" can be any enzyme involved in carbohydrate digestion, metabolism and/or hydrolysis, including amylases, cellulases, hemicellulases, cellulolytic, and amylolytic accessory enzymes, inulinases, levanases, and pentose sugar utilizing enzymes. Exemplary externally produced saccharolytic enzymes are commercially available and are known to those of skill in the art and include glucoamylases.

Glycerol Production

[0155] In some embodiments of the invention where redirected carbon flux generates increased ethanol production, the glycerol output can be decreased by growth-coupled selection. For example, continuous culture or serial dilution cultures can be performed to select for cells that produce less glycerol on a desired feedstock. Glycerol can be measured, for example, by HPLC analysis of metabolite concentrations.

[0156] In some embodiments, the recombinant microorganism produces at least about 20% to at least about 30% less glycerol than a wildtype, non-recombinant organism; at least about 15% to at least about 30% less glycerol than a wildtype, non-recombinant organism; at least about 10% to at least about 40% less glycerol than a wildtype, non-recombinant organism; at least about 10% to at least about 50% less glycerol than a wildtype, non-recombinant organism; at least about 10% to at least about 60% less glycerol than a wildtype, non-recombinant organism; at least about 10% to at least about 70% less glycerol than a wildtype, non-recombinant organism; at least about 10% to at least about 80% less glycerol than a wildtype, non-recombinant organism; at least about 10% to at least about 90% less glycerol than a wildtype, non-recombinant organism; at least about 10% to at least about 99% less glycerol than a wildtype, non-recombinant organism; at least about 10% to at least about 100% less glycerol than a wildtype, non-recombinant organism; at least about 5% to at least about 100% less glycerol than a wildtype, non-recombinant organism; at least about 1% to at least about 100% less glycerol than a wildtype, non-recombinant organism.

[0157] In some embodiments, the recombinant microorganism produces glycerol at least about 50% to at least about 99% of theoretical yield, at least about 45% to at least about 80% of theoretical yield, at least about 40% to at least about 85% of theoretical yield, at least about 30% to at least about 80% of theoretical yield, at least about 25% to at least about 50% of theoretical yield, at least about 20% to at least about 50% of theoretical yield, at least about 15% to at least about 50% of theoretical yield, at least about 10% to at least about 50% of theoretical yield, at least about 5% to at least about 50% of theoretical yield, at least about 4% to at least about 50% of theoretical yield, at least about 3% to at least about 50% of theoretical yield, at least about 2% to at least about 50% of theoretical yield, at least about 1% to at least about 50% of theoretical yield, at least about 1% to at least about 25% of theoretical yield, at least about 1% to at least about 20% of theoretical yield or at least about 1% to at least about 10% of theoretical yield. In some embodiments, the recombinant microorganism produces glycerol at less about 1% of theoretical yield. In some embodiments, the recombinant microorganism produces no glycerol.

[0158] In some embodiments, the recombinant microorganism has a growth rate at least about 1/2 to at least about equal to the growth rate of a wildtype, non-recombinant organism, at least about 1/4 to at least about equal to the growth rate of a wildtype, non-recombinant organism, at least about 1/8 to at least about equal to the growth rate of a wildtype, non-recombinant organism, at least about 1/10 to at least about equal to the growth rate of a wildtype, non-recombinant organism, at least about 1/25 to at least about equal to the growth rate of a wildtype, non-recombinant organism, at least about 1/50 to at least about equal to the growth rate of a wildtype, non-recombinant organism or at least about 1/100th to at least about equal to the growth rate of a wildtype, non-recombinant organism.

[0159] In some embodiments, glycerol production is reduced as compared to a wildtype-non-recombinant organism and the organism produces ethanol at a rate of at least about 8-11 mM glycerol per gram dry cell weight (DCW) during anaerobic growth.

EXAMPLES

Example 1

Engineering S. cerevisiae to Express Integrated PKA and PTA

[0160] Phosphoketolase (PHK) enzymes from Bifidobacterium adolescentis, Lactobacillus plantarum, or Aspergillus niger were integrated at the S. cerevisiae FCY1 site in combination with the Bifidobacterium adolescentis phosphotransacetylase (PTA) gene. The genetic modification techniques utilized to develop Saccharomyces cerevisiae strain relied upon directed integration to insert genetic material at specific and known sites within the yeast chromosome. The directed integration approach creates transgenic strains with integration events that are stable and easy to characterize.

[0161] The MX cassettes are the most commonly used engineering tool when an insertion or deletion of a genetic element is desired at a given chromosomal loci (Wach A., et al.,. Yeast. 10(13):1793-1808 (1994)). A recyclable MX cassette contains one or more markers which enable both dominant and negative selection (Goldstein, A. L., et al., Yeast. 15: 507-511 (1999); Hartzog, P. E. et al., Yeast. 22: 789-798 (2005)). The dominant marker enables selection for the modification and the counter selectable marker enables subsequent removal of the marker system via Cre-Lox mediated recombination (Guldener, U., et al., Nucleic Acids Research. 24(13): 2519-2524 (1996)) or recombination between duplicated homologous regions flanking the cassette. Because the markers are removed, they can be reused during subsequent engineering steps and ensures no undesirable foreign genetic material remains in the strain.

[0162] To create stable homozygous integrations two new HSV-thymidine kinase (TDK) based MX cassettes were developed. Expression of thymidine kinase in S. cerevisiae results in sensitivity to the compound fluoro-deoxyuracil (FUDR). The cellular toxicity of FUDR is dependent on the presence of two enzymes involved in pyrimidine metabolism: thymidine kinase (Tdk) and thymidilate synthetase (ThyA). Tdk converts FUDR to fluoro-dUMP (F-dUMP) which is a covalent inhibitor of ThyA and the basis for counter selection in a variety of eukaryotic organisms (Czako, M., and Marton, L., Plant Physiol. 104:1067-1071 (1994); Gardiner, D. M., and Howlett, B. J., Curr Genet. 45:249-255 (2004); Khang, C. H., et al., Fungal Genet Biol. 42:483-492 (2005); Szybalski, W., Bioessays. 14:495-500 (1992)).

[0163] The HSV-TDK expression cassette was independently fused to two commonly used dominant selectable markers which confer resistance to the drugs G418 (Kan) or nourseothricin (Nat) (Goldstein, A. L., et al., Yeast. 15: 507-511 (1999)). Transformation of both double expression cassettes, referred to as KT-MX and NT-MX, enabled positive selection for integration into both chromosomes as illustrated in FIG. 8A). For each round of engineering, PCR amplicons of upstream and downstream regions flanking the target site were designed to contain homologous tails for both the KT-MX and NT-MX cassettes. Both the flanks and the markers were transformed followed by selection on YPD medium containing both G418 and Nat (FIG. 8B).

[0164] After each engineering step taken in the construction, all markers were subsequently deleted and/or replaced with a desired expression cassette (Mascoma Assembly) resulting in a strain free of antibiotic markers (FIG. 9).

[0165] The integration scheme for each strain construction is shown in FIG. 10. Genotypes of the strains can be seen in Table 2.

TABLE-US-00002 TABLE 2 S. cerevisiae strains. Strain Mascoma ID Assembly ID Source Strain description M2390 Wild type Isolated from Wild type control commercial source M3293 Glycerol Constructed .DELTA.gpd1.DELTA.gpd2.DELTA.fdh1.DELTA.fdh2::adhE reduction by Mascoma background corporation M4408 M3293 Constructed .DELTA.gpd1.DELTA.gpd2.DELTA.fdh1 .DELTA.fdh2::adhE engineered by Mascoma .DELTA.fcy::B. adolescentis PHK 2/ with MA415 corporation B. adolescentis PTA expressing B. adolescentis PHK and PTA M4409 M3293 Constructed .DELTA.gpd1.DELTA.gpd2.DELTA.fdh1 .DELTA.fdh2::adhE engineered with by Mascoma .DELTA.fcy::L. plantarum PHK2/ MA415 corporation B. adolescentis PTA expressing L. plantarum PHK2 and B. adolescentis PTA M4410 M3293 Constructed .DELTA.gpd1.DELTA.gpd2.DELTA.fdh1 .DELTA.fdh2::adhE engineered by Mascoma .DELTA.fcy::A. niger PHK2/ with MA415 corporation B. adolescentis PTA expressing A. Niger PHK and B. adolescentis PTA

Example 2

PKA and PTA Expression Rescues the Anaerobic Growth Defect in the M3293 Strain

[0166] The anaerobic growth of the strains expressing PHK and PTA from Example 1 was compared to the anaerobic growth of strains that did not express PHK and PTA. M2390,which has a complete anaerobic glycerol-production pathway, was used as a wildtype control strain. M3293, which contains deletions of both GPD1, GPD2, FDH1 and FDH2 and expression of the B. adolescentis adhE, did not have the ability to grow anaerobically unless acetate was supplied. A functional heterologous phosphoketolase/phosphotransacetylase pathway complemented the anaerobic growth defect of M3293.

[0167] Cells from all strains were grown overnight in YPD and inoculated to a final concentration of 0.1 g/l DCW in 96 well plates containing 100 .mu.l YPD. The inoculated plates were allowed to equilibrate in the anaerobic chamber for 20 minutes prior to sealing the plates. As seen in FIG. 11, M3293 was not able to grow anaerobically, however, M4408, M4409, M4410, which contain a heterologous phosphoketolase pathway were able to grow. These results indicated that phosphoketolase enzymes from both B. adolescentis, Lactobacillus plantarum, and Aspergillus niger enabled anaerobic growth when expressed in combination with the phosphotransacetylase gene from B. adolescentis. The results shown in FIG. 12 indicated there were only minor differences in aerobic growth rate between M3293 and the phosphoketolase containing strains.

Example 3

Fermentation Analysis Using 31% Solids Corn Mash

[0168] In another embodiment, the ability of strains containing a deletion of both GPD1 and GPD2 expressing PHK and PTA from Example 1 to ferment corn mash was compared to the ability of strains containing deletions of both GPD1 and GPD2 but that did not express PHK and PTA to ferment corn mash (M3293). The fermentation was started by inoculating yeast to an initial concentration of 0.1 g/l DCW in 4 mls of 30% solids corn mash. The corn mash fermentation began with oxygen in both the medium and the headspace. Limited growth of M3293 was expected because the fermentation does not start anaerobically and there was acetate present. (During the corn mash fermentation, there was 0.3-0.5 g/l acetate present. The presence of the phosphoketolase/phosphotransacetylase pathway enabled growth after the supply of oxygen and acetate was depleted. All three phosphoketolase-containing strains reached a significantly higher titer than the M3293 control strain (FIG. 13). Acetate accumulation in M4408, M4409 and M4410 (FIG. 14) may have resulted in a lower pH and lower ethanol productivity.

Example 4

Fermentation Analysis in Defined Medium

[0169] All strains described in Table 2 were inoculated into defined medium without amino acids. Purged anaerobic bottles were prepared with medium as described below. The fermentations were started by inoculating these bottles with yeast strains to an initial concentration of 0.1 g/l DCW cells. After inoculation, bottles were placed in a shaking incubator at held at 35 C. Samples were withdrawn and metabolite concentrations were measured by HPLC analysis.

Defined Medium

[0170] 1.7 g/l YNB-no AA without AS (Sigma) [0171] 40 g/l glucose [0172] 2.3 g/l Urea or 5.0 g/L ammonium sulfate [0173] 6.6 g/l K.sub.2S0.sub.4 [0174] 0.5 g/l MgSO.sub.4 [0175] 3 g/l KH.sub.2PO.sub.4 [0176] 1000.times.--Tween-80/Ergosterol (Final concentration: 20 mg/L [0177] ergosterol 420 mg/L Tween) [0178] Verduyn vitamins & traces; 1 ml/L each

[0179] The phosphoketolase containing strains M4408 and M4410 had improved growth relative to M3293 (FIG. 15); however, the strains had reduced biomass formation relative to the wild type control strain, M2390. The acetate formation observed in corn mash fermentation was not observed in defined medium. The ethanol titers of M4408 and M4410 were approximately 9% higher than M2390 (FIG. 16) while glycerol was reduced by approximately 2.4 g/l (FIG. 17).

Example 5

Engineering PHK and PTA Expression Vectors for S. cerevisiae

[0180] S. cerevisiae phosphoketolase expression vectors were created using the PMU1689 vector. L. plantarum PHK1, L. plantarum PHK2, A. niger PHK, L. casei PHK, B. adolescentis PHK or N. crassa PHK were cloned into PMU1689 To create, phosphoketolase expression vectors, PCR products were amplified from genomic DNA templates using primers and templates listed in Table 3 and recombined with linearized pMU1689 such that the ORF of each gene is immediately downstream of the ADH promoter and immediately upstream of the PDC1 terminator. Schematics of the PHK expression vectors can be seen in FIGS. 18-23.

[0181] S. cerevisiae phosphotransacetylase expression vectors were created using the pMU1771 expression vector (FIG. 24). The B. adolescentis phosphotransacetylase orf was amplified from B. adolescentis genomic DNA template using primers and templates listed in Table 3 and recombined with linearized pMU1771 such that the ORF of each gene is immediately downstream of the TEF2 promoter and immediately upstream of the ADH3 terminator.

TABLE-US-00003 TABLE 3 Phosphoketolase cloning templates and vectors Donor Forward Reverse SEQ Organism Template primer primer Vector ID NO B. adolescentis B. gctataccaa TTATAA pMU1689 49, 50 adolescentis gcatacaatc AACTTT Genomic aactatctcat AACTAA DNA atacaatgac TAATTA gagtcctgtta GAGATT ttggcac AAATCG CTCACT CGTTAT CGCCAG CGGTT L. plantarum(1) L. ctataccaag AAACTT pMU1689 51, 52 plantarum catacaatca TAACTA Genomic actatctcata ATAATT DNA tacaatgaca AGAGAT acagattact TAAATC catcacca GCTTAT TTTAAA CCCTTC CATTGC CAATC( L. plantarum(2) L. gctataccaa AAAACT pMU1689 53, 54 plantarum gcatacaatc TTAACT Genomic aactatctcat AATAAT DNA atacaatgag TAGAGA tgaagcaatt TTAAAT aaatccaa CGCCTA CTTCAA TGCAGT CCATTT CCAGT N. crassa N. crassa tataccaagc AAAACT pMU1689 55, 56 Genomic atacaatcaa TTAACT DNA ctatctcatat AATAAT acaatgggc TAGAGA ggaacacag TTAAAT atcacaattc CGCCTA CTCGAA CTTGGG CAGTTG GTAGA( A. niger A. niger ctataccaag AACTTT pMU1689 57, 58 Genomic catacaatca AACTAA DNA actatctcata TAATTA tacaatgcct GAGATT ggagaggtc AAATCG atcgacagg CTTATT CAAAGG AGGGCA TATCAT ACGTA L. casei L. casei tataccaagc AAACTT pMU1689 59, 60 Genomic atacaatcaa TAACTA DNA ctatctcatat ATAATT acaatggata AGAGAT caaaagtaaa TAAATC gactgttg GCTTAC TTGATT GGTTTC CAGGTC CATTC

Example 6

Analysis of Components of the Phosphoketolase Pathway

[0182] To evaluate components of the phosphoketolase pathway, the strains listed in Table 4 were inoculated into vials containing 4 ml of YP medium (10 g/l yeast extract, 20 g/l peptone) supplemented with 225 g/L maltodexdrin. Maltodextrin hydrolysis was initiated upon addition of 0.6 AGU/gTS glucoamylase at the start of the fermentation. Vials were incubated at 32.degree. C. for 68 hrs and sampled for HPLC analysis.

TABLE-US-00004 TABLE 4 Strains used in the fermentation study. Strain Genotype M2390 WT M3293 .DELTA.gpd1::MA0606.DELTA.gpd2::MA0607.DELTA.fdh1::MA0608.DELTA.fdh1- ::MA0281 M4408 .DELTA.gpd1::MA0606.DELTA.gpd2::MA0607.DELTA.fdh1::MA0608.DELTA.fdh1- ::MA0281.DELTA.fcy::MA0449.2 M4579 .DELTA.gpd1::MA0606.DELTA.gpd2::MA0607.DELTA.fdh1::MA0608.DELTA.fdh1- ::MA0281.DELTA.fcy::MA0415.3 M4581 .DELTA.gpd1::MA0606.DELTA.gpd2::MA0607.DELTA.fdh1::MA0608.DELTA.fdh1- ::MA0281.DELTA.fcy::MA0415.4 M4582 .DELTA.gpd1::MA0606.DELTA.gpd2::MA0607.DELTA.fdh1::MA0608.DELTA.fdh1- ::MA0281.DELTA.fcy::MA0449.2 M4584 .DELTA.gpd1::MA0606.DELTA.gpd2::MA0607.DELTA.fdh1::MA0608.DELTA.fdh1- ::MA0281.DELTA.fcy::MA0449.3 M4788 .DELTA.gpd1::MA0606.DELTA.gpd2::MA0607.DELTA.fdh1::MA0608.DELTA.fdh1- ::MA0281.DELTA.fcy::MA0449.1 M4789 .DELTA.gpd1::MA0606.DELTA.gpd2::MA0607.DELTA.fdh1::MA0608.DELTA.fdh1- ::MA0281.DELTA.fcy::MA0449.1 M4790 .DELTA.gpd1::MA0606.DELTA.gpd2::MA0607.DELTA.fdh1::MA0608.DELTA.fdh1- ::MA0281.DELTA.fcy::MA0467.6 M4791 .DELTA.gpd1::MA0606.DELTA.gpd2::MA0607.DELTA.fdh1::MA0608.DELTA.fdh1- ::MA0281.DELTA.fcy::MA0467.6 M4792 .DELTA.gpd1::MA0606.DELTA.gpd2::MA0607.DELTA.fdh1::MA0608.DELTA.fdh1- ::MA0281.DELTA.fcy::MA0434.6 M4793 .DELTA.gpd1::MA0606.DELTA.gpd2::MA0607.DELTA.fdh1::MA0608.DELTA.fdh1- ::MA0281.DELTA.fcy::MA0434.6 M4794 .DELTA.gpd1::MA0606.DELTA.gpd2::MA0607.DELTA.fdh1::MA0608.DELTA.fdh1- ::MA0281.DELTA.fcy::MA0435.1

[0183] M2390 is a conventional wild type yeast. M3293 contains deletion of both glycerol-3-phosphate dehydrogenase genes (GPD1 and GPD2), both formate dehydrogenase genes (FDH1 and FDH2), and expression of the B. adolescentis bifunctional alcohol/aldehyde dehydrogenase. This strain grows poorly under anaerobic conditions and is only able to produce a titer of 42 g/l ethanol whereas M2390 was able to reach 113 g/l ethanol (FIG. 25).

[0184] The addition of components of the phosphoketolase pathway to M3293 allowed for improved ethanol production. Expression of both B. adolescentis and L. casei PHK in combination with the B. adolescentis PTA gene resulted in production of 60.7 and 72.3 g/l respectively (FIG. 25) indicating the pathway partially complements the defect observed in M3293. Expression of B. adolescentis PHK, PTA and ACK genes resulted in production of 92 g/l ethanol. This combination produced titers that were significantly higher than when the B. adolescentis PHK and PTA were the only additions.

[0185] A description of the cassettes used to create the strains is seen below and in FIGS. 26-34. Strains were created as described, for example, in Example 1.

TABLE-US-00005 MA0281 Expected Fragment Primers Template Size FDH2 5' Flank X16096/X17243 M2390 gDNA 503 bp PFK pro/HXT2 ter X16738/X14897 pMU2745 4233 bp TPI pro/FBA1 ter X14896/X16744 pMU2746 3683 bp FDH2 3' Flank X17244/X11845 M2390 gDNA 1939 bp

TABLE-US-00006 MA467.6 PHK Fragment Primers Template Expected Size FCY 5' Flank X21754/X21736 M2390 gDNA 2049 bp ADH1pro/PDC1ter X21735/X18847 pMU3397(PHK) 3728 bp FCY 3' Flank X18846/X18869 M2390 gDNA 2166 bp

TABLE-US-00007 MA449.2 4PTA/2PHK Fragment Primers Template Expected Size FCY 5' Flank X21754/X19552 M2390 gDNA 2049 bp TEF2pro/ADH3ter X19551/X19513 pMU3399(PTA) 2671 bp ADH1pro/PDC1ter X19514/X19360 pMU3397(PHK) 3728 bp TPIpro/FBA1ter X19361/X19903 pMU3402(PTA) 2621 bp FCY 3' Flank X19902/X18869 M2390 gDNA 2166 bp

TABLE-US-00008 MA449.3 2PTA/2PHK/2ACK Fragment Primers Template Expected Size FCY 5' Flank X21754/X19552 M2390 gDNA 2049 bp TEF2pro/ADH3ter X19551/X19513 pMU3399(PTA) 2671 bp ADH1pro/PDC1ter X19514/X19360 pMU3397(PHK) 3728 bp TPIpro/FBA1ter X19361/X19903 pMU3407(ACK) 2180 bp FCY 3' Flank X19902/X18869 M2390 gDNA 2166 bp

TABLE-US-00009 MA435.1 PHK/ACK Fragment Primers Template Expected Size FCY 5' Flank X21754/X21736 M2390 gDNA 2049 bp ADH1pro/PDC1ter X21735/X19855 pMU3397(PHK) 3728 bp TPIpro/FBA1ter X19854/X18861 pMU3407(ACK) 2180 bp FCY 3' Flank X18860/X18869 M2390 gDNA 2166 bp

TABLE-US-00010 MA415.4 4PTA Fragment Primers Template Expected Size FCY 5' Flank X21754/X19552 M2390 gDNA 2049 bp TEF2pro/ADH3ter X19551/X19513 pMU3399(PTA) 2671 bp ADH1pro/PDC1ter X19514/X18955 pMU3401(PTA) 2921 bp FCY 3' Flank X19950/X18869 M2390 gDNA 2166 bp

TABLE-US-00011 MA449.1 4PTA/2ACK Fragment Primers Template Expected Size FCY 5' Flank X21754/X19552 M2390 gDNA 2049 bp TEF2pro/ADH3ter X19551/X19513 pMU3399(PTA) 2671 bp ADH1pro/PDC1ter X19514/X19360 pMU3401(PTA) 2921 bp TPIpro/FBA1ter X19361/X19903 pMU3407(ACK) 2180 bp FCY 3' Flank X19902/X18869 gDNA 2166 bp

TABLE-US-00012 MA434.6 ACK Fragment Primers Template Expected Size FCY 5' Flank X21754/X18859 M2390 gDNA 2049 bp TP1pro/FBA1ter X18858/X18861 pMU3407(ACK) 2180 bp FCY 3' Flank X18860/X18869 M2390 gDNA 2166 bp

TABLE-US-00013 MA415.3 L. casei PHK Fragment Primers Template Exp Size FCY 5' Flank X21754/X19552 M2390 gDNA 2049 bp TEF2pro/ADH3ter X19551/X19513 pMU3399 (PTA) 2671 bp ADH1pro/PDC1ter X19514/X18955 pMU3416 (PHK) 3632 bp FCY 3' Flank X19950/X18869 M2390 gDNA 2166 bp

[0186] All documents cited herein, including journal articles or abstracts, published or corresponding U.S. or foreign patent applications, issued or foreign patents, or any other documents, are each entirely incorporated by reference herein, including all data, tables, figures, and text presented in the cited documents.

[0187] 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

10311671DNABifidobacterium adolescentisPTA 1gtgagtttta ccagcgtcac catcatcagt cccgaagcgg ctaacggccg taatgttgtc 60gcccttggag tcaccaagac acttgccgca gccggcaaga ccggagtgtt ccgtccggcg 120gtctgcagaa aagacacctt cacggacgtg ctgatcgaag cttccaatgc cggtttgagc 180cgggaacagt ccgtcggcgt gtgccccaag cgcgctcgta acgataagga aggttcgcgg 240gcggatatcg ttgcggccta cacccaagcc gtagaaacag cccgaccgga tgccatggtg 300attgtcggca ccgacaggtc agcagtcaat gatcccgcga tgttctcttt caacgctgac 360gtcgcggcag acctccaatc gccggtcctg ctggccgtat gcactatcga acgcacaccg 420gaacaagtca aaagcacggt tgaggcatcc accaaggtca tcgaagacgc cgggagcaaa 480gtcgtcggcg tgttcattac tgggtgtgac gacacgcagc cgaatccact caaagcatgc 540ttcgtcgact acccggtgcc ggtgtggacg ctacctgccg tcgatttcaa cgacgatgac 600gccatcagca aggcagacga agcgttcgcg acgaacgtcg acgccgtgga actgaccgtg 660gcactcgaat cgccattcga cgccccgacc acaccctatg ctttccaata cggtctgctg 720ggcaaggcga aggcggacaa gaagaccatc gtgctgccgg agggcaacga ggatcgcatc 780atcaaagccg ccgattatct gctggagcgc gacatcgtcg atctgatcat cgtcggagac 840gaaaacgcca ttctcgcccg cggccaggaa ttgggattga aatcgcttgg caaggcgaaa 900ttccaagcca aggatgatga aacggtgctg gaacccatgg tggcgaagct gtgcgagctg 960cgcgccaaga aaggcatgac cgaagagcag gcgcgcaaac agctggccga tgacagctat 1020ttcggcacca tgcttgtggt gatgggcatg gcggacggtc tcgtgtccgg ttccgtcaac 1080tccacggcca ataccgtgcg tccggccctg caggtcatca agaccaagcc aggaacgtcg 1140ctggtctccg gagcgttcct gatgtgtttc aaggaccatg ccgccgtgtt cgccgactgt 1200gccatcaacc ttaaccccaa tgccgagcag ctcgccgaaa tagcaatcca atccgccgaa 1260acggccaagg cgttcggcct tgaaccgaag gtgggcatgc tgtcctattc cacgctgggc 1320tccggaaaag gacccgacgt cgacctcgtc gaggaagcca ccaccatcgt caaagacaag 1380gctcccgacc ttgcggtggt cggatcgatc cagttcgacg cggcatggtc tccgaccgtg 1440gccgcgacca aagccaaggg cgatccggtg gcgggccatg tcaatgtgtt cgtattcccg 1500gatctgtgcg cgggcaacat cgcatacaag gcggtacagc gttcctcggg agcggccgcc 1560gtcgggccgg tgctgcaggg tctcaaccgc ccggtgaacg atctgagccg tggcgccacg 1620gtccaggaca tcatcaacac catcgccctg accgccatcg aggcccagta a 167121230DNABifidobacterium adolescentisACK 2atggcgaaaa ccgtccttgt catcaattcc ggttccagct ctatcaagta tcagctggtt 60gatcttgaaa ccggcgaagg tatcgcttcc ggtctggtcg agaagatcgg cgagccggtt 120gacggtcatt acaagcacga gtacaacggc gagaagcatg agctcgaaga gccgattcac 180gaccatgagc agggtctgaa gcgcgtgctc ggcttcttcg acgagttcgg cccgaagctc 240gccgacgcag gcatcgtggc cgtcggccat cgcgtggtgc agggtggttc catcttcccg 300aagccggctc tggtcaacga caagaccatc ggccaggtca aggacctcgc cgtgctcgcc 360ccgctgcaca acggtccgga ggccaagggc gctgaggtca tgcgctccct gctgccggat 420gttccgcaga tcttcgtgtt cgattcctcc ttcttcttcc agctgccgaa ggcttccagc 480acctacgcgc tgaacaagga agtcgcccag cagtaccata tccgccgtta cggcgcccat 540ggcacctccc acgagttcat ctcctccgtc gtgccgtccg tcatcggcaa gccggccgaa 600ggcctcaagc agatcgtgct gcacatcggc aacggtgctt ccgcctccgc cgaaatctcc 660ggcaagccgg ttgagacctc catgggtctg accccgctgg aaggcctcgt gatgggtggc 720cgtaccggcg acatcgatcc ggccgtcgtc ttccacctga ttcgcaacgc ccacatgagc 780gtggacgaac tggacaccct gttcaacaag cgttccggca tgatgggtct gaccggcttc 840ggcgacctgc gtgaagtgca tcgtctggtc gaagagggca acgaggacgc caagctggct 900ctcgacatct acgtgcaccg catcgtcggc tacatcggca actacaccgc tcagatgggt 960ggcgtcgacg tgatcacctt caccgcaggc gttggcgaga acgatgacgt cgtgcgcaag 1020atggtctgcg acaagctcgc tccgttcggc gtcaagttgg atgaggagaa gaacgcgacc 1080cgttccaagg aaccgcgcat catctccacc ccggattccg cagtcaccat ctgcgtcatc 1140ccgaccaacg aggagctggc catcgcccgc aagtccgccg ccatcgcaga agaaggcaag 1200gactcctacg gcaacgtctt ctccaagtga 123032421DNAAspergillus nigerPHK 3atgcctggag aggtcatcga caggccaaat cccaaggctg agccttcaca catccccgat 60cttgtcaatc aattgcaggt caaacttcaa gagacgcgtt tggaggaaac tgattacaat 120gcccttctga aattccgccg tgcagcggcc tacattgctg ctgcaatgat ctttctccaa 180gacaatgtgc tgctgaagca gaatctaagg cacgaggaca tcaagcccag gcttcttggc 240cactggggaa catgtcccgg gttgattctt gtatactctc acttgaacta catcatcaga 300aagcagaacc tggatatgtt gtatgtcgtc gggcctggcc acggcgcgcc agctattttg 360gcctcactgt ggcttgaggg ctctttagag aaattctacc cccactactc acgagacatg 420gatggtctcc atgagctcat ctcgaccttc agcacaagtg ctggattacc aagccatatc 480aatgcggaaa ctcccggtgc aatccatgaa ggtggtgaat tgggttatgc gttagctgtc 540tcttttggtg ctgttatgga caatcccgac atgatcgtca cctgcgtggt tggtgacggg 600gaagcagaaa ctggtcctac cgcgacgtcc tggcatgcaa tcaagtacat tgaccccgca 660gaatcaggtg ccgtcctgcc gattctccac gttaatggct ttaagatcag cgagcgcacc 720atttatggct gcatggacaa caaagagctg gtctccctct tcacgggtta tggataccag 780gtgcgcattg ttgagaacct ggatgacatc gacgcagatc tccatagctc tatgatgtgg 840gcagttgagg agatccacaa gatccaaaaa gcggcgcgtt ccggcaagcc aattatgaag 900cctagatggc caatgattgt tttgcgcaca ccgaagggtt ggtcaggacc taaagagctc 960cacgggtcat tcatagaggg atctttccac tcacatcagg ttcctctacc taatgcaaag 1020aaggataaag aggagcttca ggctctgcag aaatggctgt cctcgtataa tccgcacgaa 1080cttttcactg agacgggaga catcattgac gacatcaagt cagtgatccc tctggaagac 1140accaagaagc ttgggcagcg agcagaagcc tacaagggct atagggcacc cgatctccca 1200gactggcgca agtttggcgt agaaaagggc tcccagcaga gcgctatgaa aacaattgga 1260aagttcattg accaagtgtt tacccaaaat cctcatggcg tccgtgtatt ttcgccagac 1320gagctagaga gcaacaagct ggatgcagca ctggcgcaca cgggaaggaa ctttcagtgg 1380gatcaattct cgaatgccaa aggcggccgc gtcatcgagg tgctcagtga gcacctgtgc 1440cagggcttta tgcagggata cacgttgacg ggccgggtgg gcattttccc atcgtacgaa 1500agcttcttgg gaatcatcca taccatgatg gtgcaatatg ccaaatttaa caaaatggct 1560caagagacga cctggcataa gccggttagt agcatcaact atatcgaaac gagtacgtgg 1620gctcgtcagg agcacaatgg attctctcac cagaacccct cctttatcgg agctgtgctc 1680aggctgaagc ccaccgccgc gcgagtttat ctgccacctg atgctaacac atttttgacc 1740acccttcacc actgtctcaa gtccaagaat tatgtcaacc tcatggtagg ttcaaaacag 1800ccaactcccg tgtacttgag ccccgaggaa gcagagagcc actgccgagc cggcgcatcg 1860atctggagat tctgtagtac cgacaatggg ctgaacccgg atgtcgtgct ggttggcatt 1920ggagtagagg tgatgttcga ggtcatctac gcggcggcca tcctccgcaa gcgttgtcca 1980gaactccggg tgcgtgtggt caatgtgacc gacttgatga ttctggagaa ggaaggtcta 2040catccacatg cattgacgac cgaagctttc gacagtctgt ttggctcgga ccggccgata 2100cacttcaact accacggata cccgggcgag ctcaaaggtc tgctctttgg gcggccccgc 2160ctggaccgag tttcagtaga aggatacatg gaggaaggaa gcacgacgac gccgttcgat 2220atgatgttgc tgaaccgcgt ctcacgatac cacgtggcgc aggcagccgt gatcggggcg 2280tccagacgga atgagaaggt tcaagttcgg cagcacgaac tggtcagcga attcggccac 2340aacatcgtgg agacacgcaa atacattctg gccaaccgca aagacccgga tgatacgtat 2400gatatgccct cctttgaata a 242142448DNANeurospora crassaPHK 4atgggcggaa cacagatcac aattcccaac ccccctcctc taccatccca cctccccgac 60aatgtcctcg acctggcagt ccaaattgag cgcaagaacc tcccagacaa tgtccgcgag 120tcgctgcgcg cattccagcg tgcaggatgc tacattgccg cctccatgat cttcttgcgc 180gacaatgtcc tcctccaaga caaactccaa gtttcgcaca tcaagccccg actcctcggt 240cattggggaa catgccccgg cctaatcctg gtctggtcgc atctcaacgt cttgatccgc 300aaccatgacc tcaacatgat ttacgttatc ggccccggcc acggcgcccc tgccgccctc 360gcctctctgt ggctcgaagg ctcccttgag cgcttctacc ctggcgaata cgaccgcaat 420gctactggtt tgcgcaacct catcacccgt ttctctgtcc ccggcggttt tcctagccac 480atcaacgctg agactccagg tgcgatccat gagggtggtg agctcggata tgccctttct 540gtctcgttcg gcgccgtcat ggacaacccc gatttgatcg tcacttgtgt ggtaggcgat 600ggtgaggcgg agactggccc gacagcaacc gcctggcatg ctatcaagta catcgacccc 660aaggagtcag gtgccgtcat ccccattctg catgtcaacg gcttcaagat cagcgagcgc 720accatttttg ggtgcatgga cgacaaggag atcgcctgct tgtttagtgg ctacggatac 780caggtgcgca ttgtggaaga cctggagaac attgatgatg acctgcaaag ttcgctggaa 840tgggctctag tcgagatcaa gaagatccag aaggctgcga gggaggacaa caagccgatt 900gtgaagccga gatggccgat gattgtcctc agaacgccca agggatggac tggtcccaag 960gaggtggagg gccaaatcat cgaaggatcc ttccactccc atcaagtccc ccttcctaag 1020gctggtagtg atgagaagca actccaggct ctagacgagt ggctttccag ctacaggatc 1080agcgagctcc ttgatgatga gggcaagcct actgaggcca tcactcgtgt cttgcccaag 1140gcagaaaaga ggctcggtca actcaagatc acatatgacc catacattcc tctgaaactg 1200attgactgga agcagtttgg cgttcaaaag ggcgtcgacg aaagctgcat gaagagtgcc 1260ggcaagttcc tcgacaagtt gttccaagag aaccccaaaa ccctccggct attctcccct 1320gacgagctcg agtcgaacaa gctcaacgcc atcctggatc acacacagcg cgactttcag 1380tgggacgagt tctcgagggc acatggcggt cgcgtcatcg aggttctctc cgagcacaat 1440tgccagggct tcatgcaagg atacactctc acgggccgca ccgccctctt cccctcctat 1500gagtccttcc tgggtatcgt tcacaccatg atggtgcaat actccaagtt cgtcaagatc 1560gcgcgtgaag taccctggcg tggcgacctg gcctccatca actacatcga gacttcgacc 1620tgggcgcgtc aagaacacaa tggcttttcc catcagaacc cctccttcat tggtgctgta 1680cttaacctga aggccgaggc cgctcgcgtt tacctcccgc ccgacgccaa ctgcttcctc 1740agcaccgtgc atcatgtcat ggacagcaag aacaagacca acctcatcat cggctccaag 1800cagcccacgg ccgtctacct ctctcccgcc gaagcagccg accactgtcg ccagggtgcc 1860tcgatctggc atttcgcctc ttcctccgac cccaactcat ccgcaggaga gtcggtacaa 1920gacccagatg tagtcctcgt cggcatcggc gtcgaagtta cttttgaggt gattaaagct 1980gctgaactct tgcgctccat tgcgcccgcc ctcaagatcc gcgtcgtcaa tgtgacggat 2040ctgatgattc tcatcgaaga gtccaagcac ccgcatgccc tggccaaggc caagttcatc 2100gaaatgttca ccgccgacag accggtgttg ttcaactacc atggctaccc gacggaactg 2160cagggtctct tgtttgggag ggaaaaggcg gagcgcatgg atgtcgaggg ttaccaagaa 2220gagggaagca cgaccacgcc gtttgatatg atgttgagga accgggtcag tcggttcgat 2280gtggcgagct gggcgctgaa gaggggtgcg gagaggaatg cggaggtgaa gaatgatttg 2340gaaggtttgt tgggtgacgt ggataggaga gtgaaggagg ttcgtgagtt tattgggaat 2400gaaggaaagg atcccgatga tatctaccaa ctgcccaagt tcgagtag 244852382DNALactobacillus caseiPHK 5atggatacaa aagtaaagac tgttgattac tcatccaagg aatactttga caagatgacc 60gcttattggc gggcagccaa ctatatttca gttggtcaac tttatttgaa ggacaacccg 120ctgctggaac ggccactgaa gagcgaagac gtcaagccgc atccgatcgg ccattggggc 180acgattgctg ggcagaactt catttatacc cacttgaacc gggtgatcaa taagtatgac 240ttgaacatgt tttatattga aggcccgggt catggcggcc aagtgatggt ttccaattcc 300tatctagacg gttcgtattc ggagatttat ccgcgtgtgt cgcaggataa agaaggcatg 360aagaacctat ttacgcagtt ctcatggcca ggtggcgttg cttcacatgc gagtgcacaa 420acgccaggtt ccatccatga aggtggcgaa ttaggttatg ctttgagtca cgcaaccggc 480gcgattcttg ataatcctga ggtcatcgcg gcggttgtca cgggtgatgg tgaaactgaa 540accggcccat tggctgcatc atggttcagc aatacgttca ttaacccaat cacggatggc 600gcgattttac cgattgtgca tatgaacggg tttaagattt ccaacccaac tatcttgtca 660cgtaagtcag atgaagatct agacaaatac ttcagtggta tgggttggaa gccattcttc 720gttgaaggcg atgatccaga gaagctgaat ccggaaatgg ctaaagtttt ggatgaagca 780attgaagaca ttaaggccat tcagaaacat gcacgtgaga ctggcgacac cacgatgcca 840cactggccag tgattatttt ccggtcacca aagggctgga caggcccgaa gagttggaat 900ggcgaaccaa tcgaaggttc cttccgtgcg caccagattc cgattcctgt cgacgctgaa 960gacatggaac acgctgattc cttggcaggc tggctgaagt cctaccatcc agaggaactg 1020ttcgatgaaa acggcaagtt gatccctgaa ttggctgcat tgccgccaaa gggtgagcag 1080cggatggctg ctaacccaat tacgaacggt gggattgatc cgaagccgtt ggtattgcca 1140gattatcgca agtatgcgct ggacaacaag gaacacggca agcagattaa gcaagacatg 1200atcgtgtggt ctgattatct gcgtgatttg atcaagctga atccgcataa tttccggatc 1260tttggccctg acgaaaccat gagtaaccgg ttgtatagct tgtttgaagt gaccaaccgt 1320caatggctgg aaccaatcaa ggaaccggct gaccaatatt tggcaccggc tggccggatt 1380attgactcgc agttgtccga acatcaggcg gaaggcttta acgaaggcta cacgttgacc 1440ggccgtcatg gcttattcac cagttacgaa gctttcctcc gggtcgtcga ttccatgctc 1500actcagcact tcaagtggat tcggaaggca catgaagaac catggcataa ggcttatcca 1560agtctgaacg tggtctcaac atcgacttcc ttccagcagg atcacaatgg gtacacccat 1620caggacccag gtattttgac acacatggct gaaaagaagg ctgaatacat tcgcgagtat 1680ctaccagctg atgccaacag cttgctggcc atttctccga agttgttcag ctcacaaaat 1740accgtgaacg tgctcatcac atccaagcag ccgcggccac agttctatag cattgatgaa 1800gccactgttt tggctaattc cggcttgaag cggattgatt gggcttccaa tgatgatggc 1860gtcgaaccag atgttgtcat tgccgctgcc ggaaccgaac caaacatgga aagtcttgct 1920gccattaact tgctgcacga cgccttccca gatttgaaga ttcgcttcat taacgtggtt 1980gacctgctga agctgcgcag tccggaaatc gatccgcgcg gtttgagtga tgctgaattc 2040aacagctact tcaccaccga caaaccgatc ctcttcgctt atcatggctt tgaagggttg 2100attcgcgaca tcttcttcac tcgcccgaat cgcaatgtgc tgattcacgg ctatcgtgaa 2160gaaggtgaca ttaccacacc atttgacatg cgggtactca acgaacttga ccgttaccac 2220ttggccaagg atgttattca gcatgtgcca gcctacgctg aaaaggctgc cgcgtttgtt 2280cagaagatgg atgatacctt gcaatatcat catgacttca tccgcgcaaa cggcgatgat 2340attccagaag ttcaggaatg gacctggaaa ccaatcaagt aa 238262391DNALactobacillus plantarumPHK2 6atgagtgaag caattaaatc caaaacagtt gattactctt ctgatgaata tctaaaacgc 60gttgatgaat attggcgtgc tgctaactac atctcagttg gtcaactcta tctactaaat 120aacccgttac ttcgggaacc actaaaggcg accgacgtga aagttcatcc aatcggccat 180tggggcacga ttgctggtca aaactttatt tatgcccatt taaaccgggc aatcaataag 240tatggcttga acatgttcta cattgaaggc cctggtcatg gtggtcaagt aatggtttct 300aactcctact tagatggcac ctatacggaa acgtatccta aaatcaccca agacaaagct 360gggatgaaac gcttattcaa gcaattctca ttcccaggcg gggttgcttc ccatgccgat 420cctaagacgc ctggttcgat ccatgaaggt ggcgaacttg gctactcaat cctgcatggt 480gctggtgcag tattagataa tccaggttta attgccgcta ccgttgttgg tgatggtgaa 540tctgaaactg ggccattggc aacttcttgg caagttaaca agttccttaa cccaattaca 600gacgggacag tcttgccaat cttgaactta aacggcttca agatttctaa tccaacagtt 660ctttcacgtg aatcacatga agaacttgaa gactacttta aaggtctagg ctgggatcca 720cactttgttg aaggtacaga ccctgccaag atgcacaaaa ttatggctga agaattggat 780aaagtcattg aagaaatcca cgcaattcgt aagaacgcca aggataacaa tgatgaatct 840cgtcctaagt ggccaatgat tgttttccgg gcacctaagg gctggaccgg tcctaagagt 900tgggacggcg aaccaattga aggttcattc cgggctcacc aaattccaat tcctgtcgat 960cgcaatcaca tggaacacgc cgacaaatta gttgactggc tcaaatcata caaaccagaa 1020gaattatttg atgaaaatgg tactttaaaa ccagaaattg ccgcaatcat ccctgaaggc 1080caagctcgta tggctgctaa ccccgttact aacggcggta agttaactaa agacttaatt 1140acaccaaata tcgatgatta tgctttggac aacaagagtc acggtaagga agacggttca 1200gacatgactg aacttggtaa gtatatccgt gatttaattg agttgaacaa agacaacaag 1260aacttccgtg gctggggtcc tgacgaaacc ttatctaaca aactaggcgc tgcttttgaa 1320gataccaaac gtcagtggat ggaaccaatc cacgaaccta atgatgcttt gttagcacct 1380caaggccgga ttattgactc catgttgtca gaacacatgg atgaagggat gttggaagct 1440tacaatttaa ccggacgtta cggtttcttc gcaagttatg aatcattcct gcgcgttgtg 1500gattcaatgt taacccaaca cttcaagtgg ttacggaatt ctcacgaaga aaccccttgg 1560cgggctgatg taccttcact gaatgtgatt gcatcatcaa cagccttcca acaagatcac 1620aatggttact ctcaccaaga tccaggtatc atttcacact tggctgaaaa gaagaccgaa 1680tacgttcgtg cctatcttcc aggtgatgcc aatactttga ttgcaacctt tgataaggct 1740atccaaagca aacaattgat taatttaatc attgccagca agcaccctcg tccacaatgg 1800ttcacaatgg acgaagctaa gcgcttagtt cgtgatggcc ttggtgttgt tgattgggca 1860agcactgatc atggtgaaga acccgacgtt gtcttcgcaa ctgccggctc tgaaccaacg 1920actgaaagct tagctgccgt atcaatcttg catgcacgct tccctgaaat gaagattcgc 1980ttcattaacg ttgttgatct tctgaagctg aagaaagacg accctcgtgg tttatcagat 2040gctgaatttg atgctttctt cactaaggac aaaccagtta tctttgctta tcatgcatac 2100gacgacttag taaagaccat cttcttcgat cgccataacc ataacttaca cgttcatggt 2160taccgcgaag aaggcgacat tacaacgcca ttcgacatgc gtgttcgcaa cgaactcgat 2220cgtttccact tagtcaaagc tgccttatta gcaacgccag cttatgccga aaaaggtgcc 2280catgtcattc aagagatgaa cagcatttta gacaagcatc atgactatat ccgtgctgaa 2340ggtaccgata ttccagaagt tgaaaactgg aaatggactg cattgaagta g 239172367DNALactobacillus plantarumPHK1 7atgacaacag attactcatc accagcatat ttgcaaaaag ttgataagta ctggcgtgct 60gccaactact tatcagttgg tcaactttat ttaaaagata atccactatt acaacggcca 120ttgaaggcca gtgacgttaa ggttcatcca attggtcact gggggacgat tgccggtcaa 180aactttatct atgctcatct taaccgggtc atcaacaagt acggtttgaa gatgttctac 240gttgaaggtc caggtcatgg tggtcaagtg atggtttcaa actcttacct tgacggtact 300tacaccgata tttatccaga aattacgcag gatgttgaag ggatgcaaaa gctcttcaag 360caattctcat tcccaggtgg ggttgcttcc catgcggcac ctgaaacacc cggttcaatc 420cacgaaggtg gcgaacttgg ttactcaatt tcacacgggg ttggggcaat tcttgacaat 480cctgacgaaa tcgccgcggt tgttgttggt gatggggaat ccgaaacggg tccattagca 540acttcatggc aatcaacgaa gttcattaac ccaatcaacg acggggctgt tttaccaatc 600ttgaacttaa atggttttaa gatttctaat ccaacgattt ttggtcggac ttctgatgct 660aagattaagg aatacttcga aagcatgaat tgggaaccaa tcttcgttga aggtgacgat 720cctgaaaagg ttcacccagc cttagctaag gccatggatg aagccgttga aaagatcaag 780gcaatccaga agcatgctcg cgaaaataac gatgcaacat tgccagtatg gccaatgatc 840gtcttccgcg cacctaaggg ctggactggt ccgaagtcat gggacggtga taagatcgaa 900ggttcattcc gtgctcatca aattccgatt cctgttgatc aaaatgacat ggaacatgcg 960gatgctttag ttgattggct cgaatcatat caaccaaaag aactcttcaa tgaagatggc 1020tctttgaagg atgatattaa agaaattatt cctactgggg acagtcggat ggctgctaac 1080ccaatcacca atggtggggt cgatccgaaa gccttgaact taccaaactt ccgtgattat 1140gcggtcgata cgtccaaaga aggcgcgaat gttaagcaag atatgatcgt ttggtcagac 1200tatttgcggg atgtcatcaa gaaaaatcct gataacttcc ggttgttcgg acctgatgaa 1260accatgtcta accgtttata tggtgtcttc gaaaccacta atcgtcaatg gatggaagac 1320attcatccag atagtgacca atatgaagca ccagctggcc gggtcttaga tgctcagtta 1380tctgaacacc aagctgaagg ttggttagaa ggttacgtct taactggacg tcatgggtta 1440tttgccagtt atgaagcctt cctacgcgtt gtggactcaa tgttgacgca acacttcaag 1500tggttacgta aagccaatga acttgattgg cgtaaaaagt acccatcact taacattatc 1560gcggcttcaa ctgtattcca acaagaccat aatggttata cccaccaaga tccaggtgca 1620ttaactcatt tggccgaaaa gaaaccagaa tacattcgtg aatatttacc agccgatgcc 1680aacacgttat tagctgtcgg tgacgtcatt ttccggagcc aagaaaagat caactacgtg 1740gttacgtcaa aacacccacg tcaacaatgg ttcagcattg aagaagctaa gcaattagtt 1800gacaatggtc ttggtatcat tgattgggca agtacggacc aaggtagcga accagacatt 1860gtctttgcag ctgctgggac ggaaccaacg cttgaaacgt tggctgccat ccaattacta 1920cacgacagtt tcccagagat gaagattcgt ttcgtgaacg tggtcgacat cttgaagtta 1980cgtagtcctg aaaaggatcc gcggggcttg tcagatgctg agtttgacca ttactttact 2040aaggacaaac cagtggtctt tgctttccac ggttacgaag acttagttcg tgacatcttc 2100tttgatcgtc acaaccataa cttatacgtc cacggttacc gtgaaaatgg tgatattacc 2160acaccattcg acgtacgggt

catgaaccag atggaccgct tcgacttagc taagtcggca 2220attgcggcgc aaccagcaat ggaaaacact ggtgcggcct tcgttcaatc catggataat 2280atgcttgcta aacacaatgc ctatatccgg gatgccggaa ctgacttgcc agaagttaat 2340gattggcaat ggaagggttt aaaataa 236782478DNABifidobacterium adolescentisPHK 8atgacgagtc ctgttattgg caccccttgg aagaagctga acgctccggt ttccgaggaa 60gctatcgaag gcgtggataa gtactggcgc gcagccaact acctctccat cggccagatc 120tatctgcgta gcaacccgct gatgaaggag cctttcaccc gcgaagacgt caagcaccgt 180ctggtcggtc actggggcac caccccgggc ctgaacttcc tcatcggcca catcaaccgt 240ctcattgctg atcaccagca gaacactgtg atcatcatgg gcccgggcca cggcggcccg 300gctggtaccg ctcagtccta cctggacggc acctacaccg agtacttccc gaacatcacc 360aaggatgagg ctggcctgca gaagttcttc cgccagttct cctacccggg tggcatcccg 420tcccactacg ctccggagac cccgggctcc atccacgaag gcggcgagct gggttacgcc 480ctgtcccacg cctacggcgc tgtgatgaac aacccgagcc tgttcgtccc ggccatcgtc 540ggcgacggtg aagctgagac cggcccgctg gccaccggct ggcagtccaa caagctcatc 600aacccgcgca ccgacggtat cgtgctgccg atcctgcacc tcaacggcta caagatcgcc 660aacccgacca tcctgtcccg catctccgac gaagagctcc acgagttctt ccacggcatg 720ggctatgagc cgtacgagtt cgtcgctggc ttcgacaacg aggatcacct gtcgatccac 780cgtcgtttcg ccgagctgtt cgagaccgtc ttcgacgaga tctgcgacat caaggccgcc 840gctcagaccg acgacatgac tcgtccgttc tacccgatga tcatcttccg taccccgaag 900ggctggacct gcccgaagtt catcgacggc aagaagaccg agggctcctg gcgttcccac 960caggtgccgc tggcttccgc ccgcgatacc gaggcccact tcgaggtcct caagaactgg 1020ctcgagtcct acaagccgga agagctgttc gacgagaacg gcgccgtgaa gccggaagtc 1080accgccttca tgccgaccgg cgaactgcgc atcggtgaga acccgaacgc caacggtggc 1140cgcatccgcg aagagctgaa gctgccgaag ctggaagact acgaggtcaa ggaagtcgcc 1200gagtacggcc acggctgggg ccagctcgag gccacccgtc gtctgggcgt ctacacccgc 1260gacatcatca agaacaaccc ggactccttc cgtatcttcg gaccggatga gaccgcttcc 1320aaccgtctgc aggccgctta cgacgtcacc aacaagcagt gggacgccgg ctacctgtcc 1380gctcaggtcg acgagcacat ggctgtcacc ggccaggtca ccgagcagct ttccgagcac 1440cagatggaag gcttcctcga gggctacctg ctgaccggcc gtcacggcat ctggagctcc 1500tatgagtcct tcgtgcacgt gatcgactcc atgctgaacc agcacgccaa gtggctcgag 1560gctaccgtcc gcgagattcc gtggcgcaag ccgatctcct ccatgaacct gctcgtctcc 1620tcccacgtgt ggcgtcagga tcacaacggc ttctcccacc aggatccggg tgtcacctcc 1680gtcctgctga acaagtgctt caacaacgat cacgtgatcg gcatctactt cccggtggat 1740tccaacatgc tgctcgctgt ggctgagaag tgctacaagt ccaccaacaa gatcaacgcc 1800atcatcgccg gcaagcagcc ggccgccacc tggctgaccc tggacgaagc tcgcgccgag 1860ctcgagaagg gtgctgccga gtggaagtgg gcttccaacg tgaagtccaa cgatgaggct 1920cagatcgtgc tcgccgccac cggtgatgtt ccgactcagg aaatcatggc cgctgccgac 1980aagctggacg ccatgggcat caagttcaag gtcgtcaacg tggttgacct ggtcaagctg 2040cagtccgcca aggagaacaa cgaggccctc tccgatgagg agttcgctga gctgttcacc 2100gaggacaagc cggtcctgtt cgcttaccac tcctatgccc gcgatgtgcg tggtctgatc 2160tacgatcgcc cgaaccacga caacttcaac gttcacggct acgaggagca gggctccacc 2220accaccccgt acgacatggt tcgcgtgaac aacatcgatc gctacgagct ccaggctgaa 2280gctctgcgca tgattgacgc tgacaagtac gccgacaaga tcaacgagct cgaggccttc 2340cgtcaggaag ccttccagtt cgctgtcgac aacggctacg atcacccgga ttacaccgac 2400tgggtctact ccggtgtcaa caccaacaag cagggtgcta tctccgctac cgccgcaacc 2460gctggcgata acgagtga 24789788PRTLactobacillus plantarumPHK1 9Met Thr Thr Asp Tyr Ser Ser Pro Ala Tyr Leu Gln Lys Val Asp Lys1 5 10 15Tyr Trp Arg Ala Ala Asn Tyr Leu Ser Val Gly Gln Leu Tyr Leu Lys 20 25 30Asp Asn Pro Leu Leu Gln Arg Pro Leu Lys Ala Ser Asp Val Lys Val 35 40 45His Pro Ile Gly His Trp Gly Thr Ile Ala Gly Gln Asn Phe Ile Tyr 50 55 60Ala His Leu Asn Arg Val Ile Asn Lys Tyr Gly Leu Lys Met Phe Tyr65 70 75 80Val Glu Gly Pro Gly His Gly Gly Gln Val Met Val Ser Asn Ser Tyr 85 90 95Leu Asp Gly Thr Tyr Thr Asp Ile Tyr Pro Glu Ile Thr Gln Asp Val 100 105 110Glu Gly Met Gln Lys Leu Phe Lys Gln Phe Ser Phe Pro Gly Gly Val 115 120 125Ala Ser His Ala Ala Pro Glu Thr Pro Gly Ser Ile His Glu Gly Gly 130 135 140Glu Leu Gly Tyr Ser Ile Ser His Gly Val Gly Ala Ile Leu Asp Asn145 150 155 160Pro Asp Glu Ile Ala Ala Val Val Val Gly Asp Gly Glu Ser Glu Thr 165 170 175Gly Pro Leu Ala Thr Ser Trp Gln Ser Thr Lys Phe Ile Asn Pro Ile 180 185 190Asn Asp Gly Ala Val Leu Pro Ile Leu Asn Leu Asn Gly Phe Lys Ile 195 200 205Ser Asn Pro Thr Ile Phe Gly Arg Thr Ser Asp Ala Lys Ile Lys Glu 210 215 220Tyr Phe Glu Ser Met Asn Trp Glu Pro Ile Phe Val Glu Gly Asp Asp225 230 235 240Pro Glu Lys Val His Pro Ala Leu Ala Lys Ala Met Asp Glu Ala Val 245 250 255Glu Lys Ile Lys Ala Ile Gln Lys His Ala Arg Glu Asn Asn Asp Ala 260 265 270Thr Leu Pro Val Trp Pro Met Ile Val Phe Arg Ala Pro Lys Gly Trp 275 280 285Thr Gly Pro Lys Ser Trp Asp Gly Asp Lys Ile Glu Gly Ser Phe Arg 290 295 300Ala His Gln Ile Pro Ile Pro Val Asp Gln Asn Asp Met Glu His Ala305 310 315 320Asp Ala Leu Val Asp Trp Leu Glu Ser Tyr Gln Pro Lys Glu Leu Phe 325 330 335Asn Glu Asp Gly Ser Leu Lys Asp Asp Ile Lys Glu Ile Ile Pro Thr 340 345 350Gly Asp Ser Arg Met Ala Ala Asn Pro Ile Thr Asn Gly Gly Val Asp 355 360 365Pro Lys Ala Leu Asn Leu Pro Asn Phe Arg Asp Tyr Ala Val Asp Thr 370 375 380Ser Lys Glu Gly Ala Asn Val Lys Gln Asp Met Ile Val Trp Ser Asp385 390 395 400Tyr Leu Arg Asp Val Ile Lys Lys Asn Pro Asp Asn Phe Arg Leu Phe 405 410 415Gly Pro Asp Glu Thr Met Ser Asn Arg Leu Tyr Gly Val Phe Glu Thr 420 425 430Thr Asn Arg Gln Trp Met Glu Asp Ile His Pro Asp Ser Asp Gln Tyr 435 440 445Glu Ala Pro Ala Gly Arg Val Leu Asp Ala Gln Leu Ser Glu His Gln 450 455 460Ala Glu Gly Trp Leu Glu Gly Tyr Val Leu Thr Gly Arg His Gly Leu465 470 475 480Phe Ala Ser Tyr Glu Ala Phe Leu Arg Val Val Asp Ser Met Leu Thr 485 490 495Gln His Phe Lys Trp Leu Arg Lys Ala Asn Glu Leu Asp Trp Arg Lys 500 505 510Lys Tyr Pro Ser Leu Asn Ile Ile Ala Ala Ser Thr Val Phe Gln Gln 515 520 525Asp His Asn Gly Tyr Thr His Gln Asp Pro Gly Ala Leu Thr His Leu 530 535 540Ala Glu Lys Lys Pro Glu Tyr Ile Arg Glu Tyr Leu Pro Ala Asp Ala545 550 555 560Asn Thr Leu Leu Ala Val Gly Asp Val Ile Phe Arg Ser Gln Glu Lys 565 570 575Ile Asn Tyr Val Val Thr Ser Lys His Pro Arg Gln Gln Trp Phe Ser 580 585 590Ile Glu Glu Ala Lys Gln Leu Val Asp Asn Gly Leu Gly Ile Ile Asp 595 600 605Trp Ala Ser Thr Asp Gln Gly Ser Glu Pro Asp Ile Val Phe Ala Ala 610 615 620Ala Gly Thr Glu Pro Thr Leu Glu Thr Leu Ala Ala Ile Gln Leu Leu625 630 635 640His Asp Ser Phe Pro Glu Met Lys Ile Arg Phe Val Asn Val Val Asp 645 650 655Ile Leu Lys Leu Arg Ser Pro Glu Lys Asp Pro Arg Gly Leu Ser Asp 660 665 670Ala Glu Phe Asp His Tyr Phe Thr Lys Asp Lys Pro Val Val Phe Ala 675 680 685Phe His Gly Tyr Glu Asp Leu Val Arg Asp Ile Phe Phe Asp Arg His 690 695 700Asn His Asn Leu Tyr Val His Gly Tyr Arg Glu Asn Gly Asp Ile Thr705 710 715 720Thr Pro Phe Asp Val Arg Val Met Asn Gln Met Asp Arg Phe Asp Leu 725 730 735Ala Lys Ser Ala Ile Ala Ala Gln Pro Ala Met Glu Asn Thr Gly Ala 740 745 750Ala Phe Val Gln Ser Met Asp Asn Met Leu Ala Lys His Asn Ala Tyr 755 760 765Ile Arg Asp Ala Gly Thr Asp Leu Pro Glu Val Asn Asp Trp Gln Trp 770 775 780Lys Gly Leu Lys78510556PRTBifidobacterium adolescentisPTA 10Met Ser Phe Thr Ser Val Thr Ile Ile Ser Pro Glu Ala Ala Asn Gly1 5 10 15Arg Asn Val Val Ala Leu Gly Val Thr Lys Thr Leu Ala Ala Ala Gly 20 25 30Lys Thr Gly Val Phe Arg Pro Ala Val Cys Arg Lys Asp Thr Phe Thr 35 40 45Asp Val Leu Ile Glu Ala Ser Asn Ala Gly Leu Ser Arg Glu Gln Ser 50 55 60Val Gly Val Cys Pro Lys Arg Ala Arg Asn Asp Lys Glu Gly Ser Arg65 70 75 80Ala Asp Ile Val Ala Ala Tyr Thr Gln Ala Val Glu Thr Ala Arg Pro 85 90 95Asp Ala Met Val Ile Val Gly Thr Asp Arg Ser Ala Val Asn Asp Pro 100 105 110Ala Met Phe Ser Phe Asn Ala Asp Val Ala Ala Asp Leu Gln Ser Pro 115 120 125Val Leu Leu Ala Val Cys Thr Ile Glu Arg Thr Pro Glu Gln Val Lys 130 135 140Ser Thr Val Glu Ala Ser Thr Lys Val Ile Glu Asp Ala Gly Ser Lys145 150 155 160Val Val Gly Val Phe Ile Thr Gly Cys Asp Asp Thr Gln Pro Asn Pro 165 170 175Leu Lys Ala Cys Phe Val Asp Tyr Pro Val Pro Val Trp Thr Leu Pro 180 185 190Ala Val Asp Phe Asn Asp Asp Asp Ala Ile Ser Lys Ala Asp Glu Ala 195 200 205Phe Ala Thr Asn Val Asp Ala Val Glu Leu Thr Val Ala Leu Glu Ser 210 215 220Pro Phe Asp Ala Pro Thr Thr Pro Tyr Ala Phe Gln Tyr Gly Leu Leu225 230 235 240Gly Lys Ala Lys Ala Asp Lys Lys Thr Ile Val Leu Pro Glu Gly Asn 245 250 255Glu Asp Arg Ile Ile Lys Ala Ala Asp Tyr Leu Leu Glu Arg Asp Ile 260 265 270Val Asp Leu Ile Ile Val Gly Asp Glu Asn Ala Ile Leu Ala Arg Gly 275 280 285Gln Glu Leu Gly Leu Lys Ser Leu Gly Lys Ala Lys Phe Gln Ala Lys 290 295 300Asp Asp Glu Thr Val Leu Glu Pro Met Val Ala Lys Leu Cys Glu Leu305 310 315 320Arg Ala Lys Lys Gly Met Thr Glu Glu Gln Ala Arg Lys Gln Leu Ala 325 330 335Asp Asp Ser Tyr Phe Gly Thr Met Leu Val Val Met Gly Met Ala Asp 340 345 350Gly Leu Val Ser Gly Ser Val Asn Ser Thr Ala Asn Thr Val Arg Pro 355 360 365Ala Leu Gln Val Ile Lys Thr Lys Pro Gly Thr Ser Leu Val Ser Gly 370 375 380Ala Phe Leu Met Cys Phe Lys Asp His Ala Ala Val Phe Ala Asp Cys385 390 395 400Ala Ile Asn Leu Asn Pro Asn Ala Glu Gln Leu Ala Glu Ile Ala Ile 405 410 415Gln Ser Ala Glu Thr Ala Lys Ala Phe Gly Leu Glu Pro Lys Val Gly 420 425 430Met Leu Ser Tyr Ser Thr Leu Gly Ser Gly Lys Gly Pro Asp Val Asp 435 440 445Leu Val Glu Glu Ala Thr Thr Ile Val Lys Asp Lys Ala Pro Asp Leu 450 455 460Ala Val Val Gly Ser Ile Gln Phe Asp Ala Ala Trp Ser Pro Thr Val465 470 475 480Ala Ala Thr Lys Ala Lys Gly Asp Pro Val Ala Gly His Val Asn Val 485 490 495Phe Val Phe Pro Asp Leu Cys Ala Gly Asn Ile Ala Tyr Lys Ala Val 500 505 510Gln Arg Ser Ser Gly Ala Ala Ala Val Gly Pro Val Leu Gln Gly Leu 515 520 525Asn Arg Pro Val Asn Asp Leu Ser Arg Gly Ala Thr Val Gln Asp Ile 530 535 540Ile Asn Thr Ile Ala Leu Thr Ala Ile Glu Ala Gln545 550 55511793PRTLactobacillus caseiPHK 11Met Asp Thr Lys Val Lys Thr Val Asp Tyr Ser Ser Lys Glu Tyr Phe1 5 10 15Asp Lys Met Thr Ala Tyr Trp Arg Ala Ala Asn Tyr Ile Ser Val Gly 20 25 30Gln Leu Tyr Leu Lys Asp Asn Pro Leu Leu Glu Arg Pro Leu Lys Ser 35 40 45Glu Asp Val Lys Pro His Pro Ile Gly His Trp Gly Thr Ile Ala Gly 50 55 60Gln Asn Phe Ile Tyr Thr His Leu Asn Arg Val Ile Asn Lys Tyr Asp65 70 75 80Leu Asn Met Phe Tyr Ile Glu Gly Pro Gly His Gly Gly Gln Val Met 85 90 95Val Ser Asn Ser Tyr Leu Asp Gly Ser Tyr Ser Glu Ile Tyr Pro Arg 100 105 110Val Ser Gln Asp Lys Glu Gly Met Lys Asn Leu Phe Thr Gln Phe Ser 115 120 125Trp Pro Gly Gly Val Ala Ser His Ala Ser Ala Gln Thr Pro Gly Ser 130 135 140Ile His Glu Gly Gly Glu Leu Gly Tyr Ala Leu Ser His Ala Thr Gly145 150 155 160Ala Ile Leu Asp Asn Pro Glu Val Ile Ala Ala Val Val Thr Gly Asp 165 170 175Gly Glu Thr Glu Thr Gly Pro Leu Ala Ala Ser Trp Phe Ser Asn Thr 180 185 190Phe Ile Asn Pro Ile Thr Asp Gly Ala Ile Leu Pro Ile Val His Met 195 200 205Asn Gly Phe Lys Ile Ser Asn Pro Thr Ile Leu Ser Arg Lys Ser Asp 210 215 220Glu Asp Leu Asp Lys Tyr Phe Ser Gly Met Gly Trp Lys Pro Phe Phe225 230 235 240Val Glu Gly Asp Asp Pro Glu Lys Leu Asn Pro Glu Met Ala Lys Val 245 250 255Leu Asp Glu Ala Ile Glu Asp Ile Lys Ala Ile Gln Lys His Ala Arg 260 265 270Glu Thr Gly Asp Thr Thr Met Pro His Trp Pro Val Ile Ile Phe Arg 275 280 285Ser Pro Lys Gly Trp Thr Gly Pro Lys Ser Trp Asn Gly Glu Pro Ile 290 295 300Glu Gly Ser Phe Arg Ala His Gln Ile Pro Ile Pro Val Asp Ala Glu305 310 315 320Asp Met Glu His Ala Asp Ser Leu Ala Gly Trp Leu Lys Ser Tyr His 325 330 335Pro Glu Glu Leu Phe Asp Glu Asn Gly Lys Leu Ile Pro Glu Leu Ala 340 345 350Ala Leu Pro Pro Lys Gly Glu Gln Arg Met Ala Ala Asn Pro Ile Thr 355 360 365Asn Gly Gly Ile Asp Pro Lys Pro Leu Val Leu Pro Asp Tyr Arg Lys 370 375 380Tyr Ala Leu Asp Asn Lys Glu His Gly Lys Gln Ile Lys Gln Asp Met385 390 395 400Ile Val Trp Ser Asp Tyr Leu Arg Asp Leu Ile Lys Leu Asn Pro His 405 410 415Asn Phe Arg Ile Phe Gly Pro Asp Glu Thr Met Ser Asn Arg Leu Tyr 420 425 430Ser Leu Phe Glu Val Thr Asn Arg Gln Trp Leu Glu Pro Ile Lys Glu 435 440 445Pro Ala Asp Gln Tyr Leu Ala Pro Ala Gly Arg Ile Ile Asp Ser Gln 450 455 460Leu Ser Glu His Gln Ala Glu Gly Phe Asn Glu Gly Tyr Thr Leu Thr465 470 475 480Gly Arg His Gly Leu Phe Thr Ser Tyr Glu Ala Phe Leu Arg Val Val 485 490 495Asp Ser Met Leu Thr Gln His Phe Lys Trp Ile Arg Lys Ala His Glu 500 505 510Glu Pro Trp His Lys Ala Tyr Pro Ser Leu Asn Val Val Ser Thr Ser 515 520 525Thr Ser Phe Gln Gln Asp His Asn Gly Tyr Thr His Gln Asp Pro Gly 530 535 540Ile Leu Thr His Met Ala Glu Lys Lys Ala Glu Tyr Ile Arg Glu Tyr545 550 555 560Leu Pro Ala Asp Ala Asn Ser Leu Leu Ala Ile Ser Pro Lys Leu Phe 565 570 575Ser Ser Gln Asn Thr Val Asn Val Leu Ile Thr Ser Lys Gln Pro Arg 580 585 590Pro Gln Phe Tyr Ser Ile Asp Glu Ala Thr Val Leu Ala Asn Ser Gly 595 600 605Leu Lys Arg Ile Asp Trp Ala Ser Asn Asp Asp Gly Val Glu Pro Asp 610 615 620Val Val Ile Ala Ala Ala Gly Thr Glu Pro Asn Met Glu Ser Leu Ala625 630 635 640Ala Ile Asn Leu Leu His Asp Ala Phe Pro Asp Leu Lys Ile Arg Phe 645 650 655Ile Asn Val Val Asp Leu Leu Lys Leu Arg Ser Pro

Glu Ile Asp Pro 660 665 670Arg Gly Leu Ser Asp Ala Glu Phe Asn Ser Tyr Phe Thr Thr Asp Lys 675 680 685Pro Ile Leu Phe Ala Tyr His Gly Phe Glu Gly Leu Ile Arg Asp Ile 690 695 700Phe Phe Thr Arg Pro Asn Arg Asn Val Leu Ile His Gly Tyr Arg Glu705 710 715 720Glu Gly Asp Ile Thr Thr Pro Phe Asp Met Arg Val Leu Asn Glu Leu 725 730 735Asp Arg Tyr His Leu Ala Lys Asp Val Ile Gln His Val Pro Ala Tyr 740 745 750Ala Glu Lys Ala Ala Ala Phe Val Gln Lys Met Asp Asp Thr Leu Gln 755 760 765Tyr His His Asp Phe Ile Arg Ala Asn Gly Asp Asp Ile Pro Glu Val 770 775 780Gln Glu Trp Thr Trp Lys Pro Ile Lys785 79012825PRTBifidobacterium adolescentisPHK 12Met Thr Ser Pro Val Ile Gly Thr Pro Trp Lys Lys Leu Asn Ala Pro1 5 10 15Val Ser Glu Glu Ala Ile Glu Gly Val Asp Lys Tyr Trp Arg Ala Ala 20 25 30Asn Tyr Leu Ser Ile Gly Gln Ile Tyr Leu Arg Ser Asn Pro Leu Met 35 40 45Lys Glu Pro Phe Thr Arg Glu Asp Val Lys His Arg Leu Val Gly His 50 55 60Trp Gly Thr Thr Pro Gly Leu Asn Phe Leu Ile Gly His Ile Asn Arg65 70 75 80Leu Ile Ala Asp His Gln Gln Asn Thr Val Ile Ile Met Gly Pro Gly 85 90 95His Gly Gly Pro Ala Gly Thr Ala Gln Ser Tyr Leu Asp Gly Thr Tyr 100 105 110Thr Glu Tyr Phe Pro Asn Ile Thr Lys Asp Glu Ala Gly Leu Gln Lys 115 120 125Phe Phe Arg Gln Phe Ser Tyr Pro Gly Gly Ile Pro Ser His Tyr Ala 130 135 140Pro Glu Thr Pro Gly Ser Ile His Glu Gly Gly Glu Leu Gly Tyr Ala145 150 155 160Leu Ser His Ala Tyr Gly Ala Val Met Asn Asn Pro Ser Leu Phe Val 165 170 175Pro Ala Ile Val Gly Asp Gly Glu Ala Glu Thr Gly Pro Leu Ala Thr 180 185 190Gly Trp Gln Ser Asn Lys Leu Ile Asn Pro Arg Thr Asp Gly Ile Val 195 200 205Leu Pro Ile Leu His Leu Asn Gly Tyr Lys Ile Ala Asn Pro Thr Ile 210 215 220Leu Ser Arg Ile Ser Asp Glu Glu Leu His Glu Phe Phe His Gly Met225 230 235 240Gly Tyr Glu Pro Tyr Glu Phe Val Ala Gly Phe Asp Asn Glu Asp His 245 250 255Leu Ser Ile His Arg Arg Phe Ala Glu Leu Phe Glu Thr Val Phe Asp 260 265 270Glu Ile Cys Asp Ile Lys Ala Ala Ala Gln Thr Asp Asp Met Thr Arg 275 280 285Pro Phe Tyr Pro Met Ile Ile Phe Arg Thr Pro Lys Gly Trp Thr Cys 290 295 300Pro Lys Phe Ile Asp Gly Lys Lys Thr Glu Gly Ser Trp Arg Ser His305 310 315 320Gln Val Pro Leu Ala Ser Ala Arg Asp Thr Glu Ala His Phe Glu Val 325 330 335Leu Lys Asn Trp Leu Glu Ser Tyr Lys Pro Glu Glu Leu Phe Asp Glu 340 345 350Asn Gly Ala Val Lys Pro Glu Val Thr Ala Phe Met Pro Thr Gly Glu 355 360 365Leu Arg Ile Gly Glu Asn Pro Asn Ala Asn Gly Gly Arg Ile Arg Glu 370 375 380Glu Leu Lys Leu Pro Lys Leu Glu Asp Tyr Glu Val Lys Glu Val Ala385 390 395 400Glu Tyr Gly His Gly Trp Gly Gln Leu Glu Ala Thr Arg Arg Leu Gly 405 410 415Val Tyr Thr Arg Asp Ile Ile Lys Asn Asn Pro Asp Ser Phe Arg Ile 420 425 430Phe Gly Pro Asp Glu Thr Ala Ser Asn Arg Leu Gln Ala Ala Tyr Asp 435 440 445Val Thr Asn Lys Gln Trp Asp Ala Gly Tyr Leu Ser Ala Gln Val Asp 450 455 460Glu His Met Ala Val Thr Gly Gln Val Thr Glu Gln Leu Ser Glu His465 470 475 480Gln Met Glu Gly Phe Leu Glu Gly Tyr Leu Leu Thr Gly Arg His Gly 485 490 495Ile Trp Ser Ser Tyr Glu Ser Phe Val His Val Ile Asp Ser Met Leu 500 505 510Asn Gln His Ala Lys Trp Leu Glu Ala Thr Val Arg Glu Ile Pro Trp 515 520 525Arg Lys Pro Ile Ser Ser Met Asn Leu Leu Val Ser Ser His Val Trp 530 535 540Arg Gln Asp His Asn Gly Phe Ser His Gln Asp Pro Gly Val Thr Ser545 550 555 560Val Leu Leu Asn Lys Cys Phe Asn Asn Asp His Val Ile Gly Ile Tyr 565 570 575Phe Pro Val Asp Ser Asn Met Leu Leu Ala Val Ala Glu Lys Cys Tyr 580 585 590Lys Ser Thr Asn Lys Ile Asn Ala Ile Ile Ala Gly Lys Gln Pro Ala 595 600 605Ala Thr Trp Leu Thr Leu Asp Glu Ala Arg Ala Glu Leu Glu Lys Gly 610 615 620Ala Ala Glu Trp Lys Trp Ala Ser Asn Val Lys Ser Asn Asp Glu Ala625 630 635 640Gln Ile Val Leu Ala Ala Thr Gly Asp Val Pro Thr Gln Glu Ile Met 645 650 655Ala Ala Ala Asp Lys Leu Asp Ala Met Gly Ile Lys Phe Lys Val Val 660 665 670Asn Val Val Asp Leu Val Lys Leu Gln Ser Ala Lys Glu Asn Asn Glu 675 680 685Ala Leu Ser Asp Glu Glu Phe Ala Glu Leu Phe Thr Glu Asp Lys Pro 690 695 700Val Leu Phe Ala Tyr His Ser Tyr Ala Arg Asp Val Arg Gly Leu Ile705 710 715 720Tyr Asp Arg Pro Asn His Asp Asn Phe Asn Val His Gly Tyr Glu Glu 725 730 735Gln Gly Ser Thr Thr Thr Pro Tyr Asp Met Val Arg Val Asn Asn Ile 740 745 750Asp Arg Tyr Glu Leu Gln Ala Glu Ala Leu Arg Met Ile Asp Ala Asp 755 760 765Lys Tyr Ala Asp Lys Ile Asn Glu Leu Glu Ala Phe Arg Gln Glu Ala 770 775 780Phe Gln Phe Ala Val Asp Asn Gly Tyr Asp His Pro Asp Tyr Thr Asp785 790 795 800Trp Val Tyr Ser Gly Val Asn Thr Asn Lys Gln Gly Ala Ile Ser Ala 805 810 815Thr Ala Ala Thr Ala Gly Asp Asn Glu 820 82513806PRTAspergillus nigerPHK 13Met Pro Gly Glu Val Ile Asp Arg Pro Asn Pro Lys Ala Glu Pro Ser1 5 10 15His Ile Pro Asp Leu Val Asn Gln Leu Gln Val Lys Leu Gln Glu Thr 20 25 30Arg Leu Glu Glu Thr Asp Tyr Asn Ala Leu Leu Lys Phe Arg Arg Ala 35 40 45Ala Ala Tyr Ile Ala Ala Ala Met Ile Phe Leu Gln Asp Asn Val Leu 50 55 60Leu Lys Gln Asn Leu Arg His Glu Asp Ile Lys Pro Arg Leu Leu Gly65 70 75 80His Trp Gly Thr Cys Pro Gly Leu Ile Leu Val Tyr Ser His Leu Asn 85 90 95Tyr Ile Ile Arg Lys Gln Asn Leu Asp Met Leu Tyr Val Val Gly Pro 100 105 110Gly His Gly Ala Pro Ala Ile Leu Ala Ser Leu Trp Leu Glu Gly Ser 115 120 125Leu Glu Lys Phe Tyr Pro His Tyr Ser Arg Asp Met Asp Gly Leu His 130 135 140Glu Leu Ile Ser Thr Phe Ser Thr Ser Ala Gly Leu Pro Ser His Ile145 150 155 160Asn Ala Glu Thr Pro Gly Ala Ile His Glu Gly Gly Glu Leu Gly Tyr 165 170 175Ala Leu Ala Val Ser Phe Gly Ala Val Met Asp Asn Pro Asp Met Ile 180 185 190Val Thr Cys Val Val Gly Asp Gly Glu Ala Glu Thr Gly Pro Thr Ala 195 200 205Thr Ser Trp His Ala Ile Lys Tyr Ile Asp Pro Ala Glu Ser Gly Ala 210 215 220Val Leu Pro Ile Leu His Val Asn Gly Phe Lys Ile Ser Glu Arg Thr225 230 235 240Ile Tyr Gly Cys Met Asp Asn Lys Glu Leu Val Ser Leu Phe Thr Gly 245 250 255Tyr Gly Tyr Gln Val Arg Ile Val Glu Asn Leu Asp Asp Ile Asp Ala 260 265 270Asp Leu His Ser Ser Met Met Trp Ala Val Glu Glu Ile His Lys Ile 275 280 285Gln Lys Ala Ala Arg Ser Gly Lys Pro Ile Met Lys Pro Arg Trp Pro 290 295 300Met Ile Val Leu Arg Thr Pro Lys Gly Trp Ser Gly Pro Lys Glu Leu305 310 315 320His Gly Ser Phe Ile Glu Gly Ser Phe His Ser His Gln Val Pro Leu 325 330 335Pro Asn Ala Lys Lys Asp Lys Glu Glu Leu Gln Ala Leu Gln Lys Trp 340 345 350Leu Ser Ser Tyr Asn Pro His Glu Leu Phe Thr Glu Thr Gly Asp Ile 355 360 365Ile Asp Asp Ile Lys Ser Val Ile Pro Leu Glu Asp Thr Lys Lys Leu 370 375 380Gly Gln Arg Ala Glu Ala Tyr Lys Gly Tyr Arg Ala Pro Asp Leu Pro385 390 395 400Asp Trp Arg Lys Phe Gly Val Glu Lys Gly Ser Gln Gln Ser Ala Met 405 410 415Lys Thr Ile Gly Lys Phe Ile Asp Gln Val Phe Thr Gln Asn Pro His 420 425 430Gly Val Arg Val Phe Ser Pro Asp Glu Leu Glu Ser Asn Lys Leu Asp 435 440 445Ala Ala Leu Ala His Thr Gly Arg Asn Phe Gln Trp Asp Gln Phe Ser 450 455 460Asn Ala Lys Gly Gly Arg Val Ile Glu Val Leu Ser Glu His Leu Cys465 470 475 480Gln Gly Phe Met Gln Gly Tyr Thr Leu Thr Gly Arg Val Gly Ile Phe 485 490 495Pro Ser Tyr Glu Ser Phe Leu Gly Ile Ile His Thr Met Met Val Gln 500 505 510Tyr Ala Lys Phe Asn Lys Met Ala Gln Glu Thr Thr Trp His Lys Pro 515 520 525Val Ser Ser Ile Asn Tyr Ile Glu Thr Ser Thr Trp Ala Arg Gln Glu 530 535 540His Asn Gly Phe Ser His Gln Asn Pro Ser Phe Ile Gly Ala Val Leu545 550 555 560Arg Leu Lys Pro Thr Ala Ala Arg Val Tyr Leu Pro Pro Asp Ala Asn 565 570 575Thr Phe Leu Thr Thr Leu His His Cys Leu Lys Ser Lys Asn Tyr Val 580 585 590Asn Leu Met Val Gly Ser Lys Gln Pro Thr Pro Val Tyr Leu Ser Pro 595 600 605Glu Glu Ala Glu Ser His Cys Arg Ala Gly Ala Ser Ile Trp Arg Phe 610 615 620Cys Ser Thr Asp Asn Gly Leu Asn Pro Asp Val Val Leu Val Gly Ile625 630 635 640Gly Val Glu Val Met Phe Glu Val Ile Tyr Ala Ala Ala Ile Leu Arg 645 650 655Lys Arg Cys Pro Glu Leu Arg Val Arg Val Val Asn Val Thr Asp Leu 660 665 670Met Ile Leu Glu Lys Glu Gly Leu His Pro His Ala Leu Thr Thr Glu 675 680 685Ala Phe Asp Ser Leu Phe Gly Ser Asp Arg Pro Ile His Phe Asn Tyr 690 695 700His Gly Tyr Pro Gly Glu Leu Lys Gly Leu Leu Phe Gly Arg Pro Arg705 710 715 720Leu Asp Arg Val Ser Val Glu Gly Tyr Met Glu Glu Gly Ser Thr Thr 725 730 735Thr Pro Phe Asp Met Met Leu Leu Asn Arg Val Ser Arg Tyr His Val 740 745 750Ala Gln Ala Ala Val Ile Gly Ala Ser Arg Arg Asn Glu Lys Val Gln 755 760 765Val Arg Gln His Glu Leu Val Ser Glu Phe Gly His Asn Ile Val Glu 770 775 780Thr Arg Lys Tyr Ile Leu Ala Asn Arg Lys Asp Pro Asp Asp Thr Tyr785 790 795 800Asp Met Pro Ser Phe Glu 80514815PRTNeurospora crassaPHK 14Met Gly Gly Thr Gln Ile Thr Ile Pro Asn Pro Pro Pro Leu Pro Ser1 5 10 15His Leu Pro Asp Asn Val Leu Asp Leu Ala Val Gln Ile Glu Arg Lys 20 25 30Asn Leu Pro Asp Asn Val Arg Glu Ser Leu Arg Ala Phe Gln Arg Ala 35 40 45Gly Cys Tyr Ile Ala Ala Ser Met Ile Phe Leu Arg Asp Asn Val Leu 50 55 60Leu Gln Asp Lys Leu Gln Val Ser His Ile Lys Pro Arg Leu Leu Gly65 70 75 80His Trp Gly Thr Cys Pro Gly Leu Ile Leu Val Trp Ser His Leu Asn 85 90 95Val Leu Ile Arg Asn His Asp Leu Asn Met Ile Tyr Val Ile Gly Pro 100 105 110Gly His Gly Ala Pro Ala Ala Leu Ala Ser Leu Trp Leu Glu Gly Ser 115 120 125Leu Glu Arg Phe Tyr Pro Gly Glu Tyr Asp Arg Asn Ala Thr Gly Leu 130 135 140Arg Asn Leu Ile Thr Arg Phe Ser Val Pro Gly Gly Phe Pro Ser His145 150 155 160Ile Asn Ala Glu Thr Pro Gly Ala Ile His Glu Gly Gly Glu Leu Gly 165 170 175Tyr Ala Leu Ser Val Ser Phe Gly Ala Val Met Asp Asn Pro Asp Leu 180 185 190Ile Val Thr Cys Val Val Gly Asp Gly Glu Ala Glu Thr Gly Pro Thr 195 200 205Ala Thr Ala Trp His Ala Ile Lys Tyr Ile Asp Pro Lys Glu Ser Gly 210 215 220Ala Val Ile Pro Ile Leu His Val Asn Gly Phe Lys Ile Ser Glu Arg225 230 235 240Thr Ile Phe Gly Cys Met Asp Asp Lys Glu Ile Ala Cys Leu Phe Ser 245 250 255Gly Tyr Gly Tyr Gln Val Arg Ile Val Glu Asp Leu Glu Asn Ile Asp 260 265 270Asp Asp Leu Gln Ser Ser Leu Glu Trp Ala Leu Val Glu Ile Lys Lys 275 280 285Ile Gln Lys Ala Ala Arg Glu Asp Asn Lys Pro Ile Val Lys Pro Arg 290 295 300Trp Pro Met Ile Val Leu Arg Thr Pro Lys Gly Trp Thr Gly Pro Lys305 310 315 320Glu Val Glu Gly Gln Ile Ile Glu Gly Ser Phe His Ser His Gln Val 325 330 335Pro Leu Pro Lys Ala Gly Ser Asp Glu Lys Gln Leu Gln Ala Leu Asp 340 345 350Glu Trp Leu Ser Ser Tyr Arg Ile Ser Glu Leu Leu Asp Asp Glu Gly 355 360 365Lys Pro Thr Glu Ala Ile Thr Arg Val Leu Pro Lys Ala Glu Lys Arg 370 375 380Leu Gly Gln Leu Lys Ile Thr Tyr Asp Pro Tyr Ile Pro Leu Lys Leu385 390 395 400Ile Asp Trp Lys Gln Phe Gly Val Gln Lys Gly Val Asp Glu Ser Cys 405 410 415Met Lys Ser Ala Gly Lys Phe Leu Asp Lys Leu Phe Gln Glu Asn Pro 420 425 430Lys Thr Leu Arg Leu Phe Ser Pro Asp Glu Leu Glu Ser Asn Lys Leu 435 440 445Asn Ala Ile Leu Asp His Thr Gln Arg Asp Phe Gln Trp Asp Glu Phe 450 455 460Ser Arg Ala His Gly Gly Arg Val Ile Glu Val Leu Ser Glu His Asn465 470 475 480Cys Gln Gly Phe Met Gln Gly Tyr Thr Leu Thr Gly Arg Thr Ala Leu 485 490 495Phe Pro Ser Tyr Glu Ser Phe Leu Gly Ile Val His Thr Met Met Val 500 505 510Gln Tyr Ser Lys Phe Val Lys Ile Ala Arg Glu Val Pro Trp Arg Gly 515 520 525Asp Leu Ala Ser Ile Asn Tyr Ile Glu Thr Ser Thr Trp Ala Arg Gln 530 535 540Glu His Asn Gly Phe Ser His Gln Asn Pro Ser Phe Ile Gly Ala Val545 550 555 560Leu Asn Leu Lys Ala Glu Ala Ala Arg Val Tyr Leu Pro Pro Asp Ala 565 570 575Asn Cys Phe Leu Ser Thr Val His His Val Met Asp Ser Lys Asn Lys 580 585 590Thr Asn Leu Ile Ile Gly Ser Lys Gln Pro Thr Ala Val Tyr Leu Ser 595 600 605Pro Ala Glu Ala Ala Asp His Cys Arg Gln Gly Ala Ser Ile Trp His 610 615 620Phe Ala Ser Ser Ser Asp Pro Asn Ser Ser Ala Gly Glu Ser Val Gln625 630 635 640Asp Pro Asp Val Val Leu Val Gly Ile Gly Val Glu Val Thr Phe Glu 645 650 655Val Ile Lys Ala Ala Glu Leu Leu Arg Ser Ile Ala Pro Ala Leu Lys 660 665 670Ile Arg Val Val Asn Val Thr Asp Leu Met Ile Leu Ile Glu Glu Ser 675 680 685Lys His Pro His Ala Leu Ala Lys Ala Lys Phe Ile Glu Met Phe Thr 690 695

700Ala Asp Arg Pro Val Leu Phe Asn Tyr His Gly Tyr Pro Thr Glu Leu705 710 715 720Gln Gly Leu Leu Phe Gly Arg Glu Lys Ala Glu Arg Met Asp Val Glu 725 730 735Gly Tyr Gln Glu Glu Gly Ser Thr Thr Thr Pro Phe Asp Met Met Leu 740 745 750Arg Asn Arg Val Ser Arg Phe Asp Val Ala Ser Trp Ala Leu Lys Arg 755 760 765Gly Ala Glu Arg Asn Ala Glu Val Lys Asn Asp Leu Glu Gly Leu Leu 770 775 780Gly Asp Val Asp Arg Arg Val Lys Glu Val Arg Glu Phe Ile Gly Asn785 790 795 800Glu Gly Lys Asp Pro Asp Asp Ile Tyr Gln Leu Pro Lys Phe Glu 805 810 81515796PRTLactobacillus plantarum PHK2 15Met Ser Glu Ala Ile Lys Ser Lys Thr Val Asp Tyr Ser Ser Asp Glu1 5 10 15Tyr Leu Lys Arg Val Asp Glu Tyr Trp Arg Ala Ala Asn Tyr Ile Ser 20 25 30Val Gly Gln Leu Tyr Leu Leu Asn Asn Pro Leu Leu Arg Glu Pro Leu 35 40 45Lys Ala Thr Asp Val Lys Val His Pro Ile Gly His Trp Gly Thr Ile 50 55 60Ala Gly Gln Asn Phe Ile Tyr Ala His Leu Asn Arg Ala Ile Asn Lys65 70 75 80Tyr Gly Leu Asn Met Phe Tyr Ile Glu Gly Pro Gly His Gly Gly Gln 85 90 95Val Met Val Ser Asn Ser Tyr Leu Asp Gly Thr Tyr Thr Glu Thr Tyr 100 105 110Pro Lys Ile Thr Gln Asp Lys Ala Gly Met Lys Arg Leu Phe Lys Gln 115 120 125Phe Ser Phe Pro Gly Gly Val Ala Ser His Ala Asp Pro Lys Thr Pro 130 135 140Gly Ser Ile His Glu Gly Gly Glu Leu Gly Tyr Ser Ile Leu His Gly145 150 155 160Ala Gly Ala Val Leu Asp Asn Pro Gly Leu Ile Ala Ala Thr Val Val 165 170 175Gly Asp Gly Glu Ser Glu Thr Gly Pro Leu Ala Thr Ser Trp Gln Val 180 185 190Asn Lys Phe Leu Asn Pro Ile Thr Asp Gly Thr Val Leu Pro Ile Leu 195 200 205Asn Leu Asn Gly Phe Lys Ile Ser Asn Pro Thr Val Leu Ser Arg Glu 210 215 220Ser His Glu Glu Leu Glu Asp Tyr Phe Lys Gly Leu Gly Trp Asp Pro225 230 235 240His Phe Val Glu Gly Thr Asp Pro Ala Lys Met His Lys Ile Met Ala 245 250 255Glu Glu Leu Asp Lys Val Ile Glu Glu Ile His Ala Ile Arg Lys Asn 260 265 270Ala Lys Asp Asn Asn Asp Glu Ser Arg Pro Lys Trp Pro Met Ile Val 275 280 285Phe Arg Ala Pro Lys Gly Trp Thr Gly Pro Lys Ser Trp Asp Gly Glu 290 295 300Pro Ile Glu Gly Ser Phe Arg Ala His Gln Ile Pro Ile Pro Val Asp305 310 315 320Arg Asn His Met Glu His Ala Asp Lys Leu Val Asp Trp Leu Lys Ser 325 330 335Tyr Lys Pro Glu Glu Leu Phe Asp Glu Asn Gly Thr Leu Lys Pro Glu 340 345 350Ile Ala Ala Ile Ile Pro Glu Gly Gln Ala Arg Met Ala Ala Asn Pro 355 360 365Val Thr Asn Gly Gly Lys Leu Thr Lys Asp Leu Ile Thr Pro Asn Ile 370 375 380Asp Asp Tyr Ala Leu Asp Asn Lys Ser His Gly Lys Glu Asp Gly Ser385 390 395 400Asp Met Thr Glu Leu Gly Lys Tyr Ile Arg Asp Leu Ile Glu Leu Asn 405 410 415Lys Asp Asn Lys Asn Phe Arg Gly Trp Gly Pro Asp Glu Thr Leu Ser 420 425 430Asn Lys Leu Gly Ala Ala Phe Glu Asp Thr Lys Arg Gln Trp Met Glu 435 440 445Pro Ile His Glu Pro Asn Asp Ala Leu Leu Ala Pro Gln Gly Arg Ile 450 455 460Ile Asp Ser Met Leu Ser Glu His Met Asp Glu Gly Met Leu Glu Ala465 470 475 480Tyr Asn Leu Thr Gly Arg Tyr Gly Phe Phe Ala Ser Tyr Glu Ser Phe 485 490 495Leu Arg Val Val Asp Ser Met Leu Thr Gln His Phe Lys Trp Leu Arg 500 505 510Asn Ser His Glu Glu Thr Pro Trp Arg Ala Asp Val Pro Ser Leu Asn 515 520 525Val Ile Ala Ser Ser Thr Ala Phe Gln Gln Asp His Asn Gly Tyr Ser 530 535 540His Gln Asp Pro Gly Ile Ile Ser His Leu Ala Glu Lys Lys Thr Glu545 550 555 560Tyr Val Arg Ala Tyr Leu Pro Gly Asp Ala Asn Thr Leu Ile Ala Thr 565 570 575Phe Asp Lys Ala Ile Gln Ser Lys Gln Leu Ile Asn Leu Ile Ile Ala 580 585 590Ser Lys His Pro Arg Pro Gln Trp Phe Thr Met Asp Glu Ala Lys Arg 595 600 605Leu Val Arg Asp Gly Leu Gly Val Val Asp Trp Ala Ser Thr Asp His 610 615 620Gly Glu Glu Pro Asp Val Val Phe Ala Thr Ala Gly Ser Glu Pro Thr625 630 635 640Thr Glu Ser Leu Ala Ala Val Ser Ile Leu His Ala Arg Phe Pro Glu 645 650 655Met Lys Ile Arg Phe Ile Asn Val Val Asp Leu Leu Lys Leu Lys Lys 660 665 670Asp Asp Pro Arg Gly Leu Ser Asp Ala Glu Phe Asp Ala Phe Phe Thr 675 680 685Lys Asp Lys Pro Val Ile Phe Ala Tyr His Ala Tyr Asp Asp Leu Val 690 695 700Lys Thr Ile Phe Phe Asp Arg His Asn His Asn Leu His Val His Gly705 710 715 720Tyr Arg Glu Glu Gly Asp Ile Thr Thr Pro Phe Asp Met Arg Val Arg 725 730 735Asn Glu Leu Asp Arg Phe His Leu Val Lys Ala Ala Leu Leu Ala Thr 740 745 750Pro Ala Tyr Ala Glu Lys Gly Ala His Val Ile Gln Glu Met Asn Ser 755 760 765Ile Leu Asp Lys His His Asp Tyr Ile Arg Ala Glu Gly Thr Asp Ile 770 775 780Pro Glu Val Glu Asn Trp Lys Trp Thr Ala Leu Lys785 790 79516409PRTBifidobacterium adolescentisACK 16Met Ala Lys Thr Val Leu Val Ile Asn Ser Gly Ser Ser Ser Ile Lys1 5 10 15Tyr Gln Leu Val Asp Leu Glu Thr Gly Glu Gly Ile Ala Ser Gly Leu 20 25 30Val Glu Lys Ile Gly Glu Pro Val Asp Gly His Tyr Lys His Glu Tyr 35 40 45Asn Gly Glu Lys His Glu Leu Glu Glu Pro Ile His Asp His Glu Gln 50 55 60Gly Leu Lys Arg Val Leu Gly Phe Phe Asp Glu Phe Gly Pro Lys Leu65 70 75 80Ala Asp Ala Gly Ile Val Ala Val Gly His Arg Val Val Gln Gly Gly 85 90 95Ser Ile Phe Pro Lys Pro Ala Leu Val Asn Asp Lys Thr Ile Gly Gln 100 105 110Val Lys Asp Leu Ala Val Leu Ala Pro Leu His Asn Gly Pro Glu Ala 115 120 125Lys Gly Ala Glu Val Met Arg Ser Leu Leu Pro Asp Val Pro Gln Ile 130 135 140Phe Val Phe Asp Ser Ser Phe Phe Phe Gln Leu Pro Lys Ala Ser Ser145 150 155 160Thr Tyr Ala Leu Asn Lys Glu Val Ala Gln Gln Tyr His Ile Arg Arg 165 170 175Tyr Gly Ala His Gly Thr Ser His Glu Phe Ile Ser Ser Val Val Pro 180 185 190Ser Val Ile Gly Lys Pro Ala Glu Gly Leu Lys Gln Ile Val Leu His 195 200 205Ile Gly Asn Gly Ala Ser Ala Ser Ala Glu Ile Ser Gly Lys Pro Val 210 215 220Glu Thr Ser Met Gly Leu Thr Pro Leu Glu Gly Leu Val Met Gly Gly225 230 235 240Arg Thr Gly Asp Ile Asp Pro Ala Val Val Phe His Leu Ile Arg Asn 245 250 255Ala His Met Ser Val Asp Glu Leu Asp Thr Leu Phe Asn Lys Arg Ser 260 265 270Gly Met Met Gly Leu Thr Gly Phe Gly Asp Leu Arg Glu Val His Arg 275 280 285Leu Val Glu Glu Gly Asn Glu Asp Ala Lys Leu Ala Leu Asp Ile Tyr 290 295 300Val His Arg Ile Val Gly Tyr Ile Gly Asn Tyr Thr Ala Gln Met Gly305 310 315 320Gly Val Asp Val Ile Thr Phe Thr Ala Gly Val Gly Glu Asn Asp Asp 325 330 335Val Val Arg Lys Met Val Cys Asp Lys Leu Ala Pro Phe Gly Val Lys 340 345 350Leu Asp Glu Glu Lys Asn Ala Thr Arg Ser Lys Glu Pro Arg Ile Ile 355 360 365Ser Thr Pro Asp Ser Ala Val Thr Ile Cys Val Ile Pro Thr Asn Glu 370 375 380Glu Leu Ala Ile Ala Arg Lys Ser Ala Ala Ile Ala Glu Glu Gly Lys385 390 395 400Asp Ser Tyr Gly Asn Val Phe Ser Lys 405176671DNASaccharomyces cerevisiaeALD2 17atgtggcaag agtcacagta acgacaaaag ttaaagaaga cagatgtgca tggagagttt 60catatgcaga aggtaagatg ttggaaagcc ccaaacctgt tgctggaaaa gacggtacca 120cgatcctagt tgaagacctt tttttcaata ttccttctag attaagggcc ttgaggtccc 180ataatgatga atactctaaa atattagatg ttgtcgggcg atacgccatt cattccaagg 240acattggctt ttcttgtaaa aagttcggag actctaatta ttctttatca gttaaacctt 300catataccgt ccaggatagg attaggactg tgttcaataa atctgtggct tcgaatttaa 360ttacttttca tatcagcaaa gtagaagatt taaacctgga aagcgttgat ggaaaggtgt 420gtaatttgaa tttcatatcc aaaaagtcca tttcaccaat ttttttcatt aataatagac 480tagtgacatg tgatcttcta agaagagctt tgaacagcgt ttactccaat tatctgccaa 540agggcaacag accttttatt tatttgggaa ttgttataga tccggcggct gttgatgtta 600acgttcaccc gacaaagaga gaggttcgtt tcctgagcca agatgagatc atagagaaaa 660tcgccaatca attgcacgcc gaattatctg ccattgatac ttcacgtact ttcaaggctt 720cttcaatttc aacaaacaag ccagagtcat tgataccatt taatgacacc atagaaagtg 780ataggaatag gaagagtctc cgacaagccc aagtggtaga gaattcatat acgacagcca 840atagtcaact aaggaaagcg aaaagacaag agaataaact agtcagaata gatgcttcac 900aagctaaaat tacgtcattt ttatcctcaa gtcaacagtt caactttgaa ggatcgtcta 960caaagcgaca actgagtgaa cccaaggtaa caaatgtaag ccactcccaa gaggcagaaa 1020agctgacact aaatgaaagc gaacaaccgc gtgatgccaa tacaatcaat gataatgact 1080tgaaggatca acctaagaag aaacaaaagt tgggggatta taaagttcca agcattgccg 1140atgacgaaaa gaatgcactc ccgatttcaa aagacgggta tattagagta cctaaggagc 1200gagttaatgt taatcttacg agtatcaaga aattgcgtga aaaagtagat gattcgatac 1260atcgagaact aacagacatt tttgcaaatt tgaattacgt tggggttgta gatgaggaaa 1320gaagattagc cgctattcag catgacttaa agcttttttt aatagattac ggatctgtgt 1380gctatgagct attctatcag attggtttga cagacttcgc aaactttggt aagataaacc 1440tacagagtac aaatgtgtca gatgatatag ttttgtataa tctcctatca gaatttgacg 1500agttaaatga cgatgcttcc aaagaaaaaa taattagtaa aatatgggac atgagcagta 1560tgctaaatga gtactattcc atagaattgg tgaatgatgg tctagataat gacttaaagt 1620ctgtgaagct aaaatctcta ccactacttt taaaaggcta cattccatct ctggtcaagt 1680taccattttt tatatatcgc ctgggtaaag aagttgattg ggaggatgaa caagagtgtc 1740tagatggtat tttaagagag attgcattac tctatatacc tgatatggtt ccgaaagtcg 1800atacatctga tgcatcgttg tcagaagacg aaaaagccca gtttataaat agaaaggaac 1860acatatcctc attactagaa cacgttctct tcccttgtat caaacgaagg ttcctggccc 1920ctagacacat tctcaaggat gtcgtggaaa tagccaacct tccagatcta tacaaagttt 1980ttgagaggtg ttaactttaa aacgttttgg ctgtaatacc aaagtttttt gtttatttcc 2040tgagtgtgat tgtgtttcat ttgaaagtgt atgccctttc ctttaacgat tcatccgcga 2100gatttcaaag gatatgaaat atggttgcag ttaggaaagt atgtcagaaa tgttatattc 2160ggattgaaac tcttctacaa tagttctgaa gtcacttggt tccgtattgt tttcgtcctc 2220ttcctcaagc aacgattctt gtctaagctt attcaacggt accaaagacc cgagtccttt 2280tatgagagaa aacatttcat catttttcaa ctcaattatc ttaatatcat tttgtagtat 2340tttgaaaaca ggatggtaaa acgaatcacc tgaatctaga agctgtacct tgtcccataa 2400aagttttaat ttactgagcc tttcggtcaa gtaaactagt ttatctagtt ttgaaccgaa 2460tattgtgggc agatttgcag taagttcagt tagatctact aaaagttgtt tgacagcagc 2520cgattccaca aaaatttggt aaaaggagat gaaagagacc acgcgcgtaa tggtttgcat 2580caccatcgga tgtctgttga aaaactcact ttttgcatgg aagttattaa caataagact 2640aatgattacc ttagaataat gtataacctt tagtagtgaa tctgcggtat cataataaat 2700gagatacatc ttaaacaatt tccaggaaac agtagatatt tgaaaaagtg caataaattt 2760ttgaaatctg ctgttatttg atctaaaatc ttcttccaac gaatctactt tgtattgaaa 2820aatatcgagc tccctcctta tagcatccaa tgtttttaag ggaggtttag aactcttgtt 2880caagtattga tctaaatctc tgtctaatga gatgattttc caatacaaga cctctgtgct 2940gttttcttcc ctgtaaacat caacgttagg caaatcctgc aaatatgctg catgttgtaa 3000tagggaggat acttctgtat gaagtgcaat tggtttgcgt ggccccgatt gaagataatc 3060gcatccacac aggcgataga atattcttcc caaggttact ttagcgatgc tttcaacggg 3120gtcatcatta agtaaaggtc taaagtcatc cacatgaaaa tttttaaagg tatggatgca 3180ctgtgttagg agcgaatttg ccaacaacgg ttcatcgtag gggaaagttg tagtggctaa 3240aatcagatat gcttgaacga gccttatatc gggatgcgcc atgaaatcac attgtttcaa 3300ttggaataat gagcagcgtg caaaattttg gcacatgact aattgcatac catcttccca 3360attgagcctt ttgttgctac ccaaatattc atggagagga tatactgaaa atatttctgc 3420taacttctcg actggcatat aataaatgca catggtaaag acagaccata ttagtgcgtc 3480ccagtatttg ccgtctacac tgtgattctt gtctgccctt ctatcccagt actgttccac 3540cctcaaatat aattcactga tatctccaat ggaaccaaaa tacaaagcac ctaagttttc 3600catagcaaag ttcatcaatt ggaatgattg gtctttttga agataattca tccaaaacat 3660acattcttca gtatcagtat ccaaattagc tggtgttcta gtaagatctc tttgttttgt 3720tttgtacagc ccaatattcg ttatccaata ttcataattt tgccagaata acaataggtc 3780agggagatat tccttggaag atgaagccgt tattagtttt gttgatttca tacattcgga 3840gtcctgtcct ctcttcctgc agtttccaca cggtatcata cgatcacatt tgacttttcg 3900cctcgtacac acggaacagg gatgcttgga tttcagttga gtggtacggt taaacattat 3960tattaggatg caaactcgtg ttcgtaactt ttgtcatcac ctgctctctg aaggttggtt 4020ataatatttc attctcttac gcttagctta catggctgcg cgcacttcat ttaaagccgt 4080aaaaaataaa gatttcatag aatatgaagc tttgaatctc atgtaaacat ttatataact 4140atgtaacgaa ttatttatcc aatgaaagat ccacatggac tgattttatt tgtaaatagt 4200tatcaacgcc ggtgtcgcct gattctctac caataccact catcttaaat ccgccaaaag 4260gaactttagc ttcttcttga ttggtttgat tgatccaaac agttcctgct ttaatatcgc 4320gagcaaacat gtgcgctttc ttgacatctt tggtgaagac cgcagaggcg agcccgtagc 4380aagtatcatt agccagcttc agagcgtcat cataatttgt gaacttgcta acaaccacaa 4440ccgggccaaa tatttcatca cgcagcaact tagatgtttc tggtacatca gtgaagatgg 4500ttgggggaat gaagtagcct ttagctccac caataggaaa ttcagaggtc tggaacatgt 4560ccaacttttc ctccttttta ccacgttcta tgtaactttt gatgcggtca tactgtgtac 4620ttgatataac tggaccaacg atgcatttct catcaaacgg atcaaatttt cctgcaacat 4680cccactcctt ctttgcagtt tctttaaact tttcaacaaa cttgtcgtag atcgaacttt 4740gaacataaac tcttgagttt gcagtgcaga tctgtcccga attaaaaaaa ataccatttg 4800ctacccattc tatagcctta tcaaggtctg catcttcaaa tacaagagca ggagacttac 4860caccgcattc tagtgtgata tccttaaggt tcgattggcc ggaagcttcc aatactgagc 4920cgccaacctt agtacttccc gtaaaagata ttttgtcgat atccatgtgg gttcctaaag 4980ctttccccac aacggaacca taaccaggaa tgacattgac aacaccaggt ggaaaacctg 5040cttttttaat taaagtagca aaataaagta gagatagaga ggtattttca gcaggtttga 5100tgataaccgt gttaccggcc gctaaggcac cttgcatttt tctacaagcc atagctagag 5160gataattcca tggaacgatt tgagcaacaa cgccaaaagg aacttttaga gtatatgcaa 5220acttgttaaa agtcaatgga atggtttcac ccatattgaa cttgtcgacc gcccccgcat 5280agtatcttgt aagttctata atctgggcta agtcttgttt agcattggaa tggaaaggct 5340taccagcgtc taaagtctct aatgcggcaa gtgtgtcttg ctcctcctca ataagtttta 5400ataagtttga aagataaata ccacgttgct cagaagatgt cttcgaccaa acgttatcaa 5460aagcagccct ggcagctttc acagctttgt ctacatcctt ttcgttagct gcttggaagg 5520atgttatcgg ttcgccagta gctgggttca cagtttcgat ggtctttcca tctgatgatg 5580gacaaaactc attgttgata aacaacccta gcggttgctt taaagagatt ttcaattgtg 5640ggatttcgat atcagtatac aaggtaggca tttttctttt ggctaatttt ctaaatgtgt 5700ataatctata tctctatgat acaagtccaa ggtataatta cctggcatga gtcaaattta 5760ttcccatcca gatttgccat tttatacaat tagtttctga ggtatgatca gttaaatcta 5820cgctattcgt ttgaacaatc tccctgtctc ctccctaaac ttactaattc cttaaccctt 5880agggagttta gcaatcagct taagaagggg gctgatcaac aatccggttc aagtcttgga 5940ttagcagtaa tgaaaatccc taaatgcact aaagggcgtg gaagaaaggt actgtagagt 6000tctaatagtg atactttcct tgtactccag ggggggaagg aagaacagac gtcatatgca 6060gaaaaagaaa aaaaaaagac ggaagatcta cgtaatggtg ggaatagaga aaactaacac 6120gcaagtataa tttcttgaca caaaaataaa tctcactgag tttatagctc tattttttta 6180ccatcttaag ggatttctag gacctagtaa caacaattgt tccgcatgct gaaataatgg 6240tgcttacagt tttatatcga gtagtggact actactgttt gctagactgt caacatattc 6300atccgtcgtc aggctaagct atttaggtac ttacgcactt acataatgat aactatcaca 6360ttagttgtcc aaagagagat ttatgtgaac tgcttttgtt tgaagatagg tatcaacacc 6420actttgcccc agttctctac caataccact cattttaaac ccgccaaaag gaacggtaac 6480atcttcatcg ttagatgagt tgatccaaac agttcctgct ttaatatcgc gagcaaacat 6540gtgcgctttc ttgacatctt ttgtgaagac cgcagaggcg agcccgtagc aagtatcatt 6600agccagcttc agagcgtcat cataatttgt gaacttgcta acaaccacaa ccgggccaaa 6660tatctcatcc t 667118506PRTSaccharomyces cerevisiaeALD2 18Met Pro Thr Leu Tyr Thr Asp Ile Glu Ile Pro Gln Leu Lys Ile Ser1 5 10 15Leu Lys Gln Pro Leu Gly Leu Phe Ile Asn Asn Glu Phe Cys Pro Ser 20

25 30Ser Asp Gly Lys Thr Ile Glu Thr Val Asn Pro Ala Thr Gly Glu Pro 35 40 45Ile Thr Ser Phe Gln Ala Ala Asn Glu Lys Asp Val Asp Lys Ala Val 50 55 60Lys Ala Ala Arg Ala Ala Phe Asp Asn Val Trp Ser Lys Thr Ser Ser65 70 75 80Glu Gln Arg Gly Ile Tyr Leu Ser Asn Leu Leu Lys Leu Ile Glu Glu 85 90 95Glu Gln Asp Thr Leu Ala Ala Leu Glu Thr Leu Asp Ala Gly Lys Pro 100 105 110Phe His Ser Asn Ala Lys Gln Asp Leu Ala Gln Ile Ile Glu Leu Thr 115 120 125Arg Tyr Tyr Ala Gly Ala Val Asp Lys Phe Asn Met Gly Glu Thr Ile 130 135 140Pro Leu Thr Phe Asn Lys Phe Ala Tyr Thr Leu Lys Val Pro Phe Gly145 150 155 160Val Val Ala Gln Ile Val Pro Trp Asn Tyr Pro Leu Ala Met Ala Cys 165 170 175Arg Lys Met Gln Gly Ala Leu Ala Ala Gly Asn Thr Val Ile Ile Lys 180 185 190Pro Ala Glu Asn Thr Ser Leu Ser Leu Leu Tyr Phe Ala Thr Leu Ile 195 200 205Lys Lys Ala Gly Phe Pro Pro Gly Val Val Asn Val Ile Pro Gly Tyr 210 215 220Gly Ser Val Val Gly Lys Ala Leu Gly Thr His Met Asp Ile Asp Lys225 230 235 240Ile Ser Phe Thr Gly Ser Thr Lys Val Gly Gly Ser Val Leu Glu Ala 245 250 255Ser Gly Gln Ser Asn Leu Lys Asp Ile Thr Leu Glu Cys Gly Gly Lys 260 265 270Ser Pro Ala Leu Val Phe Glu Asp Ala Asp Leu Asp Lys Ala Ile Glu 275 280 285Trp Val Ala Asn Gly Ile Phe Phe Asn Ser Gly Gln Ile Cys Thr Ala 290 295 300Asn Ser Arg Val Tyr Val Gln Ser Ser Ile Tyr Asp Lys Phe Val Glu305 310 315 320Lys Phe Lys Glu Thr Ala Lys Lys Glu Trp Asp Val Ala Gly Lys Phe 325 330 335Asp Pro Phe Asp Glu Lys Cys Ile Val Gly Pro Val Ile Ser Ser Thr 340 345 350Gln Tyr Asp Arg Ile Lys Ser Tyr Ile Glu Arg Gly Lys Lys Glu Glu 355 360 365Lys Leu Asp Met Phe Gln Thr Ser Glu Phe Pro Ile Gly Gly Ala Lys 370 375 380Gly Tyr Phe Ile Pro Pro Thr Ile Phe Thr Asp Val Pro Glu Thr Ser385 390 395 400Lys Leu Leu Arg Asp Glu Ile Phe Gly Pro Val Val Val Val Ser Lys 405 410 415Phe Thr Asn Tyr Asp Asp Ala Leu Lys Leu Ala Asn Asp Thr Cys Tyr 420 425 430Gly Leu Ala Ser Ala Val Phe Thr Lys Asp Val Lys Lys Ala His Met 435 440 445Phe Ala Arg Asp Ile Lys Ala Gly Thr Val Trp Ile Asn Gln Thr Asn 450 455 460Gln Glu Glu Ala Lys Val Pro Phe Gly Gly Phe Lys Met Ser Gly Ile465 470 475 480Gly Arg Glu Ser Gly Asp Thr Gly Val Asp Asn Tyr Leu Gln Ile Lys 485 490 495Ser Val His Val Asp Leu Ser Leu Asp Lys 500 505191824DNASaccharomyces cerevisiaeALD3 19atgtttaacc gtaccactca actgaaatcc aagcatccct gttccgtgtg tacgaggcga 60aaagtcaaat gtgatcgtat gataccgtgt ggaaactgca ggaagagagg acaggactcc 120gaatgtatga aatcaacaaa actaataacg gcttcatctt ccaaggaata tctccctgac 180ctattgttat tctggcaaaa ttatgaatat tggataacga atattgggct gtacaaaaca 240aaacaaagag atcttactag aacaccagct aatttggata ctgatactga agaatgtatg 300ttttggatga attatcttca aaaagaccaa tcattccaat tgatgaactt tgctatggaa 360aacttaggtg ctttgtattt tggttccatt ggagatatca gtgaattata tttgagggtg 420gaacagtact gggatagaag ggcagacaag aatcacagtg tagacggcaa atactgggac 480gcactaatat ggtctgtctt taccatgtgc atttattata tgccagtcga gaagttagca 540gaaatatttt cagtatatcc tctccatgaa tatttgggta gcaacaaaag gctcaattgg 600gaagatggta tgcaattagt catgtgccaa aattttgcac gctgctcatt attccaattg 660aaacaatgtg atttcatggc gcatcccgat ataaggctcg ttcaagcata tctgatttta 720gccactacaa ctttccccta cgatgaaccg ttgttggcaa attcgctcct aacacagtgc 780atccatacct ttaaaaattt tcatgtggat gactttagac ctttacttaa tgatgacccc 840gttgaaagca tcgctaaagt aaccttggga agaatattct atcgcctgtg tggatgcgat 900tatcttcaat cggggccacg caaaccaatt gcacttcata cagaagtatc ctccctatta 960caacatgcag catatttgca ggatttgcct aacgttgatg tttacaggga agaaaacagc 1020acagaggtct tgtattggaa aatcatctca ttagacagag atttagatca atacttgaac 1080aagagttcta aacctccctt aaaaacattg gatgctataa ggagggagct cgatattttt 1140caatacaaag tagattcgtt ggaagaagat tttagatcaa ataacagcag atttcaaaaa 1200tttattgcac tttttcaaat atctactgtt tcctggaaat tgtttaagat gtatctcatt 1260tattatgata ccgcagattc actactaaag gttatacatt attctaaggt aatcattagt 1320cttattgtta ataacttcca tgcaaaaagt gagtttttca acagacatcc gatggtgatg 1380caaaccatta cgcgcgtggt ctctttcatc tccttttacc aaatttttgt ggaatcggct 1440gctgtcaaac aacttttagt agatctaact gaacttactg caaatctgcc cacaatattc 1500ggttcaaaac tagataaact agtttacttg accgaaaggc tcagtaaatt aaaactttta 1560tgggacaagg tacagcttct agattcaggt gattcgtttt accatcctgt tttcaaaata 1620ctacaaaatg atattaagat aattgagttg aaaaatgatg aaatgttttc tctcataaaa 1680ggactcgggt ctttggtacc gttgaataag cttagacaag aatcgttgct tgaggaagag 1740gacgaaaaca atacggaacc aagtgacttc agaactattg tagaagagtt tcaatccgaa 1800tataacattt ctgacatact ttcc 182420608PRTSaccharomyces cerevisiaeALD3 20Met Phe Asn Arg Thr Thr Gln Leu Lys Ser Lys His Pro Cys Ser Val1 5 10 15Cys Thr Arg Arg Lys Val Lys Cys Asp Arg Met Ile Pro Cys Gly Asn 20 25 30Cys Arg Lys Arg Gly Gln Asp Ser Glu Cys Met Lys Ser Thr Lys Leu 35 40 45Ile Thr Ala Ser Ser Ser Lys Glu Tyr Leu Pro Asp Leu Leu Leu Phe 50 55 60Trp Gln Asn Tyr Glu Tyr Trp Ile Thr Asn Ile Gly Leu Tyr Lys Thr65 70 75 80Lys Gln Arg Asp Leu Thr Arg Thr Pro Ala Asn Leu Asp Thr Asp Thr 85 90 95Glu Glu Cys Met Phe Trp Met Asn Tyr Leu Gln Lys Asp Gln Ser Phe 100 105 110Gln Leu Met Asn Phe Ala Met Glu Asn Leu Gly Ala Leu Tyr Phe Gly 115 120 125Ser Ile Gly Asp Ile Ser Glu Leu Tyr Leu Arg Val Glu Gln Tyr Trp 130 135 140Asp Arg Arg Ala Asp Lys Asn His Ser Val Asp Gly Lys Tyr Trp Asp145 150 155 160Ala Leu Ile Trp Ser Val Phe Thr Met Cys Ile Tyr Tyr Met Pro Val 165 170 175Glu Lys Leu Ala Glu Ile Phe Ser Val Tyr Pro Leu His Glu Tyr Leu 180 185 190Gly Ser Asn Lys Arg Leu Asn Trp Glu Asp Gly Met Gln Leu Val Met 195 200 205Cys Gln Asn Phe Ala Arg Cys Ser Leu Phe Gln Leu Lys Gln Cys Asp 210 215 220Phe Met Ala His Pro Asp Ile Arg Leu Val Gln Ala Tyr Leu Ile Leu225 230 235 240Ala Thr Thr Thr Phe Pro Tyr Asp Glu Pro Leu Leu Ala Asn Ser Leu 245 250 255Leu Thr Gln Cys Ile His Thr Phe Lys Asn Phe His Val Asp Asp Phe 260 265 270Arg Pro Leu Leu Asn Asp Asp Pro Val Glu Ser Ile Ala Lys Val Thr 275 280 285Leu Gly Arg Ile Phe Tyr Arg Leu Cys Gly Cys Asp Tyr Leu Gln Ser 290 295 300Gly Pro Arg Lys Pro Ile Ala Leu His Thr Glu Val Ser Ser Leu Leu305 310 315 320Gln His Ala Ala Tyr Leu Gln Asp Leu Pro Asn Val Asp Val Tyr Arg 325 330 335Glu Glu Asn Ser Thr Glu Val Leu Tyr Trp Lys Ile Ile Ser Leu Asp 340 345 350Arg Asp Leu Asp Gln Tyr Leu Asn Lys Ser Ser Lys Pro Pro Leu Lys 355 360 365Thr Leu Asp Ala Ile Arg Arg Glu Leu Asp Ile Phe Gln Tyr Lys Val 370 375 380Asp Ser Leu Glu Glu Asp Phe Arg Ser Asn Asn Ser Arg Phe Gln Lys385 390 395 400Phe Ile Ala Leu Phe Gln Ile Ser Thr Val Ser Trp Lys Leu Phe Lys 405 410 415Met Tyr Leu Ile Tyr Tyr Asp Thr Ala Asp Ser Leu Leu Lys Val Ile 420 425 430His Tyr Ser Lys Val Ile Ile Ser Leu Ile Val Asn Asn Phe His Ala 435 440 445Lys Ser Glu Phe Phe Asn Arg His Pro Met Val Met Gln Thr Ile Thr 450 455 460Arg Val Val Ser Phe Ile Ser Phe Tyr Gln Ile Phe Val Glu Ser Ala465 470 475 480Ala Val Lys Gln Leu Leu Val Asp Leu Thr Glu Leu Thr Ala Asn Leu 485 490 495Pro Thr Ile Phe Gly Ser Lys Leu Asp Lys Leu Val Tyr Leu Thr Glu 500 505 510Arg Leu Ser Lys Leu Lys Leu Leu Trp Asp Lys Val Gln Leu Leu Asp 515 520 525Ser Gly Asp Ser Phe Tyr His Pro Val Phe Lys Ile Leu Gln Asn Asp 530 535 540Ile Lys Ile Ile Glu Leu Lys Asn Asp Glu Met Phe Ser Leu Ile Lys545 550 555 560Gly Leu Gly Ser Leu Val Pro Leu Asn Lys Leu Arg Gln Glu Ser Leu 565 570 575Leu Glu Glu Glu Asp Glu Asn Asn Thr Glu Pro Ser Asp Phe Arg Thr 580 585 590Ile Val Glu Glu Phe Gln Ser Glu Tyr Asn Ile Ser Asp Ile Leu Ser 595 600 605211557DNASaccharomyces cerevisiaeALD4 21atgttcagta gatctacgct ctgcttaaag acgtctgcat cctccattgg gagacttcaa 60ttgagatatt tctcacacct tcctatgaca gtgcctatca agctgcccaa tgggttggaa 120tatgagcaac caacggggtt gttcatcaac aacaagtttg ttccttctaa acagaacaag 180accttcgaag tcattaaccc ttccacggaa gaagaaatat gtcatattta tgaaggtaga 240gaggacgatg tggaagaggc cgtgcaggcc gccgaccgtg ccttctctaa tgggtcttgg 300aacggtatcg accctattga caggggtaag gctttgtaca ggttagccga attaattgaa 360caggacaagg atgtcattgc ttccatcgag actttggata acggtaaagc tatctcttcc 420tcgagaggag atgttgattt agtcatcaac tatttgaaat cttctgctgg ctttgctgat 480aaaattgatg gtagaatgat tgatactggt agaacccatt tttcttacac taagagacag 540cctttgggtg tttgtgggca gattattcct tggaatttcc cactgttgat gtgggcctgg 600aagattgccc ctgctttggt caccggtaac accgtcgtgt tgaagactgc cgaatccacc 660ccattgtccg ctttgtatgt gtctaaatac atcccacagg cgggtattcc acctggtgtg 720atcaacattg tatccgggtt tggtaagatt gtgggtgagg ccattacaaa ccatccaaaa 780atcaaaaagg ttgccttcac agggtccacg gctacgggta gacacattta ccagtccgca 840gccgcaggct tgaaaaaagt gactttggag ctgggtggta aatcaccaaa cattgtcttc 900gcggacgccg agttgaaaaa agccgtgcaa aacattatcc ttggtatcta ctacaattct 960ggtgaggtct gttgtgcggg ttcaagggtg tatgttgaag aatctattta cgacaaattc 1020attgaagagt tcaaagccgc ttctgaatcc atcaaggtgg gcgacccatt cgatgaatct 1080actttccaag gtgcacaaac ctctcaaatg caactaaaca aaatcttgaa atacgttgac 1140attggtaaga atgaaggtgc tactttgatt accggtggtg aaagattagg tagcaagggt 1200tacttcatta agccaactgt ctttggtgac gttaaggaag acatgagaat tgtcaaagag 1260gaaatctttg gccctgttgt cactgtaacc aaattcaaat ctgccgacga agtcattaac 1320atggcgaacg attctgaata cgggttggct gctggtattc acacctctaa tattaatacc 1380gccttaaaag tggctgatag agttaatgcg ggtacggtct ggataaacac ttataacgat 1440ttccaccacg cagttccttt cggtgggttc aatgcatctg gtttgggcag ggaaatgtct 1500gttgatgctt tacaaaacta cttgcaagtt aaagcggtcc gtgccaaatt ggacgag 155722519PRTSaccharomyces cerevisiaeALD4 22Met Phe Ser Arg Ser Thr Leu Cys Leu Lys Thr Ser Ala Ser Ser Ile1 5 10 15Gly Arg Leu Gln Leu Arg Tyr Phe Ser His Leu Pro Met Thr Val Pro 20 25 30Ile Lys Leu Pro Asn Gly Leu Glu Tyr Glu Gln Pro Thr Gly Leu Phe 35 40 45Ile Asn Asn Lys Phe Val Pro Ser Lys Gln Asn Lys Thr Phe Glu Val 50 55 60Ile Asn Pro Ser Thr Glu Glu Glu Ile Cys His Ile Tyr Glu Gly Arg65 70 75 80Glu Asp Asp Val Glu Glu Ala Val Gln Ala Ala Asp Arg Ala Phe Ser 85 90 95Asn Gly Ser Trp Asn Gly Ile Asp Pro Ile Asp Arg Gly Lys Ala Leu 100 105 110Tyr Arg Leu Ala Glu Leu Ile Glu Gln Asp Lys Asp Val Ile Ala Ser 115 120 125Ile Glu Thr Leu Asp Asn Gly Lys Ala Ile Ser Ser Ser Arg Gly Asp 130 135 140Val Asp Leu Val Ile Asn Tyr Leu Lys Ser Ser Ala Gly Phe Ala Asp145 150 155 160Lys Ile Asp Gly Arg Met Ile Asp Thr Gly Arg Thr His Phe Ser Tyr 165 170 175Thr Lys Arg Gln Pro Leu Gly Val Cys Gly Gln Ile Ile Pro Trp Asn 180 185 190Phe Pro Leu Leu Met Trp Ala Trp Lys Ile Ala Pro Ala Leu Val Thr 195 200 205Gly Asn Thr Val Val Leu Lys Thr Ala Glu Ser Thr Pro Leu Ser Ala 210 215 220Leu Tyr Val Ser Lys Tyr Ile Pro Gln Ala Gly Ile Pro Pro Gly Val225 230 235 240Ile Asn Ile Val Ser Gly Phe Gly Lys Ile Val Gly Glu Ala Ile Thr 245 250 255Asn His Pro Lys Ile Lys Lys Val Ala Phe Thr Gly Ser Thr Ala Thr 260 265 270Gly Arg His Ile Tyr Gln Ser Ala Ala Ala Gly Leu Lys Lys Val Thr 275 280 285Leu Glu Leu Gly Gly Lys Ser Pro Asn Ile Val Phe Ala Asp Ala Glu 290 295 300Leu Lys Lys Ala Val Gln Asn Ile Ile Leu Gly Ile Tyr Tyr Asn Ser305 310 315 320Gly Glu Val Cys Cys Ala Gly Ser Arg Val Tyr Val Glu Glu Ser Ile 325 330 335Tyr Asp Lys Phe Ile Glu Glu Phe Lys Ala Ala Ser Glu Ser Ile Lys 340 345 350Val Gly Asp Pro Phe Asp Glu Ser Thr Phe Gln Gly Ala Gln Thr Ser 355 360 365Gln Met Gln Leu Asn Lys Ile Leu Lys Tyr Val Asp Ile Gly Lys Asn 370 375 380Glu Gly Ala Thr Leu Ile Thr Gly Gly Glu Arg Leu Gly Ser Lys Gly385 390 395 400Tyr Phe Ile Lys Pro Thr Val Phe Gly Asp Val Lys Glu Asp Met Arg 405 410 415Ile Val Lys Glu Glu Ile Phe Gly Pro Val Val Thr Val Thr Lys Phe 420 425 430Lys Ser Ala Asp Glu Val Ile Asn Met Ala Asn Asp Ser Glu Tyr Gly 435 440 445Leu Ala Ala Gly Ile His Thr Ser Asn Ile Asn Thr Ala Leu Lys Val 450 455 460Ala Asp Arg Val Asn Ala Gly Thr Val Trp Ile Asn Thr Tyr Asn Asp465 470 475 480Phe His His Ala Val Pro Phe Gly Gly Phe Asn Ala Ser Gly Leu Gly 485 490 495Arg Glu Met Ser Val Asp Ala Leu Gln Asn Tyr Leu Gln Val Lys Ala 500 505 510Val Arg Ala Lys Leu Asp Glu 515231560DNASaccharomyces cerevisiaeALD5 23atgctttctc gcacaagagc tgcagctccg aattccagaa tattcactag aagcttgtta 60cgtctttatt ctcaagcacc attacgcgtt ccaattactc ttccaaatgg tttcacctac 120gaacagccaa cagggttatt catcaatggt gaatttgttg cctcgaagca aaagaaaacg 180tttgacgtga tcaatccatc taacgaagaa aagataacaa ctgtatacaa ggctatggaa 240gatgatgttg atgaagccgt tgcagcggct aaaaaagctt ttgaaacgaa gtggtctatt 300gtagagccgg aggttcgcgc taaagcttta ttcaatctcg ctgacttggt tgagaaacac 360caagaaacac tggctgccat tgagtcaatg gataatggta agtcattgtt ttgtgcgcgc 420ggtgacgtcg ctttagtatc taaatacttg cgttcttgcg gtggttgggc agataaaatc 480tacggtaacg ttattgacac aggtaaaaac cattttacct actcaattaa ggaaccatta 540ggcgtttgcg gccaaataat cccttggaac ttccctttat tgatgtggtc atggaaaatt 600gggcctgctc tggctacagg taacaccgtc gtattgaaac ccgctgaaac aacaccttta 660tctgcccttt tcgcttccca gttgtgtcag gaagcaggca tacccgctgg tgtagtcaat 720atccttccgg gttccggtag agttgttgga gaaagattga gtgcacaccc agacgtgaag 780aagattgctt ttacaggctc tactgccacc ggccgccata ttatgaaggt cgctgccgat 840actgtcaaga aagtcacttt ggagctggga ggtaaatcac caaatattgt gtttgctgac 900gctgatctag ataaagccgt caagaacatt gccttcggta ttttttacaa ctctggtgaa 960gtttgctgcg ctggttccag aatatacatt caagatacag tatacgagga ggtgttgcaa 1020aaactaaagg attacaccga gtcactaaag gtcggtgacc catttgatga ggaagttttc 1080caaggtgctc aaacatctga caaacagctg cataaaattt tagactatgt cgatgtagca 1140aaatcagagg gggctcgtct tgtgactgga ggggccagac atggcagtaa aggttatttt 1200gtcaagccaa cagtgtttgc tgatgtcaaa gaagatatga gaattgttaa ggaggaagtg 1260tttggtccca ttgtaactgt atccaagttt tctactgttg atgaagtgat tgctatggca 1320aatgattctc aatatgggtt agccgcaggt attcacacta acgatattaa caaggctgtt 1380gatgtgtcca aaagagtgaa agctggtact gtttggataa atacctataa caacttccac 1440caaaatgttc ctttcggtgg cttcggccag tcaggtattg gccgtgaaat gggtgaggct 1500gctttaagta actacactca aacaaaatct gtcagaattg ccattgacaa gccaattcgt 156024520PRTSaccharomyces cerevisiaeALD5 24Met Leu Ser Arg Thr Arg Ala Ala Ala Pro Asn Ser Arg Ile Phe Thr1 5 10 15Arg Ser

Leu Leu Arg Leu Tyr Ser Gln Ala Pro Leu Arg Val Pro Ile 20 25 30Thr Leu Pro Asn Gly Phe Thr Tyr Glu Gln Pro Thr Gly Leu Phe Ile 35 40 45Asn Gly Glu Phe Val Ala Ser Lys Gln Lys Lys Thr Phe Asp Val Ile 50 55 60Asn Pro Ser Asn Glu Glu Lys Ile Thr Thr Val Tyr Lys Ala Met Glu65 70 75 80Asp Asp Val Asp Glu Ala Val Ala Ala Ala Lys Lys Ala Phe Glu Thr 85 90 95Lys Trp Ser Ile Val Glu Pro Glu Val Arg Ala Lys Ala Leu Phe Asn 100 105 110Leu Ala Asp Leu Val Glu Lys His Gln Glu Thr Leu Ala Ala Ile Glu 115 120 125Ser Met Asp Asn Gly Lys Ser Leu Phe Cys Ala Arg Gly Asp Val Ala 130 135 140Leu Val Ser Lys Tyr Leu Arg Ser Cys Gly Gly Trp Ala Asp Lys Ile145 150 155 160Tyr Gly Asn Val Ile Asp Thr Gly Lys Asn His Phe Thr Tyr Ser Ile 165 170 175Lys Glu Pro Leu Gly Val Cys Gly Gln Ile Ile Pro Trp Asn Phe Pro 180 185 190Leu Leu Met Trp Ser Trp Lys Ile Gly Pro Ala Leu Ala Thr Gly Asn 195 200 205Thr Val Val Leu Lys Pro Ala Glu Thr Thr Pro Leu Ser Ala Leu Phe 210 215 220Ala Ser Gln Leu Cys Gln Glu Ala Gly Ile Pro Ala Gly Val Val Asn225 230 235 240Ile Leu Pro Gly Ser Gly Arg Val Val Gly Glu Arg Leu Ser Ala His 245 250 255Pro Asp Val Lys Lys Ile Ala Phe Thr Gly Ser Thr Ala Thr Gly Arg 260 265 270His Ile Met Lys Val Ala Ala Asp Thr Val Lys Lys Val Thr Leu Glu 275 280 285Leu Gly Gly Lys Ser Pro Asn Ile Val Phe Ala Asp Ala Asp Leu Asp 290 295 300Lys Ala Val Lys Asn Ile Ala Phe Gly Ile Phe Tyr Asn Ser Gly Glu305 310 315 320Val Cys Cys Ala Gly Ser Arg Ile Tyr Ile Gln Asp Thr Val Tyr Glu 325 330 335Glu Val Leu Gln Lys Leu Lys Asp Tyr Thr Glu Ser Leu Lys Val Gly 340 345 350Asp Pro Phe Asp Glu Glu Val Phe Gln Gly Ala Gln Thr Ser Asp Lys 355 360 365Gln Leu His Lys Ile Leu Asp Tyr Val Asp Val Ala Lys Ser Glu Gly 370 375 380Ala Arg Leu Val Thr Gly Gly Ala Arg His Gly Ser Lys Gly Tyr Phe385 390 395 400Val Lys Pro Thr Val Phe Ala Asp Val Lys Glu Asp Met Arg Ile Val 405 410 415Lys Glu Glu Val Phe Gly Pro Ile Val Thr Val Ser Lys Phe Ser Thr 420 425 430Val Asp Glu Val Ile Ala Met Ala Asn Asp Ser Gln Tyr Gly Leu Ala 435 440 445Ala Gly Ile His Thr Asn Asp Ile Asn Lys Ala Val Asp Val Ser Lys 450 455 460Arg Val Lys Ala Gly Thr Val Trp Ile Asn Thr Tyr Asn Asn Phe His465 470 475 480Gln Asn Val Pro Phe Gly Gly Phe Gly Gln Ser Gly Ile Gly Arg Glu 485 490 495Met Gly Glu Ala Ala Leu Ser Asn Tyr Thr Gln Thr Lys Ser Val Arg 500 505 510Ile Ala Ile Asp Lys Pro Ile Arg 515 520251503DNASaccharomyces cerevisiaeALD6 25atgactaagc tacactttga cactgctgaa ccagtcaaga tcacacttcc aaatggtttg 60acatacgagc aaccaaccgg tctattcatt aacaacaagt ttatgaaagc tcaagacggt 120aagacctatc ccgtcgaaga tccttccact gaaaacaccg tttgtgaggt ctcttctgcc 180accactgaag atgttgaata tgctatcgaa tgtgccgacc gtgctttcca cgacactgaa 240tgggctaccc aagacccaag agaaagaggc cgtctactaa gtaagttggc tgacgaattg 300gaaagccaaa ttgacttggt ttcttccatt gaagctttgg acaatggtaa aactttggcc 360ttagcccgtg gggatgttac cattgcaatc aactgtctaa gagatgctgc tgcctatgcc 420gacaaagtca acggtagaac aatcaacacc ggtgacggct acatgaactt caccacctta 480gagccaatcg gtgtctgtgg tcaaattatt ccatggaact ttccaataat gatgttggct 540tggaagatcg ccccagcatt ggccatgggt aacgtctgta tcttgaaacc cgctgctgtc 600acacctttaa atgccctata ctttgcttct ttatgtaaga aggttggtat tccagctggt 660gtcgtcaaca tcgttccagg tcctggtaga actgttggtg ctgctttgac caacgaccca 720agaatcagaa agctggcttt taccggttct acagaagtcg gtaagagtgt tgctgtcgac 780tcttctgaat ctaacttgaa gaaaatcact ttggaactag gtggtaagtc cgcccatttg 840gtctttgacg atgctaacat taagaagact ttaccaaatc tagtaaacgg tattttcaag 900aacgctggtc aaatttgttc ctctggttct agaatttacg ttcaagaagg tatttacgac 960gaactattgg ctgctttcaa ggcttacttg gaaaccgaaa tcaaagttgg taatccattt 1020gacaaggcta acttccaagg tgctatcact aaccgtcaac aattcgacac aattatgaac 1080tacatcgata tcggtaagaa agaaggcgcc aagatcttaa ctggtggcga aaaagttggt 1140gacaagggtt acttcatcag accaaccgtt ttctacgatg ttaatgaaga catgagaatt 1200gttaaggaag aaatttttgg accagttgtc actgtcgcaa agttcaagac tttagaagaa 1260ggtgtcgaaa tggctaacag ctctgaattc ggtctaggtt ctggtatcga aacagaatct 1320ttgagcacag gtttgaaggt ggccaagatg ttgaaggccg gtaccgtctg gatcaacaca 1380tacaacgatt ttgactccag agttccattc ggtggtgtta agcaatctgg ttacggtaga 1440gaaatgggtg aagaagtcta ccatgcatac actgaagtaa aagctgtcag aattaagttg 1500taa 150326500PRTSaccharomyces cerevisiaeALD6 26Met Thr Lys Leu His Phe Asp Thr Ala Glu Pro Val Lys Ile Thr Leu1 5 10 15Pro Asn Gly Leu Thr Tyr Glu Gln Pro Thr Gly Leu Phe Ile Asn Asn 20 25 30Lys Phe Met Lys Ala Gln Asp Gly Lys Thr Tyr Pro Val Glu Asp Pro 35 40 45Ser Thr Glu Asn Thr Val Cys Glu Val Ser Ser Ala Thr Thr Glu Asp 50 55 60Val Glu Tyr Ala Ile Glu Cys Ala Asp Arg Ala Phe His Asp Thr Glu65 70 75 80Trp Ala Thr Gln Asp Pro Arg Glu Arg Gly Arg Leu Leu Ser Lys Leu 85 90 95Ala Asp Glu Leu Glu Ser Gln Ile Asp Leu Val Ser Ser Ile Glu Ala 100 105 110Leu Asp Asn Gly Lys Thr Leu Ala Leu Ala Arg Gly Asp Val Thr Ile 115 120 125Ala Ile Asn Cys Leu Arg Asp Ala Ala Ala Tyr Ala Asp Lys Val Asn 130 135 140Gly Arg Thr Ile Asn Thr Gly Asp Gly Tyr Met Asn Phe Thr Thr Leu145 150 155 160Glu Pro Ile Gly Val Cys Gly Gln Ile Ile Pro Trp Asn Phe Pro Ile 165 170 175Met Met Leu Ala Trp Lys Ile Ala Pro Ala Leu Ala Met Gly Asn Val 180 185 190Cys Ile Leu Lys Pro Ala Ala Val Thr Pro Leu Asn Ala Leu Tyr Phe 195 200 205Ala Ser Leu Cys Lys Lys Val Gly Ile Pro Ala Gly Val Val Asn Ile 210 215 220Val Pro Gly Pro Gly Arg Thr Val Gly Ala Ala Leu Thr Asn Asp Pro225 230 235 240Arg Ile Arg Lys Leu Ala Phe Thr Gly Ser Thr Glu Val Gly Lys Ser 245 250 255Val Ala Val Asp Ser Ser Glu Ser Asn Leu Lys Lys Ile Thr Leu Glu 260 265 270Leu Gly Gly Lys Ser Ala His Leu Val Phe Asp Asp Ala Asn Ile Lys 275 280 285Lys Thr Leu Pro Asn Leu Val Asn Gly Ile Phe Lys Asn Ala Gly Gln 290 295 300Ile Cys Ser Ser Gly Ser Arg Ile Tyr Val Gln Glu Gly Ile Tyr Asp305 310 315 320Glu Leu Leu Ala Ala Phe Lys Ala Tyr Leu Glu Thr Glu Ile Lys Val 325 330 335Gly Asn Pro Phe Asp Lys Ala Asn Phe Gln Gly Ala Ile Thr Asn Arg 340 345 350Gln Gln Phe Asp Thr Ile Met Asn Tyr Ile Asp Ile Gly Lys Lys Glu 355 360 365Gly Ala Lys Ile Leu Thr Gly Gly Glu Lys Val Gly Asp Lys Gly Tyr 370 375 380Phe Ile Arg Pro Thr Val Phe Tyr Asp Val Asn Glu Asp Met Arg Ile385 390 395 400Val Lys Glu Glu Ile Phe Gly Pro Val Val Thr Val Ala Lys Phe Lys 405 410 415Thr Leu Glu Glu Gly Val Glu Met Ala Asn Ser Ser Glu Phe Gly Leu 420 425 430Gly Ser Gly Ile Glu Thr Glu Ser Leu Ser Thr Gly Leu Lys Val Ala 435 440 445Lys Met Leu Lys Ala Gly Thr Val Trp Ile Asn Thr Tyr Asn Asp Phe 450 455 460Asp Ser Arg Val Pro Phe Gly Gly Val Lys Gln Ser Gly Tyr Gly Arg465 470 475 480Glu Met Gly Glu Glu Val Tyr His Ala Tyr Thr Glu Val Lys Ala Val 485 490 495Arg Ile Lys Leu 500271176DNASaccharomyces cerevisiaeGDP1 27atgtctgctg ctgctgatag attaaactta acttccggcc acttgaatgc tggtagaaag 60agaagttcct cttctgtttc tttgaaggct gccgaaaagc ctttcaaggt tactgtgatt 120ggatctggta actggggtac tactattgcc aaggtggttg ccgaaaattg taagggatac 180ccagaagttt tcgctccaat agtacaaatg tgggtgttcg aagaagagat caatggtgaa 240aaattgactg aaatcataaa tactagacat caaaacgtga aatacttgcc tggcatcact 300ctacccgaca atttggttgc taatccagac ttgattgatt cagtcaagga tgtcgacatc 360atcgttttca acattccaca tcaatttttg ccccgtatct gtagccaatt gaaaggtcat 420gttgattcac acgtcagagc tatctcctgt ctaaagggtt ttgaagttgg tgctaaaggt 480gtccaattgc tatcctctta catcactgag gaactaggta ttcaatgtgg tgctctatct 540ggtgctaaca ttgccaccga agtcgctcaa gaacactggt ctgaaacaac agttgcttac 600cacattccaa aggatttcag aggcgagggc aaggacgtcg accataaggt tctaaaggcc 660ttgttccaca gaccttactt ccacgttagt gtcatcgaag atgttgctgg tatctccatc 720tgtggtgctt tgaagaacgt tgttgcctta ggttgtggtt tcgtcgaagg tctaggctgg 780ggtaacaacg cttctgctgc catccaaaga gtcggtttgg gtgagatcat cagattcggt 840caaatgtttt tcccagaatc tagagaagaa acatactacc aagagtctgc tggtgttgct 900gatttgatca ccacctgcgc tggtggtaga aacgtcaagg ttgctaggct aatggctact 960tctggtaagg acgcctggga atgtgaaaag gagttgttga atggccaatc cgctcaaggt 1020ttaattacct gcaaagaagt tcacgaatgg ttggaaacat gtggctctgt cgaagacttc 1080ccattatttg aagccgtata ccaaatcgtt tacaacaact acccaatgaa gaacctgccg 1140gacatgattg aagaattaga tctacatgaa gattag 117628391PRTSaccharomyces cerevisiaeGDP1 28Met Ser Ala Ala Ala Asp Arg Leu Asn Leu Thr Ser Gly His Leu Asn1 5 10 15Ala Gly Arg Lys Arg Ser Ser Ser Ser Val Ser Leu Lys Ala Ala Glu 20 25 30Lys Pro Phe Lys Val Thr Val Ile Gly Ser Gly Asn Trp Gly Thr Thr 35 40 45Ile Ala Lys Val Val Ala Glu Asn Cys Lys Gly Tyr Pro Glu Val Phe 50 55 60Ala Pro Ile Val Gln Met Trp Val Phe Glu Glu Glu Ile Asn Gly Glu65 70 75 80Lys Leu Thr Glu Ile Ile Asn Thr Arg His Gln Asn Val Lys Tyr Leu 85 90 95Pro Gly Ile Thr Leu Pro Asp Asn Leu Val Ala Asn Pro Asp Leu Ile 100 105 110Asp Ser Val Lys Asp Val Asp Ile Ile Val Phe Asn Ile Pro His Gln 115 120 125Phe Leu Pro Arg Ile Cys Ser Gln Leu Lys Gly His Val Asp Ser His 130 135 140Val Arg Ala Ile Ser Cys Leu Lys Gly Phe Glu Val Gly Ala Lys Gly145 150 155 160Val Gln Leu Leu Ser Ser Tyr Ile Thr Glu Glu Leu Gly Ile Gln Cys 165 170 175Gly Ala Leu Ser Gly Ala Asn Ile Ala Thr Glu Val Ala Gln Glu His 180 185 190Trp Ser Glu Thr Thr Val Ala Tyr His Ile Pro Lys Asp Phe Arg Gly 195 200 205Glu Gly Lys Asp Val Asp His Lys Val Leu Lys Ala Leu Phe His Arg 210 215 220Pro Tyr Phe His Val Ser Val Ile Glu Asp Val Ala Gly Ile Ser Ile225 230 235 240Cys Gly Ala Leu Lys Asn Val Val Ala Leu Gly Cys Gly Phe Val Glu 245 250 255Gly Leu Gly Trp Gly Asn Asn Ala Ser Ala Ala Ile Gln Arg Val Gly 260 265 270Leu Gly Glu Ile Ile Arg Phe Gly Gln Met Phe Phe Pro Glu Ser Arg 275 280 285Glu Glu Thr Tyr Tyr Gln Glu Ser Ala Gly Val Ala Asp Leu Ile Thr 290 295 300Thr Cys Ala Gly Gly Arg Asn Val Lys Val Ala Arg Leu Met Ala Thr305 310 315 320Ser Gly Lys Asp Ala Trp Glu Cys Glu Lys Glu Leu Leu Asn Gly Gln 325 330 335Ser Ala Gln Gly Leu Ile Thr Cys Lys Glu Val His Glu Trp Leu Glu 340 345 350Thr Cys Gly Ser Val Glu Asp Phe Pro Leu Phe Glu Ala Val Tyr Gln 355 360 365Ile Val Tyr Asn Asn Tyr Pro Met Lys Asn Leu Pro Asp Met Ile Glu 370 375 380Glu Leu Asp Leu His Glu Asp385 390291320DNASaccharomyces cerevisiaeGDP2 29atgcttgctg tcagaagatt aacaagatac acattcctta agcgaacgca tccggtgtta 60tatactcgtc gtgcatataa aattttgcct tcaagatcta ctttcctaag aagatcatta 120ttacaaacac aactgcactc aaagatgact gctcatacta atatcaaaca gcacaaacac 180tgtcatgagg accatcctat cagaagatcg gactctgccg tgtcaattgt acatttgaaa 240cgtgcgccct tcaaggttac agtgattggt tctggtaact gggggaccac catcgccaaa 300gtcattgcgg aaaacacaga attgcattcc catatcttcg agccagaggt gagaatgtgg 360gtttttgatg aaaagatcgg cgacgaaaat ctgacggata tcataaatac aagacaccag 420aacgttaaat atctacccaa tattgacctg ccccataatc tagtggccga tcctgatctt 480ttacactcca tcaagggtgc tgacatcctt gttttcaaca tccctcatca atttttacca 540aacatagtca aacaattgca aggccacgtg gcccctcatg taagggccat ctcgtgtcta 600aaagggttcg agttgggctc caagggtgtg caattgctat cctcctatgt tactgatgag 660ttaggaatcc aatgtggcgc actatctggt gcaaacttgg caccggaagt ggccaaggag 720cattggtccg aaaccaccgt ggcttaccaa ctaccaaagg attatcaagg tgatggcaag 780gatgtagatc ataagatttt gaaattgctg ttccacagac cttacttcca cgtcaatgtc 840atcgatgatg ttgctggtat atccattgcc ggtgccttga agaacgtcgt ggcacttgca 900tgtggtttcg tagaaggtat gggatggggt aacaatgcct ccgcagccat tcaaaggctg 960ggtttaggtg aaattatcaa gttcggtaga atgtttttcc cagaatccaa agtcgagacc 1020tactatcaag aatccgctgg tgttgcagat ctgatcacca cctgctcagg cggtagaaac 1080gtcaaggttg ccacatacat ggccaagacc ggtaagtcag ccttggaagc agaaaaggaa 1140ttgcttaacg gtcaatccgc ccaagggata atcacatgca gagaagttca cgagtggcta 1200caaacatgtg agttgaccca agaattccca ttattcgagg cagtctacca gatagtctac 1260aacaacgtcc gcatggaaga cctaccggag atgattgaag agctagacat cgatgacgaa 132030440PRTSaccharomyces cerevisiaeGDP2 30Met Leu Ala Val Arg Arg Leu Thr Arg Tyr Thr Phe Leu Lys Arg Thr1 5 10 15His Pro Val Leu Tyr Thr Arg Arg Ala Tyr Lys Ile Leu Pro Ser Arg 20 25 30Ser Thr Phe Leu Arg Arg Ser Leu Leu Gln Thr Gln Leu His Ser Lys 35 40 45Met Thr Ala His Thr Asn Ile Lys Gln His Lys His Cys His Glu Asp 50 55 60His Pro Ile Arg Arg Ser Asp Ser Ala Val Ser Ile Val His Leu Lys65 70 75 80Arg Ala Pro Phe Lys Val Thr Val Ile Gly Ser Gly Asn Trp Gly Thr 85 90 95Thr Ile Ala Lys Val Ile Ala Glu Asn Thr Glu Leu His Ser His Ile 100 105 110Phe Glu Pro Glu Val Arg Met Trp Val Phe Asp Glu Lys Ile Gly Asp 115 120 125Glu Asn Leu Thr Asp Ile Ile Asn Thr Arg His Gln Asn Val Lys Tyr 130 135 140Leu Pro Asn Ile Asp Leu Pro His Asn Leu Val Ala Asp Pro Asp Leu145 150 155 160Leu His Ser Ile Lys Gly Ala Asp Ile Leu Val Phe Asn Ile Pro His 165 170 175Gln Phe Leu Pro Asn Ile Val Lys Gln Leu Gln Gly His Val Ala Pro 180 185 190His Val Arg Ala Ile Ser Cys Leu Lys Gly Phe Glu Leu Gly Ser Lys 195 200 205Gly Val Gln Leu Leu Ser Ser Tyr Val Thr Asp Glu Leu Gly Ile Gln 210 215 220Cys Gly Ala Leu Ser Gly Ala Asn Leu Ala Pro Glu Val Ala Lys Glu225 230 235 240His Trp Ser Glu Thr Thr Val Ala Tyr Gln Leu Pro Lys Asp Tyr Gln 245 250 255Gly Asp Gly Lys Asp Val Asp His Lys Ile Leu Lys Leu Leu Phe His 260 265 270Arg Pro Tyr Phe His Val Asn Val Ile Asp Asp Val Ala Gly Ile Ser 275 280 285Ile Ala Gly Ala Leu Lys Asn Val Val Ala Leu Ala Cys Gly Phe Val 290 295 300Glu Gly Met Gly Trp Gly Asn Asn Ala Ser Ala Ala Ile Gln Arg Leu305 310 315 320Gly Leu Gly Glu Ile Ile Lys Phe Gly Arg Met Phe Phe Pro Glu Ser 325 330 335Lys Val Glu Thr Tyr Tyr Gln Glu Ser Ala Gly Val Ala Asp Leu Ile 340 345 350Thr Thr Cys Ser Gly Gly Arg Asn Val Lys Val Ala Thr Tyr Met Ala 355 360 365Lys Thr Gly Lys Ser Ala Leu Glu Ala

Glu Lys Glu Leu Leu Asn Gly 370 375 380Gln Ser Ala Gln Gly Ile Ile Thr Cys Arg Glu Val His Glu Trp Leu385 390 395 400Gln Thr Cys Glu Leu Thr Gln Glu Phe Pro Leu Phe Glu Ala Val Tyr 405 410 415Gln Ile Val Tyr Asn Asn Val Arg Met Glu Asp Leu Pro Glu Met Ile 420 425 430Glu Glu Leu Asp Ile Asp Asp Glu 435 440311131DNASaccharomyces cerevisiaeFDH1 31atgtcgaagg gaaaggtttt gctggttctt tacgaaggtg gtaagcatgc tgaagagcag 60gaaaagttat tggggtgtat tgaaaatgaa cttggtatca gaaatttcat tgaagaacag 120ggatacgagt tggttactac cattgacaag gaccctgagc caacctcaac ggtagacagg 180gagttgaaag acgctgaaat tgtcattact acgccctttt tccccgccta catctcgaga 240aacaggattg cagaagctcc taacctgaag ctctgtgtaa ccgctggcgt cggttcagac 300catgtcgatt tagaagctgc aaatgaacgg aaaatcacgg tcaccgaagt tactggttct 360aacgtcgttt ctgtcgcaga gcacgttatg gccacaattt tggttttgat aagaaactat 420aatggtggtc atcaacaagc aattaatggt gagtgggata ttgccggcgt ggctaaaaat 480gagtatgatc tggaagacaa aataatttca acggtaggtg ccggtagaat tggatatagg 540gttctggaaa gattggtcgc atttaatccg aagaagttac tgtactacga ctaccaggaa 600ctacctgcgg aagcaatcaa tagattgaac gaggccagca agcttttcaa tggcagaggt 660gatattgttc agagagtaga gaaattggag gatatggttg ctcagtcaga tgttgttacc 720atcaactgtc cattgcacaa ggactcaagg ggtttattca ataaaaagct tatttcccac 780atgaaagatg gtgcatactt ggtgaatacc gctagaggtg ctatttgtgt cgcagaagat 840gttgccgagg cagtcaagtc tggtaaattg gctggctatg gtggtgatgt ctgggataag 900caaccagcac caaaagacca tccctggagg actatggaca ataaggacca cgtgggaaac 960gcaatgactg ttcatatcag tggcacatct ctggatgctc aaaagaggta cgctcaggga 1020gtaaagaaca tcctaaatag ttacttttcc aaaaagtttg attaccgtcc acaggatatt 1080attgtgcaga atggttctta tgccaccaga gcttatggac agaagaaata a 113132376PRTSaccharomyces cerevisiaeFDH1 32Met Ser Lys Gly Lys Val Leu Leu Val Leu Tyr Glu Gly Gly Lys His1 5 10 15Ala Glu Glu Gln Glu Lys Leu Leu Gly Cys Ile Glu Asn Glu Leu Gly 20 25 30Ile Arg Asn Phe Ile Glu Glu Gln Gly Tyr Glu Leu Val Thr Thr Ile 35 40 45Asp Lys Asp Pro Glu Pro Thr Ser Thr Val Asp Arg Glu Leu Lys Asp 50 55 60Ala Glu Ile Val Ile Thr Thr Pro Phe Phe Pro Ala Tyr Ile Ser Arg65 70 75 80Asn Arg Ile Ala Glu Ala Pro Asn Leu Lys Leu Cys Val Thr Ala Gly 85 90 95Val Gly Ser Asp His Val Asp Leu Glu Ala Ala Asn Glu Arg Lys Ile 100 105 110Thr Val Thr Glu Val Thr Gly Ser Asn Val Val Ser Val Ala Glu His 115 120 125Val Met Ala Thr Ile Leu Val Leu Ile Arg Asn Tyr Asn Gly Gly His 130 135 140Gln Gln Ala Ile Asn Gly Glu Trp Asp Ile Ala Gly Val Ala Lys Asn145 150 155 160Glu Tyr Asp Leu Glu Asp Lys Ile Ile Ser Thr Val Gly Ala Gly Arg 165 170 175Ile Gly Tyr Arg Val Leu Glu Arg Leu Val Ala Phe Asn Pro Lys Lys 180 185 190Leu Leu Tyr Tyr Asp Tyr Gln Glu Leu Pro Ala Glu Ala Ile Asn Arg 195 200 205Leu Asn Glu Ala Ser Lys Leu Phe Asn Gly Arg Gly Asp Ile Val Gln 210 215 220Arg Val Glu Lys Leu Glu Asp Met Val Ala Gln Ser Asp Val Val Thr225 230 235 240Ile Asn Cys Pro Leu His Lys Asp Ser Arg Gly Leu Phe Asn Lys Lys 245 250 255Leu Ile Ser His Met Lys Asp Gly Ala Tyr Leu Val Asn Thr Ala Arg 260 265 270Gly Ala Ile Cys Val Ala Glu Asp Val Ala Glu Ala Val Lys Ser Gly 275 280 285Lys Leu Ala Gly Tyr Gly Gly Asp Val Trp Asp Lys Gln Pro Ala Pro 290 295 300Lys Asp His Pro Trp Arg Thr Met Asp Asn Lys Asp His Val Gly Asn305 310 315 320Ala Met Thr Val His Ile Ser Gly Thr Ser Leu Asp Ala Gln Lys Arg 325 330 335Tyr Ala Gln Gly Val Lys Asn Ile Leu Asn Ser Tyr Phe Ser Lys Lys 340 345 350Phe Asp Tyr Arg Pro Gln Asp Ile Ile Val Gln Asn Gly Ser Tyr Ala 355 360 365Thr Arg Ala Tyr Gly Gln Lys Lys 370 375331129DNASaccharomyces cerevisiaeFDH2 33atgtcgaagg gaaaggtttt gctggttctt tatgaaggtg gtaagcatgc tgaagagcag 60gaaaagttat tggggtgtat tgaaaatgaa cttggtatca gaaatttcat tgaagaacag 120ggatacgagt tggttactac cattgacaag gaccctgagc caacctcaac ggtagacagg 180gagttgaaag acgctgaaat tgtcattact acgccctttt tccccgccta catctcgaga 240aacaggattg cagaagctcc taacctgaag ctctgtgtaa ccgctggcgt cggttcagac 300catgtcgatt tagaagctgc aaatgaacgg aaaatcacgg tcaccgaagt tactggttct 360aacgtcgttt ctgtcgcaga gcacgttatg gccacaattt tggttttgat aagaaactat 420aatggtggtc atcaataagc aattaatggt gagtgggata ttgccggcgt ggctaaaaaa 480tgagtatgat ctggaagaca aaataatttc aacggtaggt gccggtagaa ttggatatag 540ggttctggaa agattggtcg catttaatcc gaagaagtta ctgtactacg actaccagga 600actacctgcg gaagcaatca atagattgaa cgaggccagc aagcttttca atggcagagg 660tgatattgtt cagagagtag agaaattgga ggatatggtt gctcagtcag atgttgttac 720catcaactgt ccattgcaca aggactcaag gggtttattc aataaaaagc ttatttccca 780catgaaagat ggtgcatact tggtgaatac cgctagaggt gctatttgtg tcgcagaaga 840tgttgccgag gcagtcaagt ctggtaaatt ggctggctat ggtggtgatg tctgggataa 900gcaaccagca ccaaaagacc atccctggag gactatggac aataaggacc acgtgggaaa 960cgcaatgact gttcatatca gtggcacatc tctgcatgct caaaagaggt acgctcaggg 1020agtaaagaac atcctaaata gttacttttc caaaaagttt gattaccgtc cacaggatat 1080tattgtgcag aatggttctt atgccaccag agcttatgga cagaagaaa 112934329PRTOenococcus oeniiPTA 34Met Thr Asn Leu Phe Thr Glu Leu Ala Asp Lys Ile Lys Glu Lys Lys1 5 10 15Leu Arg Leu Val Phe Pro Glu Gly Asp Asp Ser Arg Val Gln Gly Ala 20 25 30Ala Ile Arg Leu Lys Lys Asp Gly Leu Leu Thr Pro Ile Leu Leu Gly 35 40 45Asp Gln Ala Ala Val Glu Lys Thr Ala Ala Asp Asn His Phe Asp Leu 50 55 60Ser Gly Ile Lys Leu Ile Asp Pro Leu Lys Phe Asp Glu Lys Gln Phe65 70 75 80Asn Gln Leu Ile Asp Lys Leu Val Thr Arg Arg Lys Gly Lys Thr Asp 85 90 95Pro Gln Thr Val Ala Met Trp Leu Gln Glu Val Asn Tyr Phe Gly Thr 100 105 110Met Leu Val Tyr Thr Gly Lys Ala Asp Ala Met Val Ser Gly Ala Val 115 120 125His Pro Thr Gly Asp Thr Val Arg Pro Ala Leu Gln Ile Ile Lys Thr 130 135 140Ala Pro Gly Val Ser Arg Ile Ser Gly Ala Phe Ile Met Gln Lys Asn145 150 155 160Asp Glu Arg Tyr Leu Phe Ala Asp Ser Ala Ile Asn Ile Glu Leu Asp 165 170 175Ala Gln Thr Met Ala Glu Ile Ala Val Gln Ser Ala His Thr Ala Lys 180 185 190Ile Phe Gly Ile Asp Pro Lys Val Ala Met Leu Ser Phe Ser Thr Lys 195 200 205Gly Ser Ala Lys His Ala Leu Val Asp Lys Val Ala Gln Ala Thr Lys 210 215 220Leu Ala Gln Glu Met Ala Pro Asp Leu Ala Asp Asn Ile Asp Gly Glu225 230 235 240Leu Gln Phe Asp Ala Ala Phe Val Glu Ser Val Ala Lys Leu Lys Ala 245 250 255Pro Gly Ser Lys Val Ala Gly His Ala Asn Val Phe Ile Phe Pro Ser 260 265 270Leu Glu Ala Gly Asn Ile Gly Tyr Lys Ile Ala Gln Arg Leu Gly Gly 275 280 285Tyr Glu Ala Ile Gly Pro Ile Leu Gln Gly Leu Ala Ala Pro Val Ser 290 295 300Asp Leu Ser Arg Gly Ala Ser Gln Glu Asp Val Tyr Lys Val Ala Ile305 310 315 320Ile Thr Ala Ala Gln Ala Leu Lys Asn 325351008DNASaccharomyces cerevisiaeTAL1 35atgtctgaac cagctcaaaa gaaacaaaag gttgctaaca actctctaga acaattgaaa 60gcctccggca ctgtcgttgt tgccgacact ggtgatttcg gctctattgc caagtttcaa 120cctcaagact ccacaactaa cccatcattg atcttggctg ctgccaagca accaacttac 180gccaagttga tcgatgttgc cgtggaatac ggtaagaagc atggtaagac caccgaagaa 240caagtcgaaa atgctgtgga cagattgtta gtcgaattcg gtaaggagat cttaaagatt 300gttccaggca gagtctccac cgaagttgat gctagattgt cttttgacac tcaagctacc 360attgaaaagg ctagacatat cattaaattg tttgaacaag aaggtgtctc caaggaaaga 420gtccttatta aaattgcttc cacttgggaa ggtattcaag ctgccaaaga attggaagaa 480aaggacggta tccactgtaa tttgactcta ttattctcct tcgttcaagc agttgcctgt 540gccgaggccc aagttacttt gatttcccca tttgttggta gaattctaga ctggtacaaa 600tccagcactg gtaaagatta caagggtgaa gccgacccag gtgttatttc cgtcaagaaa 660atctacaact actacaagaa gtacggttac aagactattg ttatgggtgc ttctttcaga 720agcactgacg aaatcaaaaa cttggctggt gttgactatc taacaatttc tccagcttta 780ttggacaagt tgatgaacag tactgaacct ttcccaagag ttttggaccc tgtctccgct 840aagaaggaag ccggcgacaa gatttcttac atcagcgacg aatctaaatt cagattcgac 900ttgaatgaag acgctatggc cactgaaaaa ttgtccgaag gtatcagaaa attctctgcc 960gatattgtta ctctattcga cttgattgaa aagaaagtta ccgcttaa 100836335PRTSaccharomyces cerevisiaeTAL1 36Met Ser Glu Pro Ala Gln Lys Lys Gln Lys Val Ala Asn Asn Ser Leu1 5 10 15Glu Gln Leu Lys Ala Ser Gly Thr Val Val Val Ala Asp Thr Gly Asp 20 25 30Phe Gly Ser Ile Ala Lys Phe Gln Pro Gln Asp Ser Thr Thr Asn Pro 35 40 45Ser Leu Ile Leu Ala Ala Ala Lys Gln Pro Thr Tyr Ala Lys Leu Ile 50 55 60Asp Val Ala Val Glu Tyr Gly Lys Lys His Gly Lys Thr Thr Glu Glu65 70 75 80Gln Val Glu Asn Ala Val Asp Arg Leu Leu Val Glu Phe Gly Lys Glu 85 90 95Ile Leu Lys Ile Val Pro Gly Arg Val Ser Thr Glu Val Asp Ala Arg 100 105 110Leu Ser Phe Asp Thr Gln Ala Thr Ile Glu Lys Ala Arg His Ile Ile 115 120 125Lys Leu Phe Glu Gln Glu Gly Val Ser Lys Glu Arg Val Leu Ile Lys 130 135 140Ile Ala Ser Thr Trp Glu Gly Ile Gln Ala Ala Lys Glu Leu Glu Glu145 150 155 160Lys Asp Gly Ile His Cys Asn Leu Thr Leu Leu Phe Ser Phe Val Gln 165 170 175Ala Val Ala Cys Ala Glu Ala Gln Val Thr Leu Ile Ser Pro Phe Val 180 185 190Gly Arg Ile Leu Asp Trp Tyr Lys Ser Ser Thr Gly Lys Asp Tyr Lys 195 200 205Gly Glu Ala Asp Pro Gly Val Ile Ser Val Lys Lys Ile Tyr Asn Tyr 210 215 220Tyr Lys Lys Tyr Gly Tyr Lys Thr Ile Val Met Gly Ala Ser Phe Arg225 230 235 240Ser Thr Asp Glu Ile Lys Asn Leu Ala Gly Val Asp Tyr Leu Thr Ile 245 250 255Ser Pro Ala Leu Leu Asp Lys Leu Met Asn Ser Thr Glu Pro Phe Pro 260 265 270Arg Val Leu Asp Pro Val Ser Ala Lys Lys Glu Ala Gly Asp Lys Ile 275 280 285Ser Tyr Ile Ser Asp Glu Ser Lys Phe Arg Phe Asp Leu Asn Glu Asp 290 295 300Ala Met Ala Thr Glu Lys Leu Ser Glu Gly Ile Arg Lys Phe Ser Ala305 310 315 320Asp Ile Val Thr Leu Phe Asp Leu Ile Glu Lys Lys Val Thr Ala 325 330 335372040DNASaccharomyces cerevisiaeTKL1 37atgactcaat tcactgacat tgataagcta gccgtctcca ccataagaat tttggctgtg 60gacaccgtat ccaaggccaa ctcaggtcac ccaggtgctc cattgggtat ggcaccagct 120gcacacgttc tatggagtca aatgcgcatg aacccaacca acccagactg gatcaacaga 180gatagatttg tcttgtctaa cggtcacgcg gtcgctttgt tgtattctat gctacatttg 240actggttacg atctgtctat tgaagacttg aaacagttca gacagttggg ttccagaaca 300ccaggtcatc ctgaatttga gttgccaggt gttgaagtta ctaccggtcc attaggtcaa 360ggtatctcca acgctgttgg tatggccatg gctcaagcta acctggctgc cacttacaac 420aagccgggct ttaccttgtc tgacaactac acctatgttt tcttgggtga cggttgtttg 480caagaaggta tttcttcaga agcttcctcc ttggctggtc atttgaaatt gggtaacttg 540attgccatct acgatgacaa caagatcact atcgatggtg ctaccagtat ctcattcgat 600gaagatgttg ctaagagata cgaagcctac ggttgggaag ttttgtacgt agaaaatggt 660aacgaagatc tagccggtat tgccaaggct attgctcaag ctaagttatc caaggacaaa 720ccaactttga tcaaaatgac cacaaccatt ggttacggtt ccttgcatgc cggctctcac 780tctgtgcacg gtgccccatt gaaagcagat gatgttaaac aactaaagag caaattcggt 840ttcaacccag acaagtcctt tgttgttcca caagaagttt acgaccacta ccaaaagaca 900attttaaagc caggtgtcga agccaacaac aagtggaaca agttgttcag cgaataccaa 960aagaaattcc cagaattagg tgctgaattg gctagaagat tgagcggcca actacccgca 1020aattgggaat ctaagttgcc aacttacacc gccaaggact ctgccgtggc cactagaaaa 1080ttatcagaaa ctgttcttga ggatgtttac aatcaattgc cagagttgat tggtggttct 1140gccgatttaa caccttctaa cttgaccaga tggaaggaag cccttgactt ccaacctcct 1200tcttccggtt caggtaacta ctctggtaga tacattaggt acggtattag agaacacgct 1260atgggtgcca taatgaacgg tatttcagct ttcggtgcca actacaaacc atacggtggt 1320actttcttga acttcgtttc ttatgctgct ggtgccgtta gattgtccgc tttgtctggc 1380cacccagtta tttgggttgc tacacatgac tctatcggtg tcggtgaaga tggtccaaca 1440catcaaccta ttgaaacttt agcacacttc agatccctac caaacattca agtttggaga 1500ccagctgatg gtaacgaagt ttctgccgcc tacaagaact ctttagaatc caagcatact 1560ccaagtatca ttgctttgtc cagacaaaac ttgccacaat tggaaggtag ctctattgaa 1620agcgcttcta agggtggtta cgtactacaa gatgttgcta acccagatat tattttagtg 1680gctactggtt ccgaagtgtc tttgagtgtt gaagctgcta agactttggc cgcaaagaac 1740atcaaggctc gtgttgtttc tctaccagat ttcttcactt ttgacaaaca acccctagaa 1800tacagactat cagtcttacc agacaacgtt ccaatcatgt ctgttgaagt tttggctacc 1860acatgttggg gcaaatacgc tcatcaatcc ttcggtattg acagatttgg tgcctccggt 1920aaggcaccag aagtcttcaa gttcttcggt ttcaccccag aaggtgttgc tgaaagagct 1980caaaagacca ttgcattcta taagggtgac aagctaattt ctcctttgaa aaaagctttc 204038680PRTSaccharomyces cerevisiaeTKL1 38Met Thr Gln Phe Thr Asp Ile Asp Lys Leu Ala Val Ser Thr Ile Arg1 5 10 15Ile Leu Ala Val Asp Thr Val Ser Lys Ala Asn Ser Gly His Pro Gly 20 25 30Ala Pro Leu Gly Met Ala Pro Ala Ala His Val Leu Trp Ser Gln Met 35 40 45Arg Met Asn Pro Thr Asn Pro Asp Trp Ile Asn Arg Asp Arg Phe Val 50 55 60Leu Ser Asn Gly His Ala Val Ala Leu Leu Tyr Ser Met Leu His Leu65 70 75 80Thr Gly Tyr Asp Leu Ser Ile Glu Asp Leu Lys Gln Phe Arg Gln Leu 85 90 95Gly Ser Arg Thr Pro Gly His Pro Glu Phe Glu Leu Pro Gly Val Glu 100 105 110Val Thr Thr Gly Pro Leu Gly Gln Gly Ile Ser Asn Ala Val Gly Met 115 120 125Ala Met Ala Gln Ala Asn Leu Ala Ala Thr Tyr Asn Lys Pro Gly Phe 130 135 140Thr Leu Ser Asp Asn Tyr Thr Tyr Val Phe Leu Gly Asp Gly Cys Leu145 150 155 160Gln Glu Gly Ile Ser Ser Glu Ala Ser Ser Leu Ala Gly His Leu Lys 165 170 175Leu Gly Asn Leu Ile Ala Ile Tyr Asp Asp Asn Lys Ile Thr Ile Asp 180 185 190Gly Ala Thr Ser Ile Ser Phe Asp Glu Asp Val Ala Lys Arg Tyr Glu 195 200 205Ala Tyr Gly Trp Glu Val Leu Tyr Val Glu Asn Gly Asn Glu Asp Leu 210 215 220Ala Gly Ile Ala Lys Ala Ile Ala Gln Ala Lys Leu Ser Lys Asp Lys225 230 235 240Pro Thr Leu Ile Lys Met Thr Thr Thr Ile Gly Tyr Gly Ser Leu His 245 250 255Ala Gly Ser His Ser Val His Gly Ala Pro Leu Lys Ala Asp Asp Val 260 265 270Lys Gln Leu Lys Ser Lys Phe Gly Phe Asn Pro Asp Lys Ser Phe Val 275 280 285Val Pro Gln Glu Val Tyr Asp His Tyr Gln Lys Thr Ile Leu Lys Pro 290 295 300Gly Val Glu Ala Asn Asn Lys Trp Asn Lys Leu Phe Ser Glu Tyr Gln305 310 315 320Lys Lys Phe Pro Glu Leu Gly Ala Glu Leu Ala Arg Arg Leu Ser Gly 325 330 335Gln Leu Pro Ala Asn Trp Glu Ser Lys Leu Pro Thr Tyr Thr Ala Lys 340 345 350Asp Ser Ala Val Ala Thr Arg Lys Leu Ser Glu Thr Val Leu Glu Asp 355 360 365Val Tyr Asn Gln Leu Pro Glu Leu Ile Gly Gly Ser Ala Asp Leu Thr 370 375 380Pro Ser Asn Leu Thr Arg Trp Lys Glu Ala Leu Asp Phe Gln Pro Pro385 390 395 400Ser Ser Gly Ser Gly Asn Tyr Ser Gly Arg Tyr Ile Arg Tyr Gly Ile 405 410 415Arg Glu His Ala Met Gly Ala Ile Met Asn Gly Ile Ser Ala Phe Gly 420 425 430Ala Asn Tyr Lys Pro Tyr Gly Gly Thr Phe Leu Asn Phe Val Ser Tyr

435 440 445Ala Ala Gly Ala Val Arg Leu Ser Ala Leu Ser Gly His Pro Val Ile 450 455 460Trp Val Ala Thr His Asp Ser Ile Gly Val Gly Glu Asp Gly Pro Thr465 470 475 480His Gln Pro Ile Glu Thr Leu Ala His Phe Arg Ser Leu Pro Asn Ile 485 490 495Gln Val Trp Arg Pro Ala Asp Gly Asn Glu Val Ser Ala Ala Tyr Lys 500 505 510Asn Ser Leu Glu Ser Lys His Thr Pro Ser Ile Ile Ala Leu Ser Arg 515 520 525Gln Asn Leu Pro Gln Leu Glu Gly Ser Ser Ile Glu Ser Ala Ser Lys 530 535 540Gly Gly Tyr Val Leu Gln Asp Val Ala Asn Pro Asp Ile Ile Leu Val545 550 555 560Ala Thr Gly Ser Glu Val Ser Leu Ser Val Glu Ala Ala Lys Thr Leu 565 570 575Ala Ala Lys Asn Ile Lys Ala Arg Val Val Ser Leu Pro Asp Phe Phe 580 585 590Thr Phe Asp Lys Gln Pro Leu Glu Tyr Arg Leu Ser Val Leu Pro Asp 595 600 605Asn Val Pro Ile Met Ser Val Glu Val Leu Ala Thr Thr Cys Trp Gly 610 615 620Lys Tyr Ala His Gln Ser Phe Gly Ile Asp Arg Phe Gly Ala Ser Gly625 630 635 640Lys Ala Pro Glu Val Phe Lys Phe Phe Gly Phe Thr Pro Glu Gly Val 645 650 655Ala Glu Arg Ala Gln Lys Thr Ile Ala Phe Tyr Lys Gly Asp Lys Leu 660 665 670Ile Ser Pro Leu Lys Lys Ala Phe 675 68039774DNASaccharomyces cerevisiaeRKI1 39atggctgccg gtgtcccaaa aattgatgcg ttagaatctt tgggcaatcc tttggaggat 60gccaagagag ctgcagcata cagagcagtt gatgaaaatt taaaatttga tgatcacaaa 120attattggaa ttggtagtgg tagcacagtg gtttatgttg ccgaaagaat tggacaatat 180ttgcatgacc ctaaatttta tgaagtagcg tctaaattca tttgcattcc aacaggattc 240caatcaagaa acttgatttt ggataacaag ttgcaattag gctccattga acagtatcct 300cgcattgata tagcgtttga cggtgctgat gaagtggatg agaatttaca attaattaaa 360ggtggtggtg cttgtctatt tcaagaaaaa ttggttagta ctagtgctaa aaccttcatt 420gtcgttgctg attcaagaaa aaagtcacca aaacatttag gtaagaactg gaggcaaggt 480gttcccattg aaattgtacc ttcctcatac gtgagggtca agaatgatct attagaacaa 540ttgcatgctg aaaaagttga catcagacaa ggaggttctg ctaaagcagg tcctgttgta 600actgacaata ataacttcat tatcgatgcg gatttcggtg aaatttccga tccaagaaaa 660ttgcatagag aaatcaaact gttagtgggc gtggtggaaa caggtttatt catcgacaac 720gcttcaaaag cctacttcgg taattctgac ggtagtgttg aagttaccga aaag 77440258PRTSaccharomyces cerevisiae RKI1 40Met Ala Ala Gly Val Pro Lys Ile Asp Ala Leu Glu Ser Leu Gly Asn1 5 10 15Pro Leu Glu Asp Ala Lys Arg Ala Ala Ala Tyr Arg Ala Val Asp Glu 20 25 30Asn Leu Lys Phe Asp Asp His Lys Ile Ile Gly Ile Gly Ser Gly Ser 35 40 45Thr Val Val Tyr Val Ala Glu Arg Ile Gly Gln Tyr Leu His Asp Pro 50 55 60Lys Phe Tyr Glu Val Ala Ser Lys Phe Ile Cys Ile Pro Thr Gly Phe65 70 75 80Gln Ser Arg Asn Leu Ile Leu Asp Asn Lys Leu Gln Leu Gly Ser Ile 85 90 95Glu Gln Tyr Pro Arg Ile Asp Ile Ala Phe Asp Gly Ala Asp Glu Val 100 105 110Asp Glu Asn Leu Gln Leu Ile Lys Gly Gly Gly Ala Cys Leu Phe Gln 115 120 125Glu Lys Leu Val Ser Thr Ser Ala Lys Thr Phe Ile Val Val Ala Asp 130 135 140Ser Arg Lys Lys Ser Pro Lys His Leu Gly Lys Asn Trp Arg Gln Gly145 150 155 160Val Pro Ile Glu Ile Val Pro Ser Ser Tyr Val Arg Val Lys Asn Asp 165 170 175Leu Leu Glu Gln Leu His Ala Glu Lys Val Asp Ile Arg Gln Gly Gly 180 185 190Ser Ala Lys Ala Gly Pro Val Val Thr Asp Asn Asn Asn Phe Ile Ile 195 200 205Asp Ala Asp Phe Gly Glu Ile Ser Asp Pro Arg Lys Leu His Arg Glu 210 215 220Ile Lys Leu Leu Val Gly Val Val Glu Thr Gly Leu Phe Ile Asp Asn225 230 235 240Ala Ser Lys Ala Tyr Phe Gly Asn Ser Asp Gly Ser Val Glu Val Thr 245 250 255Glu Lys41714DNASaccharomyces cerevisiaeRPE1 41atggtcaaac caattatagc tcccagtatc cttgcttctg acttcgccaa cttgggttgc 60gaatgtcata aggtcatcaa cgccggcgca gattggttac atatcgatgt catggacggc 120cattttgttc caaacattac tctgggccaa ccaattgtta cctccctacg tcgttctgtg 180ccacgccctg gcgatgctag caacacagaa aagaagccca ctgcgttctt cgattgtcac 240atgatggttg aaaatcctga aaaatgggtc gacgattttg ctaaatgtgg tgctgaccaa 300tttacgttcc actacgaggc cacacaagac cctttgcatt tagttaagtt gattaagtct 360aagggcatca aagctgcatg cgccatcaaa cctggtactt ctgttgacgt tttatttgaa 420ctagctcctc atttggatat ggctcttgtt atgactgtgg aacctgggtt tggaggccaa 480aaattcatgg aagacatgat gccaaaagtg gaaactttga gagccaagtt cccccatttg 540aatatccaag tcgatggtgg tttgggcaag gagaccatcc cgaaagccgc caaagccggt 600gccaacgtta ttgtcgctgg taccagtgtt ttcactgcag ctgacccgca cgatgttatc 660tccttcatga aagaagaagt ctcgaaggaa ttgcgttcta gagatttgct agat 71442238PRTSaccharomyces cerevisiaeRPE1 42Met Val Lys Pro Ile Ile Ala Pro Ser Ile Leu Ala Ser Asp Phe Ala1 5 10 15Asn Leu Gly Cys Glu Cys His Lys Val Ile Asn Ala Gly Ala Asp Trp 20 25 30Leu His Ile Asp Val Met Asp Gly His Phe Val Pro Asn Ile Thr Leu 35 40 45Gly Gln Pro Ile Val Thr Ser Leu Arg Arg Ser Val Pro Arg Pro Gly 50 55 60Asp Ala Ser Asn Thr Glu Lys Lys Pro Thr Ala Phe Phe Asp Cys His65 70 75 80Met Met Val Glu Asn Pro Glu Lys Trp Val Asp Asp Phe Ala Lys Cys 85 90 95Gly Ala Asp Gln Phe Thr Phe His Tyr Glu Ala Thr Gln Asp Pro Leu 100 105 110His Leu Val Lys Leu Ile Lys Ser Lys Gly Ile Lys Ala Ala Cys Ala 115 120 125Ile Lys Pro Gly Thr Ser Val Asp Val Leu Phe Glu Leu Ala Pro His 130 135 140Leu Asp Met Ala Leu Val Met Thr Val Glu Pro Gly Phe Gly Gly Gln145 150 155 160Lys Phe Met Glu Asp Met Met Pro Lys Val Glu Thr Leu Arg Ala Lys 165 170 175Phe Pro His Leu Asn Ile Gln Val Asp Gly Gly Leu Gly Lys Glu Thr 180 185 190Ile Pro Lys Ala Ala Lys Ala Gly Ala Asn Val Ile Val Ala Gly Thr 195 200 205Ser Val Phe Thr Ala Ala Asp Pro His Asp Val Ile Ser Phe Met Lys 210 215 220Glu Glu Val Ser Lys Glu Leu Arg Ser Arg Asp Leu Leu Asp225 230 235432733DNABifidobacterium adolescentisadhE 43atggcagacg caaagaagaa ggaagagccg accaagccga ctccggaaga gaagctcgcc 60gcagccgagg ctgaggtcga cgctctggtc aagaagggcc tgaaggctct tgatgaattc 120gagaagctcg atcagaagca ggttgaccac atcgtggcca aggcttccgt cgcagccctg 180aacaagcact tggtgctcgc caagatggcc gtcgaggaga cccaccgtgg tctggtcgaa 240gacaaggcca ccaagaacat cttcgcctgc gagcatgtca ccaactacct ggctggtcag 300aagaccgtcg gcatcatccg cgaggacgac gtgctgggca tcgacgaaat cgccgagccg 360gttggcgtcg tcgctggcgt gaccccggtc accaacccga cctccaccgc catcttcaag 420tcgctgatcg cactgaagac ccgctgcccg atcatcttcg gcttccaccc gggcgcacag 480aactgctccg tcgcggccgc caagatcgtt cgcgatgccg ctatcgcagc aggcgctcct 540gagaactgta ttcagtggat cgagcatccg tccatcgagg ccactggcgc cctgatgaag 600catgatggtg tcgccaccat cctcgccacc ggtggtccgg gcatggtcaa ggccgcatac 660tcctccggca agccggccct gggcgtcggc gcgggcaatg ctccggcata cgttgacaag 720aacgtcgacg tcgtgcgtgc agccaacgat ctgattcttt ccaagcactt cgattacggc 780atgatctgcg ctaccgagca ggccatcatc gccgacaagg acatctacgc tccgctcgtt 840aaggaactca agcgtcgcaa ggcctatttc gtgaacgctg acgagaaggc caagctcgag 900cagtacatgt tcggctgcac cgcttactcc ggacagaccc cgaagctcaa ctccgtggtg 960ccgggcaagt ccccgcagta catcgccaag gccgccggct tcgagattcc ggaagacgcc 1020accatccttg ccgctgagtg caaggaagtc ggcgagaacg agccgctgac catggagaag 1080cttgctccgg tccaggccgt gctgaagtcc gacaacaagg aacaggcctt cgagatgtgc 1140gaagccatgc tgaagcatgg cgccggccac accgccgcca tccacaccaa cgaccgtgac 1200ctggtccgcg agtacggcca gcgcatgcac gcctgccgta tcatctggaa ctccccgagc 1260tccctcggcg gcgtgggcga catctacaac gccatcgctc cgtccctgac cctgggctgc 1320ggctcctacg gcggcaactc cgtgtccggc aacgtccagg cagtcaacct catcaacatc 1380aagcgcatcg ctcggaggaa caacaacatg cagtggttca agattccggc caagacctac 1440ttcgagccga acgccatcaa gtacctgcgc gacatgtacg gcatcgaaaa ggccgtcatc 1500gtgtgcgata aggtcatgga gcagctcggc atcgttgaca agatcatcga tcagctgcgt 1560gcacgttcca accgcgtgac cttccgtatc atcgattatg tcgagccgga gccgagcgtg 1620gagaccgtcg aacgtggcgc cgccatgatg cgcgaggagt tcgagccgga taccatcatc 1680gccgtcggcg gtggttcccc gatggatgcg tccaagatta tgtggctgct gtacgagcac 1740ccggaaatct ccttctccga tgtgcgtgag aagttcttcg atatccgtaa gcgcgcgttc 1800aagattccgc cgctgggcaa gaaggccaag ctggtctgca ttccgacttc ttccggcacc 1860ggttccgaag tcacgccgtt cgctgtgatt accgaccaca agaccggcta taagtacccg 1920atcaccgatt acgcgctgac cccgtccgtc gctatcgtcg atccggtgct ggcacgtact 1980cagccgcgca agctggcttc cgatgctggt ttcgatgctc tgacccacgc ttttgaggct 2040tatgtgtccg tgtatgccaa cgacttcacc gatggtatgg cattgcacgc tgccaagctg 2100gtttgggaca acctcgctga gtccgtcaat ggcgagccgg gtgaggagaa gacccgtgcc 2160caggagaaga tgcataatgc cgccaccatg gccggcatgg ctttcggctc cgccttcctc 2220ggcatgtgcc acggcatggc ccacaccatt ggtgcactgt gccacgttgc ccacggtcgt 2280accaactcca tcctcctgcc gtacgtgatc cgttacaacg gttccgtccc ggaggagccg 2340accagctggc cgaagtacaa caagtacatc gctccggaac gctaccagga gatcgccaag 2400aaccttggcg tgaacccggg caagactccg gaagagggcg tcgagaacct ggccaaggct 2460gttgaggatt accgtgacaa caagctcggt atgaacaaga gcttccagga gtgcggtgtg 2520gatgaggact actattggtc catcatcgac cagatcggca tgcgcgccta cgaagaccag 2580tgcgcaccgg cgaacccgcg tatcccgcag atcgaggata tgaaggatat cgccattgcc 2640gcctactacg gcgtcagcca ggcggaaggc cacaagctgc gcgtccagcg tcagggcgaa 2700gccgctacgg aggaagcttc cgagcgcgcc tga 273344910PRTBifidobacterium adolescentisadhE 44Met Ala Asp Ala Lys Lys Lys Glu Glu Pro Thr Lys Pro Thr Pro Glu1 5 10 15Glu Lys Leu Ala Ala Ala Glu Ala Glu Val Asp Ala Leu Val Lys Lys 20 25 30Gly Leu Lys Ala Leu Asp Glu Phe Glu Lys Leu Asp Gln Lys Gln Val 35 40 45Asp His Ile Val Ala Lys Ala Ser Val Ala Ala Leu Asn Lys His Leu 50 55 60Val Leu Ala Lys Met Ala Val Glu Glu Thr His Arg Gly Leu Val Glu65 70 75 80Asp Lys Ala Thr Lys Asn Ile Phe Ala Cys Glu His Val Thr Asn Tyr 85 90 95Leu Ala Gly Gln Lys Thr Val Gly Ile Ile Arg Glu Asp Asp Val Leu 100 105 110Gly Ile Asp Glu Ile Ala Glu Pro Val Gly Val Val Ala Gly Val Thr 115 120 125Pro Val Thr Asn Pro Thr Ser Thr Ala Ile Phe Lys Ser Leu Ile Ala 130 135 140Leu Lys Thr Arg Cys Pro Ile Ile Phe Gly Phe His Pro Gly Ala Gln145 150 155 160Asn Cys Ser Val Ala Ala Ala Lys Ile Val Arg Asp Ala Ala Ile Ala 165 170 175Ala Gly Ala Pro Glu Asn Cys Ile Gln Trp Ile Glu His Pro Ser Ile 180 185 190Glu Ala Thr Gly Ala Leu Met Lys His Asp Gly Val Ala Thr Ile Leu 195 200 205Ala Thr Gly Gly Pro Gly Met Val Lys Ala Ala Tyr Ser Ser Gly Lys 210 215 220Pro Ala Leu Gly Val Gly Ala Gly Asn Ala Pro Ala Tyr Val Asp Lys225 230 235 240Asn Val Asp Val Val Arg Ala Ala Asn Asp Leu Ile Leu Ser Lys His 245 250 255Phe Asp Tyr Gly Met Ile Cys Ala Thr Glu Gln Ala Ile Ile Ala Asp 260 265 270Lys Asp Ile Tyr Ala Pro Leu Val Lys Glu Leu Lys Arg Arg Lys Ala 275 280 285Tyr Phe Val Asn Ala Asp Glu Lys Ala Lys Leu Glu Gln Tyr Met Phe 290 295 300Gly Cys Thr Ala Tyr Ser Gly Gln Thr Pro Lys Leu Asn Ser Val Val305 310 315 320Pro Gly Lys Ser Pro Gln Tyr Ile Ala Lys Ala Ala Gly Phe Glu Ile 325 330 335Pro Glu Asp Ala Thr Ile Leu Ala Ala Glu Cys Lys Glu Val Gly Glu 340 345 350Asn Glu Pro Leu Thr Met Glu Lys Leu Ala Pro Val Gln Ala Val Leu 355 360 365Lys Ser Asp Asn Lys Glu Gln Ala Phe Glu Met Cys Glu Ala Met Leu 370 375 380Lys His Gly Ala Gly His Thr Ala Ala Ile His Thr Asn Asp Arg Asp385 390 395 400Leu Val Arg Glu Tyr Gly Gln Arg Met His Ala Cys Arg Ile Ile Trp 405 410 415Asn Ser Pro Ser Ser Leu Gly Gly Val Gly Asp Ile Tyr Asn Ala Ile 420 425 430Ala Pro Ser Leu Thr Leu Gly Cys Gly Ser Tyr Gly Gly Asn Ser Val 435 440 445Ser Gly Asn Val Gln Ala Val Asn Leu Ile Asn Ile Lys Arg Ile Ala 450 455 460Arg Arg Asn Asn Asn Met Gln Trp Phe Lys Ile Pro Ala Lys Thr Tyr465 470 475 480Phe Glu Pro Asn Ala Ile Lys Tyr Leu Arg Asp Met Tyr Gly Ile Glu 485 490 495Lys Ala Val Ile Val Cys Asp Lys Val Met Glu Gln Leu Gly Ile Val 500 505 510Asp Lys Ile Ile Asp Gln Leu Arg Ala Arg Ser Asn Arg Val Thr Phe 515 520 525Arg Ile Ile Asp Tyr Val Glu Pro Glu Pro Ser Val Glu Thr Val Glu 530 535 540Arg Gly Ala Ala Met Met Arg Glu Glu Phe Glu Pro Asp Thr Ile Ile545 550 555 560Ala Val Gly Gly Gly Ser Pro Met Asp Ala Ser Lys Ile Met Trp Leu 565 570 575Leu Tyr Glu His Pro Glu Ile Ser Phe Ser Asp Val Arg Glu Lys Phe 580 585 590Phe Asp Ile Arg Lys Arg Ala Phe Lys Ile Pro Pro Leu Gly Lys Lys 595 600 605Ala Lys Leu Val Cys Ile Pro Thr Ser Ser Gly Thr Gly Ser Glu Val 610 615 620Thr Pro Phe Ala Val Ile Thr Asp His Lys Thr Gly Tyr Lys Tyr Pro625 630 635 640Ile Thr Asp Tyr Ala Leu Thr Pro Ser Val Ala Ile Val Asp Pro Val 645 650 655Leu Ala Arg Thr Gln Pro Arg Lys Leu Ala Ser Asp Ala Gly Phe Asp 660 665 670Ala Leu Thr His Ala Phe Glu Ala Tyr Val Ser Val Tyr Ala Asn Asp 675 680 685Phe Thr Asp Gly Met Ala Leu His Ala Ala Lys Leu Val Trp Asp Asn 690 695 700Leu Ala Glu Ser Val Asn Gly Glu Pro Gly Glu Glu Lys Thr Arg Ala705 710 715 720Gln Glu Lys Met His Asn Ala Ala Thr Met Ala Gly Met Ala Phe Gly 725 730 735Ser Ala Phe Leu Gly Met Cys His Gly Met Ala His Thr Ile Gly Ala 740 745 750Leu Cys His Val Ala His Gly Arg Thr Asn Ser Ile Leu Leu Pro Tyr 755 760 765Val Ile Arg Tyr Asn Gly Ser Val Pro Glu Glu Pro Thr Ser Trp Pro 770 775 780Lys Tyr Asn Lys Tyr Ile Ala Pro Glu Arg Tyr Gln Glu Ile Ala Lys785 790 795 800Asn Leu Gly Val Asn Pro Gly Lys Thr Pro Glu Glu Gly Val Glu Asn 805 810 815Leu Ala Lys Ala Val Glu Asp Tyr Arg Asp Asn Lys Leu Gly Met Asn 820 825 830Lys Ser Phe Gln Glu Cys Gly Val Asp Glu Asp Tyr Tyr Trp Ser Ile 835 840 845Ile Asp Gln Ile Gly Met Arg Ala Tyr Glu Asp Gln Cys Ala Pro Ala 850 855 860Asn Pro Arg Ile Pro Gln Ile Glu Asp Met Lys Asp Ile Ala Ile Ala865 870 875 880Ala Tyr Tyr Gly Val Ser Gln Ala Glu Gly His Lys Leu Arg Val Gln 885 890 895Arg Gln Gly Glu Ala Ala Thr Glu Glu Ala Ser Glu Arg Ala 900 905 91045879DNABifidobacterium adolescentisPFLA 45atgtctgaac atattttccg ttccacgacc agacacatgc tgagggattc caaggactac 60gtcaatcaga cgctgatggg aggcctgtcc ggattcgaat cgccaatcgg cttggaccgt 120ctcgaccgca tcaaggcgtt gaaaagcggc gatatcggtt tcgtgcactc gtgggacatc 180aacacttccg tggatggtcc tggcaccaga atgaccgtgt tcatgagcgg atgccctctg 240cgctgccagt actgccagaa tccggatact tggaagatgc gcgacggcaa gcccgtctac 300tacgaagcca tggtcaagaa aatcgagcgg tatgccgatt tattcaaggc caccggcggc 360ggcatcactt tctccggcgg cgaatccatg atgcagccgg ctttcgtgtc acgcgtgttc 420catgccgcca agcagatggg agtgcatacc tgcctcgaca cgtccggatt cctcggggcg 480agctacaccg atgacatggt ggatgacatc gacctgtgcc tgcttgacgt

caaatccggc 540gatgaggaga cctaccataa ggtgaccggc ggcatcctgc agccgaccat cgacttcgga 600cagcgtctgg ccaaggcagg caagaagatc tgggtgcgtt tcgtgctcgt gccgggcctc 660acatcctccg aagaaaacgt cgagaacgtg gcgaagatct gcgagacctt cggcgacgcg 720ttggaacata tcgacgtatt gcccttccac cagcttggcc gtccgaagtg gcacatgctg 780aacatcccat acccgttgga ggaccagaaa ggcccgtccg cggcaatgaa acaacgtgtg 840gtcgagcagt tccagtcgca cggcttcacc gtgtactaa 87946292PRTBifidobacterium adolescentisPFLA 46Met Ser Glu His Ile Phe Arg Ser Thr Thr Arg His Met Leu Arg Asp1 5 10 15Ser Lys Asp Tyr Val Asn Gln Thr Leu Met Gly Gly Leu Ser Gly Phe 20 25 30Glu Ser Pro Ile Gly Leu Asp Arg Leu Asp Arg Ile Lys Ala Leu Lys 35 40 45Ser Gly Asp Ile Gly Phe Val His Ser Trp Asp Ile Asn Thr Ser Val 50 55 60Asp Gly Pro Gly Thr Arg Met Thr Val Phe Met Ser Gly Cys Pro Leu65 70 75 80Arg Cys Gln Tyr Cys Gln Asn Pro Asp Thr Trp Lys Met Arg Asp Gly 85 90 95Lys Pro Val Tyr Tyr Glu Ala Met Val Lys Lys Ile Glu Arg Tyr Ala 100 105 110Asp Leu Phe Lys Ala Thr Gly Gly Gly Ile Thr Phe Ser Gly Gly Glu 115 120 125Ser Met Met Gln Pro Ala Phe Val Ser Arg Val Phe His Ala Ala Lys 130 135 140Gln Met Gly Val His Thr Cys Leu Asp Thr Ser Gly Phe Leu Gly Ala145 150 155 160Ser Tyr Thr Asp Asp Met Val Asp Asp Ile Asp Leu Cys Leu Leu Asp 165 170 175Val Lys Ser Gly Asp Glu Glu Thr Tyr His Lys Val Thr Gly Gly Ile 180 185 190Leu Gln Pro Thr Ile Asp Phe Gly Gln Arg Leu Ala Lys Ala Gly Lys 195 200 205Lys Ile Trp Val Arg Phe Val Leu Val Pro Gly Leu Thr Ser Ser Glu 210 215 220Glu Asn Val Glu Asn Val Ala Lys Ile Cys Glu Thr Phe Gly Asp Ala225 230 235 240Leu Glu His Ile Asp Val Leu Pro Phe His Gln Leu Gly Arg Pro Lys 245 250 255Trp His Met Leu Asn Ile Pro Tyr Pro Leu Glu Asp Gln Lys Gly Pro 260 265 270Ser Ala Ala Met Lys Gln Arg Val Val Glu Gln Phe Gln Ser His Gly 275 280 285Phe Thr Val Tyr 290472376DNABifidobacterium adolescentisPFLB 47atggcagcag ttgatgcaac ggcggtctcc caggaggaac ttgaggctaa ggcttgggaa 60ggcttcaccg agggcaactg gcagaaggac attgatgtcc gcgacttcat ccagaagaac 120tacacgccat atgagggcga cgagtccttc ctggctgacg ccaccgacaa gaccaagcac 180ctgtggaagt atctggacga caactatctg tccgtggagc gcaagcagcg cgtctacgac 240gtggacaccc acaccccggc gggcatcgac gccttcccgg ccggctacat cgattccccg 300gaagtcgaca atgtgattgt cggtctgcag accgatgtgc cgtgcaagcg cgccatgatg 360ccgaacggcg gctggcgtat ggtcgagcag gccatcaagg aagccggcaa ggagcccgat 420ccggagatca agaagatctt caccaagtac cgcaagaccc acaacgacgg cgtcttcggc 480gtctacacca agcagatcaa ggtagctcgc cacaacaaga tcctcaccgg cctgccggat 540gcctacggcc gtggccgcat catcggcgat taccgtcgtg tggccctgta cggcgtgaac 600gcgctgatca agttcaagca gcgcgacaag gactccatcc cgtaccgcaa cgacttcacc 660gagccggaga tcgagcactg gatccgcttc cgtgaggagc atgacgagca gatcaaggcc 720ctgaagcagc tgatcaacct cggcaacgag tacggcctcg acctgtcccg cccggcacag 780accgcacagg aagccgtgca gtggacctac atgggctacc tcgcctccgt caagagccag 840gacggcgccg ccatgtcctt cggccgtgtc tccaccttct tcgacgtcta cttcgagcgc 900gacctgaagg ccggcaagat caccgagacc gacgcacagg agatcatcga taacctggtc 960atgaagctgc gcatcgtgcg cttcctgcgc accaaggatt acgacgcgat cttctccggc 1020gatccgtact gggcgacttg gtccgacgcc ggcttcggcg acgacggccg taccatggtc 1080accaagacct cgttccgtct gctcaacacc ctgaccctcg agcacctcgg acctggcccg 1140gagccgaaca tcaccatctt ctgggatccg aagctgccgg aagcctacaa gcgcttctgc 1200gcccgaatct ccatcgacac ctcggccatc cagtacgagt ccgataagga aatccgctcc 1260cactggggcg acgacgccgc catcgcatgc tgcgtctccc cgatgcgcgt gggcaagcag 1320atgcagttct tcgccgcccg tgtgaactcc gccaaggccc tgctgtacgc catcaacggc 1380ggacgcgacg agatgaccgg catgcaggtc atcgacaagg gcgtcatcga cccgatcaag 1440ccggaagccg atggcacgct ggattacgag aaggtcaagg ccaactacga gaaggccctc 1500gaatggctgt ccgagaccta tgtgatggct ctgaacatca tccattacat gcatgataag 1560tacgcttacg agtccatcga gatggctctg cacgacaagg aagtgtaccg caccctcggc 1620tgcggcatgt ccggcctgtc gatcgcggcc gactccctgt ccgcatgcaa gtacgccaag 1680gtctacccga tctacaacaa ggacgccaag accacgccgg gccacgagaa cgagtacgtc 1740gaaggcgccg atgacgatct gatcgtcggc taccgcaccg aaggcgactt cccgctgtac 1800ggcaacgatg atgaccgtgc cgacgacatc gccaagtggg tcgtctccac cgtcatgggc 1860caggtcaagc gtctgccggt gtaccgcgac gccgtcccga cccagtccat cctgaccatc 1920acctccaatg tggaatacgg caaggccacc ggcgccttcc cgtccggcca caagaagggc 1980accccgtacg ctccgggcgc caacccggag aacggcatgg actcccacgg catgctgccg 2040tccatgttct ccgtcggcaa gatcgactac aacgacgctc ttgacggcat ctcgctgacc 2100aacaccatca cccctgatgg tctgggccgc gacgaggaag agcgtatcgg caacctcgtt 2160ggcatcctgg acgccggcaa cggccacggc ctgtaccacg ccaacatcaa cgtgctgcgc 2220aaggagcagc tcgaggatgc cgtcgagcat ccggagaagt acccgcacct gaccgtgcgc 2280gtctccggct acgcggtgaa cttcgtcaag ctcaccaagg aacagcagct cgacgtgatc 2340tcccgtacgt tccaccaggg cgctgtcgtc gactga 237648791PRTBifidobacterium adolescentisPFLB 48Met Ala Ala Val Asp Ala Thr Ala Val Ser Gln Glu Glu Leu Glu Ala1 5 10 15Lys Ala Trp Glu Gly Phe Thr Glu Gly Asn Trp Gln Lys Asp Ile Asp 20 25 30Val Arg Asp Phe Ile Gln Lys Asn Tyr Thr Pro Tyr Glu Gly Asp Glu 35 40 45Ser Phe Leu Ala Asp Ala Thr Asp Lys Thr Lys His Leu Trp Lys Tyr 50 55 60Leu Asp Asp Asn Tyr Leu Ser Val Glu Arg Lys Gln Arg Val Tyr Asp65 70 75 80Val Asp Thr His Thr Pro Ala Gly Ile Asp Ala Phe Pro Ala Gly Tyr 85 90 95Ile Asp Ser Pro Glu Val Asp Asn Val Ile Val Gly Leu Gln Thr Asp 100 105 110Val Pro Cys Lys Arg Ala Met Met Pro Asn Gly Gly Trp Arg Met Val 115 120 125Glu Gln Ala Ile Lys Glu Ala Gly Lys Glu Pro Asp Pro Glu Ile Lys 130 135 140Lys Ile Phe Thr Lys Tyr Arg Lys Thr His Asn Asp Gly Val Phe Gly145 150 155 160Val Tyr Thr Lys Gln Ile Lys Val Ala Arg His Asn Lys Ile Leu Thr 165 170 175Gly Leu Pro Asp Ala Tyr Gly Arg Gly Arg Ile Ile Gly Asp Tyr Arg 180 185 190Arg Val Ala Leu Tyr Gly Val Asn Ala Leu Ile Lys Phe Lys Gln Arg 195 200 205Asp Lys Asp Ser Ile Pro Tyr Arg Asn Asp Phe Thr Glu Pro Glu Ile 210 215 220Glu His Trp Ile Arg Phe Arg Glu Glu His Asp Glu Gln Ile Lys Ala225 230 235 240Leu Lys Gln Leu Ile Asn Leu Gly Asn Glu Tyr Gly Leu Asp Leu Ser 245 250 255Arg Pro Ala Gln Thr Ala Gln Glu Ala Val Gln Trp Thr Tyr Met Gly 260 265 270Tyr Leu Ala Ser Val Lys Ser Gln Asp Gly Ala Ala Met Ser Phe Gly 275 280 285Arg Val Ser Thr Phe Phe Asp Val Tyr Phe Glu Arg Asp Leu Lys Ala 290 295 300Gly Lys Ile Thr Glu Thr Asp Ala Gln Glu Ile Ile Asp Asn Leu Val305 310 315 320Met Lys Leu Arg Ile Val Arg Phe Leu Arg Thr Lys Asp Tyr Asp Ala 325 330 335Ile Phe Ser Gly Asp Pro Tyr Trp Ala Thr Trp Ser Asp Ala Gly Phe 340 345 350Gly Asp Asp Gly Arg Thr Met Val Thr Lys Thr Ser Phe Arg Leu Leu 355 360 365Asn Thr Leu Thr Leu Glu His Leu Gly Pro Gly Pro Glu Pro Asn Ile 370 375 380Thr Ile Phe Trp Asp Pro Lys Leu Pro Glu Ala Tyr Lys Arg Phe Cys385 390 395 400Ala Arg Ile Ser Ile Asp Thr Ser Ala Ile Gln Tyr Glu Ser Asp Lys 405 410 415Glu Ile Arg Ser His Trp Gly Asp Asp Ala Ala Ile Ala Cys Cys Val 420 425 430Ser Pro Met Arg Val Gly Lys Gln Met Gln Phe Phe Ala Ala Arg Val 435 440 445Asn Ser Ala Lys Ala Leu Leu Tyr Ala Ile Asn Gly Gly Arg Asp Glu 450 455 460Met Thr Gly Met Gln Val Ile Asp Lys Gly Val Ile Asp Pro Ile Lys465 470 475 480Pro Glu Ala Asp Gly Thr Leu Asp Tyr Glu Lys Val Lys Ala Asn Tyr 485 490 495Glu Lys Ala Leu Glu Trp Leu Ser Glu Thr Tyr Val Met Ala Leu Asn 500 505 510Ile Ile His Tyr Met His Asp Lys Tyr Ala Tyr Glu Ser Ile Glu Met 515 520 525Ala Leu His Asp Lys Glu Val Tyr Arg Thr Leu Gly Cys Gly Met Ser 530 535 540Gly Leu Ser Ile Ala Ala Asp Ser Leu Ser Ala Cys Lys Tyr Ala Lys545 550 555 560Val Tyr Pro Ile Tyr Asn Lys Asp Ala Lys Thr Thr Pro Gly His Glu 565 570 575Asn Glu Tyr Val Glu Gly Ala Asp Asp Asp Leu Ile Val Gly Tyr Arg 580 585 590Thr Glu Gly Asp Phe Pro Leu Tyr Gly Asn Asp Asp Asp Arg Ala Asp 595 600 605Asp Ile Ala Lys Trp Val Val Ser Thr Val Met Gly Gln Val Lys Arg 610 615 620Leu Pro Val Tyr Arg Asp Ala Val Pro Thr Gln Ser Ile Leu Thr Ile625 630 635 640Thr Ser Asn Val Glu Tyr Gly Lys Ala Thr Gly Ala Phe Pro Ser Gly 645 650 655His Lys Lys Gly Thr Pro Tyr Ala Pro Gly Ala Asn Pro Glu Asn Gly 660 665 670Met Asp Ser His Gly Met Leu Pro Ser Met Phe Ser Val Gly Lys Ile 675 680 685Asp Tyr Asn Asp Ala Leu Asp Gly Ile Ser Leu Thr Asn Thr Ile Thr 690 695 700Pro Asp Gly Leu Gly Arg Asp Glu Glu Glu Arg Ile Gly Asn Leu Val705 710 715 720Gly Ile Leu Asp Ala Gly Asn Gly His Gly Leu Tyr His Ala Asn Ile 725 730 735Asn Val Leu Arg Lys Glu Gln Leu Glu Asp Ala Val Glu His Pro Glu 740 745 750Lys Tyr Pro His Leu Thr Val Arg Val Ser Gly Tyr Ala Val Asn Phe 755 760 765Val Lys Leu Thr Lys Glu Gln Gln Leu Asp Val Ile Ser Arg Thr Phe 770 775 780His Gln Gly Ala Val Val Asp785 7904959DNABifidobacterium adolescentisForward Primer 49gctataccaa gcatacaatc aactatctca tatacaatga cgagtcctgt tattggcac 595059DNABifidobacterium adolescentisReverse Primer 50ttataaaact ttaactaata attagagatt aaatcgctca ctcgttatcg ccagcggtt 595159DNALactobacillus plantarumForward Primer 1 51ctataccaag catacaatca actatctcat atacaatgac aacagattac tcatcacca 595259DNALactobacillus plantarumReverse Primer 1 52aaactttaac taataattag agattaaatc gcttatttta aacccttcca ttgccaatc 595359DNALactobacillus plantarumForward Primer 2 53gctataccaa gcatacaatc aactatctca tatacaatga gtgaagcaat taaatccaa 595459DNALactobacillus plantarumReverse Primer 2 54aaaactttaa ctaataatta gagattaaat cgcctacttc aatgcagtcc atttccagt 595559DNANeurospora crassaForward Primer 55tataccaagc atacaatcaa ctatctcata tacaatgggc ggaacacaga tcacaattc 595659DNANeurospora crassaReverse Primer 56aaaactttaa ctaataatta gagattaaat cgcctactcg aacttgggca gttggtaga 595759DNAAspergillus nigerForward Primer 57ctataccaag catacaatca actatctcat atacaatgcc tggagaggtc atcgacagg 595859DNAAspergillus nigerReverse Primer 58aactttaact aataattaga gattaaatcg cttattcaaa ggagggcata tcatacgta 595959DNALactobacillus caseiForward Primer 59tataccaagc atacaatcaa ctatctcata tacaatggat acaaaagtaa agactgttg 596059DNALactobacillus caseiReverse Primer 60aaactttaac taataattag agattaaatc gcttacttga ttggtttcca ggtccattc 59612406DNABifidobacterium bifidumPHK 61atggacaagt actggcgcgt tgccaactac ctttccatcg gccagatcta tctgcggagc 60aacccgctga tgaaggaacc cttcacccgt gaagacgtga agcaccgtct ggtggggcac 120tggggcacca cccctggcct gaacttcctc atcggccaca tcaaccgctt catcgctgac 180cacggccaga acaccgtgtt catcatgggc ccgggccacg gtggccccgc cggtacctcc 240cagtcctacc tggacggcac ctacaccgag acgtacccca acattaccaa ggatgaggct 300ggcctgcaga agttcttccg ccagttctcc tacccgggcg gcatcccctc ccacttcgct 360ccggaaaccc cgggctccat ccacgagggt ggcgagctgg gttacgccct gtcccacgct 420tatggcgcga tcatggacaa cccgagcctg ttcgttccct gcatcgtcgg cgatggtgag 480gccgagaccg gcccgctggc caccggctgg cagtccaaca agctcgtgaa cccgcgcacc 540gacggtatcg tgctgccgat cctgcacctt aacggctaca agatcgccaa cccgaccatc 600ctgtcgcgca tctccgacga agagctccac gagttcttcc acggcatggg ctacgagccc 660tatgagttcg tcgccggctt cgacgatgag gaccacatgt ccatccaccg tcgcttcgcc 720gagctgttcg agtccgtgtg ggacgagatc tgcgacatca aggccgcggc gaacacggac 780aacatgcacc gtccgttcta cccgatgatc atcttccgca ccccgaaggg ctggacctgc 840ccgaagtaca tcgacggcaa gaagaccgag ggctcctggc gcgcccacca ggtgccgctg 900gcttccgccc gcgacaccga ggcccacttc gaggtcctca agaactggct ggagtcctac 960aagccggaag agctgttcga cgccaacggt gccgtcaagg acgacgtcct ggccttcatg 1020ccgaagggcg agctgcgcat cggcgccaac ccgaacgcca acggtggcgt gatccgcagg 1080gatctcgtgc tgccggcgct cgaggactac gaggtcaagg aagtcaagga gttcggacac 1140ggctggggcc agctcgaggc cacccgtcgt ctgggcgtct acacccgcga catcatcaag 1200aacaacatgc acgacttccg catcttcgga ccggatgaga ccgcttccaa ccgtctgcag 1260gcttcctacg aggtcaccaa caagcagtgg gacgccggct acatctccga cgaggttgac 1320gagcacatgc acgtctccgg ccaggtcgtc gagcagctgt ccgagcacca gatggaaggc 1380ttcctcgagg cctacctgct gaccggccgt cacggcatct ggagctccta cgagtccttt 1440gtgcacgtga tcgactccat gctgaaccag cacgccaagt ggcttgaggc taccgtccgc 1500gagattccgt ggcgcaagcc gattgcctcc atgaacctgc tggtctcctc gcacgtgtgg 1560cgtcaggatc acaacggttt ctcgcaccag gacccgggtg tcacctccgt cctgctgaac 1620aagtgcttcc acaacgatca cgtcatcggc atctacttcg cgaccgacgc gaacatgctg 1680ctggccatcg ccgagaagtg ctacaagtcc accaacaaga tcaacgccat catcgccggc 1740aagcagccgg ccgccacctg gctgaccctg gacgaggctc gtgccgagct cgccaagggt 1800gccgccgctt gggattgggc ttccaccgcg aagaccaacg atgaggctca ggtcgtgctc 1860gccgccgccg gcgacgtccc gacccaggag atcatggccg cttccgacaa gttgaaggcc 1920ctgggcatca agttcaaggt cgtcaacgtg gctgatctgc tctccctgca gtctgccaag 1980gagaacgacg aagcgctgac cgacgaggag ttcgccgaca tcttcaccgc cgacaagccg 2040gtgctgttcg cgtaccactc ctacgcccac gacgtgcgtg gcctgatcta cgatcgcccg 2100aaccacgata acttcaacgt ccacggctac gaggaggagg gctccaccac caccccgtac 2160gacatggttc gcgtgaacga gctggatcgc tacgagctga ccgccgaagc cctgcgcatg 2220atcgacgccg acaagtacgc cgacgagatc cagaagctgg aggacttccg tcaggaagcg 2280ttccagttcg cggtcgacaa gggctacgac cacccggact acaccgactg ggtgtactcg 2340ggcgtcaaga ccgacaagaa gggcgccgtc accgccaccg ccgccaccgc tggcgacaac 2400gagtga 240662801PRTBifidobacterium bifidumPHK 62Met Asp Lys Tyr Trp Arg Val Ala Asn Tyr Leu Ser Ile Gly Gln Ile1 5 10 15Tyr Leu Arg Ser Asn Pro Leu Met Lys Glu Pro Phe Thr Arg Glu Asp 20 25 30Val Lys His Arg Leu Val Gly His Trp Gly Thr Thr Pro Gly Leu Asn 35 40 45Phe Leu Ile Gly His Ile Asn Arg Phe Ile Ala Asp His Gly Gln Asn 50 55 60Thr Val Phe Ile Met Gly Pro Gly His Gly Gly Pro Ala Gly Thr Ser65 70 75 80Gln Ser Tyr Leu Asp Gly Thr Tyr Thr Glu Thr Tyr Pro Asn Ile Thr 85 90 95Lys Asp Glu Ala Gly Leu Gln Lys Phe Phe Arg Gln Phe Ser Tyr Pro 100 105 110Gly Gly Ile Pro Ser His Phe Ala Pro Glu Thr Pro Gly Ser Ile His 115 120 125Glu Gly Gly Glu Leu Gly Tyr Ala Leu Ser His Ala Tyr Gly Ala Ile 130 135 140Met Asp Asn Pro Ser Leu Phe Val Pro Cys Ile Val Gly Asp Gly Glu145 150 155 160Ala Glu Thr Gly Pro Leu Ala Thr Gly Trp Gln Ser Asn Lys Leu Val 165 170 175Asn Pro Arg Thr Asp Gly Ile Val Leu Pro Ile Leu His Leu Asn Gly 180 185 190Tyr Lys Ile Ala Asn Pro Thr Ile Leu Ser Arg Ile Ser Asp Glu Glu 195 200 205Leu His Glu Phe Phe His Gly Met Gly Tyr Glu Pro Tyr Glu Phe Val 210 215 220Ala Gly Phe Asp Asp Glu Asp His Met Ser Ile His Arg Arg Phe Ala225 230 235 240Glu Leu Phe Glu Ser Val Trp Asp Glu Ile Cys Asp Ile Lys Ala Ala 245 250 255Ala Asn Thr Asp Asn Met His Arg Pro Phe Tyr Pro Met Ile Ile Phe 260 265 270Arg Thr Pro Lys Gly Trp Thr Cys Pro Lys Tyr Ile Asp Gly Lys Lys 275 280 285Thr Glu Gly Ser Trp Arg Ala His Gln Val Pro Leu Ala Ser Ala Arg 290

295 300Asp Thr Glu Ala His Phe Glu Val Leu Lys Asn Trp Leu Glu Ser Tyr305 310 315 320Lys Pro Glu Glu Leu Phe Asp Ala Asn Gly Ala Val Lys Asp Asp Val 325 330 335Leu Ala Phe Met Pro Lys Gly Glu Leu Arg Ile Gly Ala Asn Pro Asn 340 345 350Ala Asn Gly Gly Val Ile Arg Arg Asp Leu Val Leu Pro Ala Leu Glu 355 360 365Asp Tyr Glu Val Lys Glu Val Lys Glu Phe Gly His Gly Trp Gly Gln 370 375 380Leu Glu Ala Thr Arg Arg Leu Gly Val Tyr Thr Arg Asp Ile Ile Lys385 390 395 400Asn Asn Met His Asp Phe Arg Ile Phe Gly Pro Asp Glu Thr Ala Ser 405 410 415Asn Arg Leu Gln Ala Ser Tyr Glu Val Thr Asn Lys Gln Trp Asp Ala 420 425 430Gly Tyr Ile Ser Asp Glu Val Asp Glu His Met His Val Ser Gly Gln 435 440 445Val Val Glu Gln Leu Ser Glu His Gln Met Glu Gly Phe Leu Glu Ala 450 455 460Tyr Leu Leu Thr Gly Arg His Gly Ile Trp Ser Ser Tyr Glu Ser Phe465 470 475 480Val His Val Ile Asp Ser Met Leu Asn Gln His Ala Lys Trp Leu Glu 485 490 495Ala Thr Val Arg Glu Ile Pro Trp Arg Lys Pro Ile Ala Ser Met Asn 500 505 510Leu Leu Val Ser Ser His Val Trp Arg Gln Asp His Asn Gly Phe Ser 515 520 525His Gln Asp Pro Gly Val Thr Ser Val Leu Leu Asn Lys Cys Phe His 530 535 540Asn Asp His Val Ile Gly Ile Tyr Phe Ala Thr Asp Ala Asn Met Leu545 550 555 560Leu Ala Ile Ala Glu Lys Cys Tyr Lys Ser Thr Asn Lys Ile Asn Ala 565 570 575Ile Ile Ala Gly Lys Gln Pro Ala Ala Thr Trp Leu Thr Leu Asp Glu 580 585 590Ala Arg Ala Glu Leu Ala Lys Gly Ala Ala Ala Trp Asp Trp Ala Ser 595 600 605Thr Ala Lys Thr Asn Asp Glu Ala Gln Val Val Leu Ala Ala Ala Gly 610 615 620Asp Val Pro Thr Gln Glu Ile Met Ala Ala Ser Asp Lys Leu Lys Ala625 630 635 640Leu Gly Ile Lys Phe Lys Val Val Asn Val Ala Asp Leu Leu Ser Leu 645 650 655Gln Ser Ala Lys Glu Asn Asp Glu Ala Leu Thr Asp Glu Glu Phe Ala 660 665 670Asp Ile Phe Thr Ala Asp Lys Pro Val Leu Phe Ala Tyr His Ser Tyr 675 680 685Ala His Asp Val Arg Gly Leu Ile Tyr Asp Arg Pro Asn His Asp Asn 690 695 700Phe Asn Val His Gly Tyr Glu Glu Glu Gly Ser Thr Thr Thr Pro Tyr705 710 715 720Asp Met Val Arg Val Asn Glu Leu Asp Arg Tyr Glu Leu Thr Ala Glu 725 730 735Ala Leu Arg Met Ile Asp Ala Asp Lys Tyr Ala Asp Glu Ile Gln Lys 740 745 750Leu Glu Asp Phe Arg Gln Glu Ala Phe Gln Phe Ala Val Asp Lys Gly 755 760 765Tyr Asp His Pro Asp Tyr Thr Asp Trp Val Tyr Ser Gly Val Lys Thr 770 775 780Asp Lys Lys Gly Ala Val Thr Ala Thr Ala Ala Thr Ala Gly Asp Asn785 790 795 800Glu632481DNABifidobacterium galliciumPHK 63atgacaagtc ctgttattgg caccccctgg cagaagctga accgtccggt ttcggaagag 60gccatcgaag gcatggacaa gtattggcgc gcgtccaact acatgtccat cggccagatc 120tacctgcgta gcaacccgct gatgaaggaa cctttcactc gcgatgacgt gaagtaccgc 180ctcgtcggcc actggggtac caccccgggc ctgaacttcc ttctggccca catcaaccgc 240ctgatcgctg accaccagca gaacaccgtg ttcatcatgg gtcctggcca cggcggtcct 300gccggtactg cacagtccta tctggacggc acgtacaccg agtactaccc gaacatcacc 360aaggatgagg aaggcctgca gaagttcttc cgtcagttct cttatccggg cggcattcct 420tcgcacttcg ctccggagac cccgggctcg atccacgaag gtggcgagct gggctatgct 480ctgtcccacg cttacggcgc tgtgatgaac aacccgagcc tgttcgtgcc ctgcatcgtc 540ggcgacggcg aagctgagac tggccctctg gccaccggct ggcagtccaa caagctcgtg 600aacccgcgca ccgatggcat cgtgctgccg atcctgcacc tcaacggcta caagatcgcc 660aacccgacca tcctcgctcg cgtgtccgat gaagagctgc atgacttctt ccgcggcctg 720ggctaccacc cgtacgagtt cgtcgccggc ttcgacaacg aggatcacct gtcgatccac 780cgtcgtttcg ctgagctctt cgagaccatc ttcgacgaga tctgcgatat caaggctgcc 840gccaacacgg atgacatgac ccgtccgttc tacccgatgc tgattttccg caccccgaag 900ggttggacct gcccgaagtt catcgacggc aagaagaccg aaggctcttg gcgtgcacac 960caggtgccgc tggcttccgc ccgcgatacc gaggctcact tcgaagtgct gaagaactgg 1020atggcttcct acaagccgga agagctcttc gacgacaagg gcgccatcaa ggatgacgtc 1080gtcgacttca tgccgaaggg cgatctgcgt atcggcgcca acccgaatgc taacggtggc 1140gtgatccgtg aagagctcga tctgccggcc ctcgagaact atgaggtcaa ggaagtcaag 1200gaattcggcc acggctgggg ccagcttgag gctacgcgta agctcggtga gtacacgcgt 1260gacatcatca agaacaaccc ggattcgttc cgcatcttcg gacctgacga gaccgcttcg 1320aaccgtctgc aggcttccta cgaagtcacc aacaagcagt gggacaacgg ctacctgtcc 1380aaggacctcg tcgatgagca catggctgtc accggccagg tcactgagca gctctccgaa 1440caccagtgcg aaggcttcct cgaggcctac ctgctcacgg gtcgtcacgg catctggagc 1500tcctacgagt ccttcgtgca cgtcatcgac tccatgctca accagcacgc caagtggctc 1560gaggctacgg tgcgtgagat tccgtggcgt aagccgatct cctcgatgaa cctgctcgtg 1620tcctcgcacg tgtggcgtca ggatcacaac ggcttctcgc accaggatcc gggtgtcacc 1680tccgtcctgt tgaacaagac gttcaacaac gatcacgtca tcggcctgta cttcgcaacc 1740gacgccaacg tgctgctggc catcgctgag aagtgctaca agtcgaccaa catgatcaac 1800gccatcgtcg ccggcaagca gcctgctgcc acctggacga cgctggacga agctcgtgag 1860ctggttgcca agggcgctgg cgaattcgag tgggcttcga acgtcaagac caacgatgag 1920gccgagattg tgctcgcttc cgctggcgat gttccgactc aggagctcat ggctgctgct 1980gatcgtctga acaagctcgg cgtcaagttc aaggtcgtca acgtggtgga tctgatcaag 2040ctgcagtccg ccaaggagaa cgatcaggct ctgtccgatg ccgagttcgc tgagctcttc 2100accgaagaca agccggtgct gttcgcgtac cactcctatg cgcacgacgt gcgcggcctg 2160atcttcgatc gcccgaacca cgacaacttc aacgtggtcg gctacaagga gcagggctcc 2220accaccacgc cgtacgacat ggtgcgtgtc aacgacatcg atcgttatga gctgaccgct 2280acggcactgc gcatgatcga cgccgacaag tacgccgacg agatcaagaa gctcgaggac 2340ttccgcatcg aggcttacca gttcgctgtc gacaacggct acgacattcc ggattacacc 2400gactgggtgt ggccgggcgt caagaccgac ctgccgggcg ccgtttccgc aacggctgcc 2460accgcaggcg acaacgagtg a 248164826PRTBifidobacterium galliciumPHK 64Met Thr Ser Pro Val Ile Gly Thr Pro Trp Gln Lys Leu Asn Arg Pro1 5 10 15Val Ser Glu Glu Ala Ile Glu Gly Met Asp Lys Tyr Trp Arg Ala Ser 20 25 30Asn Tyr Met Ser Ile Gly Gln Ile Tyr Leu Arg Ser Asn Pro Leu Met 35 40 45Lys Glu Pro Phe Thr Arg Asp Asp Val Lys Tyr Arg Leu Val Gly His 50 55 60Trp Gly Thr Thr Pro Gly Leu Asn Phe Leu Leu Ala His Ile Asn Arg65 70 75 80Leu Ile Ala Asp His Gln Gln Asn Thr Val Phe Ile Met Gly Pro Gly 85 90 95His Gly Gly Pro Ala Gly Thr Ala Gln Ser Tyr Leu Asp Gly Thr Tyr 100 105 110Thr Glu Tyr Tyr Pro Asn Ile Thr Lys Asp Glu Glu Gly Leu Gln Lys 115 120 125Phe Phe Arg Gln Phe Ser Tyr Pro Gly Gly Ile Pro Ser His Phe Ala 130 135 140Pro Glu Thr Pro Gly Ser Ile His Glu Gly Gly Glu Leu Gly Tyr Ala145 150 155 160Leu Ser His Ala Tyr Gly Ala Val Met Asn Asn Pro Ser Leu Phe Val 165 170 175Pro Cys Ile Val Gly Asp Gly Glu Ala Glu Thr Gly Pro Leu Ala Thr 180 185 190Gly Trp Gln Ser Asn Lys Leu Val Asn Pro Arg Thr Asp Gly Ile Val 195 200 205Leu Pro Ile Leu His Leu Asn Gly Tyr Lys Ile Ala Asn Pro Thr Ile 210 215 220Leu Ala Arg Val Ser Asp Glu Glu Leu His Asp Phe Phe Arg Gly Leu225 230 235 240Gly Tyr His Pro Tyr Glu Phe Val Ala Gly Phe Asp Asn Glu Asp His 245 250 255Leu Ser Ile His Arg Arg Phe Ala Glu Leu Phe Glu Thr Ile Phe Asp 260 265 270Glu Ile Cys Asp Ile Lys Ala Ala Ala Asn Thr Asp Asp Met Thr Arg 275 280 285Pro Phe Tyr Pro Met Leu Ile Phe Arg Thr Pro Lys Gly Trp Thr Cys 290 295 300Pro Lys Phe Ile Asp Gly Lys Lys Thr Glu Gly Ser Trp Arg Ala His305 310 315 320Gln Val Pro Leu Ala Ser Ala Arg Asp Thr Glu Ala His Phe Glu Val 325 330 335Leu Lys Asn Trp Met Ala Ser Tyr Lys Pro Glu Glu Leu Phe Asp Asp 340 345 350Lys Gly Ala Ile Lys Asp Asp Val Val Asp Phe Met Pro Lys Gly Asp 355 360 365Leu Arg Ile Gly Ala Asn Pro Asn Ala Asn Gly Gly Val Ile Arg Glu 370 375 380Glu Leu Asp Leu Pro Ala Leu Glu Asn Tyr Glu Val Lys Glu Val Lys385 390 395 400Glu Phe Gly His Gly Trp Gly Gln Leu Glu Ala Thr Arg Lys Leu Gly 405 410 415Glu Tyr Thr Arg Asp Ile Ile Lys Asn Asn Pro Asp Ser Phe Arg Ile 420 425 430Phe Gly Pro Asp Glu Thr Ala Ser Asn Arg Leu Gln Ala Ser Tyr Glu 435 440 445Val Thr Asn Lys Gln Trp Asp Asn Gly Tyr Leu Ser Lys Asp Leu Val 450 455 460Asp Glu His Met Ala Val Thr Gly Gln Val Thr Glu Gln Leu Ser Glu465 470 475 480His Gln Cys Glu Gly Phe Leu Glu Ala Tyr Leu Leu Thr Gly Arg His 485 490 495Gly Ile Trp Ser Ser Tyr Glu Ser Phe Val His Val Ile Asp Ser Met 500 505 510Leu Asn Gln His Ala Lys Trp Leu Glu Ala Thr Val Arg Glu Ile Pro 515 520 525Trp Arg Lys Pro Ile Ser Ser Met Asn Leu Leu Val Ser Ser His Val 530 535 540Trp Arg Gln Asp His Asn Gly Phe Ser His Gln Asp Pro Gly Val Thr545 550 555 560Ser Val Leu Leu Asn Lys Thr Phe Asn Asn Asp His Val Ile Gly Leu 565 570 575Tyr Phe Ala Thr Asp Ala Asn Val Leu Leu Ala Ile Ala Glu Lys Cys 580 585 590Tyr Lys Ser Thr Asn Met Ile Asn Ala Ile Val Ala Gly Lys Gln Pro 595 600 605Ala Ala Thr Trp Thr Thr Leu Asp Glu Ala Arg Glu Leu Val Ala Lys 610 615 620Gly Ala Gly Glu Phe Glu Trp Ala Ser Asn Val Lys Thr Asn Asp Glu625 630 635 640Ala Glu Ile Val Leu Ala Ser Ala Gly Asp Val Pro Thr Gln Glu Leu 645 650 655Met Ala Ala Ala Asp Arg Leu Asn Lys Leu Gly Val Lys Phe Lys Val 660 665 670Val Asn Val Val Asp Leu Ile Lys Leu Gln Ser Ala Lys Glu Asn Asp 675 680 685Gln Ala Leu Ser Asp Ala Glu Phe Ala Glu Leu Phe Thr Glu Asp Lys 690 695 700Pro Val Leu Phe Ala Tyr His Ser Tyr Ala His Asp Val Arg Gly Leu705 710 715 720Ile Phe Asp Arg Pro Asn His Asp Asn Phe Asn Val Val Gly Tyr Lys 725 730 735Glu Gln Gly Ser Thr Thr Thr Pro Tyr Asp Met Val Arg Val Asn Asp 740 745 750Ile Asp Arg Tyr Glu Leu Thr Ala Thr Ala Leu Arg Met Ile Asp Ala 755 760 765Asp Lys Tyr Ala Asp Glu Ile Lys Lys Leu Glu Asp Phe Arg Ile Glu 770 775 780Ala Tyr Gln Phe Ala Val Asp Asn Gly Tyr Asp Ile Pro Asp Tyr Thr785 790 795 800Asp Trp Val Trp Pro Gly Val Lys Thr Asp Leu Pro Gly Ala Val Ser 805 810 815Ala Thr Ala Ala Thr Ala Gly Asp Asn Glu 820 825652478DNABifidobacterium animalisPHK 65atgactaatc ctgttattgg taccccatgg cagaagctgg accgtccggt ttccgaagag 60gccatcgaag gcatggacaa gtactggcgc gtcgccaact acatgtccat cggccagatc 120tacctgcgta gcaacccgct gatgaaggag cccttcaccc gcgatgacgt gaagcaccgt 180ctggtcggcc actggggcac caccccgggc ctgaacttcc ttctcgccca tatcaaccgc 240ctgatcgccg atcaccagca gaacaccgtg ttcatcatgg gtcctggcca cggcggccct 300gcaggtaccg ctcagtccta catcgacggc acctacaccg agtactaccc gaacatcacc 360aaggacgagg ctggcctgca gaagttcttc cgccagttct cctacccggg tggcattcct 420tcccacttcg ctccggagac gccaggctcc atccacgaag gcggcgagct gggctacgcc 480ctgtcgcacg cctacggcgc gatcatgaac aacccgagcc tcttcgtccc gtgcatcatc 540ggtgacggcg aagccgagac cggccctctg gccaccggct ggcagtccaa caagctcgtc 600aacccgcgca ccgacggcat cgtgctgccg atcctgcacc tcaacggcta caagatcgcc 660aacccgacgc tcctcgcccg catctccgac gaggagctgc acgacttctt ccgcggtatg 720ggttaccacc cgtacgagtt cgtcgccggc ttcgacaacg aggatcacct gtcgatccac 780cgtcgcttcg ccgagctctt cgagaccatc ttcgacgaga tctgcgatat caaggctgcg 840gctcagaccg acgacatgac ccgtccgttc tacccgatgc tcatcttccg caccccgaag 900ggctggacct gcccgaagtt catcgacggc aagaagaccg aaggctcctg gcgtgcacac 960caggtcccgc tggcttccgc ccgcgacacc gaggcccact tcgaagtcct caagggctgg 1020atggaatcct acaagccgga ggagctcttc aacgccgacg gctccatcaa ggacgacgtc 1080accgcattca tgcctaaggg cgaactgcgc atcggcgcca acccgaacgc caacggtggc 1140cgcatccgcg aggatctgaa gctccctgag ctcgatcagt acgagatcac cggcgtcaag 1200gaatacggcc atggctgggg ccaggtcgag gctccgcgct ccctcggcgc gtactgccgc 1260gacatcatca agaacaaccc ggattcgttc cgcatcttcg gacctgatga gaccgcatcc 1320aaccgtctga acgcgaccta cgaggtcacc aagaagcagt gggacaacgg ctatctctcg 1380gctctcgtcg acgagaacat ggctgtcacc ggccaggttg tcgagcagct ctccgagcat 1440cagtgcgaag gcttcctcga ggcctacctg ctcacgggcc gccacggcat gtggagcacc 1500tatgagtcct tcgcccacgt gatcgactcg atgctcaacc agcatgcgaa gtggctcgag 1560gcgaccgtcc gcgagatccc gtggcgcaag ccgatctcct cggtcaacct cctcgtctcc 1620tcgcacgtgt ggcgtcagga ccacaacggc ttctcgcatc aggacccggg tgtcacctcc 1680gtcctgatca acaagacgtt caacaacgac cacgtgacga acatctactt cgcgaccgac 1740gccaacatgc tgctcgcgat cgccgagaag tgcttcaagt ccaccaacaa gatcaacgcg 1800atcttctccg gcaagcagcc ggctccgacc tggattaccc tcgacgaggc tcgtgccgag 1860ctcgaggccg gcgccgccga gtggaagtgg gcttccaacg ccaagagcaa cgacgaggtc 1920cagattgtcc tcgccgccgc aggcgatgtc ccgacccagg agatcatggc cgcttccgat 1980gccctgaaca aggatggcat caagttcaag gtcgtcaacg ttgttgacct cctgaagctg 2040cagtccccgg agaacaacga cgaggccatg tcgaacgaag acttcaccga gctcttcacc 2100gccgacaaac cggttctgtt cgcctaccac tcctatgccc aggacgttcg tggtcttatc 2160tacgaccgcc cgaaccacga caacttcaac gttgtcggct acaaggagca gggctccacg 2220accacgccgt tcgacatggt ccgcgtcaac gacatggatc gctacgcgct cgaagctcag 2280gctctcgagc tgatcgacgc cgacaagtat gccgacaaga tcgacgagct caacgcgttc 2340cgcaagaccg cgttccagtt cgccgtcgac aacggctacg acatcccgga gttcaccgac 2400tgggtgtacc cggacgtcaa ggtcgacgag acgcagatgc tctccgcgac cgcggcgacc 2460gctggcgaca acgagtga 247866825PRTBifidobacterium animalisPHK 66Met Thr Asn Pro Val Ile Gly Thr Pro Trp Gln Lys Leu Asp Arg Pro1 5 10 15Val Ser Glu Glu Ala Ile Glu Gly Met Asp Lys Tyr Trp Arg Val Ala 20 25 30Asn Tyr Met Ser Ile Gly Gln Ile Tyr Leu Arg Ser Asn Pro Leu Met 35 40 45Lys Glu Pro Phe Thr Arg Asp Asp Val Lys His Arg Leu Val Gly His 50 55 60Trp Gly Thr Thr Pro Gly Leu Asn Phe Leu Leu Ala His Ile Asn Arg65 70 75 80Leu Ile Ala Asp His Gln Gln Asn Thr Val Phe Ile Met Gly Pro Gly 85 90 95His Gly Gly Pro Ala Gly Thr Ala Gln Ser Tyr Ile Asp Gly Thr Tyr 100 105 110Thr Glu Tyr Tyr Pro Asn Ile Thr Lys Asp Glu Ala Gly Leu Gln Lys 115 120 125Phe Phe Arg Gln Phe Ser Tyr Pro Gly Gly Ile Pro Ser His Phe Ala 130 135 140Pro Glu Thr Pro Gly Ser Ile His Glu Gly Gly Glu Leu Gly Tyr Ala145 150 155 160Leu Ser His Ala Tyr Gly Ala Ile Met Asn Asn Pro Ser Leu Phe Val 165 170 175Pro Cys Ile Ile Gly Asp Gly Glu Ala Glu Thr Gly Pro Leu Ala Thr 180 185 190Gly Trp Gln Ser Asn Lys Leu Val Asn Pro Arg Thr Asp Gly Ile Val 195 200 205Leu Pro Ile Leu His Leu Asn Gly Tyr Lys Ile Ala Asn Pro Thr Leu 210 215 220Leu Ala Arg Ile Ser Asp Glu Glu Leu His Asp Phe Phe Arg Gly Met225 230 235 240Gly Tyr His Pro Tyr Glu Phe Val Ala Gly Phe Asp Asn Glu Asp His 245 250 255Leu Ser Ile His Arg Arg Phe Ala Glu Leu Phe Glu Thr Ile Phe Asp 260 265 270Glu Ile Cys Asp Ile Lys Ala Ala Ala Gln Thr Asp Asp Met Thr Arg 275 280 285Pro Phe Tyr Pro Met Leu Ile Phe Arg Thr Pro Lys

Gly Trp Thr Cys 290 295 300Pro Lys Phe Ile Asp Gly Lys Lys Thr Glu Gly Ser Trp Arg Ala His305 310 315 320Gln Val Pro Leu Ala Ser Ala Arg Asp Thr Glu Ala His Phe Glu Val 325 330 335Leu Lys Gly Trp Met Glu Ser Tyr Lys Pro Glu Glu Leu Phe Asn Ala 340 345 350Asp Gly Ser Ile Lys Asp Asp Val Thr Ala Phe Met Pro Lys Gly Glu 355 360 365Leu Arg Ile Gly Ala Asn Pro Asn Ala Asn Gly Gly Arg Ile Arg Glu 370 375 380Asp Leu Lys Leu Pro Glu Leu Asp Gln Tyr Glu Ile Thr Gly Val Lys385 390 395 400Glu Tyr Gly His Gly Trp Gly Gln Val Glu Ala Pro Arg Ser Leu Gly 405 410 415Ala Tyr Cys Arg Asp Ile Ile Lys Asn Asn Pro Asp Ser Phe Arg Ile 420 425 430Phe Gly Pro Asp Glu Thr Ala Ser Asn Arg Leu Asn Ala Thr Tyr Glu 435 440 445Val Thr Lys Lys Gln Trp Asp Asn Gly Tyr Leu Ser Ala Leu Val Asp 450 455 460Glu Asn Met Ala Val Thr Gly Gln Val Val Glu Gln Leu Ser Glu His465 470 475 480Gln Cys Glu Gly Phe Leu Glu Ala Tyr Leu Leu Thr Gly Arg His Gly 485 490 495Met Trp Ser Thr Tyr Glu Ser Phe Ala His Val Ile Asp Ser Met Leu 500 505 510Asn Gln His Ala Lys Trp Leu Glu Ala Thr Val Arg Glu Ile Pro Trp 515 520 525Arg Lys Pro Ile Ser Ser Val Asn Leu Leu Val Ser Ser His Val Trp 530 535 540Arg Gln Asp His Asn Gly Phe Ser His Gln Asp Pro Gly Val Thr Ser545 550 555 560Val Leu Ile Asn Lys Thr Phe Asn Asn Asp His Val Thr Asn Ile Tyr 565 570 575Phe Ala Thr Asp Ala Asn Met Leu Leu Ala Ile Ala Glu Lys Cys Phe 580 585 590Lys Ser Thr Asn Lys Ile Asn Ala Ile Phe Ser Gly Lys Gln Pro Ala 595 600 605Pro Thr Trp Ile Thr Leu Asp Glu Ala Arg Ala Glu Leu Glu Ala Gly 610 615 620Ala Ala Glu Trp Lys Trp Ala Ser Asn Ala Lys Ser Asn Asp Glu Val625 630 635 640Gln Ile Val Leu Ala Ala Ala Gly Asp Val Pro Thr Gln Glu Ile Met 645 650 655Ala Ala Ser Asp Ala Leu Asn Lys Asp Gly Ile Lys Phe Lys Val Val 660 665 670Asn Val Val Asp Leu Leu Lys Leu Gln Ser Pro Glu Asn Asn Asp Glu 675 680 685Ala Met Ser Asn Glu Asp Phe Thr Glu Leu Phe Thr Ala Asp Lys Pro 690 695 700Val Leu Phe Ala Tyr His Ser Tyr Ala Gln Asp Val Arg Gly Leu Ile705 710 715 720Tyr Asp Arg Pro Asn His Asp Asn Phe Asn Val Val Gly Tyr Lys Glu 725 730 735Gln Gly Ser Thr Thr Thr Pro Phe Asp Met Val Arg Val Asn Asp Met 740 745 750Asp Arg Tyr Ala Leu Glu Ala Gln Ala Leu Glu Leu Ile Asp Ala Asp 755 760 765Lys Tyr Ala Asp Lys Ile Asp Glu Leu Asn Ala Phe Arg Lys Thr Ala 770 775 780Phe Gln Phe Ala Val Asp Asn Gly Tyr Asp Ile Pro Glu Phe Thr Asp785 790 795 800Trp Val Tyr Pro Asp Val Lys Val Asp Glu Thr Gln Met Leu Ser Ala 805 810 815Thr Ala Ala Thr Ala Gly Asp Asn Glu 820 825672367DNALactobacillus pentosumPHK 67atgtctacag attactcatc accagcatat ttgcaaaaag ttgataagta ctggcgtgct 60gccaactatt tatcagttgg tcaactttat ttaaaagata atcctttatt acaacggcca 120ttaaaggcta gtgacgttaa ggttcaccca atcggtcact ggggcacgat tgccggccaa 180aacttcatct atgcgcatct taaccgggtc atcaacaagt acggtttgaa gatgttctac 240gttgaaggtc caggtcatgg tggccaagtg atggtctcca actcatacct tgatgggact 300tacacggata tttatcctga aattacgcag gatgttgaag ggatgcaaaa actcttcaag 360caattctcat tcccaggtgg cgtggcttcc catgctgctc ctgaaacacc aggctcaatc 420cacgaaggtg gcgaacttgg ttactcaatt tcacacggtg ttggggcaat ccttgacaac 480cctgatgaaa tcgccgcagt cgttgttggt gatggggaat ccgaaaccgg cccattagca 540acttcatggc aatcaacgaa gttcatcaac ccaatcaacg atggggcagt gttaccaatc 600ttgaacctta acggctttaa gatttctaac ccaacgattt ttggtcggac ttctgatgaa 660aagatcaagc aatacttcga aagcatgaac tgggaaccaa tctttgttga aggtgacgat 720cctgaaaagg ttcacccagc tttagctaag gccatggatg aagccgtcga aaagatcaaa 780gccattcaaa agaacgctcg tgaaaacgat gacgctactt taccagtatg gccgatgatc 840gtcttccgcg cacctaaggg ctggactggt cctaagtcat gggatggcga caagatcgaa 900ggttcattcc gagctcacca aattccaatt cctgttgacc aaaccgacat ggaacatgcc 960gatgcgttag ttgactggtt ggaatcatat caaccaaagg aactcttcaa tgaagatggt 1020tctttgaagg atgatatcaa agaaattatc ccaactggcg atgcacggat ggccgctaac 1080ccaatcacta atggtggggt tgatccaaag gccttgaact tacctaactt ccgtgattac 1140gccgttgata cgtctaagca tggtgccaac gttaagcaag atatgatcgt ttggtcagac 1200tacttgcgtg atgttatcaa gaagaaccca gataacttcc ggttatttgg ccctgatgaa 1260accatgtcaa accggttata tggtgtcttt gaaaccacta accgtcaatg gatggaagat 1320attcacccag atagtgacca atacgaagca cctgctggcc gggtcttgga tgctcaatta 1380tctgaacacc aagctgaagg ttggttagaa ggttacgtct taactggtcg tcatggcttg 1440tttgcaagtt acgaagcctt cttacgggtt gtcgactcaa tgttgacgca acacttcaag 1500tggttacgta aggccaacga acttgactgg cggaagaagt acccgtcact caacattatc 1560gcggcttcaa ctgtgttcca acaagaccat aatgggtaca cccaccaaga tccaggtgcc 1620ttgactcatt tggctgaaaa gaagcctgaa tatatccgcg aatatttacc agccgacgcc 1680aactccttgt tagctgttgg ggacgtcatc ttccgtagcc aagaaaagat caactacgtg 1740gttacgtcga agcacccacg tcaacaatgg ttcagcattg aagaagctaa gcaattagtt 1800gacaacggtc ttggtatcat tgactgggca agcacggacc aaggtagcga accagatatc 1860gtgtttgctg ctgccggaac ggaaccaacg cttgaaacgt tggctgcaat ccaattgctc 1920catgatagct tcccagacat gaagattcgt ttcgtgaacg tggtcgacat cttgaagtta 1980cgtagccctg aaaaggaccc tcgtggcttg tcagatgctg aatttgacca ttacttcact 2040aaggacaaac cagttgtctt cgccttccat ggttacgaag acctggttcg tgacatcttc 2100tttgatcgtc acaaccacaa cttacacgtg catggctacc gtgaaaatgg tgacattacg 2160acaccattcg atgtccgggt catgaaccaa atggaccgtt tcgacttagc aaaatctgca 2220attgcggcgc aaccagcaat ggaaaacacc ggtgcagcct ttgttcaaga catggataac 2280atgcttgcaa aacacaacgc atacatccgt gacgccggaa ccgacttgcc agaagttaac 2340gactggcaat ggaaaggttt gaaataa 236768788PRTLactobacillus pentosumPHK 68Met Ser Thr Asp Tyr Ser Ser Pro Ala Tyr Leu Gln Lys Val Asp Lys1 5 10 15Tyr Trp Arg Ala Ala Asn Tyr Leu Ser Val Gly Gln Leu Tyr Leu Lys 20 25 30Asp Asn Pro Leu Leu Gln Arg Pro Leu Lys Ala Ser Asp Val Lys Val 35 40 45His Pro Ile Gly His Trp Gly Thr Ile Ala Gly Gln Asn Phe Ile Tyr 50 55 60Ala His Leu Asn Arg Val Ile Asn Lys Tyr Gly Leu Lys Met Phe Tyr65 70 75 80Val Glu Gly Pro Gly His Gly Gly Gln Val Met Val Ser Asn Ser Tyr 85 90 95Leu Asp Gly Thr Tyr Thr Asp Ile Tyr Pro Glu Ile Thr Gln Asp Val 100 105 110Glu Gly Met Gln Lys Leu Phe Lys Gln Phe Ser Phe Pro Gly Gly Val 115 120 125Ala Ser His Ala Ala Pro Glu Thr Pro Gly Ser Ile His Glu Gly Gly 130 135 140Glu Leu Gly Tyr Ser Ile Ser His Gly Val Gly Ala Ile Leu Asp Asn145 150 155 160Pro Asp Glu Ile Ala Ala Val Val Val Gly Asp Gly Glu Ser Glu Thr 165 170 175Gly Pro Leu Ala Thr Ser Trp Gln Ser Thr Lys Phe Ile Asn Pro Ile 180 185 190Asn Asp Gly Ala Val Leu Pro Ile Leu Asn Leu Asn Gly Phe Lys Ile 195 200 205Ser Asn Pro Thr Ile Phe Gly Arg Thr Ser Asp Glu Lys Ile Lys Gln 210 215 220Tyr Phe Glu Ser Met Asn Trp Glu Pro Ile Phe Val Glu Gly Asp Asp225 230 235 240Pro Glu Lys Val His Pro Ala Leu Ala Lys Ala Met Asp Glu Ala Val 245 250 255Glu Lys Ile Lys Ala Ile Gln Lys Asn Ala Arg Glu Asn Asp Asp Ala 260 265 270Thr Leu Pro Val Trp Pro Met Ile Val Phe Arg Ala Pro Lys Gly Trp 275 280 285Thr Gly Pro Lys Ser Trp Asp Gly Asp Lys Ile Glu Gly Ser Phe Arg 290 295 300Ala His Gln Ile Pro Ile Pro Val Asp Gln Thr Asp Met Glu His Ala305 310 315 320Asp Ala Leu Val Asp Trp Leu Glu Ser Tyr Gln Pro Lys Glu Leu Phe 325 330 335Asn Glu Asp Gly Ser Leu Lys Asp Asp Ile Lys Glu Ile Ile Pro Thr 340 345 350Gly Asp Ala Arg Met Ala Ala Asn Pro Ile Thr Asn Gly Gly Val Asp 355 360 365Pro Lys Ala Leu Asn Leu Pro Asn Phe Arg Asp Tyr Ala Val Asp Thr 370 375 380Ser Lys His Gly Ala Asn Val Lys Gln Asp Met Ile Val Trp Ser Asp385 390 395 400Tyr Leu Arg Asp Val Ile Lys Lys Asn Pro Asp Asn Phe Arg Leu Phe 405 410 415Gly Pro Asp Glu Thr Met Ser Asn Arg Leu Tyr Gly Val Phe Glu Thr 420 425 430Thr Asn Arg Gln Trp Met Glu Asp Ile His Pro Asp Ser Asp Gln Tyr 435 440 445Glu Ala Pro Ala Gly Arg Val Leu Asp Ala Gln Leu Ser Glu His Gln 450 455 460Ala Glu Gly Trp Leu Glu Gly Tyr Val Leu Thr Gly Arg His Gly Leu465 470 475 480Phe Ala Ser Tyr Glu Ala Phe Leu Arg Val Val Asp Ser Met Leu Thr 485 490 495Gln His Phe Lys Trp Leu Arg Lys Ala Asn Glu Leu Asp Trp Arg Lys 500 505 510Lys Tyr Pro Ser Leu Asn Ile Ile Ala Ala Ser Thr Val Phe Gln Gln 515 520 525Asp His Asn Gly Tyr Thr His Gln Asp Pro Gly Ala Leu Thr His Leu 530 535 540Ala Glu Lys Lys Pro Glu Tyr Ile Arg Glu Tyr Leu Pro Ala Asp Ala545 550 555 560Asn Ser Leu Leu Ala Val Gly Asp Val Ile Phe Arg Ser Gln Glu Lys 565 570 575Ile Asn Tyr Val Val Thr Ser Lys His Pro Arg Gln Gln Trp Phe Ser 580 585 590Ile Glu Glu Ala Lys Gln Leu Val Asp Asn Gly Leu Gly Ile Ile Asp 595 600 605Trp Ala Ser Thr Asp Gln Gly Ser Glu Pro Asp Ile Val Phe Ala Ala 610 615 620Ala Gly Thr Glu Pro Thr Leu Glu Thr Leu Ala Ala Ile Gln Leu Leu625 630 635 640His Asp Ser Phe Pro Asp Met Lys Ile Arg Phe Val Asn Val Val Asp 645 650 655Ile Leu Lys Leu Arg Ser Pro Glu Lys Asp Pro Arg Gly Leu Ser Asp 660 665 670Ala Glu Phe Asp His Tyr Phe Thr Lys Asp Lys Pro Val Val Phe Ala 675 680 685Phe His Gly Tyr Glu Asp Leu Val Arg Asp Ile Phe Phe Asp Arg His 690 695 700Asn His Asn Leu His Val His Gly Tyr Arg Glu Asn Gly Asp Ile Thr705 710 715 720Thr Pro Phe Asp Val Arg Val Met Asn Gln Met Asp Arg Phe Asp Leu 725 730 735Ala Lys Ser Ala Ile Ala Ala Gln Pro Ala Met Glu Asn Thr Gly Ala 740 745 750Ala Phe Val Gln Asp Met Asp Asn Met Leu Ala Lys His Asn Ala Tyr 755 760 765Ile Arg Asp Ala Gly Thr Asp Leu Pro Glu Val Asn Asp Trp Gln Trp 770 775 780Lys Gly Leu Lys785692313DNALactobacillus acidophilusPHK 69atgaaaaatc cgttgttaaa gaaaccttta acagctgaag atgtaaaacc taagccaatc 60ggtcactggg gtactattgc tccacaaaac tttatttatg ctcacttaaa tcgtgcgctt 120aaaaaatatg acttggatat gttctatatt gaaggttcag gtcacggtgg ccaagtgatg 180gtttcaaatt catatcttga tggttcatat actgaacgtt atccagaaat tacccaagat 240gaaaagggta tggctaaatt gtttaagcgc tttagtttcc caggtggtgt agcttctcac 300gctgctcctg aaactccagg ttctattcat gaaggtgggg aattaggata cgcactttca 360catggggtag gtgctatttt agacaatcca gatgtaattg ctgccgttga aattggtgat 420ggtgaagcag aaactggtcc acttgcagct agctggttca gtgacaagtt tattaatcca 480attaaggatg gtgcagtttt accaattctt caaattaatg gtttcaagat ttctaaccca 540actatcgttt caagaatgag cgatgaagaa ttaactgaat acttccgtgg catgggttgg 600gatccgcact ttgtttcagt atttaagggt ggccgctttg acggtgaaaa ggatccaatg 660caagtccacg aagaaatggc taaaaccatg gacgaagtaa ttgaagaaat taaggctatt 720caaaagcatg ctcgtgaaaa taatgatgct actttgccac attggccatt gattatcttc 780caatgtccaa agggctggac cggtccaaag aaggatcttg acggcaatcc aattgaaaac 840tcatttagag cacaccaaat tccaattcct gtctcacaat acgatatgaa acatgttgat 900atgttgactg attggcttga aagttataag ccaaacgaat tattcaacga agatggttca 960ccaaaggaaa ttgttactga aaacactgct aagggtgatc aacgtatggc aatgaatccg 1020atcactaatg gtggtaagga tcctaaacga ttgaacctac cagattatcg caactttgca 1080cttaagtttg acaagccagg ttcagttgaa gcacaagaca tggttgaatg ggctaaatat 1140ttaaacgaag ttgctaaact taacccaact actttccgtg gctttggtcc tgatgaatct 1200aaatcaaacc gtttatttaa acttttagat gatcaaaagc gtcaatggga acctgaagtt 1260catgaaccaa atgatgaaaa cttggcacca agtggccgcg ttatcgattc acaattatca 1320gaacaccaag acgaaggctt ccttgaaggc tacgttttaa ctggtcgtca cggcttcttt 1380gcaacctacg aagcatttgg tcgtgtagta gattcgatgc ttactcaaca tatgaagtgg 1440cttagaaaag ctaaagaaca atattggcgt catgattatc catcacttaa ctttgttgct 1500acttcaacag tattccaaca agatcacaat ggttacactc accaagatcc aggcatttta 1560actcacttat atgaaaagaa tcgtccagat ttagttcatg aatacttgcc atcagatact 1620aatactttac ttgctgtagg taacaaggca tttactgatc gtgaatgtat taatgtttta 1680gtaacttcaa agcaacctcg tccacaatgg ttctcaattg aggaagcaca aaagttagtt 1740gataaaggtt taagttacat tgattgggct tcaactgata aaggtgtaaa accagatatt 1800gtctttgctt caacagaaac tgaaccaaca attgaaactt tggcagcaat tgatattttg 1860catgacaagt tcccagatct taagattcgc tacattaacg taattgatgt gatgaaatta 1920atgtcaccaa aggacaataa gaatggtatt tctgatgaag aatttgatcg cttattccca 1980aaggacgttc ctgtaatctt tgcatggcac ggctacaaga gtatgatgga atcaatttgg 2040tttgcacgta accgtcataa tgtacatatt cactgctacg aagaaaacgg tgatattact 2100accccatttg atatgcgtgt tttgaaccac cttgacagat ttgatcttgc caaagatgct 2160gttgaaagtg ttgataaatt gaagggcaag aacgctgact ttatcagtca tatggatgac 2220ttgcttgaaa agcaccacca atacattcgt gataatggta aagatatgcc agaagttact 2280gaatggaagt ggaagggctt gaagaatgac agt 231370799PRTLactobacillus acidophilusPHK 70Met Thr Val Asn Tyr Asp Ser Lys Asp Tyr Leu Lys Ser Val Asp Ala1 5 10 15Tyr Trp Arg Ala Ala Asn Tyr Leu Ser Val Gly Gln Leu Phe Leu Met 20 25 30Lys Asn Pro Leu Leu Lys Lys Pro Leu Thr Ala Glu Asp Val Lys Pro 35 40 45Lys Pro Ile Gly His Trp Gly Thr Ile Ala Pro Gln Asn Phe Ile Tyr 50 55 60Ala His Leu Asn Arg Ala Leu Lys Lys Tyr Asp Leu Asp Met Phe Tyr65 70 75 80Ile Glu Gly Ser Gly His Gly Gly Gln Val Met Val Ser Asn Ser Tyr 85 90 95Leu Asp Gly Ser Tyr Thr Glu Arg Tyr Pro Glu Ile Thr Gln Asp Glu 100 105 110Lys Gly Met Ala Lys Leu Phe Lys Arg Phe Ser Phe Pro Gly Gly Val 115 120 125Ala Ser His Ala Ala Pro Glu Thr Pro Gly Ser Ile His Glu Gly Gly 130 135 140Glu Leu Gly Tyr Ala Leu Ser His Gly Val Gly Ala Ile Leu Asp Asn145 150 155 160Pro Asp Val Ile Ala Ala Val Glu Ile Gly Asp Gly Glu Ala Glu Thr 165 170 175Gly Pro Leu Ala Ala Ser Trp Phe Ser Asp Lys Phe Ile Asn Pro Ile 180 185 190Lys Asp Gly Ala Val Leu Pro Ile Leu Gln Ile Asn Gly Phe Lys Ile 195 200 205Ser Asn Pro Thr Ile Val Ser Arg Met Ser Asp Glu Glu Leu Thr Glu 210 215 220Tyr Phe Arg Gly Met Gly Trp Asp Pro His Phe Val Ser Val Phe Lys225 230 235 240Gly Gly Arg Phe Asp Gly Glu Lys Asp Pro Met Gln Val His Glu Glu 245 250 255Met Ala Lys Thr Met Asp Glu Val Ile Glu Glu Ile Lys Ala Ile Gln 260 265 270Lys His Ala Arg Glu Asn Asn Asp Ala Thr Leu Pro His Trp Pro Leu 275 280 285Ile Ile Phe Gln Cys Pro Lys Gly Trp Thr Gly Pro Lys Lys Asp Leu 290 295 300Asp Gly Asn Pro Ile Glu Asn Ser Phe Arg Ala His Gln Ile Pro Ile305 310 315 320Pro Val Ser Gln Tyr Asp Met Lys His Val Asp Met Leu Thr Asp Trp 325 330 335Leu Glu Ser Tyr Lys Pro Asn Glu Leu Phe Asn Glu Asp Gly Ser Pro 340 345 350Lys Glu Ile Val Thr Glu Asn Thr Ala Lys Gly

Asp Gln Arg Met Ala 355 360 365Met Asn Pro Ile Thr Asn Gly Gly Lys Asp Pro Lys Arg Leu Asn Leu 370 375 380Pro Asp Tyr Arg Asn Phe Ala Leu Lys Phe Asp Lys Pro Gly Ser Val385 390 395 400Glu Ala Gln Asp Met Val Glu Trp Ala Lys Tyr Leu Asn Glu Val Ala 405 410 415Lys Leu Asn Pro Thr Thr Phe Arg Gly Phe Gly Pro Asp Glu Ser Lys 420 425 430Ser Asn Arg Leu Phe Lys Leu Leu Asp Asp Gln Lys Arg Gln Trp Glu 435 440 445Pro Glu Val His Glu Pro Asn Asp Glu Asn Leu Ala Pro Ser Gly Arg 450 455 460Val Ile Asp Ser Gln Leu Ser Glu His Gln Asp Glu Gly Phe Leu Glu465 470 475 480Gly Tyr Val Leu Thr Gly Arg His Gly Phe Phe Ala Thr Tyr Glu Ala 485 490 495Phe Gly Arg Val Val Asp Ser Met Leu Thr Gln His Met Lys Trp Leu 500 505 510Arg Lys Ala Lys Glu Gln Tyr Trp Arg His Asp Tyr Pro Ser Leu Asn 515 520 525Phe Val Ala Thr Ser Thr Val Phe Gln Gln Asp His Asn Gly Tyr Thr 530 535 540His Gln Asp Pro Gly Ile Leu Thr His Leu Tyr Glu Lys Asn Arg Pro545 550 555 560Asp Leu Val His Glu Tyr Leu Pro Ser Asp Thr Asn Thr Leu Leu Ala 565 570 575Val Gly Asn Lys Ala Phe Thr Asp Arg Glu Cys Ile Asn Val Leu Val 580 585 590Thr Ser Lys Gln Pro Arg Pro Gln Trp Phe Ser Ile Glu Glu Ala Gln 595 600 605Lys Leu Val Asp Lys Gly Leu Ser Tyr Ile Asp Trp Ala Ser Thr Asp 610 615 620Lys Gly Val Lys Pro Asp Ile Val Phe Ala Ser Thr Glu Thr Glu Pro625 630 635 640Thr Ile Glu Thr Leu Ala Ala Ile Asp Ile Leu His Asp Lys Phe Pro 645 650 655Asp Leu Lys Ile Arg Tyr Ile Asn Val Ile Asp Val Met Lys Leu Met 660 665 670Ser Pro Lys Asp Asn Lys Asn Gly Ile Ser Asp Glu Glu Phe Asp Arg 675 680 685Leu Phe Pro Lys Asp Val Pro Val Ile Phe Ala Trp His Gly Tyr Lys 690 695 700Ser Met Met Glu Ser Ile Trp Phe Ala Arg Asn Arg His Asn Val His705 710 715 720Ile His Cys Tyr Glu Glu Asn Gly Asp Ile Thr Thr Pro Phe Asp Met 725 730 735Arg Val Leu Asn His Leu Asp Arg Phe Asp Leu Ala Lys Asp Ala Val 740 745 750Glu Ser Val Asp Lys Leu Lys Gly Lys Asn Ala Asp Phe Ile Ser His 755 760 765Met Asp Asp Leu Leu Glu Lys His His Gln Tyr Ile Arg Asp Asn Gly 770 775 780Lys Asp Met Pro Glu Val Thr Glu Trp Lys Trp Lys Gly Leu Lys785 790 795712469DNAPenicillium chrysogenumPHK 71atgcctgcag aagttataag cgaaccaaat ccacaagctc tgccgtcgca tctgccagac 60tatcttgaga aattgagcgt cagcttggaa cgtgagaaac tagatgaaaa gacctacgat 120gccttgatca agttccgccg agcggcttgt tacattgcgg cagctatgat atttctgcaa 180gaaaatactc ttttgaagtc agaactcaca tttcaacatg tcaagcctag actactcggt 240cactggggaa catgcccggg cctaatcttt gtctactctc atttgaacta cctgatccgg 300acaatgaatc tggacatgtt gtacgttgtc ggtccaggac atggcgcacc ggcaatacta 360gccgcgctat ggttggaggg ttcgctggag aaattctatc cccactattc gcgagacgaa 420aagggcctac acagattgat ctcgaccttt agtaccacag gtggttttcc cagccacatc 480aattccgaga cccccggtgc aatccatgaa ggtggagaac tgggttacgc tctggcagtg 540tcctttggcg ctgtcatgga taaccccgat ctgattgtga cttgtattgt gggggacggg 600gaggctgaaa gcggtcccac tgctacgtcc tggcatgcga tcaagtacat tgatccaaag 660gagtccggtg cagttttacc aattctgcat ttgaatggat tcaagatcag cgagcgcact 720atcttcgggt gcatggacca caaagagctt ttgactctct tcagtggata cgggtaccag 780gttcgctttg tcgaggataa caacgatatt gatgcggatt tacacacctc tatgatctgg 840gccgtcaatg aaatccagaa gatccagaag gctgcccgtt cagggaagcc gattatgaag 900ccaaggtggc cgatgctgat tctgcgtact tccaagggct ggactgggcc caagcagctt 960catggcaagt ttcttgaggg ctcataccat tctcaccagg tgccattgcc caaagcaaag 1020accgacaagg aacaactaga tctgctgcag aattggctgt ctagttataa gccagaagag 1080ctgttcactt cgaacggtga tgtaattgat gagatcaagt ctgtgatacc cacagaagac 1140aaaaagaagc ttggccagcg catcgaagtc tacaatagtt atacaccacc gaatctgccg 1200gattggaagc ccttctgcgc ggataaaggc tctcaagaaa gcgctatgaa agcggctggc 1260accttcatta atcagacatt caaggacaac ccgaacagcg tgcgactctt ctcgccggac 1320gagttggaaa gcaacaaact cgatgcggta tttgaataca caaaccgcaa tttccaatgg 1380gacgagttcg ccaatgcccg cggcggccgt gtaattgaag tccttagcga acatatgtgt 1440caaggattca tgcagggcta taccttgacc gggcgcattg gtattttccc gtcctatgag 1500agcttcctcg gcatcattca tacgatgatg gtgcagtatg ccaagttcat caagatggga 1560ctggaaacaa cttggcactc cggcgtcagt agtgtgaact acatcgaatc gagcacctgg 1620gcacgccaag aacacaacgg cttctctcac cagaacccat ccttcatcgg cgctgtgcta 1680aagctgaagc ctagtgcagc tcgggtatac ttaccaccag atgcgaatac tttcctgaca 1740accatccacc attgtttaaa atcaaagaat tacatcaacc taatggtagg ctcaaagcag 1800ccaactccag tgtacctgac tcccaaagag gcggaaagcc actgtcgcgc aggagcatcc 1860atttggaagt tctgcagcac ggacgatgga attaatccgg atgtcgtgct ggtcggtatt 1920ggtgttgagg tgatgttcga agtaattgcg gcagcggcgc ttctgcgtaa gctcatccca 1980gagcttcgtg tttgtgtcat taacgtgacg gacctgatga ttcttgataa cgagggtgcg 2040catccgcatg ccctgtccac tgaggcattc gatggtctct ttacttccga caggcccatt 2100cacttcaatt accacggcta tccaactgag ctgcagggtt tgctctttgg acgtccccgc 2160cttgatcgcg tcagtgtcgc cggctatatc gaagagggta gcaccaccac gcctttcgac 2220atgatgcttg tcaaccgtgt ctcgcgtttc catgtcgccc agcatgctat tcgtggtgct 2280gccaaagtaa atgagaaggt cagggtgtat cagcaggagc tgaatgctca gcttgaggcg 2340agtatggtca gcacgaggaa gtatattgtg gaaaaccggg atgaccccga cggcatttat 2400gacatgccac aattccacag cttccacaaa ccggctgagt ctgagacatt ctggaatatt 2460gcccagtag 246972822PRTPenicillium chrysogenumPHK 72Met Pro Ala Glu Val Ile Ser Glu Pro Asn Pro Gln Ala Leu Pro Ser1 5 10 15His Leu Pro Asp Tyr Leu Glu Lys Leu Ser Val Ser Leu Glu Arg Glu 20 25 30Lys Leu Asp Glu Lys Thr Tyr Asp Ala Leu Ile Lys Phe Arg Arg Ala 35 40 45Ala Cys Tyr Ile Ala Ala Ala Met Ile Phe Leu Gln Glu Asn Thr Leu 50 55 60Leu Lys Ser Glu Leu Thr Phe Gln His Val Lys Pro Arg Leu Leu Gly65 70 75 80His Trp Gly Thr Cys Pro Gly Leu Ile Phe Val Tyr Ser His Leu Asn 85 90 95Tyr Leu Ile Arg Thr Met Asn Leu Asp Met Leu Tyr Val Val Gly Pro 100 105 110Gly His Gly Ala Pro Ala Ile Leu Ala Ala Leu Trp Leu Glu Gly Ser 115 120 125Leu Glu Lys Phe Tyr Pro His Tyr Ser Arg Asp Glu Lys Gly Leu His 130 135 140Arg Leu Ile Ser Thr Phe Ser Thr Thr Gly Gly Phe Pro Ser His Ile145 150 155 160Asn Ser Glu Thr Pro Gly Ala Ile His Glu Gly Gly Glu Leu Gly Tyr 165 170 175Ala Leu Ala Val Ser Phe Gly Ala Val Met Asp Asn Pro Asp Leu Ile 180 185 190Val Thr Cys Ile Val Gly Asp Gly Glu Ala Glu Ser Gly Pro Thr Ala 195 200 205Thr Ser Trp His Ala Ile Lys Tyr Ile Asp Pro Lys Glu Ser Gly Ala 210 215 220Val Leu Pro Ile Leu His Leu Asn Gly Phe Lys Ile Ser Glu Arg Thr225 230 235 240Ile Phe Gly Cys Met Asp His Lys Glu Leu Leu Thr Leu Phe Ser Gly 245 250 255Tyr Gly Tyr Gln Val Arg Phe Val Glu Asp Asn Asn Asp Ile Asp Ala 260 265 270Asp Leu His Thr Ser Met Ile Trp Ala Val Asn Glu Ile Gln Lys Ile 275 280 285Gln Lys Ala Ala Arg Ser Gly Lys Pro Ile Met Lys Pro Arg Trp Pro 290 295 300Met Leu Ile Leu Arg Thr Ser Lys Gly Trp Thr Gly Pro Lys Gln Leu305 310 315 320His Gly Lys Phe Leu Glu Gly Ser Tyr His Ser His Gln Val Pro Leu 325 330 335Pro Lys Ala Lys Thr Asp Lys Glu Gln Leu Asp Leu Leu Gln Asn Trp 340 345 350Leu Ser Ser Tyr Lys Pro Glu Glu Leu Phe Thr Ser Asn Gly Asp Val 355 360 365Ile Asp Glu Ile Lys Ser Val Ile Pro Thr Glu Asp Lys Lys Lys Leu 370 375 380Gly Gln Arg Ile Glu Val Tyr Asn Ser Tyr Thr Pro Pro Asn Leu Pro385 390 395 400Asp Trp Lys Pro Phe Cys Ala Asp Lys Gly Ser Gln Glu Ser Ala Met 405 410 415Lys Ala Ala Gly Thr Phe Ile Asn Gln Thr Phe Lys Asp Asn Pro Asn 420 425 430Ser Val Arg Leu Phe Ser Pro Asp Glu Leu Glu Ser Asn Lys Leu Asp 435 440 445Ala Val Phe Glu Tyr Thr Asn Arg Asn Phe Gln Trp Asp Glu Phe Ala 450 455 460Asn Ala Arg Gly Gly Arg Val Ile Glu Val Leu Ser Glu His Met Cys465 470 475 480Gln Gly Phe Met Gln Gly Tyr Thr Leu Thr Gly Arg Ile Gly Ile Phe 485 490 495Pro Ser Tyr Glu Ser Phe Leu Gly Ile Ile His Thr Met Met Val Gln 500 505 510Tyr Ala Lys Phe Ile Lys Met Gly Leu Glu Thr Thr Trp His Ser Gly 515 520 525Val Ser Ser Val Asn Tyr Ile Glu Ser Ser Thr Trp Ala Arg Gln Glu 530 535 540His Asn Gly Phe Ser His Gln Asn Pro Ser Phe Ile Gly Ala Val Leu545 550 555 560Lys Leu Lys Pro Ser Ala Ala Arg Val Tyr Leu Pro Pro Asp Ala Asn 565 570 575Thr Phe Leu Thr Thr Ile His His Cys Leu Lys Ser Lys Asn Tyr Ile 580 585 590Asn Leu Met Val Gly Ser Lys Gln Pro Thr Pro Val Tyr Leu Thr Pro 595 600 605Lys Glu Ala Glu Ser His Cys Arg Ala Gly Ala Ser Ile Trp Lys Phe 610 615 620Cys Ser Thr Asp Asp Gly Ile Asn Pro Asp Val Val Leu Val Gly Ile625 630 635 640Gly Val Glu Val Met Phe Glu Val Ile Ala Ala Ala Ala Leu Leu Arg 645 650 655Lys Leu Ile Pro Glu Leu Arg Val Cys Val Ile Asn Val Thr Asp Leu 660 665 670Met Ile Leu Asp Asn Glu Gly Ala His Pro His Ala Leu Ser Thr Glu 675 680 685Ala Phe Asp Gly Leu Phe Thr Ser Asp Arg Pro Ile His Phe Asn Tyr 690 695 700His Gly Tyr Pro Thr Glu Leu Gln Gly Leu Leu Phe Gly Arg Pro Arg705 710 715 720Leu Asp Arg Val Ser Val Ala Gly Tyr Ile Glu Glu Gly Ser Thr Thr 725 730 735Thr Pro Phe Asp Met Met Leu Val Asn Arg Val Ser Arg Phe His Val 740 745 750Ala Gln His Ala Ile Arg Gly Ala Ala Lys Val Asn Glu Lys Val Arg 755 760 765Val Tyr Gln Gln Glu Leu Asn Ala Gln Leu Glu Ala Ser Met Val Ser 770 775 780Thr Arg Lys Tyr Ile Val Glu Asn Arg Asp Asp Pro Asp Gly Ile Tyr785 790 795 800Asp Met Pro Gln Phe His Ser Phe His Lys Pro Ala Glu Ser Glu Thr 805 810 815Phe Trp Asn Ile Ala Gln 820732421DNAAspergillus nidulansPHK 73atgcctggag aggtcatcga caggccaaat cccaaggctg agccttcaca catccccgat 60cttgtcaatc aattgcaggt caaacttcaa gagacgcgtt tggaggaaac tgattacaat 120gcccttctga aattccgccg tgcagcggcc tacattgctg ctgcaatgat ctttctccaa 180gacaatgtgc tgctgaagca gaatctaagg cacgaggaca tcaagcccag gcttcttggc 240cactggggaa catgtcccgg gttgattctt gtatactctc acttgaacta catcatcaga 300aagcagaacc tggatatgtt gtatgtcgtc gggcctggcc acggcgcgcc agctattttg 360gcctcactgt ggcttgaggg ctctttagag aaattctacc cccactactc acgagacatg 420gatggtctcc atgagctcat ctcgaccttc agcacaagtg ctggattacc aagccatatc 480aatgcggaaa ctcccggtgc aatccatgaa ggtggtgaat tgggttatgc gttagctgtc 540tcttttggtg ctgttatgga caatcccgac atgatcgtca cctgcgtggt tggtgacggg 600gaagcagaaa ctggtcctac cgcgacgtcc tggcatgcaa tcaagtacat tgaccccgca 660gaatcaggtg ccgtcctgcc gattctccac gttaatggct ttaagatcag cgagcgcacc 720atttatggct gcatggacaa caaagagctg gtctccctct tcacgggtta tggataccag 780gtgcgcattg ttgagaacct ggatgacatc gacgcagatc tccatagctc tatgatgtgg 840gcagttgagg agatccacaa gatccaaaaa gcggcgcgtt ccggcaagcc aattatgaag 900cctagatggc caatgattgt tttgcgcaca ccgaagggtt ggtcaggacc taaagagctc 960cacgggtcat tcatagaggg atctttccac tcacatcagg ttcctctacc taatgcaaag 1020aaggataaag aggagcttca ggctctgcag aaatggctgt cctcgtataa tccgcacgaa 1080cttttcactg agacgggaga catcattgac gacatcaagt cagtgatccc tctggaagac 1140accaagaagc ttgggcagcg agcagaagcc tacaagggct atagggcacc cgatctccca 1200gactggcgca agtttggcgt agaaaagggc tcccagcaga gcgctatgaa aacaattgga 1260aagttcattg accaagtgtt tacccaaaat cctcatggcg tccgtgtatt ttcgccagac 1320gagctagaga gcaacaagct ggatgcagca ctggcgcaca cgggaaggaa ctttcagtgg 1380gatcaattct cgaatgccaa aggcggccgc gtcatcgagg tgctcagtga gcacctgtgc 1440cagggcttta tgcagggata cacgttgacg ggccgggtgg gcattttccc atcgtacgaa 1500agcttcttgg gaatcatcca taccatgatg gtgcaatatg ccaaatttaa caaaatggct 1560caagagacga cctggcataa gccggttagt agcatcaact atatcgaaac gagtacgtgg 1620gctcgtcagg agcacaatgg attctctcac cagaacccct cctttatcgg agctgtgctc 1680aggctgaagc ccaccgccgc gcgagtttat ctgccacctg atgctaacac atttttgacc 1740acccttcacc actgtctcaa gtccaagaat tatgtcaacc tcatggtagg ttcaaaacag 1800ccaactcccg tgtacttgag ccccgaggaa gcagagagcc actgccgagc cggcgcatcg 1860atctggagat tctgtagtac cgacaatggg ctgaacccgg atgtcgtgct ggttggcatt 1920ggagtagagg tgatgttcga ggtcatctac gcggcggcca tcctccgcaa gcgttgtcca 1980gaactccggg tgcgtgtggt caatgtgacc gacttgatga ttctggagaa ggaaggtcta 2040catccacatg cattgacgac cgaagctttc gacagtctgt ttggctcgga ccggccgata 2100cacttcaact accacggata cccgggcgag ctcaaaggtc tgctctttgg gcggccccgc 2160ctggaccgag tttcagtaga aggatacatg gaggaaggaa gcacgacgac gccgttcgat 2220atgatgttgc tgaaccgcgt ctcacgatac cacgtggcgc aggcagccgt gatcggggcg 2280tccagacgga atgagaaggt tcaagttcgg cagcacgaac tggtcagcga attcggccac 2340aacatcgtgg agacacgcaa atacattctg gccaaccgca aagacccgga tgatacgtat 2400gatatgccct cctttgaata a 242174780PRTAspergillus nidulansPHK 74Met Pro Gly Glu Val Ile Glu Arg Pro Asn Pro Ala Pro Lys Pro Ser1 5 10 15His Val Pro Asp Leu Val Glu Lys Leu Ile Ile Pro Ala Gln Lys Thr 20 25 30Lys Leu Glu Lys Ser Asp Cys Asp Ala Leu His Lys Tyr Arg Arg Ala 35 40 45Ala Ala Tyr Ile Ala Ala Gly His Trp Gly Thr Cys Pro Gly Leu Ile 50 55 60Leu Val Tyr Ser His Leu Asn Tyr Leu Ile Lys Lys Gln Asn Leu Asp65 70 75 80Met Leu Tyr Val Val Gly Pro Gly His Gly Ala Pro Gly Leu Leu Ala 85 90 95Ser Leu Trp Leu Glu Gly Ser Leu Gly Lys Phe Tyr Pro Gln Tyr Thr 100 105 110Lys Asp Lys Glu Gly Leu His Asn Leu Ile Ser Thr Phe Ser Thr Ser 115 120 125Ala Gly Leu Pro Ser His Ile Asn Ala Glu Thr Pro Gly Ala Ile His 130 135 140Glu Gly Gly Glu Leu Gly Tyr Ala Leu Ser Val Ser Phe Gly Ala Val145 150 155 160Met Asp Asn Pro Asp Leu Ile Val Thr Cys Val Val Gly Asp Gly Glu 165 170 175Ala Glu Thr Gly Pro Thr Ala Thr Ser Trp His Ala Ile Lys Tyr Ile 180 185 190Asp Pro Ala Glu Ser Gly Ala Val Leu Pro Ile Leu His Val Asn Gly 195 200 205Phe Lys Ile Ser Glu Arg Thr Ile Phe Gly Cys Met Asp Asn Arg Glu 210 215 220Ile Val Cys Leu Phe Thr Gly Tyr Gly Tyr Gln Val Arg Ile Val Glu225 230 235 240Asp Leu Glu Asp Ile Asp Asn Asp Leu His Ser Ala Met Ser Trp Ala 245 250 255Val Glu Glu Ile Arg Asn Ile Gln Lys Ala Ala Arg Ser Gly Lys Pro 260 265 270Ile Met Lys Pro Gln Trp Pro Met Ile Val Leu Arg Thr Pro Lys Gly 275 280 285Trp Ser Gly Pro Lys Glu Leu His Gly Gln Phe Ile Glu Gly Ser Phe 290 295 300His Ser His Gln Val Pro Leu Pro Asn Ala Lys Lys Asp Asp Glu Glu305 310 315 320Leu Gln Ala Leu Gln Lys Trp Leu Ser Ser Tyr Lys Pro Asp Glu Leu 325 330 335Phe Thr Glu Ser Gly Asp Val Ile Asp Glu Ile Leu Ser Ile Ile Pro 340 345 350Ser Asp Asp Lys Lys Leu Gly Met Arg Pro Glu Ala Tyr Lys Thr His 355 360 365Leu Pro Pro Asp Leu Pro Asp Trp Arg Gln Phe Cys Val Lys

Lys Gly 370 375 380Asp Gln Phe Ser Ala Met Lys Ala Ile Gly Ser Phe Ile Asp Gln Val385 390 395 400Phe Val Lys Asn Pro His Thr Val Arg Leu Phe Ser Pro Asp Glu Leu 405 410 415Glu Ser Asn Lys Leu Ser Ala Ala Leu Ser His Thr Gly Arg Asn Phe 420 425 430Gln Trp Asp Glu Phe Ser Asn Ala Lys Gly Gly Arg Val Ile Glu Val 435 440 445Leu Ser Glu His Leu Cys Gln Gly Phe Met Gln Gly Tyr Thr Leu Thr 450 455 460Gly Arg Thr Gly Ile Phe Pro Ser Tyr Glu Ser Phe Leu Gly Ile Ile465 470 475 480His Thr Met Met Val Gln Tyr Ala Lys Phe Ala Lys Met Ala Lys Glu 485 490 495Thr Ala Trp His His Asp Val Ser Ser Ile Asn Tyr Ile Glu Thr Ser 500 505 510Thr Trp Ala Arg Gln Glu His Asn Gly Phe Ser His Gln Asn Pro Ser 515 520 525Phe Ile Gly Ala Val Leu Lys Leu Lys Pro Tyr Ala Ala Arg Val Tyr 530 535 540Leu Pro Pro Asp Ala Asn Thr Phe Leu Thr Thr Leu His His Cys Leu545 550 555 560Lys Ser Lys Asn Tyr Ile Asn Leu Met Val Gly Ser Lys Gln Pro Thr 565 570 575Pro Val Tyr Leu Ser Pro Glu Glu Ala Glu Ser His Cys Arg Ala Gly 580 585 590Ala Ser Ile Phe Lys Phe Cys Ser Thr Asp Gly Gly Leu Arg Pro Asp 595 600 605Val Val Leu Val Gly Ile Gly Val Glu Val Met Phe Glu Val Ile Lys 610 615 620Ala Ala Ala Ile Leu Arg Glu Arg Cys Pro Glu Leu Arg Val Arg Val625 630 635 640Val Asn Val Thr Asp Leu Phe Ile Leu Glu Asn Glu Gly Ala His Pro 645 650 655His Ala Leu Lys His Glu Ala Phe Asp Asn Leu Phe Thr Glu Asp Arg 660 665 670Ser Ile His Phe Asn Tyr His Gly Tyr Val Asn Glu Leu Gln Gly Leu 675 680 685Leu Phe Gly Arg Pro Arg Leu Asp Arg Ala Thr Ile Lys Gly Tyr Lys 690 695 700Glu Glu Gly Ser Thr Thr Thr Pro Phe Asp Met Met Leu Val Asn Glu705 710 715 720Val Ser Arg Tyr His Val Ala Lys Ala Ala Val Thr Gly Gly Ala Arg 725 730 735Phe Asn Glu Lys Val Lys Leu Arg His Gln Glu Leu Cys Ser Glu Phe 740 745 750Asp His Asn Ile Ala Glu Thr Arg Lys Tyr Ile Met Asn Asn His Gln 755 760 765Asp Pro Glu Asp Thr Tyr Asn Met Pro Ser Phe Asn 770 775 780751188DNAClostridium phytofermentansACK 75atgaaagttt tagttattaa ttgcggaagt tcttccctta aatatcagtt aatcgactct 60gtgacagagc aagcattagc agtaggtctt tgtgaaagaa tcggtattga tggccgtctt 120actcacaagt cagctgacgg tgagaaggta gttcttgagg atgcacttcc aaaccatgag 180gttgctatta aaaatgtaat cgctgctctt atgaatgaaa attatggtgt gattaagtcc 240ttagatgaaa tcaacgctgt tggacataga gtagtacatg gtggtgagaa atttgctcat 300tccgtagtaa tcaatgatga agtcttaaat gcaattgaag agtgtaatga tcttgcacct 360ttacacaacc cagcaaacct tattggtatc aacgcttgta aatcaattat gccaaatgta 420ccaatggtag ctgtttttga tactgcattc catcagacaa tgccaaaaga agcttacctt 480tatggtattc catttgagta ctatgataaa tataaggtaa gaagatatgg tttccacgga 540acaagtcaca gctatgtttc taaaagagca accacgcttg ctggcttaga tgtaaataac 600tcaaaagtta tcgtttgtca ccttggtaat ggcgcatcca tttccgcagt taaaaacggt 660gagtctgtag atacaagtat gggtcttaca ccacttgaag gtttaatcat gggaacaaga 720agtggtgatc ttgatccagc aatcattgat ttcgttgcta agaaagaaaa cttatcctta 780gatgaagtaa tgaatatctt aaataagaaa tctggtgtat taggtatgtc cggagtatct 840tctgacttta gagatatcga agcagcagca aacgaaggca atgagcatgc aaaagaagct 900ttagcagttt ttgcataccg tgttgctaaa tatgtaggtt cttatatcgt agctatgaat 960ggtgtagatg ctgttgtatt tacagcagga cttggtgaga atgataagaa catcagagca 1020gcagtaagtt cacaccttga gttccttggt gtatctttag atgctgagaa gaattctcaa 1080agaggtaaag aattaatcat ctctaaccca gattctaagg ttaagattat ggttatccca 1140actaacgaag agcttgcaat ctgtagagaa gttgttgaat tagtgtag 118876395PRTClostridium phytofermentansACK 76Met Lys Val Leu Val Ile Asn Cys Gly Ser Ser Ser Leu Lys Tyr Gln1 5 10 15Leu Ile Asp Ser Val Thr Glu Gln Ala Leu Ala Val Gly Leu Cys Glu 20 25 30Arg Ile Gly Ile Asp Gly Arg Leu Thr His Lys Ser Ala Asp Gly Glu 35 40 45Lys Val Val Leu Glu Asp Ala Leu Pro Asn His Glu Val Ala Ile Lys 50 55 60Asn Val Ile Ala Ala Leu Met Asn Glu Asn Tyr Gly Val Ile Lys Ser65 70 75 80Leu Asp Glu Ile Asn Ala Val Gly His Arg Val Val His Gly Gly Glu 85 90 95Lys Phe Ala His Ser Val Val Ile Asn Asp Glu Val Leu Asn Ala Ile 100 105 110Glu Glu Cys Asn Asp Leu Ala Pro Leu His Asn Pro Ala Asn Leu Ile 115 120 125Gly Ile Asn Ala Cys Lys Ser Ile Met Pro Asn Val Pro Met Val Ala 130 135 140Val Phe Asp Thr Ala Phe His Gln Thr Met Pro Lys Glu Ala Tyr Leu145 150 155 160Tyr Gly Ile Pro Phe Glu Tyr Tyr Asp Lys Tyr Lys Val Arg Arg Tyr 165 170 175Gly Phe His Gly Thr Ser His Ser Tyr Val Ser Lys Arg Ala Thr Thr 180 185 190Leu Ala Gly Leu Asp Val Asn Asn Ser Lys Val Ile Val Cys His Leu 195 200 205Gly Asn Gly Ala Ser Ile Ser Ala Val Lys Asn Gly Glu Ser Val Asp 210 215 220Thr Ser Met Gly Leu Thr Pro Leu Glu Gly Leu Ile Met Gly Thr Arg225 230 235 240Ser Gly Asp Leu Asp Pro Ala Ile Ile Asp Phe Val Ala Lys Lys Glu 245 250 255Asn Leu Ser Leu Asp Glu Val Met Asn Ile Leu Asn Lys Lys Ser Gly 260 265 270Val Leu Gly Met Ser Gly Val Ser Ser Asp Phe Arg Asp Ile Glu Ala 275 280 285Ala Ala Asn Glu Gly Asn Glu His Ala Lys Glu Ala Leu Ala Val Phe 290 295 300Ala Tyr Arg Val Ala Lys Tyr Val Gly Ser Tyr Ile Val Ala Met Asn305 310 315 320Gly Val Asp Ala Val Val Phe Thr Ala Gly Leu Gly Glu Asn Asp Lys 325 330 335Asn Ile Arg Ala Ala Val Ser Ser His Leu Glu Phe Leu Gly Val Ser 340 345 350Leu Asp Ala Glu Lys Asn Ser Gln Arg Gly Lys Glu Leu Ile Ile Ser 355 360 365Asn Pro Asp Ser Lys Val Lys Ile Met Val Ile Pro Thr Asn Glu Glu 370 375 380Leu Ala Ile Cys Arg Glu Val Val Glu Leu Val385 390 395772421DNAAspergillus clavatusPHK 77atgcctgggg aagtcattga cagaccaaac ccgcaggcgc tgccttccca tattcccgat 60gtggttgaac agttgcaggt gaagctggat ttgacgagtt tagagcaacc ggcctgcgat 120gccttgttca aattccgtcg agctgcttca tacatcgctg cagcaatgat ctttctgcaa 180gataatgtgc tactgaaaag aagcttgacg catgacgaca taaagcctag attgcttggt 240cattggggaa catgtcccgg cctgatattg gtatactctc acctgaatta cctcattaca 300aagcgcaatc tcaatatgct gtatgtcgtg ggaccaggac atggtgcacc agccatccta 360gcctctctct ggctggaggg ctcgttggag aagttctacc cagagtattc gcgaaatgcg 420gagggcctcg ccaatctcat ctcgactttc agtacaacag ccgggctgcc tagccatatc 480aatgcggaaa caccaggtgc tatccacgaa ggaggggaac tgggatacgc acttgctgta 540tcatttggtg cagttatgga caaccccaat ctgatcgttc catgcattgt gggcgatgga 600gaagcagaaa ctggcccaac agcagcatct tggcacggca tcaaatatat cgatcctgca 660gagtcaggcg ctgtgctgcc aatcctgcac ctgaacggat tcaagatcag cgagcgcacg 720atctttggct gcatggacga caaggagctg acctcgcttt tcactgggta tgggtaccag 780gtccggattg tgcaggattt gaatgacatc gacaccgatc tccatcgatc tatgacctgg 840gctttggaag aaatccacaa gattcagcaa gcagcgcgat ccggaaagcc cattatgaag 900ccaagatggc caatgctgat tttgcgcaca ccaaagggat ggtcaggccc aaaggaaatt 960catggccagt tcattgaggg ctcattccat tcacatcaag tccccctgcc caacgcaaag 1020aaggacaagg aagagctgca ggccctcgag aaatggcttt cctcgtatgg gcccagagaa 1080cttttcactg agaatggaga tgttatagat gagattaaat ccatcatccc tacggatgat 1140gcgatgaagc ttggtcagcg tgtggaggca tacaaggctt atctgccacc aaagctcccg 1200gactggagga agttcgccgt ggataaaggt agccaacaga gcccgatgaa aacgaccgga 1260aagcttatcg acgaggtctt ccaactcaac cctcacactg ttcgattgtt ttcaccggat 1320gaattggaga gcaacaaact gagcgccgct ctggatcaca ctgggagaaa tttccagtgg 1380gatcagttct ctagcgcgaa aggtggccgg gtcatcgaag tcctgagcga acatatgtgc 1440cagggtttta tccagggtta cacgttgacc ggacgcacgg gtatcttccc atcgtatgag 1500agcttcttgg gaatcgttca taccatgatg gtgcagtact ccaaattcat caaaatggct 1560caagagacca catggcaccg tagcatagcc agcatcaatt atattgagac aagcacgtgg 1620acacggcagg agcataatgg attttcacac cagaatccat ctttcatcgg ggcagtactc 1680aaattgaagc ccacggctgc tcgggtttat ctgccgccgg atgctaacac tttcttaatc 1740actctccatc actgcttgaa gtcgacgaac tacatcaatc tcatggttag ttcgaagcag 1800cccactccgg tgtatttgac cccagaagag gcagagagcc actgcagagc aggtgcatct 1860gtctggaaat tctgcagcac cgacgatggt ctggacccag acgtagtatt ggttggaatc 1920ggcgtcgagg tgatgtttga agtcattgct gctgccgcca ttctgcgcaa acgttgcccc 1980gagcttcgcg tacgggtggt taacgtcacc gatttgatga tcttggagaa cgaaggcgca 2040cacccgcatg cgctcaccac cgacgcgttt gatagcctgt ttacatctga cagacccatc 2100catttcaact accacggata tccgactgag ttgcagggct tgctgttcgg acggccacgt 2160ctggagcgcg ctaccatcgg cggatacatg gaggagggaa gcaccacgac accatttgat 2220atgatgcttg tcaaccgcgt ttcacgcttc catgtagcac aagcggccat ttgtggggcg 2280tcaaagagaa atgaaagagt tcagatccga caacaggagc tgtacacgga gttggaccac 2340aatatggtag agaccaagaa gtacatcctc aaaaaccgaa aggatcccga tgatatgtac 2400gatatgccga aattcgaata a 242178806PRTAspergillus clavatusPHK 78Met Pro Gly Glu Val Ile Asp Arg Pro Asn Pro Gln Ala Leu Pro Ser1 5 10 15His Ile Pro Asp Val Val Glu Gln Leu Gln Val Lys Leu Asp Leu Thr 20 25 30Ser Leu Glu Gln Pro Ala Cys Asp Ala Leu Phe Lys Phe Arg Arg Ala 35 40 45Ala Ser Tyr Ile Ala Ala Ala Met Ile Phe Leu Gln Asp Asn Val Leu 50 55 60Leu Lys Arg Ser Leu Thr His Asp Asp Ile Lys Pro Arg Leu Leu Gly65 70 75 80His Trp Gly Thr Cys Pro Gly Leu Ile Leu Val Tyr Ser His Leu Asn 85 90 95Tyr Leu Ile Thr Lys Arg Asn Leu Asn Met Leu Tyr Val Val Gly Pro 100 105 110Gly His Gly Ala Pro Ala Ile Leu Ala Ser Leu Trp Leu Glu Gly Ser 115 120 125Leu Glu Lys Phe Tyr Pro Glu Tyr Ser Arg Asn Ala Glu Gly Leu Ala 130 135 140Asn Leu Ile Ser Thr Phe Ser Thr Thr Ala Gly Leu Pro Ser His Ile145 150 155 160Asn Ala Glu Thr Pro Gly Ala Ile His Glu Gly Gly Glu Leu Gly Tyr 165 170 175Ala Leu Ala Val Ser Phe Gly Ala Val Met Asp Asn Pro Asn Leu Ile 180 185 190Val Pro Cys Ile Val Gly Asp Gly Glu Ala Glu Thr Gly Pro Thr Ala 195 200 205Ala Ser Trp His Gly Ile Lys Tyr Ile Asp Pro Ala Glu Ser Gly Ala 210 215 220Val Leu Pro Ile Leu His Leu Asn Gly Phe Lys Ile Ser Glu Arg Thr225 230 235 240Ile Phe Gly Cys Met Asp Asp Lys Glu Leu Thr Ser Leu Phe Thr Gly 245 250 255Tyr Gly Tyr Gln Val Arg Ile Val Gln Asp Leu Asn Asp Ile Asp Thr 260 265 270Asp Leu His Arg Ser Met Thr Trp Ala Leu Glu Glu Ile His Lys Ile 275 280 285Gln Gln Ala Ala Arg Ser Gly Lys Pro Ile Met Lys Pro Arg Trp Pro 290 295 300Met Leu Ile Leu Arg Thr Pro Lys Gly Trp Ser Gly Pro Lys Glu Ile305 310 315 320His Gly Gln Phe Ile Glu Gly Ser Phe His Ser His Gln Val Pro Leu 325 330 335Pro Asn Ala Lys Lys Asp Lys Glu Glu Leu Gln Ala Leu Glu Lys Trp 340 345 350Leu Ser Ser Tyr Gly Pro Arg Glu Leu Phe Thr Glu Asn Gly Asp Val 355 360 365Ile Asp Glu Ile Lys Ser Ile Ile Pro Thr Asp Asp Ala Met Lys Leu 370 375 380Gly Gln Arg Val Glu Ala Tyr Lys Ala Tyr Leu Pro Pro Lys Leu Pro385 390 395 400Asp Trp Arg Lys Phe Ala Val Asp Lys Gly Ser Gln Gln Ser Pro Met 405 410 415Lys Thr Thr Gly Lys Leu Ile Asp Glu Val Phe Gln Leu Asn Pro His 420 425 430Thr Val Arg Leu Phe Ser Pro Asp Glu Leu Glu Ser Asn Lys Leu Ser 435 440 445Ala Ala Leu Asp His Thr Gly Arg Asn Phe Gln Trp Asp Gln Phe Ser 450 455 460Ser Ala Lys Gly Gly Arg Val Ile Glu Val Leu Ser Glu His Met Cys465 470 475 480Gln Gly Phe Ile Gln Gly Tyr Thr Leu Thr Gly Arg Thr Gly Ile Phe 485 490 495Pro Ser Tyr Glu Ser Phe Leu Gly Ile Val His Thr Met Met Val Gln 500 505 510Tyr Ser Lys Phe Ile Lys Met Ala Gln Glu Thr Thr Trp His Arg Ser 515 520 525Ile Ala Ser Ile Asn Tyr Ile Glu Thr Ser Thr Trp Thr Arg Gln Glu 530 535 540His Asn Gly Phe Ser His Gln Asn Pro Ser Phe Ile Gly Ala Val Leu545 550 555 560Lys Leu Lys Pro Thr Ala Ala Arg Val Tyr Leu Pro Pro Asp Ala Asn 565 570 575Thr Phe Leu Ile Thr Leu His His Cys Leu Lys Ser Thr Asn Tyr Ile 580 585 590Asn Leu Met Val Ser Ser Lys Gln Pro Thr Pro Val Tyr Leu Thr Pro 595 600 605Glu Glu Ala Glu Ser His Cys Arg Ala Gly Ala Ser Val Trp Lys Phe 610 615 620Cys Ser Thr Asp Asp Gly Leu Asp Pro Asp Val Val Leu Val Gly Ile625 630 635 640Gly Val Glu Val Met Phe Glu Val Ile Ala Ala Ala Ala Ile Leu Arg 645 650 655Lys Arg Cys Pro Glu Leu Arg Val Arg Val Val Asn Val Thr Asp Leu 660 665 670Met Ile Leu Glu Asn Glu Gly Ala His Pro His Ala Leu Thr Thr Asp 675 680 685Ala Phe Asp Ser Leu Phe Thr Ser Asp Arg Pro Ile His Phe Asn Tyr 690 695 700His Gly Tyr Pro Thr Glu Leu Gln Gly Leu Leu Phe Gly Arg Pro Arg705 710 715 720Leu Glu Arg Ala Thr Ile Gly Gly Tyr Met Glu Glu Gly Ser Thr Thr 725 730 735Thr Pro Phe Asp Met Met Leu Val Asn Arg Val Ser Arg Phe His Val 740 745 750Ala Gln Ala Ala Ile Cys Gly Ala Ser Lys Arg Asn Glu Arg Val Gln 755 760 765Ile Arg Gln Gln Glu Leu Tyr Thr Glu Leu Asp His Asn Met Val Glu 770 775 780Thr Lys Lys Tyr Ile Leu Lys Asn Arg Lys Asp Pro Asp Asp Met Tyr785 790 795 800Asp Met Pro Lys Phe Glu 80579996DNAClostridium cellulolyticumPTA 79atgaactttt tagagcagat tgtaagcaga gcaaaaaaag atatcaaaac aatagtactt 60cctgaaagta acgatatcag aactttgaag gctgctgcaa tgattcagga gcaaggcatt 120gccaatgtcg tacttgtggg caacaatgat gatattaaaa aactgggcgg tgacctggat 180atatcaaagg caaaaattgt tgatcctaca aactttgaaa gatttgacga gtatgtaaat 240accctgtacg aacttagaaa ggcaaaggga atgacacagg aagaggcaaa aaaaatattg 300tctgaaaatc ctttgtattt cggaattatg atggtaaaaa tgggtgaggc tgacggaatg 360gttgcaggag caattaattc aactgcaaat acacttaggc cggctcttca aatattaaag 420acagctcccg gagcaaaact tgtatctgca ttcttcgtaa tggttgtacc tgactgtgaa 480tacggagaaa atggagtgtt catttatgga gacagcggac tagtcgaaaa tccaaatgca 540gaagagcttt cagaaatagc gatagcatct tcaaaatcct tcaaaaacct tgtacaggct 600gaacctgttg ttgcaatgct ctcatattca acatacggca gtgcaaaaag tgtattgaca 660gaaaaggtta tagaggccac aaagcttgca aaggaaaaag ctccagatct tcagttagat 720ggagaattgc aggcagatgc agctattgtt ccttctgtag gagcatcaaa agcacccgga 780agcagtgttg caggaaaggc caatgtatta atattccctg acttgaactg tggtaatata 840tcctacaagc taacacagag acttgcaaag gcagaagcct acggccctat tattcaggga 900attgcaaaac ctgttaatga cctttcaaga ggctgtagtg ccgaggatat agtaggtgtt 960gttgcaataa cttgtgtaca agcacaaaat atataa 99680331PRTClostridium cellulolyticumPTA 80Met Asn Phe Leu Glu Gln Ile Val Ser Arg Ala Lys Lys Asp Ile Lys1 5 10 15Thr Ile Val Leu Pro Glu Ser Asn Asp Ile Arg Thr Leu Lys Ala Ala 20 25 30Ala Met Ile Gln Glu Gln Gly Ile Ala Asn Val Val Leu Val Gly Asn 35 40 45Asn Asp Asp Ile Lys Lys Leu Gly Gly Asp Leu Asp Ile Ser Lys Ala 50 55 60Lys

Ile Val Asp Pro Thr Asn Phe Glu Arg Phe Asp Glu Tyr Val Asn65 70 75 80Thr Leu Tyr Glu Leu Arg Lys Ala Lys Gly Met Thr Gln Glu Glu Ala 85 90 95Lys Lys Ile Leu Ser Glu Asn Pro Leu Tyr Phe Gly Ile Met Met Val 100 105 110Lys Met Gly Glu Ala Asp Gly Met Val Ala Gly Ala Ile Asn Ser Thr 115 120 125Ala Asn Thr Leu Arg Pro Ala Leu Gln Ile Leu Lys Thr Ala Pro Gly 130 135 140Ala Lys Leu Val Ser Ala Phe Phe Val Met Val Val Pro Asp Cys Glu145 150 155 160Tyr Gly Glu Asn Gly Val Phe Ile Tyr Gly Asp Ser Gly Leu Val Glu 165 170 175Asn Pro Asn Ala Glu Glu Leu Ser Glu Ile Ala Ile Ala Ser Ser Lys 180 185 190Ser Phe Lys Asn Leu Val Gln Ala Glu Pro Val Val Ala Met Leu Ser 195 200 205Tyr Ser Thr Tyr Gly Ser Ala Lys Ser Val Leu Thr Glu Lys Val Ile 210 215 220Glu Ala Thr Lys Leu Ala Lys Glu Lys Ala Pro Asp Leu Gln Leu Asp225 230 235 240Gly Glu Leu Gln Ala Asp Ala Ala Ile Val Pro Ser Val Gly Ala Ser 245 250 255Lys Ala Pro Gly Ser Ser Val Ala Gly Lys Ala Asn Val Leu Ile Phe 260 265 270Pro Asp Leu Asn Cys Gly Asn Ile Ser Tyr Lys Leu Thr Gln Arg Leu 275 280 285Ala Lys Ala Glu Ala Tyr Gly Pro Ile Ile Gln Gly Ile Ala Lys Pro 290 295 300Val Asn Asp Leu Ser Arg Gly Cys Ser Ala Glu Asp Ile Val Gly Val305 310 315 320Val Ala Ile Thr Cys Val Gln Ala Gln Asn Ile 325 33081996DNAClostridium phytofermentansPTA 81atgggattta ttgatgacat caaggcaaga gctaaacaaa gtattaagac tattgtttta 60cctgagagta tggacagaag aacaattgag gcagctgcta agactttaga agagggcaat 120gctaacgtaa ttattatcgg tagtgaggaa gaagttaaga agaattcaga aggtcttgac 180atttcgggag ctacaatcgt tgaccctaag acatcggaca agcttccagc ttacattaac 240aagcttgtag aacttagaca ggcaaaaggc atgacccctg aaaaagcaaa agagctttta 300acaacagact acattacata cggtgtaatg atggttaaga tgggcgatgc agatggttta 360gtatctggtg cttgtcactc tacagcagat accttaagac catgtcttca gattttaaaa 420actgctccaa atactaagtt agtttctgct ttcttcgtaa tggtagtacc taattgtgat 480atgggcgcaa atggaacttt ccttttctct gatgctggtt taaatcagaa tccaaatgct 540gaagagttag cagcaatcgc tggttccaca gcgaagagtt ttgaacaatt agttggctct 600gaacctatcg tagctatgct ttctcattca acaaagggaa gcgcaaagca tgcagatgtt 660gataaggttg tagaagcaac taagattgca aatgaattat acccagaata taagatcgac 720ggcgagttcc agttagatgc agcaatcgtt cctagtgtag gtgcttcaaa agctcctggt 780agtgatattg ctggaaaagc taacgtatta atcttcccag accttgatgc tggtaacatt 840ggatataagt taacacagcg tcttgcaaag gcagaagctt atggaccatt aactcagggt 900attgcagctc cagtaaatga tttatcaaga ggttgttctt ctgatgatat cgttggtgtt 960gttgcaatca ctgctgttca ggcacagagt aaataa 99682331PRTClostridium phytofermentansPTA 82Met Gly Phe Ile Asp Asp Ile Lys Ala Arg Ala Lys Gln Ser Ile Lys1 5 10 15Thr Ile Val Leu Pro Glu Ser Met Asp Arg Arg Thr Ile Glu Ala Ala 20 25 30Ala Lys Thr Leu Glu Glu Gly Asn Ala Asn Val Ile Ile Ile Gly Ser 35 40 45Glu Glu Glu Val Lys Lys Asn Ser Glu Gly Leu Asp Ile Ser Gly Ala 50 55 60Thr Ile Val Asp Pro Lys Thr Ser Asp Lys Leu Pro Ala Tyr Ile Asn65 70 75 80Lys Leu Val Glu Leu Arg Gln Ala Lys Gly Met Thr Pro Glu Lys Ala 85 90 95Lys Glu Leu Leu Thr Thr Asp Tyr Ile Thr Tyr Gly Val Met Met Val 100 105 110Lys Met Gly Asp Ala Asp Gly Leu Val Ser Gly Ala Cys His Ser Thr 115 120 125Ala Asp Thr Leu Arg Pro Cys Leu Gln Ile Leu Lys Thr Ala Pro Asn 130 135 140Thr Lys Leu Val Ser Ala Phe Phe Val Met Val Val Pro Asn Cys Asp145 150 155 160Met Gly Ala Asn Gly Thr Phe Leu Phe Ser Asp Ala Gly Leu Asn Gln 165 170 175Asn Pro Asn Ala Glu Glu Leu Ala Ala Ile Ala Gly Ser Thr Ala Lys 180 185 190Ser Phe Glu Gln Leu Val Gly Ser Glu Pro Ile Val Ala Met Leu Ser 195 200 205His Ser Thr Lys Gly Ser Ala Lys His Ala Asp Val Asp Lys Val Val 210 215 220Glu Ala Thr Lys Ile Ala Asn Glu Leu Tyr Pro Glu Tyr Lys Ile Asp225 230 235 240Gly Glu Phe Gln Leu Asp Ala Ala Ile Val Pro Ser Val Gly Ala Ser 245 250 255Lys Ala Pro Gly Ser Asp Ile Ala Gly Lys Ala Asn Val Leu Ile Phe 260 265 270Pro Asp Leu Asp Ala Gly Asn Ile Gly Tyr Lys Leu Thr Gln Arg Leu 275 280 285Ala Lys Ala Glu Ala Tyr Gly Pro Leu Thr Gln Gly Ile Ala Ala Pro 290 295 300Val Asn Asp Leu Ser Arg Gly Cys Ser Ser Asp Asp Ile Val Gly Val305 310 315 320Val Ala Ile Thr Ala Val Gln Ala Gln Ser Lys 325 330831815DNABifidobacterium bifidumPTA 83atgacgcacg cgcccggtcg aaccgccccg gccgcgttta ccgccatgtc aacgcccggc 60agcgaacgat cgggtactac aatgtcagta agtgtgaaac aaggcggtca cacgagcagt 120accatccccg acgaggagac aaacgtgaca acaacgaaca tctacatcgc aagtcctgaa 180ggcgcaaacg gacgcaacgt cgtggcatac ggcgtgctca aggcgctgac caccaaattc 240aagaccgccg tgttccgccc cgcggtctcc aatcacgacc acttcacccc catcctgctg 300aacgcctcca acgccggcct cggcgtcgcg ctatccaccg gaccggacat tcaccaggtg 360cgcaaggaca aggaaggctc ccgcggcgac atcgtcgcgg cgttcaacga cgcgatgaac 420gtgtcccgcg cggacgcggc gctcatcgtc ggcaccgaca agtcgcacgt caacgacccg 480accgcctacg agttcaacgc gaacatcgcc gccgacctgc aggccggcgt cttcctcgcc 540gtgtgcacca tcgatcgttg gcctgacgag ctcaaggaca ccgtccgtct ctccatcgaa 600ggcatggagg ccgccggcaa caaggtgctg ggcatcttcg tgacgggctg cgatccgcgc 660cactccgtct cggtcaagga cacgctcgcg gacttcgggc tgccggtgtg gacgatcccc 720cagatcccgt tcaccgatgc gtcacaggcc gatgcggcgc tcgccgcctt ccaggcgaac 780gtgccgaccg aagaggtcct gcaggccatc aacgtcgaga tcgacttccc gatcacctcg 840tacgccttcc agtacagcct gctcggccgt gccaaggcga acaagaaaac gatcgtgctg 900cccgaaggcg aagaggaccg catcatcaag gcggccgact acctgctgga gcgcgagatc 960gcgaacctca tcatcgtggg cgacaagggc gcgatcctcg cccggggcgc ggaactcggt 1020ctgacccatc tgggcaaggc gcgtttccag gccatggacg acgagaacat cctcaagccc 1080atggtcgcca agctgtgcga gctgcgcgcc aagaagggca tgaccgaaga gcaggcccgc 1140aagcagctga ccgacgcgag ctatttcggc acgatgctcg tcgtgctcgg ttacgccgac 1200ggtctggtgt ccggcgcggt caactccacc gcaaacaccg tgcgtccggc gctgcaggtc 1260atcaagacca agcccgggca gaggctcgtc tccggcgcct tcctgatgtg cttcaaggac 1320cacgtgaccg tgttctccga ctgcgccatc aacctcaacc cggacgccga acagctcgcg 1380gagatcgcgc tgcagtccgc cgtgaccgcc aaggcgttcg gcatcgagcc caaggtcggc 1440atgctctcct actccaccct cggctccggc aaggggccgg acgtggacat cgtcgaagag 1500gccacgcgaa tcgtcaagga gaaggctccg gacctgcccg tcgtcggctc catccagttc 1560gacgccgcat ggtcgcccga cgtggcagcc accaaggcca agggcaacga cgtggccgga 1620cacgtcaacg tgttcgtgtt ccccgacctg tgctccggca acatcggcta caaggccgtg 1680cagcgttcct ccggggcgct cgccatcggc cccgtgctgc agggcctcaa caagcccgtc 1740aacgacctct cacgcggtgc gacggtgcag gacatcatca acaccatcgc actgaccgcc 1800atcgaggccc agtga 181584604PRTBifidobacterium bifidumPTA 84Met Thr His Ala Pro Gly Arg Thr Ala Pro Ala Ala Phe Thr Ala Met1 5 10 15Ser Thr Pro Gly Ser Glu Arg Ser Gly Thr Thr Met Ser Val Ser Val 20 25 30Lys Gln Gly Gly His Thr Ser Ser Thr Ile Pro Asp Glu Glu Thr Asn 35 40 45Val Thr Thr Thr Asn Ile Tyr Ile Ala Ser Pro Glu Gly Ala Asn Gly 50 55 60Arg Asn Val Val Ala Tyr Gly Val Leu Lys Ala Leu Thr Thr Lys Phe65 70 75 80Lys Thr Ala Val Phe Arg Pro Ala Val Ser Asn His Asp His Phe Thr 85 90 95Pro Ile Leu Leu Asn Ala Ser Asn Ala Gly Leu Gly Val Ala Leu Ser 100 105 110Thr Gly Pro Asp Ile His Gln Val Arg Lys Asp Lys Glu Gly Ser Arg 115 120 125Gly Asp Ile Val Ala Ala Phe Asn Asp Ala Met Asn Val Ser Arg Ala 130 135 140Asp Ala Ala Leu Ile Val Gly Thr Asp Lys Ser His Val Asn Asp Pro145 150 155 160Thr Ala Tyr Glu Phe Asn Ala Asn Ile Ala Ala Asp Leu Gln Ala Gly 165 170 175Val Phe Leu Ala Val Cys Thr Ile Asp Arg Trp Pro Asp Glu Leu Lys 180 185 190Asp Thr Val Arg Leu Ser Ile Glu Gly Met Glu Ala Ala Gly Asn Lys 195 200 205Val Leu Gly Ile Phe Val Thr Gly Cys Asp Pro Arg His Ser Val Ser 210 215 220Val Lys Asp Thr Leu Ala Asp Phe Gly Leu Pro Val Trp Thr Ile Pro225 230 235 240Gln Ile Pro Phe Thr Asp Ala Ser Gln Ala Asp Ala Ala Leu Ala Ala 245 250 255Phe Gln Ala Asn Val Pro Thr Glu Glu Val Leu Gln Ala Ile Asn Val 260 265 270Glu Ile Asp Phe Pro Ile Thr Ser Tyr Ala Phe Gln Tyr Ser Leu Leu 275 280 285Gly Arg Ala Lys Ala Asn Lys Lys Thr Ile Val Leu Pro Glu Gly Glu 290 295 300Glu Asp Arg Ile Ile Lys Ala Ala Asp Tyr Leu Leu Glu Arg Glu Ile305 310 315 320Ala Asn Leu Ile Ile Val Gly Asp Lys Gly Ala Ile Leu Ala Arg Gly 325 330 335Ala Glu Leu Gly Leu Thr His Leu Gly Lys Ala Arg Phe Gln Ala Met 340 345 350Asp Asp Glu Asn Ile Leu Lys Pro Met Val Ala Lys Leu Cys Glu Leu 355 360 365Arg Ala Lys Lys Gly Met Thr Glu Glu Gln Ala Arg Lys Gln Leu Thr 370 375 380Asp Ala Ser Tyr Phe Gly Thr Met Leu Val Val Leu Gly Tyr Ala Asp385 390 395 400Gly Leu Val Ser Gly Ala Val Asn Ser Thr Ala Asn Thr Val Arg Pro 405 410 415Ala Leu Gln Val Ile Lys Thr Lys Pro Gly Gln Arg Leu Val Ser Gly 420 425 430Ala Phe Leu Met Cys Phe Lys Asp His Val Thr Val Phe Ser Asp Cys 435 440 445Ala Ile Asn Leu Asn Pro Asp Ala Glu Gln Leu Ala Glu Ile Ala Leu 450 455 460Gln Ser Ala Val Thr Ala Lys Ala Phe Gly Ile Glu Pro Lys Val Gly465 470 475 480Met Leu Ser Tyr Ser Thr Leu Gly Ser Gly Lys Gly Pro Asp Val Asp 485 490 495Ile Val Glu Glu Ala Thr Arg Ile Val Lys Glu Lys Ala Pro Asp Leu 500 505 510Pro Val Val Gly Ser Ile Gln Phe Asp Ala Ala Trp Ser Pro Asp Val 515 520 525Ala Ala Thr Lys Ala Lys Gly Asn Asp Val Ala Gly His Val Asn Val 530 535 540Phe Val Phe Pro Asp Leu Cys Ser Gly Asn Ile Gly Tyr Lys Ala Val545 550 555 560Gln Arg Ser Ser Gly Ala Leu Ala Ile Gly Pro Val Leu Gln Gly Leu 565 570 575Asn Lys Pro Val Asn Asp Leu Ser Arg Gly Ala Thr Val Gln Asp Ile 580 585 590Ile Asn Thr Ile Ala Leu Thr Ala Ile Glu Ala Gln 595 600851683DNABifidobacterium animalisPTA 85atgagcctca aaaacgtgag catcatcagt cccgaacctg gagacggccg gaacgtggtg 60gcattgggtg tggtggatat gctcgcccac accgcgaaga ccagcatatt ccggccggcg 120gtgcaggcag gcgaaaagct caccgatgcc ctcctctcgg tggcgagcgc gccgcagacc 180cgcgaacagg ccgttggcgt cacactgcgt gacgcacgtc tcgacaaaga caacgcccgc 240cagcagatcg tctccaaatt catcgacacg aatgccgaga tcaatccgca gatccgagtg 300atcgtcggat cagaccgcac caatgtgggc gatcccgaac ggttcacctt caatgcggac 360gtctccgccg acctgcagtc tccggtgctg ctctcagtga gctcgatggg gcgcacggcg 420gaagagattc gcgagacgat agacgcatgc cgcgaggtgg tcatcaacgc aggcacccaa 480gtgattggcg tgttcatcac cgactgcacg aattcggcgc tgcccaccct caccgacgaa 540ttcgtgagct acgacctgcc gacctggccg cttcccctcg tggaactcgg ttccgatgac 600acgaatgtca aggccgcact cgaggccttc gacgagcatg tcgacaagga atcactcctc 660aacgtgcttg acacaccttt cgtaccgccg accacaccat tcgcgttcca gtacgatctg 720ctcgcacgtg cgaagaaaga caagaagacg attgtgctgc cagaaggaga ggatgaccgc 780atcatcaccg ccgcgaacta cctgctgcag tcgaatgtgg tcgatctcgt catcatcggc 840gaccgcgacg agattcttgc ccggggtgaa aaactcggcc tcaaggcgct tgaccaggcg 900aagttcgttt ccatcgacga caggcatctg ctcgacacca tggtgcccaa actgtgtgaa 960ttgcgcgcca agaagggcat gacccccgac gtggctttga aaacgctcag agacacgaac 1020tacttcggca ccatgctcat cgtgctcggc atggccgatg gtctggtctc aggtgccatc 1080agctccaccg ccaacaccgt gcgccccgcg ctccagctca tcaagacgaa acccggtgtc 1140tcctccgtat cgggtgcctt cctcatgtgt ctcaaggacc acgtgtcggt atttgctgat 1200tgcgccatca acctcgatcc caatccgcag cagctggccg acatcgccat ccagtcagcc 1260gagaccgcca aggcattctc catcgatcct aaaatcggtt tgctctccta ttccaccttg 1320ggttccggta agggaccgga tgtcgatctg gtggtcgagg ccacttcgat cgcccagaac 1380aaacgaccgg acctacccat cgtcgggccc atccagttcg acgccgcctg gtcgaagacg 1440gttgccaagg tcaaggcctt cggcaacccg atcgccggca atgtcaccgt gttcgtgttc 1500ccggacctcg atgcggggaa catctgctac aaggcagtgc agcgcacctc cggcgccgtg 1560gcgatcggac cggtgcttca ggggctcaac cggcctgtca acgacctgag ccgaggcgcc 1620ttggtgcagg acatcatcaa taccatcgcc ttgaccgcca ttgaagcgca aagcaacgaa 1680tag 168386560PRTBifidobacterium animalisPTA 86Met Ser Leu Lys Asn Val Ser Ile Ile Ser Pro Glu Pro Gly Asp Gly1 5 10 15Arg Asn Val Val Ala Leu Gly Val Val Asp Met Leu Ala His Thr Ala 20 25 30Lys Thr Ser Ile Phe Arg Pro Ala Val Gln Ala Gly Glu Lys Leu Thr 35 40 45Asp Ala Leu Leu Ser Val Ala Ser Ala Pro Gln Thr Arg Glu Gln Ala 50 55 60Val Gly Val Thr Leu Arg Asp Ala Arg Leu Asp Lys Asp Asn Ala Arg65 70 75 80Gln Gln Ile Val Ser Lys Phe Ile Asp Thr Asn Ala Glu Ile Asn Pro 85 90 95Gln Ile Arg Val Ile Val Gly Ser Asp Arg Thr Asn Val Gly Asp Pro 100 105 110Glu Arg Phe Thr Phe Asn Ala Asp Val Ser Ala Asp Leu Gln Ser Pro 115 120 125Val Leu Leu Ser Val Ser Ser Met Gly Arg Thr Ala Glu Glu Ile Arg 130 135 140Glu Thr Ile Asp Ala Cys Arg Glu Val Val Ile Asn Ala Gly Thr Gln145 150 155 160Val Ile Gly Val Phe Ile Thr Asp Cys Thr Asn Ser Ala Leu Pro Thr 165 170 175Leu Thr Asp Glu Phe Val Ser Tyr Asp Leu Pro Thr Trp Pro Leu Pro 180 185 190Leu Val Glu Leu Gly Ser Asp Asp Thr Asn Val Lys Ala Ala Leu Glu 195 200 205Ala Phe Asp Glu His Val Asp Lys Glu Ser Leu Leu Asn Val Leu Asp 210 215 220Thr Pro Phe Val Pro Pro Thr Thr Pro Phe Ala Phe Gln Tyr Asp Leu225 230 235 240Leu Ala Arg Ala Lys Lys Asp Lys Lys Thr Ile Val Leu Pro Glu Gly 245 250 255Glu Asp Asp Arg Ile Ile Thr Ala Ala Asn Tyr Leu Leu Gln Ser Asn 260 265 270Val Val Asp Leu Val Ile Ile Gly Asp Arg Asp Glu Ile Leu Ala Arg 275 280 285Gly Glu Lys Leu Gly Leu Lys Ala Leu Asp Gln Ala Lys Phe Val Ser 290 295 300Ile Asp Asp Arg His Leu Leu Asp Thr Met Val Pro Lys Leu Cys Glu305 310 315 320Leu Arg Ala Lys Lys Gly Met Thr Pro Asp Val Ala Leu Lys Thr Leu 325 330 335Arg Asp Thr Asn Tyr Phe Gly Thr Met Leu Ile Val Leu Gly Met Ala 340 345 350Asp Gly Leu Val Ser Gly Ala Ile Ser Ser Thr Ala Asn Thr Val Arg 355 360 365Pro Ala Leu Gln Leu Ile Lys Thr Lys Pro Gly Val Ser Ser Val Ser 370 375 380Gly Ala Phe Leu Met Cys Leu Lys Asp His Val Ser Val Phe Ala Asp385 390 395 400Cys Ala Ile Asn Leu Asp Pro Asn Pro Gln Gln Leu Ala Asp Ile Ala 405 410 415Ile Gln Ser Ala Glu Thr Ala Lys Ala Phe Ser Ile Asp Pro Lys Ile 420 425 430Gly Leu Leu Ser Tyr Ser Thr Leu Gly Ser Gly Lys Gly Pro Asp Val 435 440 445Asp Leu Val Val Glu Ala Thr Ser Ile Ala Gln Asn Lys Arg Pro Asp 450 455 460Leu Pro Ile Val Gly Pro Ile Gln Phe Asp Ala Ala Trp Ser Lys Thr465 470 475

480Val Ala Lys Val Lys Ala Phe Gly Asn Pro Ile Ala Gly Asn Val Thr 485 490 495Val Phe Val Phe Pro Asp Leu Asp Ala Gly Asn Ile Cys Tyr Lys Ala 500 505 510Val Gln Arg Thr Ser Gly Ala Val Ala Ile Gly Pro Val Leu Gln Gly 515 520 525Leu Asn Arg Pro Val Asn Asp Leu Ser Arg Gly Ala Leu Val Gln Asp 530 535 540Ile Ile Asn Thr Ile Ala Leu Thr Ala Ile Glu Ala Gln Ser Asn Glu545 550 555 560871097DNAClostridium cellulolyticumACK 87atgaaggttt tagttataaa tgcggggagt tcttcattaa aatatcaatt aatagatatg 60aaaaatgaaa cagttcttgc aaagggttta tgcgacagaa tagggattga caattccttt 120ataaagcaat caaggggttc agaagaggct gttattttga ataaagagct aaagaatcac 180aaagatgcaa tagaggctgt tatttctgca ctgactgacg ataatatggg cgttataaaa 240aacatgtccg aaatatcagc agtgggacac agaatagtac acggcggtga aaaattcaac 300agttctgtag ttatagatga aaacgttatg aatgcagtaa gagagtgtat agacgttgca 360ccgcttcata atccgccgaa tattataggt atagaggctt gccagcagat tatgcccaat 420atacctatgg tagctgtatt tgataccact ttccacagct ccatgcctga ttatgcatac 480ctttacgcat tgccatatga actttatgaa aagtacggta taagaaaata tggtttccac 540ggaacatcac acaaatatgt tgcagaaaga gcttctgcaa tgcttgataa gtctttgaac 600gaattaaaga taattacatg ccatcttggg aacggttcaa gtatttgtgc tgttaacaag 660ggtaaatcaa ttgatacttc catgggcttt acacctttgc agggacttgc aatgggtaca 720agaagcggta caatagaccc tgaagttgtt acattcctta tggaaaagga aaatctggat 780gttaagggtg taagcaaact cttaaataaa aagtcaggtg ttttaggtat atcaggtgta 840agcagcgatt tcagagattt acatgctgcc gctgatgcag gaaacagcag agctgagctg 900gcaatagaaa ttttctgcta tggtgttaag aagtttatcg gagaatatat tgcagttatg 960aatggtgttg atgctatagt atttacagcc ggtgttggtg agaataattc agtagtaaga 1020aatatgatta tcagcgatat ggacttcctc ggcatcaaaa tagatgtaga aaagaataag 1080ctcacggggg caggaag 109788365PRTClostridium cellulolyticumACK 88Met Lys Val Leu Val Ile Asn Ala Gly Ser Ser Ser Leu Lys Tyr Gln1 5 10 15Leu Ile Asp Met Lys Asn Glu Thr Val Leu Ala Lys Gly Leu Cys Asp 20 25 30Arg Ile Gly Ile Asp Asn Ser Phe Ile Lys Gln Ser Arg Gly Ser Glu 35 40 45Glu Ala Val Ile Leu Asn Lys Glu Leu Lys Asn His Lys Asp Ala Ile 50 55 60Glu Ala Val Ile Ser Ala Leu Thr Asp Asp Asn Met Gly Val Ile Lys65 70 75 80Asn Met Ser Glu Ile Ser Ala Val Gly His Arg Ile Val His Gly Gly 85 90 95Glu Lys Phe Asn Ser Ser Val Val Ile Asp Glu Asn Val Met Asn Ala 100 105 110Val Arg Glu Cys Ile Asp Val Ala Pro Leu His Asn Pro Pro Asn Ile 115 120 125Ile Gly Ile Glu Ala Cys Gln Gln Ile Met Pro Asn Ile Pro Met Val 130 135 140Ala Val Phe Asp Thr Thr Phe His Ser Ser Met Pro Asp Tyr Ala Tyr145 150 155 160Leu Tyr Ala Leu Pro Tyr Glu Leu Tyr Glu Lys Tyr Gly Ile Arg Lys 165 170 175Tyr Gly Phe His Gly Thr Ser His Lys Tyr Val Ala Glu Arg Ala Ser 180 185 190Ala Met Leu Asp Lys Ser Leu Asn Glu Leu Lys Ile Ile Thr Cys His 195 200 205Leu Gly Asn Gly Ser Ser Ile Cys Ala Val Asn Lys Gly Lys Ser Ile 210 215 220Asp Thr Ser Met Gly Phe Thr Pro Leu Gln Gly Leu Ala Met Gly Thr225 230 235 240Arg Ser Gly Thr Ile Asp Pro Glu Val Val Thr Phe Leu Met Glu Lys 245 250 255Glu Asn Leu Asp Val Lys Gly Val Ser Lys Leu Leu Asn Lys Lys Ser 260 265 270Gly Val Leu Gly Ile Ser Gly Val Ser Ser Asp Phe Arg Asp Leu His 275 280 285Ala Ala Ala Asp Ala Gly Asn Ser Arg Ala Glu Leu Ala Ile Glu Ile 290 295 300Phe Cys Tyr Gly Val Lys Lys Phe Ile Gly Glu Tyr Ile Ala Val Met305 310 315 320Asn Gly Val Asp Ala Ile Val Phe Thr Ala Gly Val Gly Glu Asn Asn 325 330 335Ser Val Val Arg Asn Met Ile Ile Ser Asp Met Asp Phe Leu Gly Ile 340 345 350Lys Ile Asp Val Glu Lys Asn Lys Leu Thr Gly Ala Gly 355 360 365892703DNALeuconostoc mesenteroidesadhE 89atggcagaag caattgcaaa gaaacccgca aaaagggttt tgacccctga agaaaaagcg 60gaattacaaa cacaagctga gaagatgact gaggtattga ttgaaaaatc acaaaaggca 120ttgtctgaat tttcaacatt ttcgcaagaa caagttgata aaattgttgc agctatggcc 180ttggcaggtt ctgagaattc acttctgtta gcccatgctg ctcacgacga gactggacgt 240ggggttgtgg aagataagga tacgaaaaat cgtttcgcct cagaatcagt ttataacgct 300attaagtttg ataagactgt gggtgttatt agtgaagaca agattcaagg taaggtagaa 360ttagcagccc cacttggtat tttggctgga atcgtcccaa cgacaaatcc aacgtcgaca 420actattttca aatcaatgtt gacagcaaag acacgtaaca caattatctt tgctttccat 480ccccaggctc aaaaagcatc ggttcttgct gcaaaaattg tttatgatgc tgctgttaaa 540gcaggcgcac cggaaaactt tatccaatgg attgaaaagc cttcacttta tgcaacaagt 600gcgctgatac aaaatcctca cattgcttca attctagcta ctggtgggcc atcaatggtt 660aatgcagctt tgaagtcagg aaatccatcc atgggtgtcg gtgctggaaa cggtgcagtt 720tatattgatg caactgttga cacagatcgt gccgtgtccg atttgttgtt atcaaagcgt 780ttcgataatg gcatgatttg tgccacagaa aactcagccg ttattcaagc accaatctat 840gacgaaattt taactaagtt acaagaacaa ggtgcatacc ttgttcctaa gaaagactac 900aaaaaaattg ctgattatgt atttaagcct aacgcagagg gatttggtat tgctggtcct 960gttgctggta tgtcaggacg ttggattgct gagcaagcag gcgtaaagat tcctgatggt 1020aaagatgtac ttttgttcga attagatcag aagaacatag gtgaagcgtt atcttctgaa 1080aagttatcgc cattactttc aatttataaa gttgagaagc gtgaagaagc tattgagact 1140gttcaatcct tgttaaacta tcaaggcgcg gggcacaacg cagcaattca aattggttca 1200caagatgatc cattcattaa agagtatgct gacgctattg gtgcatcacg tattttggtt 1260aaccaacctg actcaatcgg tggtgttgga gatatttaca cagatgctat gcgtccatcg 1320ttgacacttg gtaccggatc atgggggaag aattcattgt ctcataactt atcaacatac 1380gacttactta atattaagac cgtggctcgc cgccgtaatc gtcctcaatg ggttcgttta 1440cctaaggaag tttactacga agccaatgcc attacttact tacaagactt gcctactata 1500aaccgtgcat ttattgtcgc tgatcctggt atggttcagt tcggatttgt tggcagagta 1560ctaggtcaac ttgagttacg tcaagaacag gttgaaacaa atatctatgg ttcagttaag 1620cctgacccaa ctttgtcaca agctgttgaa attgctcgcc aaatggcaga cttcaaacca 1680gatacagtta ttttacttgg cggtggttcg gcacttgacg ctggtaaaat tggtcggttc 1740ttgtacgaat actcgacacg ccatgaagga attttagaag atgacgaggc gattaaagat 1800ctattcttag aactacaaca aaagtttatg gatattcgta agcgaatcgt taagttttac 1860cacgcacgtt tgacacaaat ggttgcgatt ccaacaactt caggtactgg atcagaagtc 1920acaccatttg ccgttattac agatgatgaa acacatgtaa agtatccact agccgattat 1980gaattgacac cggaagttgc tattgttgat ccagaatttg ttatgaccgt accacaacac 2040acggtatctt ggtcaggatt agatgctttg tcacatgctt tggaatcgta tgtctcagtg 2100atggcttctg aattcacacg tccttgggca ttacaagcta ttaagttgat ttttgataac 2160ttaacaaatt catacaatta tgatcctaaa cacccaacta aggaaggtca gaatgcacgc 2220acaaagatgc actatgcgtc aacattggct ggtatgtcat ttgcgaatgc cttcttggga 2280cttaaccact cactagcaca caaaactggt ggagaattcg gactacctca cggtatggca 2340atcgctattg caatgccaca tgtgattaag tttaatgcgg taacaggaaa tgtaaagcgc 2400acaccatacc cacgttacga aacctataca gcacaaaaag attatgctga tattgcacgt 2460tacttaggtt tgaaaggtga aacagatgct gaattggtcg atgtattgat tgcagaaatc 2520aagaagttgg ctgcatcagt gggtgtcaat caaacactat ctggcaacgg tgtttcaaag 2580catgactttg atacaaagtt agaaaagatg attgacttag tttacaatga ccaatgcacg 2640ccgggaaacc ctcgccaacc aagcttggca gaaattcgtc aattgttgaa agatcagttt 2700taa 270390900PRTLeuconostoc mesenteroidesadhE 90Met Ala Glu Ala Ile Ala Lys Lys Pro Ala Lys Arg Val Leu Thr Pro1 5 10 15Glu Glu Lys Ala Glu Leu Gln Thr Gln Ala Glu Lys Met Thr Glu Val 20 25 30Leu Ile Glu Lys Ser Gln Lys Ala Leu Ser Glu Phe Ser Thr Phe Ser 35 40 45Gln Glu Gln Val Asp Lys Ile Val Ala Ala Met Ala Leu Ala Gly Ser 50 55 60Glu Asn Ser Leu Leu Leu Ala His Ala Ala His Asp Glu Thr Gly Arg65 70 75 80Gly Val Val Glu Asp Lys Asp Thr Lys Asn Arg Phe Ala Ser Glu Ser 85 90 95Val Tyr Asn Ala Ile Lys Phe Asp Lys Thr Val Gly Val Ile Ser Glu 100 105 110Asp Lys Ile Gln Gly Lys Val Glu Leu Ala Ala Pro Leu Gly Ile Leu 115 120 125Ala Gly Ile Val Pro Thr Thr Asn Pro Thr Ser Thr Thr Ile Phe Lys 130 135 140Ser Met Leu Thr Ala Lys Thr Arg Asn Thr Ile Ile Phe Ala Phe His145 150 155 160Pro Gln Ala Gln Lys Ala Ser Val Leu Ala Ala Lys Ile Val Tyr Asp 165 170 175Ala Ala Val Lys Ala Gly Ala Pro Glu Asn Phe Ile Gln Trp Ile Glu 180 185 190Lys Pro Ser Leu Tyr Ala Thr Ser Ala Leu Ile Gln Asn Pro His Ile 195 200 205Ala Ser Ile Leu Ala Thr Gly Gly Pro Ser Met Val Asn Ala Ala Leu 210 215 220Lys Ser Gly Asn Pro Ser Met Gly Val Gly Ala Gly Asn Gly Ala Val225 230 235 240Tyr Ile Asp Ala Thr Val Asp Thr Asp Arg Ala Val Ser Asp Leu Leu 245 250 255Leu Ser Lys Arg Phe Asp Asn Gly Met Ile Cys Ala Thr Glu Asn Ser 260 265 270Ala Val Ile Gln Ala Pro Ile Tyr Asp Glu Ile Leu Thr Lys Leu Gln 275 280 285Glu Gln Gly Ala Tyr Leu Val Pro Lys Lys Asp Tyr Lys Lys Ile Ala 290 295 300Asp Tyr Val Phe Lys Pro Asn Ala Glu Gly Phe Gly Ile Ala Gly Pro305 310 315 320Val Ala Gly Met Ser Gly Arg Trp Ile Ala Glu Gln Ala Gly Val Lys 325 330 335Ile Pro Asp Gly Lys Asp Val Leu Leu Phe Glu Leu Asp Gln Lys Asn 340 345 350Ile Gly Glu Ala Leu Ser Ser Glu Lys Leu Ser Pro Leu Leu Ser Ile 355 360 365Tyr Lys Val Glu Lys Arg Glu Glu Ala Ile Glu Thr Val Gln Ser Leu 370 375 380Leu Asn Tyr Gln Gly Ala Gly His Asn Ala Ala Ile Gln Ile Gly Ser385 390 395 400Gln Asp Asp Pro Phe Ile Lys Glu Tyr Ala Asp Ala Ile Gly Ala Ser 405 410 415Arg Ile Leu Val Asn Gln Pro Asp Ser Ile Gly Gly Val Gly Asp Ile 420 425 430Tyr Thr Asp Ala Met Arg Pro Ser Leu Thr Leu Gly Thr Gly Ser Trp 435 440 445Gly Lys Asn Ser Leu Ser His Asn Leu Ser Thr Tyr Asp Leu Leu Asn 450 455 460Ile Lys Thr Val Ala Arg Arg Arg Asn Arg Pro Gln Trp Val Arg Leu465 470 475 480Pro Lys Glu Val Tyr Tyr Glu Ala Asn Ala Ile Thr Tyr Leu Gln Asp 485 490 495Leu Pro Thr Ile Asn Arg Ala Phe Ile Val Ala Asp Pro Gly Met Val 500 505 510Gln Phe Gly Phe Val Gly Arg Val Leu Gly Gln Leu Glu Leu Arg Gln 515 520 525Glu Gln Val Glu Thr Asn Ile Tyr Gly Ser Val Lys Pro Asp Pro Thr 530 535 540Leu Ser Gln Ala Val Glu Ile Ala Arg Gln Met Ala Asp Phe Lys Pro545 550 555 560Asp Thr Val Ile Leu Leu Gly Gly Gly Ser Ala Leu Asp Ala Gly Lys 565 570 575Ile Gly Arg Phe Leu Tyr Glu Tyr Ser Thr Arg His Glu Gly Ile Leu 580 585 590Glu Asp Asp Glu Ala Ile Lys Asp Leu Phe Leu Glu Leu Gln Gln Lys 595 600 605Phe Met Asp Ile Arg Lys Arg Ile Val Lys Phe Tyr His Ala Arg Leu 610 615 620Thr Gln Met Val Ala Ile Pro Thr Thr Ser Gly Thr Gly Ser Glu Val625 630 635 640Thr Pro Phe Ala Val Ile Thr Asp Asp Glu Thr His Val Lys Tyr Pro 645 650 655Leu Ala Asp Tyr Glu Leu Thr Pro Glu Val Ala Ile Val Asp Pro Glu 660 665 670Phe Val Met Thr Val Pro Gln His Thr Val Ser Trp Ser Gly Leu Asp 675 680 685Ala Leu Ser His Ala Leu Glu Ser Tyr Val Ser Val Met Ala Ser Glu 690 695 700Phe Thr Arg Pro Trp Ala Leu Gln Ala Ile Lys Leu Ile Phe Asp Asn705 710 715 720Leu Thr Asn Ser Tyr Asn Tyr Asp Pro Lys His Pro Thr Lys Glu Gly 725 730 735Gln Asn Ala Arg Thr Lys Met His Tyr Ala Ser Thr Leu Ala Gly Met 740 745 750Ser Phe Ala Asn Ala Phe Leu Gly Leu Asn His Ser Leu Ala His Lys 755 760 765Thr Gly Gly Glu Phe Gly Leu Pro His Gly Met Ala Ile Ala Ile Ala 770 775 780Met Pro His Val Ile Lys Phe Asn Ala Val Thr Gly Asn Val Lys Arg785 790 795 800Thr Pro Tyr Pro Arg Tyr Glu Thr Tyr Thr Ala Gln Lys Asp Tyr Ala 805 810 815Asp Ile Ala Arg Tyr Leu Gly Leu Lys Gly Glu Thr Asp Ala Glu Leu 820 825 830Val Asp Val Leu Ile Ala Glu Ile Lys Lys Leu Ala Ala Ser Val Gly 835 840 845Val Asn Gln Thr Leu Ser Gly Asn Gly Val Ser Lys His Asp Phe Asp 850 855 860Thr Lys Leu Glu Lys Met Ile Asp Leu Val Tyr Asn Asp Gln Cys Thr865 870 875 880Pro Gly Asn Pro Arg Gln Pro Ser Leu Ala Glu Ile Arg Gln Leu Leu 885 890 895Lys Asp Gln Phe 900911260DNALeuconostoc mesenteroidesACK 91atggctaaaa caatggctgt taacgcaggt agttcatcac taaagtttca attattagag 60atgcctgcag aacaagtaat tgcacaaggt gttatcgaac gcattggtat ggacgatgca 120atagtaacga ttaaatacgg tgcagaaatc aatgcagagc gcttggttgg ccattcagaa 180gatgacgaac atattacttt gtccaaagac ggcaaaggta agaagtacga aaacgttact 240gcaattaaaa accaccaaca ggccattaac ttcatgttgc aaaagttaac agatctgggt 300attattaagg actttaatga aattactggt gtcggacatc gtgttgttgc tggtggtgaa 360tggtttaatc attccgtggt ggtaacagat gaggtgctgg ctaaaattga tcgtcttgca 420gattatgcac cacttcataa cccagccaat gctatgggaa ttcgtgcttt ccaaaagctc 480ctccctgatg cattgtcagt ggcggttttt gatacttctt tccatcaaac aatgcctgat 540gaaaactttc tttatgcctt gccatatgaa tactacacgc gttatggcgc ccgcaagtat 600ggtgctcatg gtacatcgca ccgatatgtt gcatcgcgtg cagccgagtt gctcggtaaa 660gatttaaaaa gcttgaaatt aatcacgctg catttaggtg caggtgcctc aattacggct 720attaaggatg gtaagtcatt agatacttct atgggatttt ctcccttagc tggtgttgct 780atggcaacgc gttcagggga tgtcgatgct tcgcttgttt attatattca agaacgcgaa 840ggactatcta acgaagagat gttaaatata ttgaacaaaa agtccggttt gcttggtatt 900tcgacaattt caagtgatat gcgtgacttg ttagatgtcc aagacactaa tgagcatgct 960gacatggcga ttagaatatt tattaatcgt gttgttaaat atgtgggaca gtatgttgcg 1020gaaatgggag acgttgatgc tatcgtattt acagcaggaa ttggtgaaaa ctcagtccca 1080gttcgtaaaa tgattattga taaattaaac tactttggtg tgaaattaga tgaagaaaaa 1140aataatgttc gcggcaaaga agttgaaata tctacgaatg actccaaggt aaaagccttg 1200ttgatcccaa ccaatgaaga actaatgatt gctagagatg ttgaatcttt gaaagcatag 126092419PRTLeuconostoc mesenteroidesACK 92Met Ala Lys Thr Met Ala Val Asn Ala Gly Ser Ser Ser Leu Lys Phe1 5 10 15Gln Leu Leu Glu Met Pro Ala Glu Gln Val Ile Ala Gln Gly Val Ile 20 25 30Glu Arg Ile Gly Met Asp Asp Ala Ile Val Thr Ile Lys Tyr Gly Ala 35 40 45Glu Ile Asn Ala Glu Arg Leu Val Gly His Ser Glu Asp Asp Glu His 50 55 60Ile Thr Leu Ser Lys Asp Gly Lys Gly Lys Lys Tyr Glu Asn Val Thr65 70 75 80Ala Ile Lys Asn His Gln Gln Ala Ile Asn Phe Met Leu Gln Lys Leu 85 90 95Thr Asp Leu Gly Ile Ile Lys Asp Phe Asn Glu Ile Thr Gly Val Gly 100 105 110His Arg Val Val Ala Gly Gly Glu Trp Phe Asn His Ser Val Val Val 115 120 125Thr Asp Glu Val Leu Ala Lys Ile Asp Arg Leu Ala Asp Tyr Ala Pro 130 135 140Leu His Asn Pro Ala Asn Ala Met Gly Ile Arg Ala Phe Gln Lys Leu145 150 155 160Leu Pro Asp Ala Leu Ser Val Ala Val Phe Asp Thr Ser Phe His Gln 165 170 175Thr Met Pro Asp Glu Asn Phe Leu Tyr Ala Leu Pro Tyr Glu Tyr Tyr 180 185 190Thr Arg Tyr Gly Ala Arg Lys Tyr Gly Ala His Gly Thr Ser His Arg 195 200 205Tyr Val Ala Ser Arg Ala Ala Glu Leu Leu Gly Lys Asp Leu Lys Ser 210 215 220Leu Lys Leu Ile Thr Leu His Leu Gly Ala Gly Ala Ser Ile Thr Ala225 230 235

240Ile Lys Asp Gly Lys Ser Leu Asp Thr Ser Met Gly Phe Ser Pro Leu 245 250 255Ala Gly Val Ala Met Ala Thr Arg Ser Gly Asp Val Asp Ala Ser Leu 260 265 270Val Tyr Tyr Ile Gln Glu Arg Glu Gly Leu Ser Asn Glu Glu Met Leu 275 280 285Asn Ile Leu Asn Lys Lys Ser Gly Leu Leu Gly Ile Ser Thr Ile Ser 290 295 300Ser Asp Met Arg Asp Leu Leu Asp Val Gln Asp Thr Asn Glu His Ala305 310 315 320Asp Met Ala Ile Arg Ile Phe Ile Asn Arg Val Val Lys Tyr Val Gly 325 330 335Gln Tyr Val Ala Glu Met Gly Asp Val Asp Ala Ile Val Phe Thr Ala 340 345 350Gly Ile Gly Glu Asn Ser Val Pro Val Arg Lys Met Ile Ile Asp Lys 355 360 365Leu Asn Tyr Phe Gly Val Lys Leu Asp Glu Glu Lys Asn Asn Val Arg 370 375 380Gly Lys Glu Val Glu Ile Ser Thr Asn Asp Ser Lys Val Lys Ala Leu385 390 395 400Leu Ile Pro Thr Asn Glu Glu Leu Met Ile Ala Arg Asp Val Glu Ser 405 410 415Leu Lys Ala932367DNALeuconostoc mesenteroidesPHK 93atgaatatcg attcaacaga ttacctgaac aatttagatg cctactggcg agctaccaat 60tatttgtcag ttggacaact ctatttgtta gataacccat tgcttaagga aaagttaacg 120gcagaacaag ttaaaattca cccaatcggg cactggggca cgattcctag tcaaaacttt 180atttatgccc atttaaatcg tgctattaat aaatttaatt tgaatatgtt ttacattgag 240gggcctggcc atggcggaca agtaatgatt tcaaatgctt atcttgatgg tagttatacc 300gaagcatttc cagaaatcac acaggatgaa gcgggtatgc agaaaatgtt taagcgtttc 360tctttccccg gtggcgttgc ttcacatgct gatcctaaag tacctggatc gattcatgaa 420ggtggtgctt taggctattc tatcttgcac ggtgctggtg ctgtattgga taatccagac 480ctcattgctg cggttgttgt tggcgatggt gaagcagaaa cagcacccct agccacttca 540tggcatgtta ataaattttt gaatcctaaa aatgatggta ctgtattacc aattttgaat 600ttaaatggct tcaagattgc gaatcccaca gttctctctc gagaatctga tgaaacacta 660accgattatt tccacagtct gggatggcac ccttactttg taagttcttt tgacaaaccg 720attatgcaag ttcatgaaga aatggcaaaa accatggaca cagtatttac tgagattaaa 780gacattagag aaaaagccgt tcagcagaca aatgaagaga ttacacgacc actttggcca 840atgattgttt tacgttcacc taaaggctgg acagggccaa aaacttggga tgacaaacca 900attgaaaatt catttcgtgc acaccaaatt ccaattcctg ctgaccaaaa tcatcctgaa 960tatataccac aacttgttga ttggttacaa tcttacaaac cagatgaatt atttgatgaa 1020aacggtcagt taacacaatc tatccaagaa gtattaccga aaaaagagct acgtatggcc 1080aataattcgg tcaccaatgc tggtattatc aaacctttga ttttacctga cattgacaat 1140tacttggtag aaaataatca gccagacaat aacttagctc aagatgctat tttgttaggt 1200gactatttgc gtgatatcat caaacttaat cctactaact tccgtggctt tggcccagat 1260gaaacagctt ctaatcgttt ccaagacatt tttgaaacaa ctaatcgcca atggctatta 1320ccaatcaaag aacctaatga tcaattcatg gcacctgaag gccgtattat tgattctatg 1380ttatcagagc attatgatga ggggatgctt gaagcttata cgctaaccgg tcgtcatgga 1440ttctttgcca gttatgaggt ctttattcgt gaagttgatg acatgattgt gcaacatttt 1500aagtggttga atcattctca tgacgtttca tggcgtaagg acgtaccggc attaaatatt 1560attgctgatt caaccgtttt ccaacaagac cataatggtt actcacacca agatcctggt 1620gtaacaacaa tgttgtatga aaagcaacca gacttcattc gcgagttctt cccagcggat 1680gctaatagct tggtcgcaac atttgaacat gctgcgcaag caacgcaaca gattaattat 1740attgtggcca gcaaacatcc ccgcttacag tggttctcac ctactgaggc caaacagctt 1800gttacgcagg gattacgtgt cattgattgg gcaagtactg acaaaggtga aaaacctgat 1860attattatta cttcagctgg ttctgaacca acgacagaaa gtttagcagc aatacaaatt 1920ttgcacgaac acataccatc attaaaaata cgttacatca atgtattgga tctatttaaa 1980ctacgggccg atgccagcta tggcttaagt gatgatgaat ttgatgctta cttcactact 2040gacacaccag tattatttgc attccatggc tatgaaccaa tgattgaatc tattttcttc 2100aaacgacaca atcaccactt agcagttcat ggttaccgag aagttggtga cattacaaca 2160ccatttgata tgcgtgttct taacaaaatt gatcgattta atttagttaa agctgctatt 2220aatctcttgc cagaaaatat tcgcaccaag caagcagccc tcgtgcaaga aatgaccgat 2280aagctcgacc tacacgttgc gtatactagg tcaaagggaa cggacttgcc cgaagtagaa 2340gattggcgct ggcgaccact taagtga 236794788PRTLeuconostoc mesenteroidesPHK 94Met Asn Ile Asp Ser Thr Asp Tyr Leu Asn Asn Leu Asp Ala Tyr Trp1 5 10 15Arg Ala Thr Asn Tyr Leu Ser Val Gly Gln Leu Tyr Leu Leu Asp Asn 20 25 30Pro Leu Leu Lys Glu Lys Leu Thr Ala Glu Gln Val Lys Ile His Pro 35 40 45Ile Gly His Trp Gly Thr Ile Pro Ser Gln Asn Phe Ile Tyr Ala His 50 55 60Leu Asn Arg Ala Ile Asn Lys Phe Asn Leu Asn Met Phe Tyr Ile Glu65 70 75 80Gly Pro Gly His Gly Gly Gln Val Met Ile Ser Asn Ala Tyr Leu Asp 85 90 95Gly Ser Tyr Thr Glu Ala Phe Pro Glu Ile Thr Gln Asp Glu Ala Gly 100 105 110Met Gln Lys Met Phe Lys Arg Phe Ser Phe Pro Gly Gly Val Ala Ser 115 120 125His Ala Asp Pro Lys Val Pro Gly Ser Ile His Glu Gly Gly Ala Leu 130 135 140Gly Tyr Ser Ile Leu His Gly Ala Gly Ala Val Leu Asp Asn Pro Asp145 150 155 160Leu Ile Ala Ala Val Val Val Gly Asp Gly Glu Ala Glu Thr Ala Pro 165 170 175Leu Ala Thr Ser Trp His Val Asn Lys Phe Leu Asn Pro Lys Asn Asp 180 185 190Gly Thr Val Leu Pro Ile Leu Asn Leu Asn Gly Phe Lys Ile Ala Asn 195 200 205Pro Thr Val Leu Ser Arg Glu Ser Asp Glu Thr Leu Thr Asp Tyr Phe 210 215 220His Ser Leu Gly Trp His Pro Tyr Phe Val Ser Ser Phe Asp Lys Pro225 230 235 240Ile Met Gln Val His Glu Glu Met Ala Lys Thr Met Asp Thr Val Phe 245 250 255Thr Glu Ile Lys Asp Ile Arg Glu Lys Ala Val Gln Gln Thr Asn Glu 260 265 270Glu Ile Thr Arg Pro Leu Trp Pro Met Ile Val Leu Arg Ser Pro Lys 275 280 285Gly Trp Thr Gly Pro Lys Thr Trp Asp Asp Lys Pro Ile Glu Asn Ser 290 295 300Phe Arg Ala His Gln Ile Pro Ile Pro Ala Asp Gln Asn His Pro Glu305 310 315 320Tyr Ile Pro Gln Leu Val Asp Trp Leu Gln Ser Tyr Lys Pro Asp Glu 325 330 335Leu Phe Asp Glu Asn Gly Gln Leu Thr Gln Ser Ile Gln Glu Val Leu 340 345 350Pro Lys Lys Glu Leu Arg Met Ala Asn Asn Ser Val Thr Asn Ala Gly 355 360 365Ile Ile Lys Pro Leu Ile Leu Pro Asp Ile Asp Asn Tyr Leu Val Glu 370 375 380Asn Asn Gln Pro Asp Asn Asn Leu Ala Gln Asp Ala Ile Leu Leu Gly385 390 395 400Asp Tyr Leu Arg Asp Ile Ile Lys Leu Asn Pro Thr Asn Phe Arg Gly 405 410 415Phe Gly Pro Asp Glu Thr Ala Ser Asn Arg Phe Gln Asp Ile Phe Glu 420 425 430Thr Thr Asn Arg Gln Trp Leu Leu Pro Ile Lys Glu Pro Asn Asp Gln 435 440 445Phe Met Ala Pro Glu Gly Arg Ile Ile Asp Ser Met Leu Ser Glu His 450 455 460Tyr Asp Glu Gly Met Leu Glu Ala Tyr Thr Leu Thr Gly Arg His Gly465 470 475 480Phe Phe Ala Ser Tyr Glu Val Phe Ile Arg Glu Val Asp Asp Met Ile 485 490 495Val Gln His Phe Lys Trp Leu Asn His Ser His Asp Val Ser Trp Arg 500 505 510Lys Asp Val Pro Ala Leu Asn Ile Ile Ala Asp Ser Thr Val Phe Gln 515 520 525Gln Asp His Asn Gly Tyr Ser His Gln Asp Pro Gly Val Thr Thr Met 530 535 540Leu Tyr Glu Lys Gln Pro Asp Phe Ile Arg Glu Phe Phe Pro Ala Asp545 550 555 560Ala Asn Ser Leu Val Ala Thr Phe Glu His Ala Ala Gln Ala Thr Gln 565 570 575Gln Ile Asn Tyr Ile Val Ala Ser Lys His Pro Arg Leu Gln Trp Phe 580 585 590Ser Pro Thr Glu Ala Lys Gln Leu Val Thr Gln Gly Leu Arg Val Ile 595 600 605Asp Trp Ala Ser Thr Asp Lys Gly Glu Lys Pro Asp Ile Ile Ile Thr 610 615 620Ser Ala Gly Ser Glu Pro Thr Thr Glu Ser Leu Ala Ala Ile Gln Ile625 630 635 640Leu His Glu His Ile Pro Ser Leu Lys Ile Arg Tyr Ile Asn Val Leu 645 650 655Asp Leu Phe Lys Leu Arg Ala Asp Ala Ser Tyr Gly Leu Ser Asp Asp 660 665 670Glu Phe Asp Ala Tyr Phe Thr Thr Asp Thr Pro Val Leu Phe Ala Phe 675 680 685His Gly Tyr Glu Pro Met Ile Glu Ser Ile Phe Phe Lys Arg His Asn 690 695 700His His Leu Ala Val His Gly Tyr Arg Glu Val Gly Asp Ile Thr Thr705 710 715 720Pro Phe Asp Met Arg Val Leu Asn Lys Ile Asp Arg Phe Asn Leu Val 725 730 735Lys Ala Ala Ile Asn Leu Leu Pro Glu Asn Ile Arg Thr Lys Gln Ala 740 745 750Ala Leu Val Gln Glu Met Thr Asp Lys Leu Asp Leu His Val Ala Tyr 755 760 765Thr Arg Ser Lys Gly Thr Asp Leu Pro Glu Val Glu Asp Trp Arg Trp 770 775 780Arg Pro Leu Lys78595981DNALeuconostoc mesenteroidesPTA 95atggaacttt ttgagcaatt aaaaaataaa attaccggtc aaaataagac aattgtattt 60ccggaaggtg aagatccccg tatccagggc gcagctatca gattagcagc tgataatttg 120atcgagccaa tattgttggg ggatgcacag gaaatttcaa aaactgccca agcccataac 180tttgatttat cgaacataga gacgattgat cctgcttcat atgatgagaa tgaattggca 240aaattaaatg cgacacttgt cgaaaggcgc aaagggaaaa cagacgctga gacagctgcc 300aaatggctac aaaatgttaa ctattttggg acaatgctgg tttatactgg caaagctgac 360ggtatggtgt caggcgcagt acatccaact ggtgatacag ttcgtcctgc gttgcagatc 420attaaaacgg cgcctggctc atcacgtatt tctggtgcat tcattatgca aaaaggcgaa 480gagcgttatg tatttgcgga cgctgccatt aacattgaca ttgattcgga cacgatggct 540gaaattgcaa tacaatcagc ccacacagct caagtgtttg atattgaacc aaaagttgcg 600atgctttcat tttcaactaa gggctccgcg aaatcaccct tggtcgataa ggtggcaact 660gccactgcat tggctaagaa gttagcacct gaactggcgg actcaattga tggagagtta 720cagtttgatg ctgcgtttgt tgaatctgtt gctgccgcaa aggcgcctga ttcaaaagtc 780gcaggtaaag cgaacacatt tattttccca agtctagaag caggtaatat tggctataaa 840attgcacaac gcttgggtgg gtttgaagcg attggaccta ttttacaagg cttagcaaaa 900cctgtttctg atttatcgcg aggagccaat gaagaggatg tttataaagt agctattatt 960actgctgcgc aagcattata a 98196326PRTLeuconostoc mesenteroidesPTA 96Met Glu Leu Phe Glu Gln Leu Lys Asn Lys Ile Thr Gly Gln Asn Lys1 5 10 15Thr Ile Val Phe Pro Glu Gly Glu Asp Pro Arg Ile Gln Gly Ala Ala 20 25 30Ile Arg Leu Ala Ala Asp Asn Leu Ile Glu Pro Ile Leu Leu Gly Asp 35 40 45Ala Gln Glu Ile Ser Lys Thr Ala Gln Ala His Asn Phe Asp Leu Ser 50 55 60Asn Ile Glu Thr Ile Asp Pro Ala Ser Tyr Asp Glu Asn Glu Leu Ala65 70 75 80Lys Leu Asn Ala Thr Leu Val Glu Arg Arg Lys Gly Lys Thr Asp Ala 85 90 95Glu Thr Ala Ala Lys Trp Leu Gln Asn Val Asn Tyr Phe Gly Thr Met 100 105 110Leu Val Tyr Thr Gly Lys Ala Asp Gly Met Val Ser Gly Ala Val His 115 120 125Pro Thr Gly Asp Thr Val Arg Pro Ala Leu Gln Ile Ile Lys Thr Ala 130 135 140Pro Gly Ser Ser Arg Ile Ser Gly Ala Phe Ile Met Gln Lys Gly Glu145 150 155 160Glu Arg Tyr Val Phe Ala Asp Ala Ala Ile Asn Ile Asp Ile Asp Ser 165 170 175Asp Thr Met Ala Glu Ile Ala Ile Gln Ser Ala His Thr Ala Gln Val 180 185 190Phe Asp Ile Glu Pro Lys Val Ala Met Leu Ser Phe Ser Thr Lys Gly 195 200 205Ser Ala Lys Ser Pro Leu Val Asp Lys Val Ala Thr Ala Thr Ala Leu 210 215 220Ala Lys Lys Leu Ala Pro Glu Leu Ala Asp Ser Ile Asp Gly Glu Leu225 230 235 240Gln Phe Asp Ala Ala Phe Val Glu Ser Val Ala Ala Ala Lys Ala Pro 245 250 255Asp Ser Lys Val Ala Gly Lys Ala Asn Thr Phe Ile Phe Pro Ser Leu 260 265 270Glu Ala Gly Asn Ile Gly Tyr Lys Ile Ala Gln Arg Leu Gly Gly Phe 275 280 285Glu Ala Ile Gly Pro Ile Leu Gln Gly Leu Ala Lys Pro Val Ser Asp 290 295 300Leu Ser Arg Gly Ala Asn Glu Glu Asp Val Tyr Lys Val Ala Ile Ile305 310 315 320Thr Ala Ala Gln Ala Leu 325972697DNAOenococcus oeniiadhE 97atggcagttg aaacgaagaa gattactaaa aagatactga cttccgacga acttaaagac 60aaaaaggaga cagcaaaaaa atacattgga gaattagtcg ataaatcaag acaggcatta 120ttagaattct ctcaatatag tcaatcacaa gttgacaaaa ttgttgccgc aatggcttta 180gccggttccg aacattcttt ggaattagca cactttgctc gaaatgaaac tggaagagga 240gttgttgaag acaaagatac gaaaaatcgt tttgcctcgg aatctgtata taacacgatt 300aaaaatgaca aaacagttgg cgtaattgat gaagacccaa tcactggcaa agttcagtta 360gctgcaccac ttgggatttt agccggaatt gttccaacga caaatccaac gtctaccacg 420atttttaaat cattgttaac cgctaagaca cgtaatacga tcatttttgc ttttcacccg 480caagcacaaa aatcgtctgt agcggctgcg aaaattgttt accaggcggc tagaaaagct 540ggagcaccga aagatttcat tcagtggatc gaaacgcctt ccttggaaaa tacgactgct 600ttgatgcaaa atcctgaaat agcttcaatt ttggctactg gaggcccttc tatggttcat 660gccgcattaa catcgggaaa cccttcaatg ggtgttgggg caggaaatgg agctgttttc 720attgaccata cggctcgtgt tcgtcgagcg gcagaagact tgttgctatc aaaacgtttt 780gataatggaa tgatttgcgc tacagagaat tccgcagtta tcgaagcttc ggtttatgac 840gaatttatga aattaatgca ggaaaagggt acttatctcg ttccgaagga tgattacaaa 900aagatcgctg attttgtttt cagcgataaa catgcagtta atggtccggt tgccgggatg 960agtggccgtt ggattgctga acatgccggt gttactttgc cgaagggcaa agatgttttg 1020ttgtttgagt tggatcaaga cgacattggc gaaaaacttt cctcggaaaa acttagcccc 1080ttgctttcgg tctataaagc tgctgatcgt aaagaagcga taaaagttgt tcaaagactt 1140ttgaactatc aaggagctgg tcataatgcg gcaatccaga ttggcgctca agacgatccc 1200tttgttaaag aatatgccga tgcagtttct gcatctagaa ttttggttaa ccagcccgat 1260tcaataggtg gagtcggcga tatttataca gatgcattgc gtccaagtat gacgcttggt 1320actggttcat gggggaagaa ttcattgtca cataatttat caacttacga tttgttgaat 1380gtcaaaacgg ttgcccgcag acgtaatcgt cctcaatggg ttcgtcttcc aaaagatatc 1440tactatgaga aaaatgccat tacctatctg caggaattac cgaacgtcaa tcgggctttc 1500gttattaccg atcattcgtt ggtcaaatat ggattcgtcg atcggattct tgagcaattc 1560gaattgcgtc ccgaccaggt taaaacaagt atttacagtt ctgttcaacc ggatccttat 1620ttgagtcagg ctgttgaaat tgcaaaacaa atgcaggagt tcgaaccgga tacagtaatt 1680gctcttggag gaggttcctc tttggatgtt gccaagattt cccgttttct ttatgaatat 1740tctcaagaac cggaccacat tggttttttg gaaaacattg acgatattaa agaattgttt 1800aaaggattgc agcaaaagtt tatggatatt cggaaacgga ttgttaaatt cgaacatcag 1860aatttgactc aactggttgc tattccaaca acttcgggta ccggttcgga agtaacaccg 1920ttttcggtta tcaccgatga tgaaactcac gttaaatacc ctttggccga ttatgaatta 1980actccgcaag ttgcaatcgt tgatcctgaa ttggttatga ctgtaccaaa acggaccctg 2040gcttggtctg gactggacgc cttatctcac tcgcttgaat cctacgtgtc ggttatgagt 2100tctgaattta cacggccgtg ggctttacag gcaatcaaac tgatttttga aaacatcgtt 2160gattcctata attacgatcc aaagcaccca actaatcggg gcgccgaggc acgtgaaaaa 2220atgcattatg ctgcaacttt ggccggcatg tctttcggaa acgccttctt gggtataaac 2280cattcactgg cccataaaac cggtggagag ttcggtcttc ctcatggttt ggcaatcagc 2340atcgcgatgc cacatgtaat tcgatttaat gcggttaccg gaaacgtcaa acggactcct 2400tttcctcgtt atgaggttta tacagcccaa aaagattatg ccgatattgc gcgccatatt 2460ggcctcagcg gtaagaacga tgccgaattg gttgaaaaac ttatcgccaa aatcaaggaa 2520ttaaccgatg ccttggatgt caatattact ttgagcggca atggagtcga taaaaagagc 2580tttgaacatt ctttggatca actggttgat cttgtttacg atgaccagtg cacgccagga 2640aatccgcgac aaccgaattt agctgaaatt aggcagttat tgatcgacca attttaa 269798898PRTOenococcus oeniiadhE 98Met Ala Val Glu Thr Lys Lys Ile Thr Lys Lys Ile Leu Thr Ser Asp1 5 10 15Glu Leu Lys Asp Lys Lys Glu Thr Ala Lys Lys Tyr Ile Gly Glu Leu 20 25 30Val Asp Lys Ser Arg Gln Ala Leu Leu Glu Phe Ser Gln Tyr Ser Gln 35 40 45Ser Gln Val Asp Lys Ile Val Ala Ala Met Ala Leu Ala Gly Ser Glu 50 55 60His Ser Leu Glu Leu Ala His Phe Ala Arg Asn Glu Thr Gly Arg Gly65 70 75 80Val Val Glu Asp Lys Asp Thr Lys Asn Arg Phe Ala Ser Glu Ser Val 85 90 95Tyr Asn Thr Ile Lys Asn Asp Lys Thr Val Gly Val Ile Asp Glu Asp 100 105 110Pro Ile Thr Gly Lys Val Gln Leu Ala Ala Pro Leu Gly Ile Leu Ala 115 120 125Gly Ile Val Pro Thr Thr Asn Pro Thr Ser Thr Thr Ile Phe Lys Ser 130 135 140Leu Leu Thr Ala Lys Thr Arg

Asn Thr Ile Ile Phe Ala Phe His Pro145 150 155 160Gln Ala Gln Lys Ser Ser Val Ala Ala Ala Lys Ile Val Tyr Gln Ala 165 170 175Ala Arg Lys Ala Gly Ala Pro Lys Asp Phe Ile Gln Trp Ile Glu Thr 180 185 190Pro Ser Leu Glu Asn Thr Thr Ala Leu Met Gln Asn Pro Glu Ile Ala 195 200 205Ser Ile Leu Ala Thr Gly Gly Pro Ser Met Val His Ala Ala Leu Thr 210 215 220Ser Gly Asn Pro Ser Met Gly Val Gly Ala Gly Asn Gly Ala Val Phe225 230 235 240Ile Asp His Thr Ala Arg Val Arg Arg Ala Ala Glu Asp Leu Leu Leu 245 250 255Ser Lys Arg Phe Asp Asn Gly Met Ile Cys Ala Thr Glu Asn Ser Ala 260 265 270Val Ile Glu Ala Ser Val Tyr Asp Glu Phe Met Lys Leu Met Gln Glu 275 280 285Lys Gly Thr Tyr Leu Val Pro Lys Asp Asp Tyr Lys Lys Ile Ala Asp 290 295 300Phe Val Phe Ser Asp Lys His Ala Val Asn Gly Pro Val Ala Gly Met305 310 315 320Ser Gly Arg Trp Ile Ala Glu His Ala Gly Val Thr Leu Pro Lys Gly 325 330 335Lys Asp Val Leu Leu Phe Glu Leu Asp Gln Asp Asp Ile Gly Glu Lys 340 345 350Leu Ser Ser Glu Lys Leu Ser Pro Leu Leu Ser Val Tyr Lys Ala Ala 355 360 365Asp Arg Lys Glu Ala Ile Lys Val Val Gln Arg Leu Leu Asn Tyr Gln 370 375 380Gly Ala Gly His Asn Ala Ala Ile Gln Ile Gly Ala Gln Asp Asp Pro385 390 395 400Phe Val Lys Glu Tyr Ala Asp Ala Val Ser Ala Ser Arg Ile Leu Val 405 410 415Asn Gln Pro Asp Ser Ile Gly Gly Val Gly Asp Ile Tyr Thr Asp Ala 420 425 430Leu Arg Pro Ser Met Thr Leu Gly Thr Gly Ser Trp Gly Lys Asn Ser 435 440 445Leu Ser His Asn Leu Ser Thr Tyr Asp Leu Leu Asn Val Lys Thr Val 450 455 460Ala Arg Arg Arg Asn Arg Pro Gln Trp Val Arg Leu Pro Lys Asp Ile465 470 475 480Tyr Tyr Glu Lys Asn Ala Ile Thr Tyr Leu Gln Glu Leu Pro Asn Val 485 490 495Asn Arg Ala Phe Val Ile Thr Asp His Ser Leu Val Lys Tyr Gly Phe 500 505 510Val Asp Arg Ile Leu Glu Gln Phe Glu Leu Arg Pro Asp Gln Val Lys 515 520 525Thr Ser Ile Tyr Ser Ser Val Gln Pro Asp Pro Tyr Leu Ser Gln Ala 530 535 540Val Glu Ile Ala Lys Gln Met Gln Glu Phe Glu Pro Asp Thr Val Ile545 550 555 560Ala Leu Gly Gly Gly Ser Ser Leu Asp Val Ala Lys Ile Ser Arg Phe 565 570 575Leu Tyr Glu Tyr Ser Gln Glu Pro Asp His Ile Gly Phe Leu Glu Asn 580 585 590Ile Asp Asp Ile Lys Glu Leu Phe Lys Gly Leu Gln Gln Lys Phe Met 595 600 605Asp Ile Arg Lys Arg Ile Val Lys Phe Glu His Gln Asn Leu Thr Gln 610 615 620Leu Val Ala Ile Pro Thr Thr Ser Gly Thr Gly Ser Glu Val Thr Pro625 630 635 640Phe Ser Val Ile Thr Asp Asp Glu Thr His Val Lys Tyr Pro Leu Ala 645 650 655Asp Tyr Glu Leu Thr Pro Gln Val Ala Ile Val Asp Pro Glu Leu Val 660 665 670Met Thr Val Pro Lys Arg Thr Leu Ala Trp Ser Gly Leu Asp Ala Leu 675 680 685Ser His Ser Leu Glu Ser Tyr Val Ser Val Met Ser Ser Glu Phe Thr 690 695 700Arg Pro Trp Ala Leu Gln Ala Ile Lys Leu Ile Phe Glu Asn Ile Val705 710 715 720Asp Ser Tyr Asn Tyr Asp Pro Lys His Pro Thr Asn Arg Gly Ala Glu 725 730 735Ala Arg Glu Lys Met His Tyr Ala Ala Thr Leu Ala Gly Met Ser Phe 740 745 750Gly Asn Ala Phe Leu Gly Ile Asn His Ser Leu Ala His Lys Thr Gly 755 760 765Gly Glu Phe Gly Leu Pro His Gly Leu Ala Ile Ser Ile Ala Met Pro 770 775 780His Val Ile Arg Phe Asn Ala Val Thr Gly Asn Val Lys Arg Thr Pro785 790 795 800Phe Pro Arg Tyr Glu Val Tyr Thr Ala Gln Lys Asp Tyr Ala Asp Ile 805 810 815Ala Arg His Ile Gly Leu Ser Gly Lys Asn Asp Ala Glu Leu Val Glu 820 825 830Lys Leu Ile Ala Lys Ile Lys Glu Leu Thr Asp Ala Leu Asp Val Asn 835 840 845Ile Thr Leu Ser Gly Asn Gly Val Asp Lys Lys Ser Phe Glu His Ser 850 855 860Leu Asp Gln Leu Val Asp Leu Val Tyr Asp Asp Gln Cys Thr Pro Gly865 870 875 880Asn Pro Arg Gln Pro Asn Leu Ala Glu Ile Arg Gln Leu Leu Ile Asp 885 890 895Gln Phe991194DNAOenococcus oeniiACK 99atgacaaaaa ttttatcggt aaatgccggc tcgtcttcat tgaagttcaa attgcttaat 60atgcccgaag agagtgttat cgccgaagga ctgattgaaa gaatcggtgg cagcctggat 120caagacgata atgtcacgat taaattcaat ggcgaaaaac ataagagccg tcaggctttt 180aaaaatcacg aggatgctat caacttgatg ctttctcaat ttaaaaagtt tggaatcgtt 240gataatttta atgagatcaa aggaattggc caccgtgttg ttgccggggg tgaatggttc 300aataaatctg tattaattaa cgatgaagtg ctgaataaaa ttgaacgttt ggctgcttac 360gctccactcc ataatcctgc gaacgctatg ggcatcaggg ctttttccaa aataatcccc 420gatgcaaccg aagttgccgt atttgataca gctttccatc aaacgatgcc gaaagagaat 480tacttgtatg cggttcctta tgagtactat acgaaatacg gtgttcgtag gtatggagct 540cacggaactt cccataaata cgtttccgaa caggctgcta aattaattgg gaaaccccta 600gaagaactta aactgattac tttgcattta ggagccggag cgtctgtaac ggccattaag 660aatggtcgat catttgatac ctcgatggga ttttcgcctt tggccggatt ggttatggcg 720actcgttcag gggatgttga cgtctcattg attgattacg ttaaagcaaa agaagatatt 780tccgatcagg aaatgctaaa tgttcttaac caaaaatccg gattactggg tgtgtcaaca 840atttcaagtg atatgcgtga tttgttggat gtttatgata ctaacgatca tgctaaattg 900gctattgata tgtttgtcgg tcgtgtagtt gattatatcg gtcaatatta ttttgaatta 960aagggagctg atgcgcttgt tttcacggcc ggaattggag agaattcggt tccaatcagg 1020aaaatgatta tcgatcgtct ggctttcttg ggcattaaac tcgatgaaga tgcaaatgat 1080aagcatggag tcgaaacaaa gattacaact gatgattcca aaatggctgc atatgtaatt 1140ccaaccaatg aggaactcga aattgctcgt gatgtacaaa atttgatgaa gtaa 1194100397PRTOenococcus oeniiACK 100Met Thr Lys Ile Leu Ser Val Asn Ala Gly Ser Ser Ser Leu Lys Phe1 5 10 15Lys Leu Leu Asn Met Pro Glu Glu Ser Val Ile Ala Glu Gly Leu Ile 20 25 30Glu Arg Ile Gly Gly Ser Leu Asp Gln Asp Asp Asn Val Thr Ile Lys 35 40 45Phe Asn Gly Glu Lys His Lys Ser Arg Gln Ala Phe Lys Asn His Glu 50 55 60Asp Ala Ile Asn Leu Met Leu Ser Gln Phe Lys Lys Phe Gly Ile Val65 70 75 80Asp Asn Phe Asn Glu Ile Lys Gly Ile Gly His Arg Val Val Ala Gly 85 90 95Gly Glu Trp Phe Asn Lys Ser Val Leu Ile Asn Asp Glu Val Leu Asn 100 105 110Lys Ile Glu Arg Leu Ala Ala Tyr Ala Pro Leu His Asn Pro Ala Asn 115 120 125Ala Met Gly Ile Arg Ala Phe Ser Lys Ile Ile Pro Asp Ala Thr Glu 130 135 140Val Ala Val Phe Asp Thr Ala Phe His Gln Thr Met Pro Lys Glu Asn145 150 155 160Tyr Leu Tyr Ala Val Pro Tyr Glu Tyr Tyr Thr Lys Tyr Gly Val Arg 165 170 175Arg Tyr Gly Ala His Gly Thr Ser His Lys Tyr Val Ser Glu Gln Ala 180 185 190Ala Lys Leu Ile Gly Lys Pro Leu Glu Glu Leu Lys Leu Ile Thr Leu 195 200 205His Leu Gly Ala Gly Ala Ser Val Thr Ala Ile Lys Asn Gly Arg Ser 210 215 220Phe Asp Thr Ser Met Gly Phe Ser Pro Leu Ala Gly Leu Val Met Ala225 230 235 240Thr Arg Ser Gly Asp Val Asp Val Ser Leu Ile Asp Tyr Val Lys Ala 245 250 255Lys Glu Asp Ile Ser Asp Gln Glu Met Leu Asn Val Leu Asn Gln Lys 260 265 270Ser Gly Leu Leu Gly Val Ser Thr Ile Ser Ser Asp Met Arg Asp Leu 275 280 285Leu Asp Val Tyr Asp Thr Asn Asp His Ala Lys Leu Ala Ile Asp Met 290 295 300Phe Val Gly Arg Val Val Asp Tyr Ile Gly Gln Tyr Tyr Phe Glu Leu305 310 315 320Lys Gly Ala Asp Ala Leu Val Phe Thr Ala Gly Ile Gly Glu Asn Ser 325 330 335Val Pro Ile Arg Lys Met Ile Ile Asp Arg Leu Ala Phe Leu Gly Ile 340 345 350Lys Leu Asp Glu Asp Ala Asn Asp Lys His Gly Val Glu Thr Lys Ile 355 360 365Thr Thr Asp Asp Ser Lys Met Ala Ala Tyr Val Ile Pro Thr Asn Glu 370 375 380Glu Leu Glu Ile Ala Arg Asp Val Gln Asn Leu Met Lys385 390 3951012385DNAOenococcus oeniiPHK 101atgactatta attatgattc aaaggattat ttgaaatatg ttgatgccta ttggcgggct 60gccaattatc tttcagtcgg tcaacttttc ttgagaaaca atccgttact taaagacgaa 120cttcagtcca aagacgttaa gatcaagccg atcggtcatt ggggaacggt tgcgccgcag 180aattttattt atgctcattt aaatcgagca attttaaaat acgatttaaa tatgttttat 240attgaaggct ctggccatgg tggccaggtg atggtttcca attcatattt ggatgggtct 300tataccgaaa cttatccaaa agtaactcag gatattcaag gaatgcaacg tttattcaag 360caattctctt ttcctggcgg aattgcttca catgctgctc cggaaactcc tggatcgatt 420cacgaaggag gagagctggg ttattcgatt tcgcatggag ttggtgctat tttggacaat 480cccgatgtga ttgctgctgt tgaaatcggg gatggtgaat cagaaacggg gcctttggcc 540gcatcgtggt tttccgacaa attcattaat ccgattcatg atggtgctgt gttgccgatt 600gttcagatta atggatttaa aatttccaat ccaacaattc tttctcgcat gagcgatcgt 660gatttgacca attattatca tggaatgggt tgggagcctt tgttcgttga aacggatggt 720tcagataatt ttaaagtgca tgccgagatg gccgatgctg ttgataaggc cattgaaaaa 780attaaagcta ttcaaaaaaa cgcccgtaat aataatgatg attcattgcc aatctggccg 840atgattgtat tacgtgctcc taaaggttgg acaggtccaa aaaaagattt ggatggtaat 900ccgattgaaa attcatttcg ggcacatcag gtgccgattc cagtcgatgc aaatcatttg 960gaacataaag atatgctgat tgattggatg aagtcctaca aaccggaaga attgttcaat 1020gaggatggca gcttaaaaga aattgttaag gtaaatcagc caaaaggcca acggcgaatg 1080gcaatgaacc caatcacgaa tggtggaata aagccaagga cattgaatat gcctgatatg 1140gaacgatttg cttttccgaa aaatagcttg aaaaacaata aaaaaccagg gatggacctt 1200caggttgttt caacatttat tgccgaaata ataaagaaaa atcctatcaa tttccgtcag 1260ttcggtcccg atgaaacaat gtccaatcgt ctctgggacg agtttaaagt cactaatcgt 1320cagtggatgc aagcagtcca tgaaccaaat gatcaatata tggctcattc tggtcgtatt 1380ctagatgctc aattgtctga acatcaggcc gagggatgga tggaaggata cgttttgacc 1440ggccgacacg ctttctttgc gtcctacgaa gcctttacgc gtgtaattga ttcgatgctg 1500actcaatatt ataagtggtt gcgaaaagcg gtcgagcaag attggcgtca cgattatcct 1560tcgttaaatg tcattaatgc ttcacatgct tttcaacagg accataatgg ttatactcat 1620caagatcccg gtatgcttac tcatatggct gaaaaaggtc atgagtttgt caatgaattc 1680ttgccagccg atgctaattc ccttttggca gttatgaata agtctctgca ggttagaaac 1740aaaattaaca ttattgtcgc ttccaaacac ccacgaactc aatggtttac gatcgacgaa 1800gccaaggaat tggttgacaa cggtttggga attattccat gggcttccaa cgatgacgga 1860gtggaaccgg acgttgtttt tgcagccggt ggaacggaag cgactatgga aagtttggct 1920gctatttcgc tcttgcatga aagcttccct gaattgaagt tccgctttat taatgtaatc 1980gatttgttga aactgcgcaa aaaaggggat aacgatgatt atcgcggtct aagcgatttg 2040gaattcgacc attactttac gcgtgaaaaa ccagtagtct tttccttcca cggctttgaa 2100agtttggcac gcgacttgtt ctatgaccgc cacaatcata atgttatttt tcatggatat 2160cgtgaaaacg gagacattac aactccgttc gatatgcgtg tcctgaatca tcttgatcgt 2220ttccaccttg ccaaagacgc aatcaacgca actaagtacg ctgatgtggc tggccaattt 2280gaccaacgaa tggatgatat gttggccaaa cataccgctt atatttgcga tcagggaaca 2340gacctgcctg aagtaacctc ttggcagtgg caggatatta aataa 2385102794PRTOenococcus oeniiPHK 102Met Thr Ile Asn Tyr Asp Ser Lys Asp Tyr Leu Lys Tyr Val Asp Ala1 5 10 15Tyr Trp Arg Ala Ala Asn Tyr Leu Ser Val Gly Gln Leu Phe Leu Arg 20 25 30Asn Asn Pro Leu Leu Lys Asp Glu Leu Gln Ser Lys Asp Val Lys Ile 35 40 45Lys Pro Ile Gly His Trp Gly Thr Val Ala Pro Gln Asn Phe Ile Tyr 50 55 60Ala His Leu Asn Arg Ala Ile Leu Lys Tyr Asp Leu Asn Met Phe Tyr65 70 75 80Ile Glu Gly Ser Gly His Gly Gly Gln Val Met Val Ser Asn Ser Tyr 85 90 95Leu Asp Gly Ser Tyr Thr Glu Thr Tyr Pro Lys Val Thr Gln Asp Ile 100 105 110Gln Gly Met Gln Arg Leu Phe Lys Gln Phe Ser Phe Pro Gly Gly Ile 115 120 125Ala Ser His Ala Ala Pro Glu Thr Pro Gly Ser Ile His Glu Gly Gly 130 135 140Glu Leu Gly Tyr Ser Ile Ser His Gly Val Gly Ala Ile Leu Asp Asn145 150 155 160Pro Asp Val Ile Ala Ala Val Glu Ile Gly Asp Gly Glu Ser Glu Thr 165 170 175Gly Pro Leu Ala Ala Ser Trp Phe Ser Asp Lys Phe Ile Asn Pro Ile 180 185 190His Asp Gly Ala Val Leu Pro Ile Val Gln Ile Asn Gly Phe Lys Ile 195 200 205Ser Asn Pro Thr Ile Leu Ser Arg Met Ser Asp Arg Asp Leu Thr Asn 210 215 220Tyr Tyr His Gly Met Gly Trp Glu Pro Leu Phe Val Glu Thr Asp Gly225 230 235 240Ser Asp Asn Phe Lys Val His Ala Glu Met Ala Asp Ala Val Asp Lys 245 250 255Ala Ile Glu Lys Ile Lys Ala Ile Gln Lys Asn Ala Arg Asn Asn Asn 260 265 270Asp Asp Ser Leu Pro Ile Trp Pro Met Ile Val Leu Arg Ala Pro Lys 275 280 285Gly Trp Thr Gly Pro Lys Lys Asp Leu Asp Gly Asn Pro Ile Glu Asn 290 295 300Ser Phe Arg Ala His Gln Val Pro Ile Pro Val Asp Ala Asn His Leu305 310 315 320Glu His Lys Asp Met Leu Ile Asp Trp Met Lys Ser Tyr Lys Pro Glu 325 330 335Glu Leu Phe Asn Glu Asp Gly Ser Leu Lys Glu Ile Val Lys Val Asn 340 345 350Gln Pro Lys Gly Gln Arg Arg Met Ala Met Asn Pro Ile Thr Asn Gly 355 360 365Gly Ile Lys Pro Arg Thr Leu Asn Met Pro Asp Met Glu Arg Phe Ala 370 375 380Phe Pro Lys Asn Ser Leu Lys Asn Asn Lys Lys Pro Gly Met Asp Leu385 390 395 400Gln Val Val Ser Thr Phe Ile Ala Glu Ile Ile Lys Lys Asn Pro Ile 405 410 415Asn Phe Arg Gln Phe Gly Pro Asp Glu Thr Met Ser Asn Arg Leu Trp 420 425 430Asp Glu Phe Lys Val Thr Asn Arg Gln Trp Met Gln Ala Val His Glu 435 440 445Pro Asn Asp Gln Tyr Met Ala His Ser Gly Arg Ile Leu Asp Ala Gln 450 455 460Leu Ser Glu His Gln Ala Glu Gly Trp Met Glu Gly Tyr Val Leu Thr465 470 475 480Gly Arg His Ala Phe Phe Ala Ser Tyr Glu Ala Phe Thr Arg Val Ile 485 490 495Asp Ser Met Leu Thr Gln Tyr Tyr Lys Trp Leu Arg Lys Ala Val Glu 500 505 510Gln Asp Trp Arg His Asp Tyr Pro Ser Leu Asn Val Ile Asn Ala Ser 515 520 525His Ala Phe Gln Gln Asp His Asn Gly Tyr Thr His Gln Asp Pro Gly 530 535 540Met Leu Thr His Met Ala Glu Lys Gly His Glu Phe Val Asn Glu Phe545 550 555 560Leu Pro Ala Asp Ala Asn Ser Leu Leu Ala Val Met Asn Lys Ser Leu 565 570 575Gln Val Arg Asn Lys Ile Asn Ile Ile Val Ala Ser Lys His Pro Arg 580 585 590Thr Gln Trp Phe Thr Ile Asp Glu Ala Lys Glu Leu Val Asp Asn Gly 595 600 605Leu Gly Ile Ile Pro Trp Ala Ser Asn Asp Asp Gly Val Glu Pro Asp 610 615 620Val Val Phe Ala Ala Gly Gly Thr Glu Ala Thr Met Glu Ser Leu Ala625 630 635 640Ala Ile Ser Leu Leu His Glu Ser Phe Pro Glu Leu Lys Phe Arg Phe 645 650 655Ile Asn Val Ile Asp Leu Leu Lys Leu Arg Lys Lys Gly Asp Asn Asp 660 665 670Asp Tyr Arg Gly Leu Ser Asp Leu Glu Phe Asp His Tyr Phe Thr Arg 675 680 685Glu Lys Pro Val Val Phe Ser Phe His Gly Phe Glu Ser Leu Ala Arg 690 695 700Asp Leu Phe Tyr Asp Arg His Asn His Asn Val Ile Phe His Gly Tyr705 710

715 720Arg Glu Asn Gly Asp Ile Thr Thr Pro Phe Asp Met Arg Val Leu Asn 725 730 735His Leu Asp Arg Phe His Leu Ala Lys Asp Ala Ile Asn Ala Thr Lys 740 745 750Tyr Ala Asp Val Ala Gly Gln Phe Asp Gln Arg Met Asp Asp Met Leu 755 760 765Ala Lys His Thr Ala Tyr Ile Cys Asp Gln Gly Thr Asp Leu Pro Glu 770 775 780Val Thr Ser Trp Gln Trp Gln Asp Ile Lys785 790103990DNAOenococcus oeniiPTA 103atgacaaatt tatttactga gttggctgat aaaattaagg aaaaaaaact gagactcgtt 60tttccggaag gcgatgattc aagagtccag ggggcggcca tccgtttaaa aaaagatggt 120ttactaacgc cgatattatt gggagatcag gcagcggtcg aaaagacagc tgcagataat 180catttcgacc tgagtggcat caagcttatc gatccgctta aattcgatga gaagcagttc 240aatcaattga tcgataaact ggttacccga cgaaaaggga aaactgaccc gcaaacggta 300gctatgtggc tgcaggaagt aaattatttc ggtaccatgc ttgtttatac cggcaaggcc 360gacgcaatgg tttccggtgc tgttcatcca accggcgata cggttcgtcc cgctttgcag 420attatcaaga cagctcctgg tgtcagtcgt atttccggtg cttttattat gcaaaaaaat 480gatgaacgct atctgtttgc cgattcggca attaatatcg aacttgatgc acaaaccatg 540gctgagattg ccgttcaatc agcccatacg gctaaaatat ttggaatcga tccaaaagtt 600gccatgctga gtttttcaac caaaggatcg gcaaaacatg cattggtcga taaagttgct 660caggcaacta agctggcgca ggagatggca ccggatttag ctgataatat tgatggtgaa 720ttacaattcg atgcagcttt tgtcgaatcg gttgccaaat taaaagctcc gggatcaaaa 780gtggccggtc atgcgaacgt tttcattttt ccaagtttgg aagccggaaa tattggttat 840aaaattgccc aacgcttggg tggatacgaa gcaattggtc ctattttgca aggattggca 900gcacctgttt ccgatctgtc gcgtggagca agtcaagaag acgtatataa agttgcaatt 960attactgctg ctcaggcttt aaaaaattag 990

Claims

1. A co-culture comprising at least two host cells wherein (a) one of the host cells comprises: (i) a heterologous nucleic acid encoding a phosphoketolase; (ii) at least one heterologous nucleic acid encoding an enzyme in an acetyl-CoA production pathway; (iii) a heterologous nucleic acid encoding a bifunctional acetaldehyde-alcohol dehydrogenase; and, (iv) at least one genetic modification that leads to the down-regulation of an enzyme in a glycerol-production pathway; and, (b) another host cell that is genetically distinct from (a).

2. The co-culture of claim 1, wherein the host cell is a yeast and the genetically distinct host cell is a yeast or bacterium.

3. The recombinant microorganism of claim 1, wherein said phosphoketolase is a single-specificity phosphoketolase with the Enzyme Commission Number 4.1.2.9.

4. The recombinant microorganism of claim 1, wherein said phosphoketolase is dual-specificity phosphoketolase with the Enzyme Commission Number 4.1.2.22.

5. The recombinant microorganism of claim 1, wherein said phosphoketolase is from a genus selected from the group consisting of Aspergillus, Neurospora, Lactobacillus, Bificlobacterium, Penicillium, Leuconostoc, and Oenococcus.

6. The recombinant microorganism of claim 1, wherein said phosphoketolase corresponds to a polypeptide selected from a group consisting of SEQ ID NOs: 9, 11, 12, 13, 14, 15, 62, 64, 66, 68, 70, 72, 74, 78, 94, and 102.

7. The recombinant microorganism of claim 1, wherein said enzyme in the acetyl-CoA production pathway is a phosphotransacetylase with Enzyme Commission Number 2.3.1.8 or an acetate kinase with Enzyme Commission Number 2.7.2.12.

8. The recombinant microorganism of claim 7, wherein said phosphotransacetylase or said acetate kinase is from a genus selected from the group consisting of Bifidobacterium, Lactobacillus, Clostridium, Leuconostoc and Oenococcus.

9. The recombinant microorganism of claim 8, wherein said phosphotransacetylase corresponds to a polypeptide selected from a group consisting of SEQ ID NOs: 10, 34, 80, 82, 84, 86, and 96.

10. The recombinant microorganism of claim 7, wherein said acetate kinase corresponds to a polypeptide selected from a group consisting of SEQ ID NOs: 16, 76, 88, 92, and 100.

11. The recombinant microorganism of claim 1, wherein said bifunctional acetaldehyde-alcohol dehydrogenase is selected from a group of enzymes having both of the following Enzyme Commission Numbers: EC 1.2.1.10 and 1.1.1.1.

12. The recombinant microorganism of claim 11, wherein said bifunctional acetaldehyde-alcohol dehydrogenase is from B. adolescentis and corresponds to a polypeptide encoded by SEQ ID NO: 44.

13. The recombinant microorganism of claim 1, wherein said bifunctional acetaldehyde-alcohol dehydrogenase is an NADPH dependent bifunctional acetaldehyde-alcohol dehydrogenase selected from a group of enzymes having the following Enzyme Commission Numbers: EC 1.2.1.10 and 1.1.1.2.

14. The recombinant microorganism of claim 13, wherein said NADPH dependent bifunctional acetaldehyde-alcohol dehydrogenase is derived from a genus selected from the group consisting of Leuconostoc and Oenococcus.

15. The recombinant microorganism of claim 13, wherein said NADPH dependent bifunctional acetaldehyde-alcohol dehydrogenase corresponds to polypeptide selected from a group consisting of SEQ ID NOs: 90 and 98.

16. The recombinant microorganism of claim 1, wherein said enzyme in the glycerol-production pathway is a glycerol-3-phosphate dehydrogenase with Enzyme Commission Number 1.1.1.8 or a glycerol-3-phosphate phosphatase with Enzyme Commission Number 3.1.3.21.

17. The recombinant microorganism of claim 16, wherein said glycerol-3-phosphate dehydrogenase is selected from the group consisting of glycerol-3-phosphate dehydrogenase 1, glycerol-3-phosphate dehydrogenase 2, and combinations thereof.

18. The recombinant microorganism of claim 17, wherein said glycerol-3-phosphate dehydrogenase 1 is from S. cerevisiae and corresponds to a polypeptide encoded by SEQ ID NO: 28.

19. The recombinant microorganism of claim 17, wherein said glycerol-3-phosphate dehydrogenase 2 is from S. cerevisiae and corresponds to a polypeptide encoded by SEQ ID NO: 30.

20. The recombinant microorganism of claim 1, wherein said microorganism further comprises at least one additional up-regulated enzyme, wherein the up-regulated enzyme is a transaldolase with Enzyme Commission Number 2.2.1.2, a transketolase with Enzyme Commission Number 2.2.1.1, a ribose-5-P isomerase with Enzyme Commission Number 5.3.1.6, or a ribulose-5-P 3-epimerase with Enzyme Commission Number 5.1.3.1.

21. The recombinant microorganism of claim 20, wherein said transaldolase is from S. cerevisiae and corresponds to a polypeptide encoded by SEQ ID NO: 36, said transketolase is from S. cerevisiae and corresponds to a polypeptide encoded by SEQ ID NO: 38, said ribose-5-P isomerase is from S. cerevisiae and corresponds to a polypeptide encoded by SEQ ID NO: 40, or said ribulose-5-P 3-epimerase is from S. cerevisiae and corresponds to a polypeptide encoded by SEQ ID NO: 42.

22. The recombinant microorganism of claim 1, wherein at least one enzyme in a glycolysis pathway is up-regulated.

23. The recombinant microorganism of claim 22, wherein said enzyme in the glycolysis pathway is a pyruvate decarboxylase with Enzyme Commission Number 4.1.1.1, or is an alcohol dehydrogenase selected from a group of enzymes having the following Enzyme Commission Numbers: 1.1.1.1 and 1.1.1.2.

24. The recombinant microorganism of claim 1, additionally comprising at least one genetic modification that leads to the down-regulation of aldehyde dehydrogenase selected from a group of enzymes having the following Enzyme Commission Numbers: 1.2.1.3, 1.2.1.4 and 1.2.1.10 or formate dehydrogenase selected from a group of enzymes having the following Enzyme Commission Numbers: 1.2.1.43 and 1.2.1.2.

25. The recombinant microorganism of claim 1, additionally comprising at least one genetic modification that leads to the up-regulation of aldehyde dehydrogenase selected from a group of enzymes having the following Enzyme Commission Numbers: 1.2.1.3, 1.2.1.4 and 1.2.1.10 or a pyruvate formate lyase with Enzyme Commission Number: 2.3.1.54.

26. The recombinant microorganism of claim 24, wherein said aldehyde dehydrogenase is acetaldehyde dehydrogenase from S. cerevisiae and corresponds to a polypeptide sequence selected from the group consisting of: SEQ ID NO: 18, SEQ ID NO: 20, SEQ ID NO: 22, SEQ ID NO: 24, and SEQ ID NO: 26.

27. The recombinant microorganism of claim 24, wherein said formate dehydrogenase is from S. cerevisiae and corresponds to a sequence selected from the group consisting of: a polypeptide sequence corresponding to SEQ ID NO: 32 and a polynucleotide sequence corresponding to SEQ ID NO: 33.

28. The recombinant microorganism of claim 25, wherein said pyruvate formate lyase corresponds to a polypeptide encoded by SEQ ID NO: 48.

29. The recombinant microorganism of claim 1, additionally comprising at least one genetic modification that leads to the up-regulation of a pyruvate formate lyase activating enzyme with Enzyme Commission Number 1.97.1.4.

30. The recombinant microorganism of claim 29, wherein said pyruvate formate lyase activating enzyme corresponds to a polypeptide encoded by SEQ ID NO: 46.

31. A method of producing a fermentation product using the co-culture of claim 1, wherein the co-culture is capable of fermenting the carbon containing feedstock to yield the fermentation product.

32. A method of producing ethanol comprising: (a) providing the co-culture of claim 1; (b) culturing said co-culture in the presence of a carbon containing feedstock for sufficient time to produce ethanol; and, optionally, (c) extracting the ethanol.

33. A method of reducing glycerol production comprising providing a co-culture of claim 1, wherein said glycerol titer is about 10 to about 100% less than the rate compared to an otherwise identical host cell lacking said heterologous nucleic acids and genetic modification, when the co-culture is cultured in the presence of a carbon containing feedstock for a sufficient time to produce ethanol.

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