Desizing Process

Abstract

A process for desizing of a sized fabric containing starch or starch derivatives during manufacture of a fabric, which process comprises incubating said sized fabric in an aqueous treating solution having a pH in the range between 1 and 5 which aqueous treating solution comprises an alpha-amylase.

Description

REFERENCE TO A SEQUENCE LISTING

[0001] This application contains a Sequence Listing in computer readable form. The computer readable form is incorporated herein by reference.

FIELD OF THE INVENTION

[0002] The present invention relates to a process of desizing sized fabrics during manufacture of especially new fabrics.

BACKGROUND OF THE INVENTION

[0003] The processing of a fabric, such as of a cellulosic material, into material ready for garment manufacture involves several steps: spinning of the fiber into a yarn; construction of woven or knit fabric from the yarn; and subsequent preparation, dyeing and finishing operations. The preparation process, which may involve desizing (for woven goods), scouring, and bleaching, produces a fabric suitable for dyeing or finishing.

[0004] Alkaline alpha-amylases are used as auxiliaries in desizing processes to facilitate the removal of starch-containing size which has served as a protective coating on yarns during weaving. Complete removal of the size coating after weaving is important to ensure optimum results in the subsequent processes in which the fabric is generally scoured, bleached, dyed and/or printed.

[0005] After the desizing step it is often desirable to include a demineralization step in order to remove metal ions, such as Mn.sup.2+, Fe.sup.2+/Fe.sup.3+ Cu.sup.2+ etc., which--if present on the fabric--may result in an uneven bleaching in a later process step or might even make pin-holes in the bleached fabric. Demineralization is typically accomplished by acid precipitation and typically involves addition of acids such as acetic acid or sulphuric acid.

[0006] It is desirable to provide improved processes for desizing of sized fabrics during manufacture of especially new fabrics.

BRIEF DISCLOSURE OF THE INVENTION

[0007] The present invention is directed towards providing a process of desizing a sized fabric during manufacture of especially new fabrics.

[0008] In the first aspect the invention relates to a process for desizing a sized fabric containing starch or starch derivatives during manufacture of a fabric, which process comprises incubating said sized fabric in an aqueous treating solution having a pH in the range between 1 and 5 which aqueous treating solution comprises an alpha-amylase.

[0009] The present inventors have found that when carrying out a desizing process of the invention, as defined in the claims, no demineralization is needed. The demineralization takes place simultaneously and/or after the desizing of the sized fabric in the same treating solution. Compared to traditional processes involving an alkaline desizing step and a demineralization step a pH adjusting step is avoided. Another advantage of the invention is that process time is saved/reduced as desizing and demineralization may be carried out simultaneously. Even if the desizing and demineralization are not carried out as a one step process, i.e., simultaneously, costs of, e.g., acids and manpower for adding acid(s) are saved/reduced as the pH adjustment step between the traditional alkaline desizing step and the demineralization step is avoided.

[0010] In context of the invention the term "fabric" is used interchangeable with the term "textile" and means, in contrast to "used" laundry fabric, newly manufactured, preferably undyed, fabrics, garments, fibres, yarns or other types of processed fabrics. Fabrics can be constructed from fibers by weaving, knitting or non-woven operations. Weaving and knitting require yarn as the input whereas the non-woven fabric is the result of random bonding of fibers (paper can be thought of as non-woven).

[0011] Woven fabric is constructed by weaving "filling" or weft yarns between warp yarns stretched in the longitudinal direction on the loom. The wrap yarns must be sized before weaving in order to lubricate and protect them from abrasion at the high speed insertion of the filling yarns during weaving. The filling yarn can be woven through the warp yarns in a "over one--under the next" fashion (plain weave) or by "over one--under two" (twill) or any other myriad of permutations. Strength, texture and pattern are related not only to the type/quality of the yarn but also the type of weave. Generally, dresses, shirts, pants, sheeting's, towels, draperies, etc. are produced from woven fabric.

[0012] Knitting is forming a fabric by joining together interlocking loops of yarn. As opposed to weaving, which is constructed from two types of yarn and has many "ends", knitted fabric is produced from a single continuous strand of yarn. As with weaving, there are many different ways to loop yarn together and the final fabric properties are dependent both upon the yarn and the type of knit. Underwear, sweaters, socks, sport shirts, sweat shirts, etc. are derived from knit fabrics.

[0013] Non-woven fabrics are sheets of fabric made by bonding and/or interlocking fibers and filaments by mechanical, thermal, chemical or solvent mediated processes. The resultant fabric can be in the form of web-like structures, laminates or films. Typical examples are disposable baby diapers, towels, wipes, surgical gowns, fibers for the "environmental friendly" fashion, filter media, bedding, roofing materials, backing for two-dimensional fabrics and many others.

[0014] According to the invention, the process may be applied to any sized fabric known in the art (woven, knitted, or non-woven). The process is applied to newly manufactured sized fabric, as opposed to used and/or soiled fabric to be cleaned during laundry washing. In an embodiment the fabric is made of fibres of natural and/or man-made origin. In another embodiment the fabric is made of fibres from animal origin. In particular, the process of the invention may be applied to cellulose-containing or cellulosic fabrics, such as cotton, viscose, rayon, ramie, linen, cellulose acetate, denim, lyocell (Tencel.TM., e.g., produced by Courtaulds Fibers), or mixtures thereof, or mixtures of any of these fibers together with synthetic fibres (e.g., polyester, polyamid, acrylic, or polyurethane, nylon, poly(ethylene terephthalate) or poly(lactic acid) or other natural fibers, such as wool and silk, such as viscose/cotton blends, lyocell/cotton blends, viscose/wool blends, lyocell/wool blends, cotton/wool blends; flax (linen), ramie and other fabrics based on cellulose fibers, including all blends of cellulosic fibers with other fibers such as wool, polyamide, acrylic and polyester fibers, e.g., viscose/cotton/polyester blends, wool/cotton/polyester blends, flax/cotton blends etc. The process may also be used on synthetic fabric, e.g., consisting of essentially 100% polyester, polyamid, nylon, respectively. The term "wool," means any commercially useful animal hair product, for example, wool from sheep, camel, rabbit, goat, lama, and known as merino wool, Shetland wool, cashmere wool, alpaca wool, mohair, etc. and includes wool fiber and animal hair. The process of the invention can be used with wool or animal hair material in the form of top, fiber, yarn, or woven or knitted fabric.

[0015] The alpha-amylase used in accordance with the process of the invention may be any alpha-amylase, but is preferably of either bacterial or fungal origin. Preferably the alpha-amylase is an acid alpha-amylase, such as an acid alpha-amylase derived from a filamentous fungus, especially a strain of the genera Aspergillus, Rhizomucor or Merpillus.

[0016] The term "acid alpha-amylase" means an alpha-amylase (E.C. 3.2.1.1) which has an optimum activity at a pH in the range of 1 to 7, preferably from 1 to 5 at a temperature of 50.degree. C.

[0017] The term "desizing" is intended to be understood in a conventional manner, i.e., the degradation and/or removal of sizing agents from fabric, such as warp yarns in a woven fabric.

[0018] The term "fabric containing starch or starch derivatives" is intended to indicate any type of fabric, in particular woven fabric prepared from a cellulose-containing material, containing starch or starch derivatives. The fabric is normally undyed and made of cotton, viscose, flax, and the like. The main part of the starch or starch derivatives present on the fabric is normally size with which the yarns, normally warp yarns, have been coated prior to weaving.

[0019] The term "carbohydrate-binding module (CBM)", or as often referred to a "carbohydrate-binding domain (CBD)", is a polypeptide amino acid sequence which binds preferentially to a poly- or oligosaccharide (carbohydrate), frequently--but not necessarily exclusively--to a water-insoluble (including crystalline) form thereof.

[0020] Even if not specifically mentioned in connection with the process of the invention, it is to be understood that the enzyme(s) or agent(s) is(are) used in an "effective amount". The term "effective amount" means an amount of, e.g., alpha-amylase that is capable of providing the desired effect, i.e., desizing of the fabric, as compared to a fabric which has not been treated with said enzyme(s).

BRIEF DESCRIPTION OF THE DRAWING

[0021] FIG. 1 shows the desizing performance of Alpha-Amylase D on Vlisco fabric at pH 4.0.

[0022] FIG. 2 shows the desizing performance of Alpha-Amylase C on Vlisco fabric at pH 4.0.

DETAILED DISCLOSURE OF THE INVENTION

[0023] The present invention is directed towards providing a process of desizing a sized fabric during manufacture of especially new fabrics.

[0024] The desizing step of the invention is in a preferred embodiment followed by a scouring step, preferable an enzymatic scouring step, preferably with a scouring enzyme such as a pectinase, e.g., a pectate lyase, a lipase, a protease, or combination thereof, and a bleaching step, preferably involving bleaching with hydrogen peroxide and/or a hydrogen peroxide generating agent. Relevant scouring processes are described in U.S. Pat. No. 5,578,489, U.S. Pat. No. 5,912,407, and U.S. Pat. No. 6,630,342. Relevant bleach processes are described in U.S. Pat. No. 5,851,233, U.S. Pat. No. 5,752,980, and U.S. Pat. No. 5,928,380. Relevant combined scouring and bleach processes are described in WO 2003/002810 (Novozymes) and WO 2003/002705 (Novozymes), respectively.

[0025] According to the present invention, fabric may be desized and demineralised simultaneously in the same aqueous treating solution or subsequently in the same or two separate treating solutions. In a preferred embodiment the desizing and demineralization are carried out simultaneously in the same treating solution. The process of the invention may be carried out using traditional sizing/desizing equipment, e.g., pad systems, J-boxes, jets, jiggers, etc. In general, no additional process equipment is needed.

[0026] According to the invention simultaneous desizing and demineralisation are carried out by incubating sized fabric in an aqueous treating solution having a pH in the range between 1 and 5 which aqueous treating solution comprises an alpha-amylase. In a preferred embodiment the pH during incubation is in the range between 1 and 4, especially between pH 2 and 4.

[0027] Woven goods are the prevalent form of fabric construction. The weaving process demands a "sizing" of the warp yarn to protect it from abrasion. Starches, unmodified and modified, polyvinyl alcohol (PVA), carboxy methyl cellulose (CMC), waxes and acrylic binders, and mixtures thereof, are examples of typically used sizing agents. The sizing agent may according to the invention be a starch-based or starch derivative-based sizing agent, but may also contain one or more non-starch or starch derivative-based sizing agents. The sizing agent(s) are in general removed after the weaving process as the first step in preparing the woven goods.

[0028] One or more other agents including stabilizers, surfactants, wetting agents, dispersing agent, sequestering agents and emulsifying agents, or mixtures thereof, may be present during a desizing process of the invention. The sized fabric is allowed to incubate in the aqueous treating solution for a sufficiently long period of time to accomplish desizing of the sized fabric. The optimal period is dependent upon the type of processing regime and the temperature and can vary from about 15 minutes to several days, e.g., 48 hours. A process of the invention is preferably carried out at a temperature in the range from 5 to 90.degree. C., in particular 20 to 90.degree. C. dependent on the processing regime.

[0029] The processing regime can be either batch or continuous with the fabric being contacted by the aqueous treating stream in open width or rope form.

[0030] Continuous operations may use a saturator whereby an approximate equal weight of treating solution per weight of fabric is applied to the fabric, followed by a heated dwell chamber where the chemical reaction takes place. A washing section then prepares the fabric for the next processing step. In order to ensure a high whiteness or a good wettability and resulting dyeability, the desizing enzyme(s) and other agents must be thoroughly removed.

[0031] Batch processes may take place in one bath (treating solution) whereby the fabric is contacted with, e.g., approximately 8-15 times its weight of aqueous treating solution. After an incubation period, the aqueous treating solution is drained, the fabric is rinsed, and the next processing step is initiated. Discontinuous PB-processes (i.e., pad-batch processes) involves a saturator whereby an approximate equal weight of aqueous treating solution per weight of fabric is applied to the fabric, followed by a dwell period, which in the case of CPB-process (i.e., cold pad-batch process) might be one or more days. For instance, a CPB-process may be carried out at between 20-40.degree. C. for 8-24 hours or more at a pH in the range between 1 and 5, preferably at a pH in the range between around 1 and 4, especially between pH 2 and 4. Further, a PB-process may be carried out at between 40-90.degree. C. for 1-6 hours at a pH in the range between around 1 and 5, preferably between around pH 1 and 4, especially between pH 2 and 4.

[0032] In one embodiment the desizing process of the invention may be carried out using an effective amount of alpha-amylase, preferably acid alpha-amylase, and an acid such as acetic acid or sulphuric acid or the like.

Detergents

[0033] In the context of this invention, a detergent is synonymous with a surfactant, and it may in particular be a non-ionic surfactant, an anionic surfactant, a cationic surfactant, an ampholytic surfactant, a zwitterionic surfactant, and a semi-polar surfactant, or a mixture hereof.

[0034] The surfactant is typically present in a composition of the invention at a level from 0.1% to 60% by weight.

[0035] The surfactant is preferably formulated to be compatible with enzyme components present. In liquid or gel compositions the surfactant is most preferably formulated in such a way that it promotes, or at least does not degrade, the stability of any enzyme in these compositions.

[0036] Preferred systems to be used according to the present invention comprise as a surfactant one or more of the non-ionic and/or anionic surfactants described herein.

[0037] Polyethylene, polypropylene, and polybutylene oxide condensates of alkyl phenols are suitable for use as the non-ionic surfactant of the surfactant systems of the present invention, with the polyethylene oxide condensates being preferred. These compounds include the condensation products of alkyl phenols having an alkyl group containing from about 6 to about 14 carbon atoms, preferably from about 8 to about 14 carbon atoms, in either a straight chain or branched-chain configuration with the alkylene oxide. In a preferred embodiment, the ethylene oxide is present in an amount equal to from about 2 to about 25 moles, more preferably from about 3 to about 15 moles, of ethylene oxide per mole of alkyl phenol.

[0038] Commercially available non-ionic surfactants of this type include Igepal.TM. CO-630, marketed by the GAF Corporation; and TRITON.TM. X-45, X-114, X-100 and X-102, all marketed by the Rohm & Haas Company. These surfactants are commonly referred to as alkylphenol alkoxylates (e.g., alkyl phenol ethoxylates).

[0039] The condensation products of primary and secondary aliphatic alcohols with about 1 to about 25 moles of ethylene oxide are suitable for use as the nonionic surfactant of the non-ionic surfactant system. The alkyl chain of the aliphatic alcohol can either be straight or branched, primary or secondary, and generally contains from about 8 to about 22 carbon atoms. Preferred are the condensation products of alcohols having an alkyl group containing from about 8 to about 20 carbon atoms, more preferably from about 10 to about 18 carbon atoms, with from about 2 to about 10 moles of ethylene oxide per mole of alcohol. About 2 to about 7 moles of ethylene oxide and most preferably from 2 to 5 moles of ethylene oxide per mole of alcohol are present in said condensation products. Examples of commercially available non-ionic surfactants of this type include TERGITOL.TM. 15-S-9 (The condensation product of C.sub.11-C.sub.15 linear alcohol with 9 moles ethylene oxide), TERGITOL.TM. 24-L-6 NMW (the condensation product of C.sub.12-C.sub.14 primary alcohol with 6 moles ethylene oxide with a narrow molecular weight distribution), both marketed by Union Carbide Corporation; NEODOL.TM. 45-9 (the condensation product of C.sub.14-C.sub.15 linear alcohol with 9 moles of ethylene oxide), NEODOL.TM. 23-3 (the condensation product of C.sub.12-C.sub.13 linear alcohol with 3.0 moles of ethylene oxide), NEODOL.TM. 45-7 (the condensation product of C.sub.14-C.sub.15 linear alcohol with 7 moles of ethylene oxide), NEODOL.TM. 45-5 (the condensation product of C.sub.14-C.sub.15 linear alcohol with 5 moles of ethylene oxide) marketed by Shell Chemical Company, KYRO.TM. EOB (the condensation product of C.sub.13-C.sub.15 alcohol with 9 moles ethylene oxide), marketed by The Procter & Gamble Company, and Genapol LA 050 (the condensation product of C.sub.12-C.sub.14 alcohol with 5 moles of ethylene oxide) marketed by Hoechst. Preferred range of HLB in these products is from 8-11 and most preferred from 8-10.

[0040] Also useful as the nonionic surfactant of the surfactant system are alkylpolysaccharides disclosed in U.S. Pat. No. 4,565,647, having a hydrophobic group containing from about 6 to about 30 carbon atoms, preferably from about 10 to about 16 carbon atoms and a polysaccharide, e.g., a polyglycoside, hydrophilic group containing from about 1.3 to about 10, preferably from about 1.3 to about 3, most preferably from about 1.3 to about 2.7 saccharide units. Any reducing saccharide containing 5 or 6 carbon atoms can be used, e.g., glucose, galactose and galactosyl moieties can be substituted for the glucosyl moieties (optionally the hydrophobic group is attached at the 2-, 3-, 4-, etc. positions thus giving a glucose or galactose as opposed to a glucoside or galactoside). The intersaccharide bonds can be, e.g., between the one position of the additional saccharide units and the 2-, 3-, 4, and/or 6-positions on the preceding saccharide units.

[0041] The preferred alkylpolyglycosides have the formula

R.sup.2O(C.sub.nH.sub.2nO).sub.t(glycosyl).sub.x

wherein R.sup.2 is selected from the group consisting of alkyl, alkylphenyl, hydroxyalkyl, hydroxyalkylphenyl, and mixtures thereof in which the alkyl groups contain from about 10 to about 18, preferably from about 12 to about 14, carbon atoms; n is 2 or 3, preferably 2; t is from 0 to about 10, preferably 0; and x is from about 1.3 to about 10, preferably from about 1.3 to about 3, most preferably from about 1.3 to about 2.7. The glycosyl is preferably derived from glucose. To prepare these compounds, the alcohol or alkylpolyethoxy alcohol is formed first and then reacted with glucose, or a source of glucose, to form the glucoside (attachment at the 1-position). The additional glycosyl units can then be attached between their 1-position and the preceding glycosyl units 2-, 3-, 4-, and/or 6-position, preferably predominantly the 2-position.

[0042] The condensation products of ethylene oxide with a hydrophobic base formed by the condensation of propylene oxide with propylene glycol are also suitable for use as the additional non-ionic surfactant system. The hydrophobic portion of these compounds will preferably have a molecular weight from about 1500 to about 1800 and will exhibit water insolubility. The addition of polyoxyethylene moieties to this hydrophobic portion tends to increase the water solubility of the molecule as a whole, and the liquid character of the product is retained up to the point where the polyoxyethylene content is about 50% of the total weight of the condensation product, which corresponds to condensation with up to about 40 moles of ethylene oxide. Examples of compounds of this type include certain of the commercially available PLURONIC.TM. surfactants, marketed by BASF.

[0043] Also suitable for use as the non-ionic surfactant of the non-ionic surfactant system are the condensation products of ethylene oxide with the product resulting from the reaction of propylene oxide and ethylenediamine. The hydrophobic moiety of these products consists of the reaction product of ethylenediamine and excess propylene oxide, and generally has a molecular weight of from about 2,500 to about 3,000. This hydrophobic moiety is condensed with ethylene oxide to the extent that the condensation product contains from about 40% to about 80% by weight of polyoxyethylene and has a molecular weight of from about 5,000 to about 11,000. Examples of this type of non-ionic surfactant include certain of the commercially available TETRONIC.TM. compounds, marketed by BASF.

[0044] Preferred for use as the non-ionic surfactant of the surfactant system are polyethylene oxide condensates of alkyl phenols, condensation products of primary and secondary aliphatic alcohols with from about 1 to about 25 moles of ethyleneoxide, alkylpolysaccharides, and mixtures hereof. Most preferred are C.sub.8-C.sub.14 alkyl phenol ethoxylates having from 3 to 15 ethoxy groups and C.sub.8-C.sub.18 alcohol ethoxylates (preferably C.sub.10 avg.) having from 2 to 10 ethoxy groups, and mixtures thereof.

[0045] Highly preferred non-ionic surfactants are polyhydroxy fatty acid amide surfactants of the formula

##STR00001##

wherein R.sup.1 is H, or R.sup.1 is C.sub.1-4 hydrocarbyl, 2-hydroxyethyl, 2-hydroxypropyl or a mixture thereof, R.sup.2 is C.sub.5-31 hydrocarbyl, and Z is a polyhydroxyhydrocarbyl having a linear hydrocarbyl chain with at least 3 hydroxyls directly connected to the chain, or an alkoxylated derivative thereof. Preferably, R.sup.1 is methyl, R.sup.2 is straight C.sub.11-15 alkyl or C.sub.16-18 alkyl or alkenyl chain such as coconut alkyl or mixtures thereof, and Z is derived from a reducing sugar such as glucose, fructose, maltose or lactose, in a reductive amination reaction.

[0046] Highly preferred anionic surfactants include alkyl alkoxylated sulfate surfactants. Examples hereof are water soluble salts or acids of the formula RO(A).sub.mSO3M wherein R is an unsubstituted C.sub.10-C-.sub.24 alkyl or hydroxyalkyl group having a C.sub.10-C.sub.24 alkyl component, preferably a C.sub.12-C.sub.20 alkyl or hydroxyalkyl, more preferably C.sub.12-C.sub.18 alkyl or hydroxyalkyl, A is an ethoxy or propoxy unit, m is greater than zero, typically between about 0.5 and about 6, more preferably between about 0.5 and about 3, and M is H or a cation which can be, for example, a metal cation (e.g., sodium, potassium, lithium, calcium, magnesium, etc.), ammonium or substituted-ammonium cation. Alkyl ethoxylated sulfates as well as alkyl propoxylated sulfates are contemplated herein. Specific examples of substituted ammonium cations include methyl-, dimethyl, trimethyl-ammonium cations and quaternary ammonium cations such as tetramethyl-ammonium and dimethyl piperidinium cations and those derived from alkylamines such as ethylamine, diethylamine, triethylamine, mixtures thereof, and the like. Exemplary surfactants are C.sub.12-C.sub.18 alkyl polyethoxylate (1.0) sulfate (C.sub.12-C.sub.18E(1.0)M), C.sub.12-C.sub.18 alkyl polyethoxylate (2.25) sulfate (C.sub.12-C.sub.18(2.25)M, and C.sub.12-C.sub.18 alkyl polyethoxylate (3.0) sulfate (C.sub.12-C.sub.18E(3.0)M), and C.sub.12-C.sub.18 alkyl polyethoxylate (4.0) sulfate (C.sub.12-C.sub.18E(4.0)M), wherein M is conveniently selected from sodium and potassium.

[0047] Suitable anionic surfactants to be used are alkyl ester sulfonate surfactants including linear esters of C.sub.8-C.sub.20 carboxylic acids (i.e., fatty acids) which are sulfonated with gaseous SO.sub.3 according to "The Journal of the American Oil Chemists Society", 52 (1975), pp. 323-329. Suitable starting materials would include natural fatty substances as derived from tallow, palm oil, etc.

[0048] The preferred alkyl ester sulfonate surfactant comprise alkyl ester sulfonate surfactants of the structural formula:

##STR00002##

wherein R.sup.3 is a C.sub.8-C.sub.20 hydrocarbyl, preferably an alkyl, or combination thereof, R.sup.4 is a C.sub.1-C.sub.6 hydrocarbyl, preferably an alkyl, or combination thereof, and M is a cation which forms a water soluble salt with the alkyl ester sulfonate. Suitable salt-forming cations include metals such as sodium, potassium, and lithium, and substituted or unsubstituted ammonium cations, such as monoethanolamine, diethonolamine, and triethanolamine. Preferably, R.sup.3 is C.sub.10-C.sub.16 alkyl, and R.sup.4 is methyl, ethyl or isopropyl. Especially preferred are the methyl ester sulfonates wherein R.sup.3 is C.sub.10-C.sub.16 alkyl.

[0049] Other suitable anionic surfactants include the alkyl sulfate surfactants which are water soluble salts or acids of the formula ROSO.sub.3M wherein R preferably is a C.sub.10-C.sub.24 hydrocarbyl, preferably an alkyl or hydroxyalkyl having a C.sub.10-C.sub.20 alkyl component, more preferably a C.sub.12-C.sub.16 alkyl or hydroxyalkyl, and M is H or a cation, e.g., an alkali metal cation (e.g., sodium, potassium, lithium), or ammonium or substituted ammonium (e.g., methyl-, dimethyl-, and trimethyl ammonium cations and quaternary ammonium cations such as tetramethyl-ammonium and dimethyl piperidinium cations and quaternary ammonium cations derived from alkylamines such as ethylamine, diethylamine, triethylamine, and mixtures thereof, and the like). Typically, alkyl chains of C.sub.12-C.sub.16 are preferred for lower wash temperatures (e.g. below about 50.degree. C.) and C.sub.16-C.sub.18 alkyl chains are preferred for higher wash temperatures (e.g. above about 50.degree. C.).

[0050] Other anionic surfactants useful for detersive purposes include salts (including, for example, sodium, potassium, ammonium, and substituted ammonium salts such as mono- di- and triethanolamine salts) of soap, C.sub.8-C.sub.22 primary or secondary alkanesulfonates, C.sub.8-C.sub.24 olefinsulfonates, sulfonated polycarboxylic acids prepared by sulfonation of the pyrolyzed product of alkaline earth metal citrates, e.g., as described in British patent specification No. 1,082,179, C.sub.8-C.sub.24 alkylpolyglycolethersulfates (containing up to 10 moles of ethylene oxide); alkyl glycerol sulfonates, fatty acyl glycerol sulfonates, fatty oleyl glycerol sulfates, alkyl phenol ethylene oxide ether sulfates, paraffin sulfonates, alkyl phosphates, isethionates such as the acyl isethionates, N-acyl taurates, alkyl succinamates and sulfosuccinates, monoesters of sulfosuccinates (especially saturated and unsaturated C.sub.12-C.sub.18 monoesters) and diesters of sulfosuccinates (especially saturated and unsaturated C.sub.6-C.sub.12 diesters), acyl sarcosinates, sulfates of alkylpolysaccharides such as the sulfates of alkylpolyglucoside (the nonionic nonsulfated compounds being described below), branched primary alkyl sulfates, and alkyl polyethoxy carboxylates such as those of the formula RO(CH.sub.2CH.sub.2O).sub.k--CH.sub.2C00-M+ wherein R is a C.sub.8-C.sub.22 alkyl, k is an integer from 1 to 10, and M is a soluble salt forming cation. Resin acids and hydrogenated resin acids are also suitable, such as rosin, hydrogenated rosin, and resin acids and hydrogenated resin acids present in or derived from tall oil.

[0051] Alkylbenzene sulfonates are highly preferred. Especially preferred are linear (straight-chain) alkyl benzene sulfonates (LAS) wherein the alkyl group preferably contains from 10 to 18 carbon atoms.

[0052] Further examples are described in "Surface Active Agents and Detergents" (Vol. I and II by Schwartz, Perrry and Berch). A variety of such surfactants are also generally disclosed in U.S. Pat. No. 3,929,678, (Column 23, line 58 through Column 29, line 23, herein incorporated by reference).

[0053] When included therein the compositions of the present invention typically comprise from about 1% to about 40%, preferably from about 3% to about 20% by weight of such anionic surfactants.

[0054] The compositions of the present invention may also contain cationic, ampholytic, zwitterionic, and semi-polar surfactants, as well as the nonionic and/or anionic surfactants other than those already described herein.

[0055] Cationic detersive surfactants suitable for use in the compositions of the present invention are those having one long-chain hydrocarbyl group. Examples of such cationic surfactants include the ammonium surfactants such as alkyltrimethylammonium halogenides, and those surfactants having the formula:

[R.sup.2(OR.sup.3).sub.y][R.sup.4(OR.sup.3).sub.y].sub.2R.sup.5N+X--

wherein R.sup.2 is an alkyl or alkyl benzyl group having from about 8 to about 18 carbon atoms in the alkyl chain, each R.sup.3 is selected form the group consisting of --CH.sub.2CH.sub.2--, --CH.sub.2CH(CH.sub.3)--, --CH.sub.2CH(CH.sub.2OH)--, --CH.sub.2CH.sub.2CH.sub.2--, and mixtures thereof; each R.sup.4 is selected from the group consisting of C.sub.1-C.sub.4 alkyl, C.sub.1-C.sub.4 hydroxyalkyl, benzyl ring structures formed by joining the two R.sup.4 groups, --CH.sub.2CHOHCHOHCOR.sup.6CHOHCH.sub.-2OH, wherein R.sup.6 is any hexose or hexose polymer having a molecular weight less than about 1000, and hydrogen when y is not 0; R.sup.5 is the same as R.sup.4 or is an alkyl chain, wherein the total number of carbon atoms or R.sup.2 plus R.sup.5 is not more than about 18; each y is from 0 to about 10, and the sum of the y values is from 0 to about 15; and X is any compatible anion.

[0056] Highly preferred cationic surfactants are the water soluble quaternary ammonium compounds useful in the present composition having the formula:

R.sub.1R.sub.2R.sub.3R.sub.4N.sup.+X.sup.- (i)

wherein R.sub.1 is C.sub.8-C.sub.16 alkyl, each of R.sub.2, R.sub.3 and R.sub.4 is independently C.sub.1-C.sub.4 alkyl, C.sub.1-C.sub.4 hydroxy alkyl, benzyl, and --(C.sub.2H.sub.40).sub.xH where x has a value from 2 to 5, and X is an anion. Not more than one of R.sub.2, R.sub.3 or R.sub.4 should be benzyl.

[0057] The preferred alkyl chain length for R.sub.1 is Cl.sub.2-C.sub.15, particularly where the alkyl group is a mixture of chain lengths derived from coconut or palm kernel fat or is derived synthetically by olefin build up or OXO alcohols synthesis.

[0058] Preferred groups for R.sub.2R.sub.3 and R.sub.4 are methyl and hydroxyethyl groups and the anion X may be selected from halide, methosulphate, acetate and phosphate ions.

[0059] Examples of suitable quaternary ammonium compounds of formulae (i) for use herein are:

[0060] coconut trimethyl ammonium chloride or bromide;

[0061] coconut methyl dihydroxyethyl ammonium chloride or bromide;

[0062] decyl triethyl ammonium chloride;

[0063] decyl dimethyl hydroxyethyl ammonium chloride or bromide;

[0064] C.sub.12-15 dimethyl hydroxyethyl ammonium chloride or bromide;

[0065] coconut dimethyl hydroxyethyl ammonium chloride or bromide;

[0066] myristyl trimethyl ammonium methyl sulphate;

[0067] lauryl dimethyl benzyl ammonium chloride or bromide;

[0068] lauryl dimethyl (ethenoxy).sub.4 ammonium chloride or bromide;

[0069] choline esters (compounds of formula (I) wherein R.sub.1 is

##STR00003##

di-alkyl imidazolines [compounds of formula (i)].

[0070] Other cationic surfactants useful herein are also described in U.S. Pat. No. 4,228,044 and in EP 000 224.

[0071] When included therein, the compositions of the present invention typically comprise from 0.2% to about 25%, preferably from about 1% to about 8% by weight of such cationic surfactants.

[0072] Ampholytic surfactants are also suitable for use in the compositions of the present invention. These surfactants can be broadly described as aliphatic derivatives of secondary or tertiary amines, or aliphatic derivatives of heterocyclic secondary and tertiary amines in which the aliphatic radical can be straight- or branched-chain. One of the aliphatic substituents contains at least about 8 carbon atoms, typically from about 8 to about 18 carbon atoms, and at least one contains an anionic water-solubilizing group, e.g., carboxy, sulfonate, sulfate. See U.S. Pat. No. 3,929,678 (column 19, lines 18-35) for examples of ampholytic surfactants.

[0073] When included therein, the compositions of the present invention typically comprise from 0.2% to about 15%, preferably from about 1% to about 10% by weight of such ampholytic surfactants.

[0074] Zwitterionic surfactants are also suitable for use in the composition of the invention. These surfactants can be broadly described as derivatives of secondary and tertiary amines, derivatives of heterocyclic secondary and tertiary amines, or derivatives of quaternary ammonium, quaternary phosphonium or tertiary sulfonium compounds. See U.S. Pat. No. 3,929,678 (column 19, line 38 through column 22, line 48) for examples of zwitterionic surfactants.

[0075] When included therein, the compositions of the present invention typically comprise from 0.2% to about 15%, preferably from about 1% to about 10% by weight of such zwitterionic surfactants.

[0076] Semi-polar nonionic surfactants are a special category of nonionic surfactants which include water-soluble amine oxides containing one alkyl moiety of from about 10 to about 18 carbon atoms and 2 moieties selected from the group consisting of alkyl groups and hydroxyalkyl groups containing from about 1 to about 3 carbon atoms; watersoluble phosphine oxides containing one alkyl moiety of from about 10 to about 18 carbon atoms and 2 moieties selected from the group consisting of alkyl groups and hydroxyalkyl groups containing from about 1 to about 3 carbon atoms; and water-soluble sulfoxides containing one alkyl moiety from about 10 to about 18 carbon atoms and a moiety selected from the group consisting of alkyl and hydroxyalkyl moieties of from about 1 to about 3 carbon atoms.

[0077] Semi-polar nonionic detergent surfactants include the amine oxide surfactants having the formula:

##STR00004##

wherein R.sup.3 is an alkyl, hydroxyalkyl, or alkyl phenyl group or mixtures thereof containing from about 8 to about 22 carbon atoms; R.sup.4 is an alkylene or hydroxyalkylene group containing from about 2 to about 3 carbon atoms or mixtures thereof; x is from 0 to about 3: and each R.sup.5 is an alkyl or hydroxyalkyl group containing from about 1 to about 3 carbon atoms or a polyethylene oxide group containing from about 1 to about 3 ethylene oxide groups. The R.sup.5 groups can be attached to each other, e.g., through an oxygen or nitrogen atom, to form a ring structure.

[0078] These amine oxide surfactants in particular include C.sub.10-C.sub.18 alkyl dimethyl amine oxides and C.sub.8-C.sub.12 alkoxy ethyl dihydroxy ethyl amine oxides.

[0079] When included therein, the composition of the present invention typically comprises from 0.2% to about 15%, preferably from about 1% to about 10% by weight of such semi-polar nonionic surfactants.

Enzymes

Alpha-Amylases

[0080] The alpha-amylase(s) used in the process of the invention may be any alpha-amylase, preferably of bacterial or fungal origin. In a preferred embodiment the alpha-amylase is an acid alpha-amylase, such as an alpha-amylase or hybrid alpha-amylase disclosed in WO 2005/003311 which is hereby incorporated by reference.

[0081] In a preferred embodiment the alpha-amylase include a carbohydrate-binding module (CBM) as defined in WO 2005/003311, preferably a family 20 CBM as defined in WO 2005/003311.

[0082] Specifically contemplated are CBMs include the ones selected from the group consisting of Aspergillus kawachli disclosed in SEQ ID NO: 2; Bacillus flavothermus disclosed in SEQ ID NO: 5; Bacillus sp. disclosed in SEQ ID NO: 6; Alcaliphilic Bacillus disclosed in SEQ ID NO: 7; Hormoconis resinae disclosed in SEQ ID NO: 8; Lentinula edodes disclosed in SEQ ID NO: 9; Neurospora crassa disclosed in SEQ ID NO: 10; Talaromyces byssochlamydiodes disclosed in SEQ ID NO: 11; Geosmithia cylindrospora disclosed in SEQ ID NO: 12; Scorias spodiosa disclosed in SEQ ID NO: 13; Eupenicillium ludwigii disclosed in SEQ ID NO: 14; Aspergillus japonicus disclosed in SEQ ID NO: 15; Penicillium cf. miczynskii disclosed in SEQ ID NO: 16; Mz1 Penicillium sp. disclosed in SEQ ID NO: 17; Thysanospora sp. disclosed in SEQ ID NO: 18; Humicola grisea var. thermoidea disclosed in SEQ ID NO: 19; Aspergillus niger disclosed in SEQ ID NO: 20; or Althea rolfsii disclosed in SEQ ID NO: 21.

[0083] Fungal Alpha-Amylases

[0084] In an embodiment the fungal alpha-amylase is of yeast or filamentous fungus origin. In a preferred embodiment the fungal alpha-amylase is an acid alpha-amylase.

[0085] Preferred alpha-amylases include, for example, alpha-amylases obtainable from Aspergillus species, in particular from Aspergillus niger, A. oryzae, and A. awamori, A. kawachii, such as the acid alpha-amylase disclosed as SWISSPROT P56271, or described in more detail in WO 89/01969 (Example 3). The mature acid alpha-amylase has the amino acid sequence shown as 22-511 of SEQ ID NO: 4, encoded by the DNA sequence shown in SEQ ID NO: 3, or the amino acid sequence shown in SEQ ID NO: 38. Also preferred are alpha-amylase sequences having more than 50%, such as more than 60%, more than 70%, more than 80% or more than 90%, more than 95%, more than 96%, more than 97%, more than 98%, or even more than 99% identity to the amino acid sequence shown in SEQ ID NOS: 4 or 38, respectively.

[0086] In another preferred embodiment the alpha-amylase sequence is derived from an A. oryzae acid alpha-amylase. More preferably the alpha-amylase sequence has more than 50%, such as more than 60%, more than 70%, more than 80% or more than 90%, more than 95%, more than 96%, more than 97%, more than 98%, or more than 99% identity to the amino acid sequence shown in SEQ ID NO: 39.

[0087] In one embodiment the alpha-amylase is the Aspergillus kawachii alpha-amylase disclosed in SEQ ID NO: 37, which in wild-type form contains a carbohydrate-binding domain (CBD) also shown in SEQ ID NO: 2.

[0088] In a preferred embodiment the alpha-amylase is an alpha-amylase having more than 50%, such as more than 60%, more than 70%, more than 80% or more than 90%, more than 95%, more than 96%, more than 97%, more than 98%, or even more than 99% identity to the amino acid sequence shown in SEQ ID NOS: 43, 44, 46 or 47, respectively.

[0089] The alpha-amylase may be present in a concentration of 1-3,000 AFAU/kg fabric, preferably 10-1,000 AFAU/kg fabric, especially 100-500 AFAU/kg fabric or 1-3,000 AFAU/L treating solution, preferably 10-1,000 AFAU/L treating solution, especially 100-500 AFAU/L treating solution.

Bacterial Alpha-Amylases

[0090] In an embodiment the alpha-amylase is of bacterial origin. In a preferred embodiment the bacterial alpha-amylase is an acid alpha-amylase.

[0091] The bacterial alpha-amylase is preferably derived from a strain of Bacillus, such as Bacillus licheniformis, Bacillus amyloliquefaciens, Bacillus stearothermophilus, Bacillus subtilis, or other Bacillus sp., such as Bacillus sp. NCIB 12289, NCIB 12512 (WO 95/26397), NCIB 12513 (WO 95126397), DSM 9375 (WO 95/26397), DSMZ 12648 (WO 00/60060), DSMZ 12649 (WO 00/60060), KSM AP1378 (WO 97/00324), KSM K36 or KSM K38 (EP 1,022,334). Preferred are the Bacillus sp. alpha-amylases disclosed in WO 95/26397 as SEQ ID NOS. 1 and 2, respectively, the AA560 alpha-amylase disclosed as SEQ ID NO: 2 in WO 00/60060 (i.e., SEQ ID NO: 40 herein), and the #707 alpha-amylase disclosed by Tsukamoto et al., Biochemical and Biophysical Research Communications, 151 (1988), pp. 25-31.

[0092] In an embodiment of the invention the bacterial alpha-amylase is the SP722 alpha-amylase disclosed as SEQ ID NO: 2 in WO 95/26397 or the M560 alpha-amylase (SEQ ID NO: 40 herein).

[0093] In a preferred embodiment the parent alpha-amylase has one or more deletions in positions or corresponding to the following positions: D183 and G184, preferably wherein said alpha-amylase variant further has a substitution in position or corresponding to position N195F (using the SEQ ID NO: 40 numbering).

[0094] In another preferred embodiment the parent alpha-amylase has one or more of the following deletions/substitutions or corresponding to the following deletions/substitutions: Delta (R81-G182); Delta (D183-G184); Delta (D183-G184)+N195F; R181Q+N445Q+K446N; Delta (D183-G184)+R181Q, Delta (D183-G184) and one or more of the following substitutions or corresponding to: R118K, N195F, R320K, R458K, especially wherein the variant has the following mutations: .DELTA.(D183+G184)+R118K+N195F+R320K+R458K (using the SEQ ID NO: 40 numbering).

[0095] In another preferred embodiment the alpha-amylase is the AA560 alpha-amylase shown in SEQ ID NO: 40 further comprising one or more of the following substitutions M9L, M202L, V214T, M323T, M382Y, E345R or the A560 alpha-amylase with all of the following substitutions: M9L, M202L, V214T, M323T, M382Y or M9L, M202L, V214T, M323T and E345R.

[0096] Commercially available alpha-amylase products or products comprising alpha-amylases include product sold under the following tradenames: NATALASE.TM., STAINZYME.TM. (Novozymes A/S), Bioamylase--D(G), BIOAMYLASE.TM. L (Biocon India Ltd.), KEMZYM.TM. AT 9000(Biozym Ges. m.b.H, Austria), PURASTAR.TM. ST, PURASTAR.TM. HPAmL, PURAFECT.TM. OxAm, RAPIDASE.TM. TEX (Genencor Int. Inc, USA), KAM (KAO, Japan)

[0097] The alpha-amylase may be present in a concentration of from about 0.05-150 KNU/L treating solution, preferably 1-100 KNU/L treating solution, especially 2-20 KNU/L treating solution or 0.05-150 KNU/Kg fabric, preferably, 1-100 KNU/kg fabric, especially 2-20-KNU/kg fabric

Hybrid Enzyme

[0098] The alpha-amylase may in a preferred embodiment be an alpha-amylase comprising a carbohydrate-binding domain (CBD). Such alpha-amylase with a CBD may be a wild type enzyme (see e.g., Aspergillus kawachii above) or a hybrid enzyme (fusion protein) as will be described further below. Hybrid enzymes or a genetically modified wild type enzymes as referred to herein include species comprising an amino acid sequence of an alpha-amylase enzyme (EC 3.2.1.1) linked (i.e. covalently bound) to an amino acid sequence comprising a carbohydrate-binding domain (CBD).

[0099] CBD-containing hybrid enzymes, as well as detailed descriptions of the preparation and purification thereof, are known in the art [see, e.g. WO 90/00609, WO 94/24158 and WO 95/16782, as well as Greenwood et al. Biotechnology and Bioengineering 44 (1994) pp. 1295-1305]. They may, e.g., be prepared by transforming into a host cell a DNA construct comprising at least a fragment of DNA encoding the carbohydrate-binding domain ligated, with or without a linker, to a DNA sequence encoding the enzyme of interest, and growing the transformed host cell to express the fused gene. The resulting recombinant product (hybrid enzyme)--often referred to in the art as a "fusion protein--may be described by the following general formula:

A-CBD-MR-X

[0100] In the latter formula, A-CBD is the N-terminal or the C-terminal region of an amino acid sequence comprising at least the carbohydrate-binding domain (CBD) per se. MR is the middle region (the "linker"), and X is the sequence of amino acid residues of a polypeptide encoded by a DNA sequence encoding the enzyme (or other protein) to which the CBD is to be linked.

[0101] The moiety A may either be absent (such that A-CBD is a CBD per se, i.e. comprises no amino acid residues other than those constituting the CBD) or may be a sequence of one or more amino acid residues (functioning as a terminal extension of the CBD per se). The linker (MR) may be a bond, or a short linking group comprising from about 2 to about 100 carbon atoms, in particular of from 2 to 40 carbon atoms. However, MR is preferably a sequence of from about 2 to about 100 amino acid residues, more preferably of from 2 to 40 amino acid residues, such as from 2 to 15 amino acid residues.

[0102] The moiety X may constitute either the N-terminal or the C-terminal region of the overall hybrid enzyme.

[0103] It will thus be apparent from the above that the CBD in a hybrid enzyme of the type in question may be positioned C-terminally, N-terminally or internally in the hybrid enzyme.

Linker Sequence

[0104] The linker sequence may be any suitable linker sequence. In preferred embodiments the linker sequence is derived from the Athelia roffsii glucoamylase, the A. niger glucoamylase, the A. kawachii alpha-amylase such as a linker sequence selected from the group consisting of A. niger glucoamylase linker: TGGTTTTATPTGSGSVTSTSKTTATASKTSTSTSSTSA (SEQ ID NO:22), A. kawachii alpha-amylase linker: T T T T T T A A A T S T S K A T T S S S S S S A A A T T S S S (SEQ ID NO:23), Athelia roffsii glucoamylase linker: G A T S P G G S S G S (SEQ ID NO:24), and the PEPT linker: P E P T P E P T (SEQ ID NO:25). In another preferred embodiment the hybrid enzymes has a linker sequence which differs from the amino acid sequence shown in SEQ ID NO:22, SEQ ID NO:23, SEQ ID NO:24, or SEQ ID NO:25 in no more than 10 positions, no more than 9 positions, no more than 8 positions, no more than 7 positions, no more than 6 positions, no more than 5 positions, no more than 4 positions, no more than 3 positions, no more than 2 positions, or even no more than 1 position.

Carbohydrate-Binding Domain

[0105] A carbohydrate-binding domains (CBD), or as often referred to, a carbohydrate-binding modules (CBM), is a polypeptide amino acid sequence which binds preferentially to a poly- or oligosaccharide (carbohydrate), frequently--but not necessarily exclusively--to a water-insoluble (including crystalline) form thereof.

[0106] CBDs derived from starch degrading enzymes are often referred to as starch-binding domains (SBD) or starch-binding modules (SBM). SBDs are CBDs which may occur in certain amylolytic enzymes, such as certain glucoamylases, or in enzymes such as cyclodextrin glucanotransferases, or in alpha-amylases. Likewise, other sub-classes of CBDs would embrace, e.g. cellulose-binding domains (CBDs from cellulolytic enzymes), chitin-binding domains (CBDs which typically occur in chitinases), xylan-binding domains (CBDs which typically occur in xylanases), mannan-binding domains (CBDs which typically occur in mannanases).

[0107] CBDs are found as integral parts of large polypeptides or proteins consisting of two or more polypeptide amino acid sequence regions, especially in hydrolytic enzymes (hydrolases) which typically comprise a catalytic domain containing the active site for substrate hydrolysis and a carbohydrate-binding domain (CBD) for binding to the carbohydrate substrate in question. Such enzymes can comprise more than one catalytic domain and one, two or three CBDs, and optionally further comprise one or more polypeptide amino acid sequence regions linking the CBD(s) with the catalytic domain(s), a region of the latter type usually being denoted a "linker". Examples of hydrolytic enzymes comprising a CBD--some of which have already been mentioned above--are cellulases, xylanases, mannanases, arabinofuranosidases, acetylesterases and chitinases. CBDs have also been found in algae, e.g., in the red alga Porphyra purpurea in the form of a non-hydrolytic polysaccharide-binding protein.

[0108] In proteins/polypeptides in which CBDs occur (e.g. enzymes, typically hydrolytic enzymes), a CBD may be located at the N or C terminus or at an internal position.

[0109] That part of a polypeptide or protein (e.g. hydrolytic enzyme) which constitutes a CBD per se typically consists of more than about 30 and less than about 250 amino acid residues.

[0110] The "Carbohydrate-Binding Module of Family 20" or a CBM-20 module is in the context of this invention defined as a sequence of approximately 100 amino acids having at least 45% homology to the Carbohydrate-Binding Module (CBM) of the polypeptide disclosed in FIG. 1 by Joergensen et al (1997) in Biotechnol. Lett. 19:1027-1031. The CBM comprises the last 102 amino acids of the polypeptide, i.e. the subsequence from amino acid 582 to amino acid 683. The numbering of Glycoside Hydrolase Families applied in this disclosure follows the concept of Coutinho, P. M. & Henrissat, B. (1999) CAZy--Carbohydrate-Active Enzymes server at URL: http)://afmb.cnrs-mrs.fr/.about.cazy/CAZY/index.html or alternatively Coutinho, P. M. & Henrissat, B. 1999; The modular structure of cellulases and other carbohydrate-active enzymes: an integrated database approach. In "Genetics, Biochemistry and Ecology of Cellulose Degradation", K. Ohmiya, K. Hayashi, K. Sakka, Y. Kobayashi, S. Karita and T. Kimura eds., Uni Publishers Co., Tokyo, pp. 15-23, and Bourne, Y. & Henrissat, B. 2001; Glycoside hydrolases and glycosyltransferases: families and functional modules, Current Opinion in Structural Biology 11:593-600.

[0111] Examples of enzymes which comprise a CBD suitable for use in the context of the invention are alpha-amylases, maltogenic alpha-amylases, cellulases, xylanases, mannanases, arabinofuranosidases, acetylesterases and chitinases. Further CBDs of interest in relation to the present invention include CBDs derived from glucoamylases (EC 3.2.1.3) or from CGTases (EC 2.4.1.19).

[0112] CBDs derived from fungal, bacterial or plant sources will generally be suitable for use in the context of the invention. Preferred are CBDs of fungal origin, more preferably from Aspergillus sp., Bacillus sp., Klebsiella sp., or Rhizopus sp. In this connection, techniques suitable for isolating the relevant genes are well known in the art.

[0113] Preferred for the invention is CBDs of Carbohydrate-Binding Module Family 20. CBDs of Carbohydrate-Binding Module Family 20 suitable for the invention may be derived from glucoamylases of Aspergillus awamori (SWISSPROT Q12537), Aspergillus kawachii (SWISSPROT P23176), Aspergillus niger (SWISSPROT P04064), Aspergillus oryzae (SWISSPROT P36914), from alpha-amylases of Aspergillus kawachii (EMBL:#AB008370), Aspergillus nidulans (NCBI AA17100.1), from beta-amylases of Bacillus cereus (SWISSPROT P36924), or from CGTases of Bacillus circulans (SWISSPROT P43379). Preferred is a CBD from the alpha-amylase of Aspergillus kawachii (EMBL:#AB008370) as well as CBDs having at least 50%, 60%, 70%, 80% or even at least 90%, 95%, 96%, 97%, 98%, or 99% identity with the CBD of the alpha-amylase of Aspergillus kawachii (EMBL:#AB008370), i.e. a CBD having at least 50%, 60%, 70%, 80% or even at least 90%, 95%, 96%, 97%, 98%, or 99% identity with the amino acid sequence of SEQ ID NO:2. Also preferred for the invention are the CBDs of Carbohydrate-Binding Module Family 20 having the amino acid sequences shown in SEQ ID NO:5, SEQ ID NO:6, and SEQ ID NO:7 and disclosed in PCT application no. PCT/DK2004/000456 (or Danish patent application PA 2003 00949) as SEQ ID NO:1, SEQ ID NO:2 and SEQ ID NO:3 respectively. Further preferred CBDs include the CBDs of the glucoamylase from Hormoconis sp. such as from Hormoconis resinae (Syn. Creosote fungus or Amorphotheca resinae) such as the CBD in SWISSPROT:Q03045 (SEQ ID NO:8), from Lentinula sp. such as from Lentinula edodes (shiitake mushroom) such as the CBD of SPTREMBL:Q9P4C5 (SEQ ID NO:9), from Neurospora sp. such as from Neurospora crassa such as the CBD of SWISSPROT:P14804 (SEQ ID NO:10), from Talaromyces sp. such as from Talaromyces byssochlamydioides such as the CBD from NN005220 (SEQ ID NO:11), from Geosmithia sp. such as from Geosmithia cylindrospora, such as the CBD of NN48286 (SEQ ID NO:12), from Scorias sp. such as from Scorias spongiosa such as the CBD of NN007096 (SEQ ID NO:13), from Eupenicillium sp. such as from Eupenicillium ludwigii such as the CBD of NN005968 (SEQ ID NO:14), from Aspergillus sp. such as from Aspergillus japonicus such as the CBD from NN001136 (SEQ ID NO:15), from Penicillium sp. such as from Penicillium cf. miczynskii such as the CBD of NN48691 (SEQ ID NO:16), from Mz1 Penicillium sp. such as the CBD of NN48690 (SEQ ID NO:17), from Thysanophora sp. such as the CBD of NN48711 (SEQ ID NO:18), and from Humicola sp. such as from Humicola grisea var. thermoidea such as the CBD of SPTREMBL:Q12623 (SEQ ID NO: 19). Most preferred CBDs include the CBDs of the glucoamylase from Aspergillus sp. such as from Aspergillus niger, such as SEQ ID NO:20, and Athelia sp. such as from Athelia rolfsii, such as SEQ ID NO:21. Also preferred according to the invention are any CBD having at least 50%, 60%, 70%, 80% or even at least 90%, 95%, 96%, 97%, 98%, or 99% identity to any of the afore mentioned CBD amino acid sequences.

[0114] Further suitable CBDs of Carbohydrate-Binding Module Family 20 may be found at URL: http://afmb.cnrs-mrs.fr/.about.cazy/CAZY/index.html).

[0115] Once a nucleotide sequence encoding the substrate-binding (carbohydrate-binding) region has been identified, either as cDNA or chromosomal DNA, it may then be manipulated in a variety of ways to fuse it to a DNA sequence encoding the enzyme of interest. The DNA fragment encoding the carbohydrate-binding amino acid sequence and the DNA encoding the enzyme of interest are then ligated with or without a linker. The resulting ligated DNA may then be manipulated in a variety of ways to achieve expression.

[0116] In an embodiment the alpha-amylase comprised in the hybrid is an alpha-amylase described above in the "Alpha-amylase"-section. In a preferred embodiment the alpha-amylase is of fungal origin. In a more preferred embodiment the alpha-amylase is an acid alpha-amylase.

[0117] In a preferred embodiment the carbohydrate-binding domain and/or linker sequence is of fungal origin. The carbohydrate-binding domain may be derived from an alpha-amylase, but may also be derived from of proteins, e.g., enzymes having glucoamylase activity.

[0118] In an embodiment the alpha-amylase is derived from a strain of Aspergillus, or Athelia. In an embodiment the alpha-amylase is derived from a strain of Aspergillus oryzae or Aspergillus niger. In a specific embodiment the alpha-amylase is the A. oryzae acid alpha-amylase disclosed in SEQ ID NO:39. In a specific embodiment the linker sequence may be derived from a strain of Aspergillus, such as the A. kawachii alpha-amylase (SEQ ID NO: 23) or the A. rolfsii glucoamylase (SEQ ID NO: 24). In an embodiment the CBD is derived from a strain of Aspergillus or Athelia. In a specific embodiment the CBD is the A. kawachii alpha-amylase shown in SEQ ID NO:1 or the A. rolfsii glucoamylase shown in SEQ ID NO:21.

[0119] Preferred is the embodiment wherein the hybrid enzyme comprises an alpha-amylase sequence derived from the A. niger acid alpha-amylase catalytic domain having the sequence shown in SEQ ID NO:38, and/or a linker sequence derived from the A. kawachii alpha-amylase shown in SEQ ID NO: 23 or the A. rolfsii glucoamylase shown in SEQ ID NO: 24, and/or the CBD is derived from the A. kawachii alpha-amylase shown in SEQ ID NO: 2, the A. rolfsii glucoamylase shown in SEQ ID NO: 21 or the A. niger glucoamylase shown in SEQ ID NO: 22.

[0120] In a preferred embodiment the hybrid enzyme comprises the A. niger acid alpha-amylase catalytic domain having the sequence shown in SEQ ID NO:38, the A. kawachii alpha-amylase linker shown in SEQ ID NO: 23, and A. kawachii alpha-amylase CBD shown in SEQ ID NO:2.

[0121] In a specific embodiment the hybrid enzyme is the mature part of the amino acid sequence shown in SEQ ID NO: 28 (A. niger acid alpha-amylase catalytic domain-A. kawachii alpha-amylase linker-A. niger glucoamylase CBD), SEQ ID NO:30 (A. niger acid alpha-amylase catalytic domain-A. kawachii alpha-amylase linker-A. rolfsii glucoamylase CBD), or SEQ ID NO:32 (A. oryzae acid alpha-amylase catalytic domain-A. kawachii alpha-amylase linker-A. kawachii alpha-amylase CBD), or SEQ ID NO:34 (A. niger acid alpha-amylase catalytic domain-A. rolfsii glucoamylase linker-A. roffsii glucoamylase CBD), or SEQ ID NO: 36 (A. oryzae acid alpha-amylase catalytic domain-A. rolfsii glucoamylase linker-A. rolfsii glucoamylase CBD) or the hybrid consisting of A. niger acid alpha-amylase catalytic domain (SEQ ID NOS: 4 or 38, respectively)-A. kawachii glucoamylase linker (SEQ ID NO: 23)-A. kawachi glucoamylase CBD (SEQ ID NO: 2) or a hybrid enzyme that has an amino acid sequence having at least 50%, 60%, 70%, 80% or even at least 90%, 95%, 96%, 97%, 98%, or 99% identity to any of the afore mentioned amino acid sequences.

[0122] In another preferred embodiment the hybrid enzyme has an amino acid sequence which differs from the amino acid sequence amino acid sequence shown in SEQ ID NO:28 (A. niger acid alpha-amylase catalytic domain-A. kawachii alpha-amylase linker-A. niger glucoamylase CBD), SEQ ID NO:30 (A. niger acid alpha-amylase catalytic domain-A. kawachii alpha-amylase linker-A. rolfsii glucoamylase CBD), SEQ ID NO:32 (A. oryzae acid alpha-amylase catalytic domain-A. kawachii alpha-amylase linker-A. kawachii alpha-amylase CBD), SEQ ID NO:34 (A. niger acid alpha-amylase catalytic domain-A. rolfsii glucoamylase linker-A. rolfsii glucoamylase CBD) or SEQ ID NO:36 (A. oryzae acid alpha-amylase catalytic domain-A. rolfsii glucoamylase linker-A. rolfsii glucoamylase CBD) or the hybrid consisting of A. niger acid alpha-amylase catalytic domain (SEQ ID NOS: 4 or 38, respectively)-A. kawachii glucoamylase linker (SEQ ID NO: 23)-A. kawachi glucoamylase CBD (SEQ ID NO: 2) in no more than 10 positions, no more than 9 positions, no more than 8 positions, no more than 7 positions, no more than 6 positions, no more than 5 positions, no more than 4 positions, no more than 3 positions, no more than 2 positions, or even no more than 1 position.

[0123] Preferably the hybrid enzyme comprises a CBD sequence having at least 50%, 60%, 70%, 80% or even at least 90%, 95%, 96%, 97%, 98%, or 99% identity to any of the amino acid sequences shown in SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:9, SEQ ID NO:10, SEQ ID NO:11, SEQ ID NO:12, SEQ ID NO:13, SEQ ID NO:14, SEQ ID NO:15, SEQ ID NO:16, SEQ ID NO:17, SEQ ID NO:18, SEQ ID NO:19, SEQ ID NO:20 or SEQ ID NO:21. Even more preferred the hybrid enzyme comprises a CBD sequence having an amino acid sequence shown in SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:9, SEQ ID NO:10, SEQ ID NO:11, SEQ ID NO:12, SEQ ID NO:13, SEQ ID NO:14, SEQ ID NO:15, SEQ ID NO:16, SEQ ID NO:17, SEQ ID NO:18, SEQ ID NO:19, SEQ ID NO:20 or SEQ ID NO:21. In yet another preferred embodiment the CBD sequence has an amino acid sequence which differs from the amino acid sequence amino acid sequence shown in SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:9, SEQ ID NO:10, SEQ ID NO:11, SEQ ID NO:12, SEQ ID NO:13, SEQ ID NO:14, SEQ ID NO:15, SEQ ID NO:16, SEQ ID NO:17, SEQ ID NO:18, SEQ ID NO:19, SEQ ID NO:20 or SEQ ID NO:21 in no more than 10 amino acid positions, no more than 9 positions, no more than 8 positions, no more than 7 positions, no more than 6 positions, no more than 5 positions, no more than 4 positions, no more than 3 positions, no more than 2 positions, or even no more than 1 position.

[0124] In a most preferred embodiment the hybrid enzyme comprises a CBD derived from a glucoamylase from A. rolfsii, such as the glucoamylase from A. rolfsii AHU 9627 disclosed in U.S. Pat. No. 4,727,026.

[0125] The invention described and claimed herein is not to be limited in scope by the specific embodiments herein disclosed, since these embodiments are intended as illustrations of several aspects of the invention. Any equivalent embodiments are intended to be within the scope of this invention. Indeed, various modifications of the invention in addition to those shown and described herein will become apparent to those skilled in the art from the foregoing description. Such modifications are also intended to fall within the scope of the appended claims. In the case of conflict, the present disclosure including definitions will control.

[0126] Various references are cited herein, the disclosures of which are incorporated by reference in their entireties.

Materials & Methods

Enzymes

[0127] Acid Alpha-Amylase A: Wild-type acid alpha-amylase derived from Aspergillus niger disclosed in SEQ ID NO: 38.

[0128] Acid Alpha-Amylase B: hybrid acid alpha-amylase consisting of A. niger acid alpha-amylase catalytic domain (SEQ ID NO: 38)-A. kawachii glucoamylase linker (SEQ ID NO: 23)-A. kawachii glucoamylase CBD (SEQ ID NO: 2).

[0129] Alpha-Amylase C: Hybrid alpha-amylase shown in SEQ ID NO: 44 comprising a catalytic domain (CD) from Rhizomucor pusillus alpha-amylase having a carbohydrate-binding domain (CBD) from the Athelia rolfsii glucoamylase.

[0130] Alpha-Amylase D: wild-type Rhizomucor pusillus alpha-amylase disclosed in SEQ ID NO: 43.

[0131] Enzyme classification numbers (EC numbers) referred to in the present specification with claims are in accordance with the Recommendations (1992) of the Nomenclature Committee of the International Union of Biochemistry and Molecular Biology, Academic Press Inc, 1992.

Fabric

[0132] Vlisco fabric from Vlisco BV, NL

Buffer

Citrate Buffer

[0133] 10 mM Citrate buffer (pH4.0) 1.376 g of Citric acid monohydrate and 1.015 g of Sodium Citrate dihydrate are dissolved in 1 L of de-ionized water.

Methods:

Determination of Homology/Identity

[0134] For purposes of the present invention, the degree of homology is determined as the degree of identity between two amino acid sequences as determined by the Clustal method (Higgins, 1989, CABIOS 5: 151-153) using the LASERGENE.TM. MEGALIGN.TM. software (DNASTAR, Inc., Madison, Wis.) with an identity table and the following multiple alignment parameters: Gap penalty of 10, and gap length penalty of 10. Pairwise alignment parameters were Ktuple=1, gap penalty=3, windows=5, and diago-nals=5].

Acid Alpha-Amylase Activity (AFAU Assay)

[0135] When used according to the present invention the activity of any acid alpha-amylase may be measured in AFAU (Acid Fungal Alpha-amylase Units), which are determined relative to an enzyme standard. 1 FAU is defined as the amount of enzyme which degrades 5.260 mg starch dry matter per hour under the below mentioned standard conditions.

[0136] Acid alpha-amylase, an endo-alpha-amylase (1,4-alpha-D-glucan-glucano-hydrolase, E.C. 3.2.1.1) hydrolyzes alpha-1,4-glucosidic bonds in the inner regions of the starch molecule to form dextrins and oligosaccharides with different chain lengths. The intensity of color formed with iodine is directly proportional to the concentration of starch. Amylase activity is determined using reverse colorimetry as a reduction in the concentration of starch under the specified analytical conditions.

##STR00005##

[0137] Standard Conditions/Reaction Conditions:

TABLE-US-00001 Substrate: Soluble starch, approx. 0.17 g/L Buffer: Citrate, approx. 0.03 M Iodine (I.sub.2): 0.03 g/L CaCl.sub.2: 1.85 mM pH: 2.50 .+-. 0.05 Incubation temperature: 40.degree. C. Reaction time: 23 seconds Wavelength: 590 nm Enzyme concentration: 0.025 AFAU/mL Enzyme working range: 0.01-0.04 AFAU/mL

[0138] A folder EB-SM-0259.02/01 describing this analytical method in more detail is available upon request to Novozymes A/S, Denmark, which folder is hereby included by reference.

Alpha-Amylase Activity (KNU)

[0139] The amylolytic activity may be determined using potato starch as substrate. This method is based on the break-down of modified potato starch by the enzyme, and the reaction is followed by mixing samples of the starch/enzyme solution with an iodine solution. Initially, a blackish-blue color is formed, but during the break-down of the starch the blue color gets weaker and gradually turns into a reddish-brown, which is compared to a colored glass standard.

[0140] One Kilo Novo alpha amylase Unit (KNU) is defined as the amount of enzyme which, under standard conditions (i.e. at 37.degree. C.+/-0.05; 0.0003 M Ca.sup.2+; and pH 5.6) dextrinizes 5260 mg starch dry substance Merck Amylum solubile.

[0141] A folder EB-SM-0009.02/01 describing this analytical method in more detail is available upon request to Novozymes A/S, Denmark, which folder is hereby included by reference.

Determination of Acid Amylolytic Activity (FAU)

[0142] One Fungal Alpha-Amylase Unit (1 FAU) is defined as the amount of enzyme, which breaks down 5.26 g starch (Merck Amylum solubile Erg. B.6, Batch 9947275) per hour at Novozymes' standard method for determination of alpha-amylase based upon the following standard conditions:

TABLE-US-00002 Substrate Soluble starch Temperature 37.degree. C. pH 4.7 Reaction time 7-20 minutes

A detailed description of Novozymes' method for determining KNU and FAU is available on request as standard method EB-SM-0009.02/01.

Desizing (Tegewa Method)

[0143] The starch size residue is determined visually by comparing an iodine stained fabric swatch to a standard set of photos with 1-9 scale where 1 is dark blue and 9 has no color stain. The iodine stain solution is made by dissolving 10 g KI in 10 ml water, add 0.635 g I.sub.2, and 200 mL ethanol in deionized water to make total 1 L solution. A fabric sample is cut and immersed in the iodine solution for 60 seconds and rinsed in deionized water for about 5 seconds. The fabric sample is rated by at least two professionals after excess water in the sample is pressed out. An average number is given. Method and standard scales obtainable from Verband TEGEWA, Karlstrasse 21, Frankfurt a.M., Germany.

EXAMPLES

Example 1

Desizing cotton fabric with wild-type acid Alpha-Amylase A

[0144] A 100% cotton fabric (270 g/m.sup.2) was from Bor{dot over (a)}s Wafveri Kungsfors AB, Sweden. It was made in 2003 with Cupper 3/1 construction. The fabric contained 28 thread/cm warp yarn and 14 thread/cm weft yarn. The warp yarn has Ne 11 and the weft has Ne 8. Both yarns were open end. The dry size pick up on the warp yarn was 8%. The size contained mainly Kollotex 5, Solvitose XO, and beef tallow wax with emulsifier. Kollotex 5 is a low viscous potato starch ester. Solvitose XO is a high viscous starch ether with DS about 0.07. Fabric swatches were cut to about 25 g each.

[0145] Three buffers were made for this study. Buffer pH2 was made by dissolving 11.53 g 85% phosphoric acid in 4.5 liter pure water, titrating with 5N HCl to pH 1.95, then adding water to 5 liter. Buffer pH3 was made by dissolving 11.53 g 85% phosphoric acid in 4.5 liter pure water, titrating with 5N NaOH to pH 2.95, then adding water to 10 liter. Buffer pH4 was made by dissolving 6.005 g acetic acid in 4.5 liter pure water, titrating with 5N HCl to pH 3.95, then adding water to 10 liter. A 2 g/l nonionic surfactant (wetting agent) was added in each buffer. The measured final pH values of these three buffers were 2, 2.99, and 3.98, respectively.

[0146] For each treatment, a 1 liter buffer solution was taken. A given amount of wild type Acid Alpha-Amylase A was added in the buffer. A fabric swatch was immersed in the solution for 30 seconds and then padded through a padder (Werner Mathis) to achieve 85% wet pickup. The fabric swatch was rolled and sealed in a plastic bag. It was then incubated at 50.degree. C. oven for 2 hours. The fabric swatch was washed in a pad-steam range (Werner Mathis) which contained four small rinsing boxes. The water temperature in rinsing boxes is 95, 95, 90, and 90.degree. C., respectively.

[0147] After drying overnight in air, the fabric swatch was stained with an iodine solution. The stained fabric sample was visually compared to TEGEWA standard photos with 1-9 scale where 1 is dark and 9 has no color stain. Thus higher number indicates a better starch removal. The visual evaluation was done by at least three professionals and an average TEGEWA value was given for each fabric sample. The results are shown in Table 1. The wild type acid Alpha-Amylase A treated fabric gave higher TEGEWA value than the fabric treated with no enzyme control at all three pH conditions, indicating that acid Alpha-Amylase A hydrolyzed and removed starch size on fabric. Increased enzyme activity resulted in higher TEGEWA indicating increased starch removal.

Example 2

Desizing Cotton Fabric with Acid Alpha-Amylase B

[0148] The same fabric swatch and buffers were prepared as in Example 1. Acid Alpha-Amylase B was different from the wild-type Acid Alpha-Amylase A used in Example 1 in that the enzyme protein was constructed to comprise the wild-type Acid Alpha-Amylase A from A. niger (Example 1) linked with a CBD from Aspergillus kawachii acid alpha-amylase. The enzyme used in this study has an activity of 316 AFAU/g. The same treatments and measurements were conducted as in Example 1. The results are shown in Table 1. Acid Alpha-Amylase B with CBD gave higher TEGEWA value than wild-type acid Alpha-Amylase A at the same activity at all conditions, indicating that Acid Alpha-Amylase B with CBD improves alpha-amylase desizing performance.

TABLE-US-00003 TABLE 1 [Enzyme] (AFAU/kg TEGEWA Value pH Enzyme type fabric) (average) 2 No enzyme 0 1.0 Acid Alpha-amylase A 45.4 2.5 Acid Alpha-Amylase B (with CBD) 45.4 2.8 3 No enzyme 0 1.0 Acid Alpha-Amylase A 11.4 1.7 45.4 2.3 Acid Alpha-Amylase B 11.4 2.0 (with CBD) 45.4 2.7 4 No enzyme 0 1.0 Acid Alpha-Amylase A 11.4 1.7 45.4 2.5 Acid Alpha-amylase B 11.4 1.8 (with CBD) 45.4 3.7

Example 3

[0149] The same fabric swatches were prepared as in Example 1. Buffer pH3 was made by dissolving 11.53 g 85% phosphoric acid in 4.5 liter pure water, titrating with 5N NaOH to pH 2.95, then adding water to 5 liter. After adding 2 g/l nonionic surfactant (a wetting agent) in the buffer, the buffer pH was measured as 3.05 at 25.degree. C. The same enzyme as in Example 1 was used.

[0150] The desizing treatment was conducted in a Lab-o-mat (Werner Mathis). A 250 mL buffer solution was added in each beaker. A given amount of alpha-amylase enzyme was added. One fabric swatch (25 g) was placed in each beaker. The beaker was closed and placed in the Lab-o-mat. Beakers were heated at 5.degree. C./min to 50.degree. C. by an infrared heating system equipped within the Lab-o-mat. Beakers were rotated at 30 rpm, 50.degree. C. for 45 minutes. After the enzyme treatment, the fabric swatch was sequentially washed with water in the same beaker three times at 95, 75, and 40.degree. C., respectively.

[0151] After dry overnight in air, the swatch was evaluated the same way as in Example 1. The results are shown in Table 2. The same conclusions in Example 1 can be drawn. A TEGEWA value of above 5 was achieved in lab equipment, indicating desizing at acidic condition is viable approach.

[0152] The residue of metal ions on fabric was also evaluated. The fabric was first cut through 1 mm sieve with a Thomas-Wiley mill. Fabric mash 4.00 (+/-0.01) g was mixed with 80 mL 1 g/L EDTA solution. The mixture was incubated at 70.degree. C. and 200 rpm in a shaker (new Brunswick Scientific Co. Inc, Series 25) for 15 hours. After cooled down for about 30 minutes, the mixture was centrifuged at 2500 rpm at 20.degree. C. for 10 minutes. The supernatant was collected for metal content analysis with a Perkinelmer atomic absorption spectrophometer.

Example 4

[0153] Essentially the same set of experiments was conducted in the same way as in the Example 3, except the alpha-amylase in Example 2 was used instead. The same experimental conditions and evaluation were carried out. The results are shown in Table 2. The same conclusions as in Example 2 can be drawn from this example. Much higher TEGEWA value was obtained in this experiment, and the highest value was 5.8.

TABLE-US-00004 TABLE 2 [Enzyme] (AFAU/kg TEGEWA Value Metal content (mg/L) Enzyme Type fabric) (average) Mn Fe No enzyme 0 1.3 0.23 3.91 Acid Alpha- 27.5 2.3 n/a n/a Amylase A 275 3.8 0.20 2.72 1100 5.2 n/a n/a Acid Alpha- 27.5 3.7 n/a n/a Amylase B 275 5.3 0.175 2.85 (with CBD) 1100 5.8 n/a n/a n/a = not measured

Example 5

[0154] The same type of fabric from Bor{dot over (a)}s Wafveri Kungsfors AB (Sweden) as in Example 1 was used and the swatch was cut to about 62 g each. Buffer pH4 was made by dissolving 12.01 g acetic acid in 4.5 liter pure water, titrating with 5N HCl to pH 3.95, then adding water to 10 liter. A 2 g/l nonionic surfactant (wetting agent) was added the buffer. The final pH value of the buffer was 3.99.

[0155] For each treatment, a 1.2 liter buffer solution was taken. A given amount of alpha-amylase enzyme was added in the buffer. A fabric swatch was immersed in the solution for 30 seconds and then padded through a padder (Werner Mathis) to achieve 90% wet pickup. The fabric swatch was cut into two equal size swatches. Both were rolled and sealed in plastic bags. One bag was incubated at 40.degree. C. oven and the other at room temperature (20.degree. C.). Both swatches were treated for 16 hours. The fabric swatch was washed in a pad-steam range (Werner Mathis) which contained four small rinsing boxes. The water temperature in rinsing boxes is 95, 95, 90, and 90.degree. C., respectively.

[0156] The same evaluation as in Example 1 was conducted. The results are shown in Table 3. Starch size on cotton fabric were hydrolyzed by alpha-amylase and removed during desizing process. At pH4, both amylase enzymes were able to perform desizing at both 20.degree. C. and 40.degree. C.

TABLE-US-00005 TABLE 3 Activity TEGEWA Enzyme (AFAU/kg fabric) 40.degree. C. 20.degree. C. control 0 1.5 1.8 Acid Alpha-Amylase A 17 3.2 3.3 (AKF0018) 56 4.8 4.0 111 4.8 4.7 Acid Alpha-Amylase B 17 3.0 3.0 (with CBD) 56 4.3 4.0 (AKP0001) 111 5.2 4.5

Example 6

Alpha-Amylase D for Desizing Fabric at pH 4.0

[0157] De-ionized water was filled into beakers, up to 13 cm from the bottom. The water temperature was adjusted to 60.degree. C. 200 ml of 10 mM Citrate buffer (pH 4.0) was added to each beaker and placed in a Lab-o-Mat. The solutions were heated to 60.degree. C. Different amounts of Alpha-Amylase D were added to the buffer. The final enzyme concentrations were 50 FAU/L, 100 FAU/L, 200 FAU/L and 400 FAU/L, respectively. Two swatches of Vlisco fabric were fixed in a pair of forceps and dipped in the impregnation bath for 30 seconds and then padded. The dipping and squeezing was repeated. The wet pick up was about 100%.

[0158] One swatch was placed in a two-layer plastic bag, the air was squeezed out and the bag was placed in water bath at 60.degree. C. for 2 hours. The other swatch was placed in a two-layer plastic bag, the air was squeezed out, and the bag was kept at room temperature for 24 hours. The samples were removed from the water bath after the required time was reached. The samples were fixed in the forceps, dipped in a hot rinse solution (90.degree. C.) for 30 seconds and squeezed. The dipping and squeezing was repeated twice.

[0159] The fabric was dipped in cold tap water for around 60 seconds and water was squeezed off with hands. The swatches were placed on a shelf and dried at room temperature. The residual starch content on the fabric was checked by TEGEWA rating. The result of the tests is shown in FIG. 1

Example 7

Alpha-Amylase C Desizing at pH 4.0

[0160] The desizing procedure described in Example 6 was repeated, except that Alpha-Amylase C was used. The result of the tests is shown in FIG. 2.

Sequence CWU 1

1

471396DNAAspergillus kawachiiCDS(1)..(396)A. kawachii CBD 1act agt aca tcc aaa gcc acc acc tcc tct tct tct tct tct gct gct 48Thr Ser Thr Ser Lys Ala Thr Thr Ser Ser Ser Ser Ser Ser Ala Ala1 5 10 15gct act act tct tca tca tgc acc gca aca agc acc acc ctc ccc atc 96Ala Thr Thr Ser Ser Ser Cys Thr Ala Thr Ser Thr Thr Leu Pro Ile 20 25 30acc ttc gaa gaa ctc gtc acc act acc tac ggg gaa gaa gtc tac ctc 144Thr Phe Glu Glu Leu Val Thr Thr Thr Tyr Gly Glu Glu Val Tyr Leu 35 40 45agc gga tct atc tcc cag ctc gga gag tgg gat acg agt gac gcg gtg 192Ser Gly Ser Ile Ser Gln Leu Gly Glu Trp Asp Thr Ser Asp Ala Val 50 55 60aag ttg tcc gcg gat gat tat acc tcg agt aac ccc gag tgg tct gtt 240Lys Leu Ser Ala Asp Asp Tyr Thr Ser Ser Asn Pro Glu Trp Ser Val65 70 75 80act gtg tcg ttg ccg gtg ggg acg acc ttc gag tat aag ttt att aag 288Thr Val Ser Leu Pro Val Gly Thr Thr Phe Glu Tyr Lys Phe Ile Lys 85 90 95gtc gat gag ggt gga agt gtg act tgg gaa agt gat ccg aat agg gag 336Val Asp Glu Gly Gly Ser Val Thr Trp Glu Ser Asp Pro Asn Arg Glu 100 105 110tat act gtg cct gaa tgt ggg aat ggg agt ggg gag acg gtg gtt gat 384Tyr Thr Val Pro Glu Cys Gly Asn Gly Ser Gly Glu Thr Val Val Asp 115 120 125acg tgg agg tag 396Thr Trp Arg 1302131PRTAspergillus kawachii 2Thr Ser Thr Ser Lys Ala Thr Thr Ser Ser Ser Ser Ser Ser Ala Ala1 5 10 15Ala Thr Thr Ser Ser Ser Cys Thr Ala Thr Ser Thr Thr Leu Pro Ile 20 25 30Thr Phe Glu Glu Leu Val Thr Thr Thr Tyr Gly Glu Glu Val Tyr Leu 35 40 45Ser Gly Ser Ile Ser Gln Leu Gly Glu Trp Asp Thr Ser Asp Ala Val 50 55 60Lys Leu Ser Ala Asp Asp Tyr Thr Ser Ser Asn Pro Glu Trp Ser Val65 70 75 80Thr Val Ser Leu Pro Val Gly Thr Thr Phe Glu Tyr Lys Phe Ile Lys 85 90 95Val Asp Glu Gly Gly Ser Val Thr Trp Glu Ser Asp Pro Asn Arg Glu 100 105 110Tyr Thr Val Pro Glu Cys Gly Asn Gly Ser Gly Glu Thr Val Val Asp 115 120 125Thr Trp Arg 13031533DNAAspergillus nigerCDS(1)..(1533)Aspergillus niger acid alpha-amylase 3atg aga tta tcg act tcg agt ctc ttc ctt tcc gtg tct ctg ctg ggg 48Met Arg Leu Ser Thr Ser Ser Leu Phe Leu Ser Val Ser Leu Leu Gly -20 -15 -10aag ctg gcc ctc ggg ctg tcg gct gca gaa tgg cgc act cag tcg att 96Lys Leu Ala Leu Gly Leu Ser Ala Ala Glu Trp Arg Thr Gln Ser Ile-5 -1 1 5 10tac ttc cta ttg acg gat cgg ttc ggt agg acg gac aat tcg acg aca 144Tyr Phe Leu Leu Thr Asp Arg Phe Gly Arg Thr Asp Asn Ser Thr Thr 15 20 25gct aca tgc gat acg ggt gac caa atc tat tgt ggt ggc agt tgg caa 192Ala Thr Cys Asp Thr Gly Asp Gln Ile Tyr Cys Gly Gly Ser Trp Gln 30 35 40gga atc atc aac cat ctg gat tat atc cag ggc atg gga ttc acg gcc 240Gly Ile Ile Asn His Leu Asp Tyr Ile Gln Gly Met Gly Phe Thr Ala 45 50 55atc tgg atc tcg cct atc act gaa cag ctg ccc cag gat act gct gat 288Ile Trp Ile Ser Pro Ile Thr Glu Gln Leu Pro Gln Asp Thr Ala Asp60 65 70 75ggt gaa gct tac cat gga tat tgg cag cag aag ata tac gac gtg aac 336Gly Glu Ala Tyr His Gly Tyr Trp Gln Gln Lys Ile Tyr Asp Val Asn 80 85 90tcc aac ttc ggc act gca gat gac ctc aag tcc ctc tca gat gcg ctt 384Ser Asn Phe Gly Thr Ala Asp Asp Leu Lys Ser Leu Ser Asp Ala Leu 95 100 105cat gcc cgc gga atg tac ctc atg gtg gac gtc gtc cct aac cac atg 432His Ala Arg Gly Met Tyr Leu Met Val Asp Val Val Pro Asn His Met 110 115 120ggc tac gcc ggc aac ggc aac gat gta gac tac agc gtc ttc gac ccc 480Gly Tyr Ala Gly Asn Gly Asn Asp Val Asp Tyr Ser Val Phe Asp Pro 125 130 135ttc gat tcc tcc tcc tac ttc cac cca tac tgc ctg atc aca gat tgg 528Phe Asp Ser Ser Ser Tyr Phe His Pro Tyr Cys Leu Ile Thr Asp Trp140 145 150 155gac aac ttg acc atg gtc caa gat tgt tgg gag ggt gac acc atc gta 576Asp Asn Leu Thr Met Val Gln Asp Cys Trp Glu Gly Asp Thr Ile Val 160 165 170tct ctg cca gac cta aac acc acc gaa act gcc gtg aga aca atc tgg 624Ser Leu Pro Asp Leu Asn Thr Thr Glu Thr Ala Val Arg Thr Ile Trp 175 180 185tat gac tgg gta gcc gac ctg gta tcc aat tat tca gtc gac gga ctc 672Tyr Asp Trp Val Ala Asp Leu Val Ser Asn Tyr Ser Val Asp Gly Leu 190 195 200cgc atc gac agt gtc ctc gaa gtc gaa cca gac ttc ttc ccg ggc tac 720Arg Ile Asp Ser Val Leu Glu Val Glu Pro Asp Phe Phe Pro Gly Tyr 205 210 215cag gaa gca gca ggt gtc tac tgc gtc ggc gaa gtc gac aac ggc aac 768Gln Glu Ala Ala Gly Val Tyr Cys Val Gly Glu Val Asp Asn Gly Asn220 225 230 235cct gcc ctc gac tgc cca tac cag aag gtc ctg gac ggc gtc ctc aac 816Pro Ala Leu Asp Cys Pro Tyr Gln Lys Val Leu Asp Gly Val Leu Asn 240 245 250tat ccg atc tac tgg caa ctc ctc tac gcc ttc gaa tcc tcc agc ggc 864Tyr Pro Ile Tyr Trp Gln Leu Leu Tyr Ala Phe Glu Ser Ser Ser Gly 255 260 265agc atc agc aat ctc tac aac atg atc aaa tcc gtc gca agc gac tgc 912Ser Ile Ser Asn Leu Tyr Asn Met Ile Lys Ser Val Ala Ser Asp Cys 270 275 280tcc gat ccg aca cta ctc ggc aac ttc atc gaa aac cac gac aat ccc 960Ser Asp Pro Thr Leu Leu Gly Asn Phe Ile Glu Asn His Asp Asn Pro 285 290 295cgt ttc gcc tcc tac acc tcc gac tac tcg caa gcc aaa aac gtc ctc 1008Arg Phe Ala Ser Tyr Thr Ser Asp Tyr Ser Gln Ala Lys Asn Val Leu300 305 310 315agc tac atc ttc ctc tcc gac ggc atc ccc atc gtc tac gcc ggc gaa 1056Ser Tyr Ile Phe Leu Ser Asp Gly Ile Pro Ile Val Tyr Ala Gly Glu 320 325 330gaa cag cac tac tcc ggc ggc aag gtg ccc tac aac cgc gaa gcg acc 1104Glu Gln His Tyr Ser Gly Gly Lys Val Pro Tyr Asn Arg Glu Ala Thr 335 340 345tgg ctt tca ggc tac gac acc tcc gca gag ctg tac acc tgg ata gcc 1152Trp Leu Ser Gly Tyr Asp Thr Ser Ala Glu Leu Tyr Thr Trp Ile Ala 350 355 360acc acg aac gcg atc cgc aaa cta gcc atc tca gct gac tcg gcc tac 1200Thr Thr Asn Ala Ile Arg Lys Leu Ala Ile Ser Ala Asp Ser Ala Tyr 365 370 375att acc tac gcg aat gat gca ttc tac act gac agc aac acc atc gca 1248Ile Thr Tyr Ala Asn Asp Ala Phe Tyr Thr Asp Ser Asn Thr Ile Ala380 385 390 395atg cgc aaa ggc acc tca ggg agc caa gtc atc acc gtc ctc tcc aac 1296Met Arg Lys Gly Thr Ser Gly Ser Gln Val Ile Thr Val Leu Ser Asn 400 405 410aaa ggc tcc tca gga agc agc tac acc ctg acc ctc agc gga agc ggc 1344Lys Gly Ser Ser Gly Ser Ser Tyr Thr Leu Thr Leu Ser Gly Ser Gly 415 420 425tac aca tcc ggc acg aag ctg atc gaa gcg tac aca tgc aca tcc gtg 1392Tyr Thr Ser Gly Thr Lys Leu Ile Glu Ala Tyr Thr Cys Thr Ser Val 430 435 440acc gtg gac tcg agc ggc gat att ccc gtg ccg atg gcg tcg gga tta 1440Thr Val Asp Ser Ser Gly Asp Ile Pro Val Pro Met Ala Ser Gly Leu 445 450 455ccg aga gtt ctt ctg ccc gcg tcc gtc gtc gat agc tct tcg ctc tgt 1488Pro Arg Val Leu Leu Pro Ala Ser Val Val Asp Ser Ser Ser Leu Cys460 465 470 475ggc ggg agc gga aga aca acc acg acc aca act gct gct act agt 1533Gly Gly Ser Gly Arg Thr Thr Thr Thr Thr Thr Ala Ala Thr Ser 480 485 4904511PRTAspergillus niger 4Met Arg Leu Ser Thr Ser Ser Leu Phe Leu Ser Val Ser Leu Leu Gly -20 -15 -10Lys Leu Ala Leu Gly Leu Ser Ala Ala Glu Trp Arg Thr Gln Ser Ile-5 -1 1 5 10Tyr Phe Leu Leu Thr Asp Arg Phe Gly Arg Thr Asp Asn Ser Thr Thr 15 20 25Ala Thr Cys Asp Thr Gly Asp Gln Ile Tyr Cys Gly Gly Ser Trp Gln 30 35 40Gly Ile Ile Asn His Leu Asp Tyr Ile Gln Gly Met Gly Phe Thr Ala 45 50 55Ile Trp Ile Ser Pro Ile Thr Glu Gln Leu Pro Gln Asp Thr Ala Asp60 65 70 75Gly Glu Ala Tyr His Gly Tyr Trp Gln Gln Lys Ile Tyr Asp Val Asn 80 85 90Ser Asn Phe Gly Thr Ala Asp Asp Leu Lys Ser Leu Ser Asp Ala Leu 95 100 105His Ala Arg Gly Met Tyr Leu Met Val Asp Val Val Pro Asn His Met 110 115 120Gly Tyr Ala Gly Asn Gly Asn Asp Val Asp Tyr Ser Val Phe Asp Pro 125 130 135Phe Asp Ser Ser Ser Tyr Phe His Pro Tyr Cys Leu Ile Thr Asp Trp140 145 150 155Asp Asn Leu Thr Met Val Gln Asp Cys Trp Glu Gly Asp Thr Ile Val 160 165 170Ser Leu Pro Asp Leu Asn Thr Thr Glu Thr Ala Val Arg Thr Ile Trp 175 180 185Tyr Asp Trp Val Ala Asp Leu Val Ser Asn Tyr Ser Val Asp Gly Leu 190 195 200Arg Ile Asp Ser Val Leu Glu Val Glu Pro Asp Phe Phe Pro Gly Tyr 205 210 215Gln Glu Ala Ala Gly Val Tyr Cys Val Gly Glu Val Asp Asn Gly Asn220 225 230 235Pro Ala Leu Asp Cys Pro Tyr Gln Lys Val Leu Asp Gly Val Leu Asn 240 245 250Tyr Pro Ile Tyr Trp Gln Leu Leu Tyr Ala Phe Glu Ser Ser Ser Gly 255 260 265Ser Ile Ser Asn Leu Tyr Asn Met Ile Lys Ser Val Ala Ser Asp Cys 270 275 280Ser Asp Pro Thr Leu Leu Gly Asn Phe Ile Glu Asn His Asp Asn Pro 285 290 295Arg Phe Ala Ser Tyr Thr Ser Asp Tyr Ser Gln Ala Lys Asn Val Leu300 305 310 315Ser Tyr Ile Phe Leu Ser Asp Gly Ile Pro Ile Val Tyr Ala Gly Glu 320 325 330Glu Gln His Tyr Ser Gly Gly Lys Val Pro Tyr Asn Arg Glu Ala Thr 335 340 345Trp Leu Ser Gly Tyr Asp Thr Ser Ala Glu Leu Tyr Thr Trp Ile Ala 350 355 360Thr Thr Asn Ala Ile Arg Lys Leu Ala Ile Ser Ala Asp Ser Ala Tyr 365 370 375Ile Thr Tyr Ala Asn Asp Ala Phe Tyr Thr Asp Ser Asn Thr Ile Ala380 385 390 395Met Arg Lys Gly Thr Ser Gly Ser Gln Val Ile Thr Val Leu Ser Asn 400 405 410Lys Gly Ser Ser Gly Ser Ser Tyr Thr Leu Thr Leu Ser Gly Ser Gly 415 420 425Tyr Thr Ser Gly Thr Lys Leu Ile Glu Ala Tyr Thr Cys Thr Ser Val 430 435 440Thr Val Asp Ser Ser Gly Asp Ile Pro Val Pro Met Ala Ser Gly Leu 445 450 455Pro Arg Val Leu Leu Pro Ala Ser Val Val Asp Ser Ser Ser Leu Cys460 465 470 475Gly Gly Ser Gly Arg Thr Thr Thr Thr Thr Thr Ala Ala Thr Ser 480 485 4905102PRTBacillus flavothermusDOMAIN(1)..(102)CBD 5Ile Ser Thr Thr Ser Gln Ile Thr Phe Thr Val Asn Asn Ala Thr Thr1 5 10 15Val Trp Gly Gln Asn Val Tyr Val Val Gly Asn Ile Ser Gln Leu Gly 20 25 30Asn Trp Asp Pro Val His Ala Val Gln Met Thr Pro Ser Ser Tyr Pro 35 40 45Thr Trp Thr Val Thr Ile Pro Leu Leu Gln Gly Gln Asn Ile Gln Phe 50 55 60Lys Phe Ile Lys Lys Asp Ser Ala Gly Asn Val Ile Trp Glu Asp Ile65 70 75 80Ser Asn Arg Thr Tyr Thr Val Pro Thr Ala Ala Ser Gly Ala Tyr Thr 85 90 95Ala Ser Trp Asn Val Pro 100699PRTBacillus speciesDOMAIN(1)..(99)CBD 6Thr Ser Asn Val Thr Phe Thr Val Asn Asn Ala Thr Thr Val Tyr Gly1 5 10 15Gln Asn Val Tyr Val Val Gly Asn Ile Pro Glu Leu Gly Asn Trp Asn 20 25 30Ile Ala Asn Ala Ile Gln Met Thr Pro Ser Ser Tyr Pro Thr Trp Lys 35 40 45Thr Thr Val Ser Leu Pro Gln Gly Lys Ala Ile Glu Phe Lys Phe Ile 50 55 60Lys Lys Asp Ser Ala Gly Asn Val Ile Trp Glu Asn Ile Ala Asn Arg65 70 75 80Thr Tyr Thr Val Pro Phe Ser Ser Thr Gly Ser Tyr Thr Ala Asn Trp 85 90 95Asn Val Pro7102PRTAlcaliphilic BacillusDOMAIN(1)..(102)CBD 7Thr Ser Thr Thr Ser Gln Ile Thr Phe Thr Val Asn Asn Ala Thr Thr1 5 10 15Val Trp Gly Gln Asn Val Tyr Val Val Gly Asn Ile Ser Gln Leu Gly 20 25 30Asn Trp Asp Pro Val Asn Ala Val Gln Met Thr Pro Ser Ser Tyr Pro 35 40 45Thr Trp Val Val Thr Val Pro Leu Pro Gln Ser Gln Asn Ile Gln Phe 50 55 60Lys Phe Ile Lys Lys Asp Gly Ser Gly Asn Val Ile Trp Glu Asn Ile65 70 75 80Ser Asn Arg Thr Tyr Thr Val Pro Thr Ala Ala Ser Gly Ala Tyr Thr 85 90 95Ala Asn Trp Asn Val Pro 1008112PRTHormoconis resinaeDOMAIN(1)..(112)CBD 8Cys Gln Val Ser Ile Thr Phe Asn Ile Asn Ala Thr Thr Tyr Tyr Gly1 5 10 15Glu Asn Leu Tyr Val Ile Gly Asn Ser Ser Asp Leu Gly Ala Trp Asn 20 25 30Ile Ala Asp Ala Tyr Pro Leu Ser Ala Ser Ala Tyr Thr Gln Asp Arg 35 40 45Pro Leu Trp Ser Ala Ala Ile Pro Leu Asn Ala Gly Glu Val Ile Ser 50 55 60Tyr Gln Tyr Val Arg Gln Glu Asp Cys Asp Gln Pro Tyr Ile Tyr Glu65 70 75 80Thr Val Asn Arg Thr Leu Thr Val Pro Ala Cys Gly Gly Ala Ala Val 85 90 95Thr Thr Asp Asp Ala Trp Met Gly Pro Val Gly Ser Ser Gly Asn Cys 100 105 110995PRTLentinula edodesDOMAIN(1)..(95)CBD 9Val Ser Val Thr Phe Asn Val Asp Ala Ser Thr Leu Glu Gly Gln Asn1 5 10 15Val Tyr Leu Thr Gly Ala Val Asp Ala Leu Glu Asp Trp Ser Thr Asp 20 25 30Asn Ala Ile Leu Leu Ser Ser Ala Asn Tyr Pro Thr Trp Ser Val Thr 35 40 45Val Asp Leu Pro Gly Ser Thr Asp Val Gln Tyr Lys Tyr Ile Lys Lys 50 55 60Asp Gly Ser Gly Thr Val Thr Trp Glu Ser Asp Pro Asn Met Glu Ile65 70 75 80Thr Thr Pro Ala Asn Gly Thr Tyr Ala Thr Asn Asp Thr Trp Arg 85 90 9510107PRTNeurospora crassaDOMAIN(1)..(107)CBD 10Cys Ala Ala Asp His Glu Val Leu Val Thr Phe Asn Glu Lys Val Thr1 5 10 15Thr Ser Tyr Gly Gln Thr Val Lys Val Val Gly Ser Ile Ala Ala Leu 20 25 30Gly Asn Trp Ala Pro Ala Ser Gly Val Thr Leu Ser Ala Lys Gln Tyr 35 40 45Ser Ser Ser Asn Pro Leu Trp Ser Thr Thr Ile Ala Leu Pro Gln Gly 50 55 60Thr Ser Phe Lys Tyr Lys Tyr Val Val Val Asn Ser Asp Gly Ser Val65 70 75 80Lys Trp Glu Asn Asp Pro Asp Arg Ser Tyr Ala Val Gly Thr Asp Cys 85 90 95Ala Ser Thr Ala Thr Leu Asp Asp Thr Trp Arg 100 10511115PRTTalaromyces byssochlamydioidesDOMAIN(1)..(115)CBD 11Thr Thr Thr Gly Ala Ala Pro Cys Thr Thr Pro Thr Thr Val Ala Val1 5 10 15Thr Phe Asp Glu Ile Val Thr Thr Thr Tyr Gly Glu Thr Val Tyr Leu 20 25 30Ser Gly Ser Ile Pro Ala Leu Gly Asn Trp Asp Thr Ser Ser Ala Ile 35 40 45Ala Leu Ser Ala Val Asp Tyr Thr Ser Ser Asn Pro Leu Trp Tyr Val 50 55 60Thr Val Asn Leu Pro Ala Gly Thr Ser Phe Glu Tyr Lys Phe Phe Val65 70 75 80Gln Gln Thr Asp Gly Thr Ile Val Trp Glu Asp Asp Pro Asn Arg Ser 85 90 95Tyr Thr Val Pro Ala Asn Cys Gly Gln Thr Thr Ala Ile

Ile Asp Asp 100 105 110Ser Trp Gln 11512115PRTGeosmithia cylindrosporaDOMAIN(1)..(115)CBD 12Thr Ser Thr Gly Ser Ala Pro Cys Thr Thr Pro Thr Thr Val Ala Val1 5 10 15Thr Phe Asp Glu Ile Val Thr Thr Ser Tyr Gly Glu Thr Val Tyr Leu 20 25 30Ala Gly Ser Ile Ala Ala Leu Gly Asn Trp Asp Thr Asn Ser Ala Ile 35 40 45Ala Leu Ser Ala Ala Asp Tyr Thr Ser Asn Asn Asn Leu Trp Tyr Val 50 55 60Thr Val Asn Leu Ala Ala Gly Thr Ser Phe Gln Tyr Lys Phe Phe Val65 70 75 80Lys Glu Thr Asp Ser Thr Ile Val Trp Glu Asp Asp Pro Asn Arg Ser 85 90 95Tyr Thr Val Pro Ala Asn Cys Gly Gln Thr Thr Ala Ile Ile Asp Asp 100 105 110Thr Trp Gln 11513139PRTScorias spongiosaDOMAIN(1)..(139)CBD 13Ala Lys Val Pro Ser Thr Cys Ser Ala Ser Ser Ala Thr Gly Thr Cys1 5 10 15Thr Thr Ala Thr Ser Thr Phe Gly Gly Ser Thr Pro Thr Thr Ser Cys 20 25 30Ala Thr Thr Pro Thr Leu Thr Thr Val Leu Phe Asn Glu Arg Ala Thr 35 40 45Thr Asn Phe Gly Gln Asn Val His Leu Thr Gly Ser Ile Ser Gln Leu 50 55 60Gly Ser Trp Asp Thr Asp Ser Ala Val Ala Leu Ser Ala Val Asn Tyr65 70 75 80Thr Ser Ser Asp Pro Leu Trp Phe Val Arg Val Gln Leu Pro Ala Gly 85 90 95Thr Ser Phe Gln Tyr Lys Tyr Phe Lys Lys Asp Ser Ser Asn Ala Val 100 105 110Ala Trp Glu Ser Asp Pro Asn Arg Ser Tyr Thr Val Pro Leu Asn Cys 115 120 125Ala Gly Thr Ala Thr Glu Asn Asp Thr Trp Arg 130 13514126PRTEupenicillium ludwigiiDOMAIN(1)..(126)CBD 14Ser Thr Thr Thr Thr Ser Thr Thr Lys Thr Thr Thr Thr Ser Thr Thr1 5 10 15Thr Ser Cys Thr Thr Pro Thr Ala Val Ala Val Thr Phe Asp Leu Ile 20 25 30Ala Thr Thr Tyr Tyr Gly Glu Asn Ile Lys Ile Ala Gly Ser Ile Ser 35 40 45Gln Leu Gly Asp Trp Asp Thr Ser Asn Ala Val Ala Leu Ser Ala Ala 50 55 60Asp Tyr Thr Ser Ser Asp His Leu Trp Phe Val Asp Ile Asp Leu Pro65 70 75 80Ala Gly Thr Val Phe Glu Tyr Lys Tyr Ile Arg Ile Glu Ser Asp Gly 85 90 95Ser Ile Glu Trp Glu Ser Asp Pro Asn Arg Ser Tyr Thr Val Pro Ala 100 105 110Ala Cys Ala Thr Thr Ala Val Thr Glu Asn Asp Thr Trp Arg 115 120 12515116PRTAspergillus japonicusDOMAIN(1)..(116)CBD 15Lys Thr Ser Thr Thr Thr Ser Ser Cys Ser Thr Pro Thr Ser Val Ala1 5 10 15Val Thr Phe Asp Val Ile Ala Thr Thr Thr Tyr Gly Glu Asn Val Tyr 20 25 30Ile Ser Gly Ser Ile Ser Gln Leu Gly Ser Trp Asp Thr Ser Ser Ala 35 40 45Ile Ala Leu Ser Ala Ser Gln Tyr Thr Ser Ser Asn Asn Leu Trp Tyr 50 55 60Ala Thr Val His Leu Pro Ala Gly Thr Thr Phe Gln Tyr Lys Tyr Ile65 70 75 80Arg Lys Glu Thr Asp Gly Ser Val Thr Trp Glu Ser Asp Pro Asn Arg 85 90 95Ser Tyr Thr Val Pro Ser Ser Cys Gly Val Ser Ser Ala Thr Glu Ser 100 105 110Asp Thr Trp Arg 11516133PRTPenicillium cf. miczynskiiDOMAIN(1)..(133)CBD 16Thr Thr Thr Gly Gly Thr Thr Thr Ser Gln Gly Ser Thr Thr Thr Thr1 5 10 15Ser Lys Thr Ser Thr Thr Thr Ser Ser Cys Thr Ala Pro Thr Ser Val 20 25 30Ala Val Thr Phe Asp Leu Ile Ala Thr Thr Val Tyr Asp Glu Asn Val 35 40 45Gln Leu Ala Gly Ser Ile Ser Ala Leu Gly Ser Trp Asp Thr Ser Ser 50 55 60Ala Ile Arg Leu Ser Ala Ser Gln Tyr Thr Ser Ser Asn His Leu Trp65 70 75 80Tyr Val Ala Val Ser Leu Pro Ala Gly Gln Val Phe Gln Tyr Lys Tyr 85 90 95Ile Arg Val Ala Ser Ser Gly Thr Ile Thr Trp Glu Ser Asp Pro Asn 100 105 110Leu Ser Tyr Thr Val Pro Val Ala Cys Ala Ala Thr Ala Val Thr Ile 115 120 125Ser Asp Thr Trp Arg 13017116PRTMz1 Penicillium speciesDOMAIN(1)..(116)CBD 17Thr Lys Thr Ser Thr Ser Thr Ser Cys Thr Thr Pro Thr Ala Val Ala1 5 10 15Val Thr Phe Asp Leu Ile Ala Thr Thr Thr Tyr Gly Glu Asn Ile Lys 20 25 30Ile Ala Gly Ser Ile Ala Ala Leu Gly Ala Trp Asp Thr Asp Asp Ala 35 40 45Val Ala Leu Ser Ala Ala Asp Tyr Thr Asp Ser Asp His Leu Trp Phe 50 55 60Val Thr Gln Ser Ile Pro Ala Gly Thr Val Phe Glu Tyr Lys Tyr Ile65 70 75 80Arg Val Glu Ser Asp Gly Thr Ile Glu Trp Glu Ser Asp Pro Asn Arg 85 90 95Ser Tyr Thr Val Pro Ala Ala Cys Ala Thr Thr Ala Val Thr Glu Ser 100 105 110Asp Thr Trp Arg 11518114PRTThysanophora speciesDOMAIN(1)..(114)CBD 18Phe Thr Ser Thr Thr Lys Thr Ser Cys Thr Thr Pro Thr Ser Val Ala1 5 10 15Val Thr Phe Asp Leu Ile Ala Thr Thr Thr Tyr Gly Glu Ser Ile Arg 20 25 30Leu Val Gly Ser Ile Ser Glu Leu Gly Asp Trp Asp Thr Gly Ser Ala 35 40 45Ile Ala Leu His Ala Thr Asp Tyr Thr Asp Ser Asp His Leu Trp Phe 50 55 60Val Thr Val Gly Leu Pro Ala Gly Ala Ser Phe Glu Tyr Lys Tyr Ile65 70 75 80Arg Val Glu Ser Ser Gly Thr Ile Glu Trp Glu Ser Asp Pro Asn Arg 85 90 95Ser Tyr Thr Val Pro Ala Ala Cys Ala Thr Thr Ala Val Thr Glu Ser 100 105 110Asp Thr19111PRTHumicola grisea var. thermoideaDOMAIN(1)..(111)CBD 19Ala Asp Ala Ser Glu Val Tyr Val Thr Phe Asn Glu Arg Val Ser Thr1 5 10 15Ala Trp Gly Glu Thr Ile Lys Val Val Gly Asn Val Pro Ala Leu Gly 20 25 30Asn Trp Asp Thr Ser Lys Ala Val Thr Leu Ser Ala Ser Gly Tyr Lys 35 40 45Ser Asn Asp Pro Leu Trp Ser Ile Thr Val Pro Ile Lys Ala Thr Gly 50 55 60Ser Ala Val Gln Tyr Lys Tyr Ile Lys Val Gly Thr Asn Gly Lys Ile65 70 75 80Thr Trp Glu Ser Asp Pro Asn Arg Ser Ile Thr Leu Gln Thr Ala Ser 85 90 95Ser Ala Gly Lys Cys Ala Ala Gln Thr Val Asn Asp Ser Trp Arg 100 105 11020108PRTAspergillus nigerDOMAIN(1)..(108)CBD 20Cys Thr Thr Pro Thr Ala Val Ala Val Thr Phe Asp Leu Thr Ala Thr1 5 10 15Thr Thr Tyr Gly Glu Asn Ile Tyr Leu Val Gly Ser Ile Ser Gln Leu 20 25 30Gly Asp Trp Glu Thr Ser Asp Gly Ile Ala Leu Ser Ala Asp Lys Tyr 35 40 45Thr Ser Ser Asp Pro Leu Trp Tyr Val Thr Val Thr Leu Pro Ala Gly 50 55 60Glu Ser Phe Glu Tyr Lys Phe Ile Arg Ile Glu Ser Asp Asp Ser Val65 70 75 80Glu Trp Glu Ser Asp Pro Asn Arg Glu Tyr Thr Val Pro Gln Ala Cys 85 90 95Gly Thr Ser Thr Ala Thr Val Thr Asp Thr Trp Arg 100 1052197PRTAlthea rolfsiiDOMAIN(1)..(97)CBD 21Val Glu Val Thr Phe Asp Val Tyr Ala Thr Thr Val Tyr Gly Gln Asn1 5 10 15Ile Tyr Ile Thr Gly Asp Val Ser Glu Leu Gly Asn Trp Thr Pro Ala 20 25 30Asn Gly Val Ala Leu Ser Ser Ala Asn Tyr Pro Thr Trp Ser Ala Thr 35 40 45Ile Ala Leu Pro Ala Asp Thr Thr Ile Gln Tyr Lys Tyr Val Asn Ile 50 55 60Asp Gly Ser Thr Val Ile Trp Glu Asp Ala Ile Ser Asn Arg Glu Ile65 70 75 80Thr Thr Pro Ala Ser Gly Thr Tyr Thr Glu Lys Asp Thr Trp Asp Glu 85 90 95Ser 2238PRTAspergillus nigerDOMAIN(1)..(38)Linker 22Thr Gly Gly Thr Thr Thr Thr Ala Thr Pro Thr Gly Ser Gly Ser Val1 5 10 15Thr Ser Thr Ser Lys Thr Thr Ala Thr Ala Ser Lys Thr Ser Thr Ser 20 25 30Thr Ser Ser Thr Ser Ala 352331PRTAspergillus kawachiiDOMAIN(1)..(31)Linker 23Thr Thr Thr Thr Thr Thr Ala Ala Ala Thr Ser Thr Ser Lys Ala Thr1 5 10 15Thr Ser Ser Ser Ser Ser Ser Ala Ala Ala Thr Thr Ser Ser Ser 20 25 302411PRTAthelia rolfsiiDOMAIN(1)..(11)Linker 24Gly Ala Thr Ser Pro Gly Gly Ser Ser Gly Ser1 5 10258PRTArtificial SequenceSynthetic construct 25Pro Glu Pro Thr Pro Glu Pro Thr1 526498PRTAspergillus oryzaeSIGNAL(1)..(20)mat_peptide(20)..(498) 26Met Val Ala Trp Trp Ser Leu Phe Leu Tyr Gly Leu Gln Val Ala Ala -15 -10 -5Pro Ala Leu Ala Ala Thr Pro Ala Asp Trp Arg Ser Gln Ser Ile Tyr -1 1 5 10Phe Leu Leu Thr Asp Arg Phe Ala Arg Thr Asp Gly Ser Thr Thr Ala 15 20 25Thr Cys Asn Thr Ala Asp Gln Lys Tyr Cys Gly Gly Thr Trp Gln Gly30 35 40 45Ile Ile Asp Lys Leu Asp Tyr Ile Gln Gly Met Gly Phe Thr Ala Ile 50 55 60Trp Ile Thr Pro Val Thr Ala Gln Leu Pro Gln Thr Thr Ala Tyr Gly 65 70 75Asp Ala Tyr His Gly Tyr Trp Gln Gln Asp Ile Tyr Ser Leu Asn Glu 80 85 90Asn Tyr Gly Thr Ala Asp Asp Leu Lys Ala Leu Ser Ser Ala Leu His 95 100 105Glu Arg Gly Met Tyr Leu Met Val Asp Val Val Ala Asn His Met Gly110 115 120 125Tyr Asp Gly Ala Gly Ser Ser Val Asp Tyr Ser Val Phe Lys Pro Phe 130 135 140Ser Ser Gln Asp Tyr Phe His Pro Phe Cys Phe Ile Gln Asn Tyr Glu 145 150 155Asp Gln Thr Gln Val Glu Asp Cys Trp Leu Gly Asp Asn Thr Val Ser 160 165 170Leu Pro Asp Leu Asp Thr Thr Lys Asp Val Val Lys Asn Glu Trp Tyr 175 180 185Asp Trp Val Gly Ser Leu Val Ser Asn Tyr Ser Ile Asp Gly Leu Arg190 195 200 205Ile Asp Thr Val Lys His Val Gln Lys Asp Phe Trp Pro Gly Tyr Asn 210 215 220Lys Ala Ala Gly Val Tyr Cys Ile Gly Glu Val Leu Asp Gly Asp Pro 225 230 235Ala Tyr Thr Cys Pro Tyr Gln Asn Val Met Asp Gly Val Leu Asn Tyr 240 245 250Pro Ile Tyr Tyr Pro Leu Leu Asn Ala Phe Lys Ser Thr Ser Gly Ser 255 260 265Met Asp Asp Leu Tyr Asn Met Ile Asn Thr Val Lys Ser Asp Cys Pro270 275 280 285Asp Ser Thr Leu Leu Gly Thr Phe Val Glu Asn His Asp Asn Pro Arg 290 295 300Phe Ala Ser Tyr Thr Asn Asp Ile Ala Leu Ala Lys Asn Val Ala Ala 305 310 315Phe Ile Ile Leu Asn Asp Gly Ile Pro Ile Ile Tyr Ala Gly Gln Glu 320 325 330Gln His Tyr Ala Gly Gly Asn Asp Pro Ala Asn Arg Glu Ala Thr Trp 335 340 345Leu Ser Gly Tyr Pro Thr Asp Ser Glu Leu Tyr Lys Leu Ile Ala Ser350 355 360 365Ala Asn Ala Ile Arg Asn Tyr Ala Ile Ser Lys Asp Thr Gly Phe Val 370 375 380Thr Tyr Lys Asn Trp Pro Ile Tyr Lys Asp Asp Thr Thr Ile Ala Met 385 390 395Arg Lys Gly Thr Asp Gly Ser Gln Ile Val Thr Ile Leu Ser Asn Lys 400 405 410Gly Ala Ser Gly Asp Ser Tyr Thr Leu Ser Leu Ser Gly Ala Gly Tyr 415 420 425Thr Ala Gly Gln Gln Leu Thr Glu Val Ile Gly Cys Thr Thr Val Thr430 435 440 445Val Gly Ser Asp Gly Asn Val Pro Val Pro Met Ala Gly Gly Leu Pro 450 455 460Arg Val Leu Tyr Pro Thr Glu Lys Leu Ala Gly Ser Lys Ile Cys Ser 465 470 475Ser Ser271860DNAArtificial sequenceSynthetic construct 27ctg tcg gct gca gaa tgg cgc act cag tcg att tac ttc cta ttg acg 48Leu Ser Ala Ala Glu Trp Arg Thr Gln Ser Ile Tyr Phe Leu Leu Thr1 5 10 15gat cgg ttc ggt agg acg gac aat tcg acg aca gct aca tgc gat acg 96Asp Arg Phe Gly Arg Thr Asp Asn Ser Thr Thr Ala Thr Cys Asp Thr 20 25 30ggt gac caa atc tat tgt ggt ggc agt tgg caa gga atc atc aac cat 144Gly Asp Gln Ile Tyr Cys Gly Gly Ser Trp Gln Gly Ile Ile Asn His 35 40 45ctg gat tat atc cag ggc atg gga ttc acg gcc atc tgg atc tcg cct 192Leu Asp Tyr Ile Gln Gly Met Gly Phe Thr Ala Ile Trp Ile Ser Pro 50 55 60atc act gaa cag ctg ccc cag gat act gct gat ggt gaa gct tac cat 240Ile Thr Glu Gln Leu Pro Gln Asp Thr Ala Asp Gly Glu Ala Tyr His65 70 75 80gga tat tgg cag cag aag ata tac gac gtg aac tcc aac ttc ggc act 288Gly Tyr Trp Gln Gln Lys Ile Tyr Asp Val Asn Ser Asn Phe Gly Thr 85 90 95gca gat gac ctc aag tcc ctc tca gat gcg ctt cat gcc cgc gga atg 336Ala Asp Asp Leu Lys Ser Leu Ser Asp Ala Leu His Ala Arg Gly Met 100 105 110tac ctc atg gtg gac gtc gtc cct aac cac atg ggc tac gcc ggc aac 384Tyr Leu Met Val Asp Val Val Pro Asn His Met Gly Tyr Ala Gly Asn 115 120 125ggc aac gat gta gac tac agc gtc ttc gac ccc ttc gat tcc tcc tcc 432Gly Asn Asp Val Asp Tyr Ser Val Phe Asp Pro Phe Asp Ser Ser Ser 130 135 140tac ttc cac cca tac tgc ctg atc aca gat tgg gac aac ttg acc atg 480Tyr Phe His Pro Tyr Cys Leu Ile Thr Asp Trp Asp Asn Leu Thr Met145 150 155 160gtc caa gat tgt tgg gag ggt gac acc atc gta tct ctg cca gac cta 528Val Gln Asp Cys Trp Glu Gly Asp Thr Ile Val Ser Leu Pro Asp Leu 165 170 175aac acc acc gaa act gcc gtg aga aca atc tgg tat gac tgg gta gcc 576Asn Thr Thr Glu Thr Ala Val Arg Thr Ile Trp Tyr Asp Trp Val Ala 180 185 190gac ctg gta tcc aat tat tca gtc gac gga ctc cgc atc gac agt gtc 624Asp Leu Val Ser Asn Tyr Ser Val Asp Gly Leu Arg Ile Asp Ser Val 195 200 205ctc gaa gtc gaa cca gac ttc ttc ccg ggc tac cag gaa gca gca ggt 672Leu Glu Val Glu Pro Asp Phe Phe Pro Gly Tyr Gln Glu Ala Ala Gly 210 215 220gtc tac tgc gtc ggc gaa gtc gac aac ggc aac cct gcc ctc gac tgc 720Val Tyr Cys Val Gly Glu Val Asp Asn Gly Asn Pro Ala Leu Asp Cys225 230 235 240cca tac cag aag gtc ctg gac ggc gtc ctc aac tat ccg atc tac tgg 768Pro Tyr Gln Lys Val Leu Asp Gly Val Leu Asn Tyr Pro Ile Tyr Trp 245 250 255caa ctc ctc tac gcc ttc gaa tcc tcc agc ggc agc atc agc aat ctc 816Gln Leu Leu Tyr Ala Phe Glu Ser Ser Ser Gly Ser Ile Ser Asn Leu 260 265 270tac aac atg atc aaa tcc gtc gca agc gac tgc tcc gat ccg aca cta 864Tyr Asn Met Ile Lys Ser Val Ala Ser Asp Cys Ser Asp Pro Thr Leu 275 280 285ctc ggc aac ttc atc gaa aac cac gac aat ccc cgt ttc gcc tcc tac 912Leu Gly Asn Phe Ile Glu Asn His Asp Asn Pro Arg Phe Ala Ser Tyr 290 295 300acc tcc gac tac tcg caa gcc aaa aac gtc ctc agc tac atc ttc ctc 960Thr Ser Asp Tyr Ser Gln Ala Lys Asn Val Leu Ser Tyr Ile Phe Leu305 310 315 320tcc gac ggc atc ccc atc gtc tac gcc ggc gaa gaa cag cac tac tcc 1008Ser Asp Gly Ile Pro Ile Val Tyr Ala Gly Glu Glu Gln His Tyr Ser 325 330 335ggc ggc aag gtg ccc tac aac cgc gaa gcg acc tgg ctt tca ggc tac 1056Gly Gly Lys Val Pro Tyr Asn Arg Glu Ala Thr Trp Leu Ser Gly Tyr 340 345 350gac acc tcc gca gag ctg tac acc tgg ata gcc acc acg aac gcg atc 1104Asp Thr Ser Ala Glu Leu Tyr Thr Trp Ile Ala Thr Thr Asn Ala Ile 355 360 365cgc aaa

cta gcc atc tca gct gac tcg gcc tac att acc tac gcg aat 1152Arg Lys Leu Ala Ile Ser Ala Asp Ser Ala Tyr Ile Thr Tyr Ala Asn 370 375 380gat gca ttc tac act gac agc aac acc atc gca atg cgc aaa ggc acc 1200Asp Ala Phe Tyr Thr Asp Ser Asn Thr Ile Ala Met Arg Lys Gly Thr385 390 395 400tca ggg agc caa gtc atc acc gtc ctc tcc aac aaa ggc tcc tca gga 1248Ser Gly Ser Gln Val Ile Thr Val Leu Ser Asn Lys Gly Ser Ser Gly 405 410 415agc agc tac acc ctg acc ctc agc gga agc ggc tac aca tcc ggc acg 1296Ser Ser Tyr Thr Leu Thr Leu Ser Gly Ser Gly Tyr Thr Ser Gly Thr 420 425 430aag ctg atc gaa gcg tac aca tgc aca tcc gtg acc gtg gac tcg agc 1344Lys Leu Ile Glu Ala Tyr Thr Cys Thr Ser Val Thr Val Asp Ser Ser 435 440 445ggc gat att ccc gtg ccg atg gcg tcg gga tta ccg aga gtt ctt ctg 1392Gly Asp Ile Pro Val Pro Met Ala Ser Gly Leu Pro Arg Val Leu Leu 450 455 460ccc gcg tcc gtc gtc gat agc tct tcg ctc tgt ggc ggg agc gga aga 1440Pro Ala Ser Val Val Asp Ser Ser Ser Leu Cys Gly Gly Ser Gly Arg465 470 475 480aca acc acg acc aca act gct gct gct act agt aca tcc aaa gcc acc 1488Thr Thr Thr Thr Thr Thr Ala Ala Ala Thr Ser Thr Ser Lys Ala Thr 485 490 495acc tcc tct tct tct tct tct gct gct gct act act tct tca tca tgt 1536Thr Ser Ser Ser Ser Ser Ser Ala Ala Ala Thr Thr Ser Ser Ser Cys 500 505 510acc act ccc acc gcc gtg gct gtg act ttc gat ctg aca gct acc acc 1584Thr Thr Pro Thr Ala Val Ala Val Thr Phe Asp Leu Thr Ala Thr Thr 515 520 525acc tac ggc gag aac atc tac ctg gtc gga tcg atc tct cag ctg ggt 1632Thr Tyr Gly Glu Asn Ile Tyr Leu Val Gly Ser Ile Ser Gln Leu Gly 530 535 540gac tgg gaa acc agc gac ggc ata gct ctg agt gct gac aag tac act 1680Asp Trp Glu Thr Ser Asp Gly Ile Ala Leu Ser Ala Asp Lys Tyr Thr545 550 555 560tcc agc gac ccg ctc tgg tat gtc act gtg act ctg ccg gct ggt gag 1728Ser Ser Asp Pro Leu Trp Tyr Val Thr Val Thr Leu Pro Ala Gly Glu 565 570 575tcg ttt gag tac aag ttt atc cgc att gag agc gat gac tcc gtg gag 1776Ser Phe Glu Tyr Lys Phe Ile Arg Ile Glu Ser Asp Asp Ser Val Glu 580 585 590tgg gag agt gat ccc aac cga gaa tac acc gtt cct cag gcg tgc gga 1824Trp Glu Ser Asp Pro Asn Arg Glu Tyr Thr Val Pro Gln Ala Cys Gly 595 600 605acg tcg acc gcg acg gtg act gac acc tgg cgg tag 1860Thr Ser Thr Ala Thr Val Thr Asp Thr Trp Arg 610 61528619PRTArtificial sequenceSynthetic Construct 28Leu Ser Ala Ala Glu Trp Arg Thr Gln Ser Ile Tyr Phe Leu Leu Thr1 5 10 15Asp Arg Phe Gly Arg Thr Asp Asn Ser Thr Thr Ala Thr Cys Asp Thr 20 25 30Gly Asp Gln Ile Tyr Cys Gly Gly Ser Trp Gln Gly Ile Ile Asn His 35 40 45Leu Asp Tyr Ile Gln Gly Met Gly Phe Thr Ala Ile Trp Ile Ser Pro 50 55 60Ile Thr Glu Gln Leu Pro Gln Asp Thr Ala Asp Gly Glu Ala Tyr His65 70 75 80Gly Tyr Trp Gln Gln Lys Ile Tyr Asp Val Asn Ser Asn Phe Gly Thr 85 90 95Ala Asp Asp Leu Lys Ser Leu Ser Asp Ala Leu His Ala Arg Gly Met 100 105 110Tyr Leu Met Val Asp Val Val Pro Asn His Met Gly Tyr Ala Gly Asn 115 120 125Gly Asn Asp Val Asp Tyr Ser Val Phe Asp Pro Phe Asp Ser Ser Ser 130 135 140Tyr Phe His Pro Tyr Cys Leu Ile Thr Asp Trp Asp Asn Leu Thr Met145 150 155 160Val Gln Asp Cys Trp Glu Gly Asp Thr Ile Val Ser Leu Pro Asp Leu 165 170 175Asn Thr Thr Glu Thr Ala Val Arg Thr Ile Trp Tyr Asp Trp Val Ala 180 185 190Asp Leu Val Ser Asn Tyr Ser Val Asp Gly Leu Arg Ile Asp Ser Val 195 200 205Leu Glu Val Glu Pro Asp Phe Phe Pro Gly Tyr Gln Glu Ala Ala Gly 210 215 220Val Tyr Cys Val Gly Glu Val Asp Asn Gly Asn Pro Ala Leu Asp Cys225 230 235 240Pro Tyr Gln Lys Val Leu Asp Gly Val Leu Asn Tyr Pro Ile Tyr Trp 245 250 255Gln Leu Leu Tyr Ala Phe Glu Ser Ser Ser Gly Ser Ile Ser Asn Leu 260 265 270Tyr Asn Met Ile Lys Ser Val Ala Ser Asp Cys Ser Asp Pro Thr Leu 275 280 285Leu Gly Asn Phe Ile Glu Asn His Asp Asn Pro Arg Phe Ala Ser Tyr 290 295 300Thr Ser Asp Tyr Ser Gln Ala Lys Asn Val Leu Ser Tyr Ile Phe Leu305 310 315 320Ser Asp Gly Ile Pro Ile Val Tyr Ala Gly Glu Glu Gln His Tyr Ser 325 330 335Gly Gly Lys Val Pro Tyr Asn Arg Glu Ala Thr Trp Leu Ser Gly Tyr 340 345 350Asp Thr Ser Ala Glu Leu Tyr Thr Trp Ile Ala Thr Thr Asn Ala Ile 355 360 365Arg Lys Leu Ala Ile Ser Ala Asp Ser Ala Tyr Ile Thr Tyr Ala Asn 370 375 380Asp Ala Phe Tyr Thr Asp Ser Asn Thr Ile Ala Met Arg Lys Gly Thr385 390 395 400Ser Gly Ser Gln Val Ile Thr Val Leu Ser Asn Lys Gly Ser Ser Gly 405 410 415Ser Ser Tyr Thr Leu Thr Leu Ser Gly Ser Gly Tyr Thr Ser Gly Thr 420 425 430Lys Leu Ile Glu Ala Tyr Thr Cys Thr Ser Val Thr Val Asp Ser Ser 435 440 445Gly Asp Ile Pro Val Pro Met Ala Ser Gly Leu Pro Arg Val Leu Leu 450 455 460Pro Ala Ser Val Val Asp Ser Ser Ser Leu Cys Gly Gly Ser Gly Arg465 470 475 480Thr Thr Thr Thr Thr Thr Ala Ala Ala Thr Ser Thr Ser Lys Ala Thr 485 490 495Thr Ser Ser Ser Ser Ser Ser Ala Ala Ala Thr Thr Ser Ser Ser Cys 500 505 510Thr Thr Pro Thr Ala Val Ala Val Thr Phe Asp Leu Thr Ala Thr Thr 515 520 525Thr Tyr Gly Glu Asn Ile Tyr Leu Val Gly Ser Ile Ser Gln Leu Gly 530 535 540Asp Trp Glu Thr Ser Asp Gly Ile Ala Leu Ser Ala Asp Lys Tyr Thr545 550 555 560Ser Ser Asp Pro Leu Trp Tyr Val Thr Val Thr Leu Pro Ala Gly Glu 565 570 575Ser Phe Glu Tyr Lys Phe Ile Arg Ile Glu Ser Asp Asp Ser Val Glu 580 585 590Trp Glu Ser Asp Pro Asn Arg Glu Tyr Thr Val Pro Gln Ala Cys Gly 595 600 605Thr Ser Thr Ala Thr Val Thr Asp Thr Trp Arg 610 615291827DNAArtificial sequenceSynthetic construct 29ctg tcg gct gca gaa tgg cgc act cag tcg att tac ttc cta ttg acg 48Leu Ser Ala Ala Glu Trp Arg Thr Gln Ser Ile Tyr Phe Leu Leu Thr1 5 10 15gat cgg ttc ggt agg acg gac aat tcg acg aca gct aca tgc gat acg 96Asp Arg Phe Gly Arg Thr Asp Asn Ser Thr Thr Ala Thr Cys Asp Thr 20 25 30ggt gac caa atc tat tgt ggt ggc agt tgg caa gga atc atc aac cat 144Gly Asp Gln Ile Tyr Cys Gly Gly Ser Trp Gln Gly Ile Ile Asn His 35 40 45ctg gat tat atc cag ggc atg gga ttc acg gcc atc tgg atc tcg cct 192Leu Asp Tyr Ile Gln Gly Met Gly Phe Thr Ala Ile Trp Ile Ser Pro 50 55 60atc act gaa cag ctg ccc cag gat act gct gat ggt gaa gct tac cat 240Ile Thr Glu Gln Leu Pro Gln Asp Thr Ala Asp Gly Glu Ala Tyr His65 70 75 80gga tat tgg cag cag aag ata tac gac gtg aac tcc aac ttc ggc act 288Gly Tyr Trp Gln Gln Lys Ile Tyr Asp Val Asn Ser Asn Phe Gly Thr 85 90 95gca gat gac ctc aag tcc ctc tca gat gcg ctt cat gcc cgc gga atg 336Ala Asp Asp Leu Lys Ser Leu Ser Asp Ala Leu His Ala Arg Gly Met 100 105 110tac ctc atg gtg gac gtc gtc cct aac cac atg ggc tac gcc ggc aac 384Tyr Leu Met Val Asp Val Val Pro Asn His Met Gly Tyr Ala Gly Asn 115 120 125ggc aac gat gta gac tac agc gtc ttc gac ccc ttc gat tcc tcc tcc 432Gly Asn Asp Val Asp Tyr Ser Val Phe Asp Pro Phe Asp Ser Ser Ser 130 135 140tac ttc cac cca tac tgc ctg atc aca gat tgg gac aac ttg acc atg 480Tyr Phe His Pro Tyr Cys Leu Ile Thr Asp Trp Asp Asn Leu Thr Met145 150 155 160gtc caa gat tgt tgg gag ggt gac acc atc gta tct ctg cca gac cta 528Val Gln Asp Cys Trp Glu Gly Asp Thr Ile Val Ser Leu Pro Asp Leu 165 170 175aac acc acc gaa act gcc gtg aga aca atc tgg tat gac tgg gta gcc 576Asn Thr Thr Glu Thr Ala Val Arg Thr Ile Trp Tyr Asp Trp Val Ala 180 185 190gac ctg gta tcc aat tat tca gtc gac gga ctc cgc atc gac agt gtc 624Asp Leu Val Ser Asn Tyr Ser Val Asp Gly Leu Arg Ile Asp Ser Val 195 200 205ctc gaa gtc gaa cca gac ttc ttc ccg ggc tac cag gaa gca gca ggt 672Leu Glu Val Glu Pro Asp Phe Phe Pro Gly Tyr Gln Glu Ala Ala Gly 210 215 220gtc tac tgc gtc ggc gaa gtc gac aac ggc aac cct gcc ctc gac tgc 720Val Tyr Cys Val Gly Glu Val Asp Asn Gly Asn Pro Ala Leu Asp Cys225 230 235 240cca tac cag aag gtc ctg gac ggc gtc ctc aac tat ccg atc tac tgg 768Pro Tyr Gln Lys Val Leu Asp Gly Val Leu Asn Tyr Pro Ile Tyr Trp 245 250 255caa ctc ctc tac gcc ttc gaa tcc tcc agc ggc agc atc agc aat ctc 816Gln Leu Leu Tyr Ala Phe Glu Ser Ser Ser Gly Ser Ile Ser Asn Leu 260 265 270tac aac atg atc aaa tcc gtc gca agc gac tgc tcc gat ccg aca cta 864Tyr Asn Met Ile Lys Ser Val Ala Ser Asp Cys Ser Asp Pro Thr Leu 275 280 285ctc ggc aac ttc atc gaa aac cac gac aat ccc cgt ttc gcc tcc tac 912Leu Gly Asn Phe Ile Glu Asn His Asp Asn Pro Arg Phe Ala Ser Tyr 290 295 300acc tcc gac tac tcg caa gcc aaa aac gtc ctc agc tac atc ttc ctc 960Thr Ser Asp Tyr Ser Gln Ala Lys Asn Val Leu Ser Tyr Ile Phe Leu305 310 315 320tcc gac ggc atc ccc atc gtc tac gcc ggc gaa gaa cag cac tac tcc 1008Ser Asp Gly Ile Pro Ile Val Tyr Ala Gly Glu Glu Gln His Tyr Ser 325 330 335ggc ggc aag gtg ccc tac aac cgc gaa gcg acc tgg ctt tca ggc tac 1056Gly Gly Lys Val Pro Tyr Asn Arg Glu Ala Thr Trp Leu Ser Gly Tyr 340 345 350gac acc tcc gca gag ctg tac acc tgg ata gcc acc acg aac gcg atc 1104Asp Thr Ser Ala Glu Leu Tyr Thr Trp Ile Ala Thr Thr Asn Ala Ile 355 360 365cgc aaa cta gcc atc tca gct gac tcg gcc tac att acc tac gcg aat 1152Arg Lys Leu Ala Ile Ser Ala Asp Ser Ala Tyr Ile Thr Tyr Ala Asn 370 375 380gat gca ttc tac act gac agc aac acc atc gca atg cgc aaa ggc acc 1200Asp Ala Phe Tyr Thr Asp Ser Asn Thr Ile Ala Met Arg Lys Gly Thr385 390 395 400tca ggg agc caa gtc atc acc gtc ctc tcc aac aaa ggc tcc tca gga 1248Ser Gly Ser Gln Val Ile Thr Val Leu Ser Asn Lys Gly Ser Ser Gly 405 410 415agc agc tac acc ctg acc ctc agc gga agc ggc tac aca tcc ggc acg 1296Ser Ser Tyr Thr Leu Thr Leu Ser Gly Ser Gly Tyr Thr Ser Gly Thr 420 425 430aag ctg atc gaa gcg tac aca tgc aca tcc gtg acc gtg gac tcg agc 1344Lys Leu Ile Glu Ala Tyr Thr Cys Thr Ser Val Thr Val Asp Ser Ser 435 440 445ggc gat att ccc gtg ccg atg gcg tcg gga tta ccg aga gtt ctt ctg 1392Gly Asp Ile Pro Val Pro Met Ala Ser Gly Leu Pro Arg Val Leu Leu 450 455 460ccc gcg tcc gtc gtc gat agc tct tcg ctc tgt ggc ggg agc gga aga 1440Pro Ala Ser Val Val Asp Ser Ser Ser Leu Cys Gly Gly Ser Gly Arg465 470 475 480aca acc acg acc aca act gct gct gct act agt aca tcc aaa gcc acc 1488Thr Thr Thr Thr Thr Thr Ala Ala Ala Thr Ser Thr Ser Lys Ala Thr 485 490 495acc tcc tct tct tct tct tct gct gct gct act act tct tca tca gtc 1536Thr Ser Ser Ser Ser Ser Ser Ala Ala Ala Thr Thr Ser Ser Ser Val 500 505 510gag gtc act ttc gac gtt tac gct acc aca gta tat ggc cag aac atc 1584Glu Val Thr Phe Asp Val Tyr Ala Thr Thr Val Tyr Gly Gln Asn Ile 515 520 525tat atc acc ggt gat gtg agt gag ctc ggc aac tgg aca ccc gcc aat 1632Tyr Ile Thr Gly Asp Val Ser Glu Leu Gly Asn Trp Thr Pro Ala Asn 530 535 540ggt gtt gca ctc tct tct gct aac tac ccc acc tgg agt gcc acg atc 1680Gly Val Ala Leu Ser Ser Ala Asn Tyr Pro Thr Trp Ser Ala Thr Ile545 550 555 560gct ctc ccc gct gac acg aca atc cag tac aag tat gtc aac att gac 1728Ala Leu Pro Ala Asp Thr Thr Ile Gln Tyr Lys Tyr Val Asn Ile Asp 565 570 575ggc agc acc gtc atc tgg gag gat gct atc agc aat cgc gag atc acg 1776Gly Ser Thr Val Ile Trp Glu Asp Ala Ile Ser Asn Arg Glu Ile Thr 580 585 590acg ccc gcc agc ggc aca tac acc gaa aaa gac act tgg gat gaa tct 1824Thr Pro Ala Ser Gly Thr Tyr Thr Glu Lys Asp Thr Trp Asp Glu Ser 595 600 605tag 182730608PRTArtificial sequenceSynthetic Construct 30Leu Ser Ala Ala Glu Trp Arg Thr Gln Ser Ile Tyr Phe Leu Leu Thr1 5 10 15Asp Arg Phe Gly Arg Thr Asp Asn Ser Thr Thr Ala Thr Cys Asp Thr 20 25 30Gly Asp Gln Ile Tyr Cys Gly Gly Ser Trp Gln Gly Ile Ile Asn His 35 40 45Leu Asp Tyr Ile Gln Gly Met Gly Phe Thr Ala Ile Trp Ile Ser Pro 50 55 60Ile Thr Glu Gln Leu Pro Gln Asp Thr Ala Asp Gly Glu Ala Tyr His65 70 75 80Gly Tyr Trp Gln Gln Lys Ile Tyr Asp Val Asn Ser Asn Phe Gly Thr 85 90 95Ala Asp Asp Leu Lys Ser Leu Ser Asp Ala Leu His Ala Arg Gly Met 100 105 110Tyr Leu Met Val Asp Val Val Pro Asn His Met Gly Tyr Ala Gly Asn 115 120 125Gly Asn Asp Val Asp Tyr Ser Val Phe Asp Pro Phe Asp Ser Ser Ser 130 135 140Tyr Phe His Pro Tyr Cys Leu Ile Thr Asp Trp Asp Asn Leu Thr Met145 150 155 160Val Gln Asp Cys Trp Glu Gly Asp Thr Ile Val Ser Leu Pro Asp Leu 165 170 175Asn Thr Thr Glu Thr Ala Val Arg Thr Ile Trp Tyr Asp Trp Val Ala 180 185 190Asp Leu Val Ser Asn Tyr Ser Val Asp Gly Leu Arg Ile Asp Ser Val 195 200 205Leu Glu Val Glu Pro Asp Phe Phe Pro Gly Tyr Gln Glu Ala Ala Gly 210 215 220Val Tyr Cys Val Gly Glu Val Asp Asn Gly Asn Pro Ala Leu Asp Cys225 230 235 240Pro Tyr Gln Lys Val Leu Asp Gly Val Leu Asn Tyr Pro Ile Tyr Trp 245 250 255Gln Leu Leu Tyr Ala Phe Glu Ser Ser Ser Gly Ser Ile Ser Asn Leu 260 265 270Tyr Asn Met Ile Lys Ser Val Ala Ser Asp Cys Ser Asp Pro Thr Leu 275 280 285Leu Gly Asn Phe Ile Glu Asn His Asp Asn Pro Arg Phe Ala Ser Tyr 290 295 300Thr Ser Asp Tyr Ser Gln Ala Lys Asn Val Leu Ser Tyr Ile Phe Leu305 310 315 320Ser Asp Gly Ile Pro Ile Val Tyr Ala Gly Glu Glu Gln His Tyr Ser 325 330 335Gly Gly Lys Val Pro Tyr Asn Arg Glu Ala Thr Trp Leu Ser Gly Tyr 340 345 350Asp Thr Ser Ala Glu Leu Tyr Thr Trp Ile Ala Thr Thr Asn Ala Ile 355 360 365Arg Lys Leu Ala Ile Ser Ala Asp Ser Ala Tyr Ile Thr Tyr Ala Asn 370 375 380Asp Ala Phe Tyr Thr Asp Ser Asn Thr Ile Ala Met Arg Lys Gly Thr385 390 395 400Ser Gly Ser Gln Val Ile Thr Val Leu Ser Asn Lys Gly Ser Ser Gly 405 410 415Ser Ser Tyr Thr Leu Thr Leu Ser Gly Ser Gly Tyr Thr Ser Gly Thr 420 425 430Lys Leu Ile Glu Ala Tyr Thr Cys Thr Ser Val Thr Val Asp Ser Ser 435 440

445Gly Asp Ile Pro Val Pro Met Ala Ser Gly Leu Pro Arg Val Leu Leu 450 455 460Pro Ala Ser Val Val Asp Ser Ser Ser Leu Cys Gly Gly Ser Gly Arg465 470 475 480Thr Thr Thr Thr Thr Thr Ala Ala Ala Thr Ser Thr Ser Lys Ala Thr 485 490 495Thr Ser Ser Ser Ser Ser Ser Ala Ala Ala Thr Thr Ser Ser Ser Val 500 505 510Glu Val Thr Phe Asp Val Tyr Ala Thr Thr Val Tyr Gly Gln Asn Ile 515 520 525Tyr Ile Thr Gly Asp Val Ser Glu Leu Gly Asn Trp Thr Pro Ala Asn 530 535 540Gly Val Ala Leu Ser Ser Ala Asn Tyr Pro Thr Trp Ser Ala Thr Ile545 550 555 560Ala Leu Pro Ala Asp Thr Thr Ile Gln Tyr Lys Tyr Val Asn Ile Asp 565 570 575Gly Ser Thr Val Ile Trp Glu Asp Ala Ile Ser Asn Arg Glu Ile Thr 580 585 590Thr Pro Ala Ser Gly Thr Tyr Thr Glu Lys Asp Thr Trp Asp Glu Ser 595 600 605311863DNAArtificial sequenceSynthetic construct 31gca acg cct gcg gac tgg cga tcg caa tcc att tat ttc ctt ctc acg 48Ala Thr Pro Ala Asp Trp Arg Ser Gln Ser Ile Tyr Phe Leu Leu Thr1 5 10 15gat cga ttt gca agg acg gat ggg tcg acg act gcg act tgt aat act 96Asp Arg Phe Ala Arg Thr Asp Gly Ser Thr Thr Ala Thr Cys Asn Thr 20 25 30gcg gat cag aaa tac tgt ggt gga aca tgg cag ggc atc atc gac aag 144Ala Asp Gln Lys Tyr Cys Gly Gly Thr Trp Gln Gly Ile Ile Asp Lys 35 40 45ttg gac tat atc cag gga atg ggc ttc aca gcc atc tgg atc acc ccc 192Leu Asp Tyr Ile Gln Gly Met Gly Phe Thr Ala Ile Trp Ile Thr Pro 50 55 60gtt aca gcc cag ctg ccc cag acc acc gca tat gga gat gcc tac cat 240Val Thr Ala Gln Leu Pro Gln Thr Thr Ala Tyr Gly Asp Ala Tyr His65 70 75 80ggc tac tgg cag cag gat ata tac tct ctg aac gaa aac tac ggc act 288Gly Tyr Trp Gln Gln Asp Ile Tyr Ser Leu Asn Glu Asn Tyr Gly Thr 85 90 95gca gat gac ttg aag gcg ctc tct tcg gcc ctt cat gag agg ggg atg 336Ala Asp Asp Leu Lys Ala Leu Ser Ser Ala Leu His Glu Arg Gly Met 100 105 110tat ctt atg gtc gat gtg gtt gct aac cat atg ggc tat gat gga gcg 384Tyr Leu Met Val Asp Val Val Ala Asn His Met Gly Tyr Asp Gly Ala 115 120 125ggt agc tca gtc gat tac agt gtg ttt aaa ccg ttc agt tcc caa gac 432Gly Ser Ser Val Asp Tyr Ser Val Phe Lys Pro Phe Ser Ser Gln Asp 130 135 140tac ttc cac ccg ttc tgt ttc att caa aac tat gaa gat cag act cag 480Tyr Phe His Pro Phe Cys Phe Ile Gln Asn Tyr Glu Asp Gln Thr Gln145 150 155 160gtt gag gat tgc tgg cta gga gat aac act gtc tcc ttg cct gat ctc 528Val Glu Asp Cys Trp Leu Gly Asp Asn Thr Val Ser Leu Pro Asp Leu 165 170 175gat acc acc aag gat gtg gtc aag aat gaa tgg tac gac tgg gtg gga 576Asp Thr Thr Lys Asp Val Val Lys Asn Glu Trp Tyr Asp Trp Val Gly 180 185 190tca ttg gta tcg aac tac tcc att gac ggc ctc cgt atc gac aca gta 624Ser Leu Val Ser Asn Tyr Ser Ile Asp Gly Leu Arg Ile Asp Thr Val 195 200 205aaa cac gtc cag aag gac ttc tgg ccc ggg tac aac aaa gcc gca ggc 672Lys His Val Gln Lys Asp Phe Trp Pro Gly Tyr Asn Lys Ala Ala Gly 210 215 220gtg tac tgt atc ggc gag gtg ctc gac ggt gat ccg gcc tac act tgt 720Val Tyr Cys Ile Gly Glu Val Leu Asp Gly Asp Pro Ala Tyr Thr Cys225 230 235 240ccc tac cag aac gtc atg gac ggc gta ctg aac tat ccc att tac tat 768Pro Tyr Gln Asn Val Met Asp Gly Val Leu Asn Tyr Pro Ile Tyr Tyr 245 250 255cca ctc ctc aac gcc ttc aag tca acc tcc ggc agc atg gac gac ctc 816Pro Leu Leu Asn Ala Phe Lys Ser Thr Ser Gly Ser Met Asp Asp Leu 260 265 270tac aac atg atc aac acc gtc aaa tcc gac tgt cca gac tca aca ctc 864Tyr Asn Met Ile Asn Thr Val Lys Ser Asp Cys Pro Asp Ser Thr Leu 275 280 285ctg ggc aca ttc gtc gag aac cac gac aac cca cgg ttc gct tct tac 912Leu Gly Thr Phe Val Glu Asn His Asp Asn Pro Arg Phe Ala Ser Tyr 290 295 300acc aac gac ata gcc ctc gcc aag aac gtc gca gca ttc atc atc ctc 960Thr Asn Asp Ile Ala Leu Ala Lys Asn Val Ala Ala Phe Ile Ile Leu305 310 315 320aac gac gga atc ccc atc atc tac gcc ggc caa gaa cag cac tac gcc 1008Asn Asp Gly Ile Pro Ile Ile Tyr Ala Gly Gln Glu Gln His Tyr Ala 325 330 335ggc gga aac gac ccc gcg aac cgc gaa gca acc tgg ctc tcg ggc tac 1056Gly Gly Asn Asp Pro Ala Asn Arg Glu Ala Thr Trp Leu Ser Gly Tyr 340 345 350ccg acc gac agc gag ctg tac aag tta att gcc tcc gcg aac gca atc 1104Pro Thr Asp Ser Glu Leu Tyr Lys Leu Ile Ala Ser Ala Asn Ala Ile 355 360 365cgg aac tat gcc att agc aaa gat aca gga ttc gtg acc tac aag aac 1152Arg Asn Tyr Ala Ile Ser Lys Asp Thr Gly Phe Val Thr Tyr Lys Asn 370 375 380tgg ccc atc tac aaa gac gac aca acg atc gcc atg cgc aag ggc aca 1200Trp Pro Ile Tyr Lys Asp Asp Thr Thr Ile Ala Met Arg Lys Gly Thr385 390 395 400gat ggg tcg cag atc gtg act atc ttg tcc aac aag ggt gct tcg ggt 1248Asp Gly Ser Gln Ile Val Thr Ile Leu Ser Asn Lys Gly Ala Ser Gly 405 410 415gat tcg tat acc ctc tcc ttg agt ggt gcg ggt tac aca gcc ggc cag 1296Asp Ser Tyr Thr Leu Ser Leu Ser Gly Ala Gly Tyr Thr Ala Gly Gln 420 425 430caa ttg acg gag gtc att ggc tgc acg acc gtg acg gtt ggt tcg gat 1344Gln Leu Thr Glu Val Ile Gly Cys Thr Thr Val Thr Val Gly Ser Asp 435 440 445gga aat gtg cct gtt cct atg gca ggt ggg cta cct agg gta ttg tat 1392Gly Asn Val Pro Val Pro Met Ala Gly Gly Leu Pro Arg Val Leu Tyr 450 455 460ccg act gag aag ttg gca ggt agc aag atc tgt agt agc tcg gga aga 1440Pro Thr Glu Lys Leu Ala Gly Ser Lys Ile Cys Ser Ser Ser Gly Arg465 470 475 480aca acc acg acc aca act gct gct gct act agt aca tcc aaa gcc acc 1488Thr Thr Thr Thr Thr Thr Ala Ala Ala Thr Ser Thr Ser Lys Ala Thr 485 490 495acc tcc tct tct tct tct tct gct gct gct act act tct tca tca tgc 1536Thr Ser Ser Ser Ser Ser Ser Ala Ala Ala Thr Thr Ser Ser Ser Cys 500 505 510acc gca aca agc acc acc ctc ccc atc acc ttc gaa gaa ctc gtc acc 1584Thr Ala Thr Ser Thr Thr Leu Pro Ile Thr Phe Glu Glu Leu Val Thr 515 520 525act acc tac ggg gaa gaa gtc tac ctc agc gga tct atc tcc cag ctc 1632Thr Thr Tyr Gly Glu Glu Val Tyr Leu Ser Gly Ser Ile Ser Gln Leu 530 535 540gga gag tgg gat acg agt gac gcg gtg aag ttg tcc gcg gat gat tat 1680Gly Glu Trp Asp Thr Ser Asp Ala Val Lys Leu Ser Ala Asp Asp Tyr545 550 555 560acc tcg agt aac ccc gag tgg tct gtt act gtg tcg ttg ccg gtg ggg 1728Thr Ser Ser Asn Pro Glu Trp Ser Val Thr Val Ser Leu Pro Val Gly 565 570 575acg acc ttc gag tat aag ttt att aag gtc gat gag ggt gga agt gtg 1776Thr Thr Phe Glu Tyr Lys Phe Ile Lys Val Asp Glu Gly Gly Ser Val 580 585 590act tgg gaa agt gat ccg aat agg gag tat act gtg cct gaa tgt ggg 1824Thr Trp Glu Ser Asp Pro Asn Arg Glu Tyr Thr Val Pro Glu Cys Gly 595 600 605aat ggg agt ggg gag acg gtg gtt gat acg tgg agg tag 1863Asn Gly Ser Gly Glu Thr Val Val Asp Thr Trp Arg 610 615 62032620PRTArtificial sequenceSynthetic Construct 32Ala Thr Pro Ala Asp Trp Arg Ser Gln Ser Ile Tyr Phe Leu Leu Thr1 5 10 15Asp Arg Phe Ala Arg Thr Asp Gly Ser Thr Thr Ala Thr Cys Asn Thr 20 25 30Ala Asp Gln Lys Tyr Cys Gly Gly Thr Trp Gln Gly Ile Ile Asp Lys 35 40 45Leu Asp Tyr Ile Gln Gly Met Gly Phe Thr Ala Ile Trp Ile Thr Pro 50 55 60Val Thr Ala Gln Leu Pro Gln Thr Thr Ala Tyr Gly Asp Ala Tyr His65 70 75 80Gly Tyr Trp Gln Gln Asp Ile Tyr Ser Leu Asn Glu Asn Tyr Gly Thr 85 90 95Ala Asp Asp Leu Lys Ala Leu Ser Ser Ala Leu His Glu Arg Gly Met 100 105 110Tyr Leu Met Val Asp Val Val Ala Asn His Met Gly Tyr Asp Gly Ala 115 120 125Gly Ser Ser Val Asp Tyr Ser Val Phe Lys Pro Phe Ser Ser Gln Asp 130 135 140Tyr Phe His Pro Phe Cys Phe Ile Gln Asn Tyr Glu Asp Gln Thr Gln145 150 155 160Val Glu Asp Cys Trp Leu Gly Asp Asn Thr Val Ser Leu Pro Asp Leu 165 170 175Asp Thr Thr Lys Asp Val Val Lys Asn Glu Trp Tyr Asp Trp Val Gly 180 185 190Ser Leu Val Ser Asn Tyr Ser Ile Asp Gly Leu Arg Ile Asp Thr Val 195 200 205Lys His Val Gln Lys Asp Phe Trp Pro Gly Tyr Asn Lys Ala Ala Gly 210 215 220Val Tyr Cys Ile Gly Glu Val Leu Asp Gly Asp Pro Ala Tyr Thr Cys225 230 235 240Pro Tyr Gln Asn Val Met Asp Gly Val Leu Asn Tyr Pro Ile Tyr Tyr 245 250 255Pro Leu Leu Asn Ala Phe Lys Ser Thr Ser Gly Ser Met Asp Asp Leu 260 265 270Tyr Asn Met Ile Asn Thr Val Lys Ser Asp Cys Pro Asp Ser Thr Leu 275 280 285Leu Gly Thr Phe Val Glu Asn His Asp Asn Pro Arg Phe Ala Ser Tyr 290 295 300Thr Asn Asp Ile Ala Leu Ala Lys Asn Val Ala Ala Phe Ile Ile Leu305 310 315 320Asn Asp Gly Ile Pro Ile Ile Tyr Ala Gly Gln Glu Gln His Tyr Ala 325 330 335Gly Gly Asn Asp Pro Ala Asn Arg Glu Ala Thr Trp Leu Ser Gly Tyr 340 345 350Pro Thr Asp Ser Glu Leu Tyr Lys Leu Ile Ala Ser Ala Asn Ala Ile 355 360 365Arg Asn Tyr Ala Ile Ser Lys Asp Thr Gly Phe Val Thr Tyr Lys Asn 370 375 380Trp Pro Ile Tyr Lys Asp Asp Thr Thr Ile Ala Met Arg Lys Gly Thr385 390 395 400Asp Gly Ser Gln Ile Val Thr Ile Leu Ser Asn Lys Gly Ala Ser Gly 405 410 415Asp Ser Tyr Thr Leu Ser Leu Ser Gly Ala Gly Tyr Thr Ala Gly Gln 420 425 430Gln Leu Thr Glu Val Ile Gly Cys Thr Thr Val Thr Val Gly Ser Asp 435 440 445Gly Asn Val Pro Val Pro Met Ala Gly Gly Leu Pro Arg Val Leu Tyr 450 455 460Pro Thr Glu Lys Leu Ala Gly Ser Lys Ile Cys Ser Ser Ser Gly Arg465 470 475 480Thr Thr Thr Thr Thr Thr Ala Ala Ala Thr Ser Thr Ser Lys Ala Thr 485 490 495Thr Ser Ser Ser Ser Ser Ser Ala Ala Ala Thr Thr Ser Ser Ser Cys 500 505 510Thr Ala Thr Ser Thr Thr Leu Pro Ile Thr Phe Glu Glu Leu Val Thr 515 520 525Thr Thr Tyr Gly Glu Glu Val Tyr Leu Ser Gly Ser Ile Ser Gln Leu 530 535 540Gly Glu Trp Asp Thr Ser Asp Ala Val Lys Leu Ser Ala Asp Asp Tyr545 550 555 560Thr Ser Ser Asn Pro Glu Trp Ser Val Thr Val Ser Leu Pro Val Gly 565 570 575Thr Thr Phe Glu Tyr Lys Phe Ile Lys Val Asp Glu Gly Gly Ser Val 580 585 590Thr Trp Glu Ser Asp Pro Asn Arg Glu Tyr Thr Val Pro Glu Cys Gly 595 600 605Asn Gly Ser Gly Glu Thr Val Val Asp Thr Trp Arg 610 615 620331767DNAArtificial sequenceSynthetic construct 33ctg tcg gct gca gaa tgg cgc act cag tcg att tac ttc cta ttg acg 48Leu Ser Ala Ala Glu Trp Arg Thr Gln Ser Ile Tyr Phe Leu Leu Thr1 5 10 15gat cgg ttc ggt agg acg gac aat tcg acg aca gct aca tgc gat acg 96Asp Arg Phe Gly Arg Thr Asp Asn Ser Thr Thr Ala Thr Cys Asp Thr 20 25 30ggt gac caa atc tat tgt ggt ggc agt tgg caa gga atc atc aac cat 144Gly Asp Gln Ile Tyr Cys Gly Gly Ser Trp Gln Gly Ile Ile Asn His 35 40 45ctg gat tat atc cag ggc atg gga ttc acg gcc atc tgg atc tcg cct 192Leu Asp Tyr Ile Gln Gly Met Gly Phe Thr Ala Ile Trp Ile Ser Pro 50 55 60atc act gaa cag ctg ccc cag gat act gct gat ggt gaa gct tac cat 240Ile Thr Glu Gln Leu Pro Gln Asp Thr Ala Asp Gly Glu Ala Tyr His65 70 75 80gga tat tgg cag cag aag ata tac gac gtg aac tcc aac ttc ggc act 288Gly Tyr Trp Gln Gln Lys Ile Tyr Asp Val Asn Ser Asn Phe Gly Thr 85 90 95gca gat gac ctc aag tcc ctc tca gat gcg ctt cat gcc cgc gga atg 336Ala Asp Asp Leu Lys Ser Leu Ser Asp Ala Leu His Ala Arg Gly Met 100 105 110tac ctc atg gtg gac gtc gtc cct aac cac atg ggc tac gcc ggc aac 384Tyr Leu Met Val Asp Val Val Pro Asn His Met Gly Tyr Ala Gly Asn 115 120 125ggc aac gat gta gac tac agc gtc ttc gac ccc ttc gat tcc tcc tcc 432Gly Asn Asp Val Asp Tyr Ser Val Phe Asp Pro Phe Asp Ser Ser Ser 130 135 140tac ttc cac cca tac tgc ctg atc aca gat tgg gac aac ttg acc atg 480Tyr Phe His Pro Tyr Cys Leu Ile Thr Asp Trp Asp Asn Leu Thr Met145 150 155 160gtc caa gat tgt tgg gag ggt gac acc atc gta tct ctg cca gac cta 528Val Gln Asp Cys Trp Glu Gly Asp Thr Ile Val Ser Leu Pro Asp Leu 165 170 175aac acc acc gaa act gcc gtg aga aca atc tgg tat gac tgg gta gcc 576Asn Thr Thr Glu Thr Ala Val Arg Thr Ile Trp Tyr Asp Trp Val Ala 180 185 190gac ctg gta tcc aat tat tca gtc gac gga ctc cgc atc gac agt gtc 624Asp Leu Val Ser Asn Tyr Ser Val Asp Gly Leu Arg Ile Asp Ser Val 195 200 205ctc gaa gtc gaa cca gac ttc ttc ccg ggc tac cag gaa gca gca ggt 672Leu Glu Val Glu Pro Asp Phe Phe Pro Gly Tyr Gln Glu Ala Ala Gly 210 215 220gtc tac tgc gtc ggc gaa gtc gac aac ggc aac cct gcc ctc gac tgc 720Val Tyr Cys Val Gly Glu Val Asp Asn Gly Asn Pro Ala Leu Asp Cys225 230 235 240cca tac cag aag gtc ctg gac ggc gtc ctc aac tat ccg atc tac tgg 768Pro Tyr Gln Lys Val Leu Asp Gly Val Leu Asn Tyr Pro Ile Tyr Trp 245 250 255caa ctc ctc tac gcc ttc gaa tcc tcc agc ggc agc atc agc aat ctc 816Gln Leu Leu Tyr Ala Phe Glu Ser Ser Ser Gly Ser Ile Ser Asn Leu 260 265 270tac aac atg atc aaa tcc gtc gca agc gac tgc tcc gat ccg aca cta 864Tyr Asn Met Ile Lys Ser Val Ala Ser Asp Cys Ser Asp Pro Thr Leu 275 280 285ctc ggc aac ttc atc gaa aac cac gac aat ccc cgt ttc gcc tcc tac 912Leu Gly Asn Phe Ile Glu Asn His Asp Asn Pro Arg Phe Ala Ser Tyr 290 295 300acc tcc gac tac tcg caa gcc aaa aac gtc ctc agc tac atc ttc ctc 960Thr Ser Asp Tyr Ser Gln Ala Lys Asn Val Leu Ser Tyr Ile Phe Leu305 310 315 320tcc gac ggc atc ccc atc gtc tac gcc ggc gaa gaa cag cac tac tcc 1008Ser Asp Gly Ile Pro Ile Val Tyr Ala Gly Glu Glu Gln His Tyr Ser 325 330 335ggc ggc aag gtg ccc tac aac cgc gaa gcg acc tgg ctt tca ggc tac 1056Gly Gly Lys Val Pro Tyr Asn Arg Glu Ala Thr Trp Leu Ser Gly Tyr 340 345 350gac acc tcc gca gag ctg tac acc tgg ata gcc acc acg aac gcg atc 1104Asp Thr Ser Ala Glu Leu Tyr Thr Trp Ile Ala Thr Thr Asn Ala Ile 355 360 365cgc aaa cta gcc atc tca gct gac tcg gcc tac att acc tac gcg aat 1152Arg Lys Leu Ala Ile Ser Ala Asp Ser Ala Tyr Ile Thr Tyr Ala Asn 370 375 380gat gca ttc tac act gac agc aac acc atc gca atg cgc aaa ggc acc 1200Asp Ala Phe Tyr Thr Asp Ser Asn Thr Ile Ala Met Arg Lys Gly Thr385 390 395 400tca ggg agc caa gtc atc acc gtc ctc tcc aac aaa ggc tcc tca gga 1248Ser Gly Ser Gln Val Ile Thr Val Leu Ser Asn Lys Gly Ser Ser Gly 405 410 415agc agc tac acc ctg acc ctc agc gga agc ggc tac aca tcc ggc acg

1296Ser Ser Tyr Thr Leu Thr Leu Ser Gly Ser Gly Tyr Thr Ser Gly Thr 420 425 430aag ctg atc gaa gcg tac aca tgc aca tcc gtg acc gtg gac tcg agc 1344Lys Leu Ile Glu Ala Tyr Thr Cys Thr Ser Val Thr Val Asp Ser Ser 435 440 445ggc gat att ccc gtg ccg atg gcg tcg gga tta ccg aga gtt ctt ctg 1392Gly Asp Ile Pro Val Pro Met Ala Ser Gly Leu Pro Arg Val Leu Leu 450 455 460ccc gcg tcc gtc gtc gat agc tct tcg ctc tgt ggc ggg agc gga aga 1440Pro Ala Ser Val Val Asp Ser Ser Ser Leu Cys Gly Gly Ser Gly Arg465 470 475 480ggt gct aca agc ccg ggt ggc tcc tcg ggt agt gtc gag gtc act ttc 1488Gly Ala Thr Ser Pro Gly Gly Ser Ser Gly Ser Val Glu Val Thr Phe 485 490 495 gac gtt tac gct acc aca gta tat ggc cag aac atc tat atc acc ggt 1536Asp Val Tyr Ala Thr Thr Val Tyr Gly Gln Asn Ile Tyr Ile Thr Gly 500 505 510gat gtg agt gag ctc ggc aac tgg aca ccc gcc aat ggt gtt gca ctc 1584Asp Val Ser Glu Leu Gly Asn Trp Thr Pro Ala Asn Gly Val Ala Leu 515 520 525tct tct gct aac tac ccc acc tgg agt gcc acg atc gct ctc ccc gct 1632Ser Ser Ala Asn Tyr Pro Thr Trp Ser Ala Thr Ile Ala Leu Pro Ala 530 535 540gac acg aca atc cag tac aag tat gtc aac att gac ggc agc acc gtc 1680Asp Thr Thr Ile Gln Tyr Lys Tyr Val Asn Ile Asp Gly Ser Thr Val545 550 555 560atc tgg gag gat gct atc agc aat cgc gag atc acg acg ccc gcc agc 1728Ile Trp Glu Asp Ala Ile Ser Asn Arg Glu Ile Thr Thr Pro Ala Ser 565 570 575ggc aca tac acc gaa aaa gac act tgg gat gaa tct tag 1767Gly Thr Tyr Thr Glu Lys Asp Thr Trp Asp Glu Ser 580 58534588PRTArtificial sequenceSynthetic Construct 34Leu Ser Ala Ala Glu Trp Arg Thr Gln Ser Ile Tyr Phe Leu Leu Thr1 5 10 15Asp Arg Phe Gly Arg Thr Asp Asn Ser Thr Thr Ala Thr Cys Asp Thr 20 25 30Gly Asp Gln Ile Tyr Cys Gly Gly Ser Trp Gln Gly Ile Ile Asn His 35 40 45Leu Asp Tyr Ile Gln Gly Met Gly Phe Thr Ala Ile Trp Ile Ser Pro 50 55 60Ile Thr Glu Gln Leu Pro Gln Asp Thr Ala Asp Gly Glu Ala Tyr His65 70 75 80Gly Tyr Trp Gln Gln Lys Ile Tyr Asp Val Asn Ser Asn Phe Gly Thr 85 90 95Ala Asp Asp Leu Lys Ser Leu Ser Asp Ala Leu His Ala Arg Gly Met 100 105 110Tyr Leu Met Val Asp Val Val Pro Asn His Met Gly Tyr Ala Gly Asn 115 120 125Gly Asn Asp Val Asp Tyr Ser Val Phe Asp Pro Phe Asp Ser Ser Ser 130 135 140Tyr Phe His Pro Tyr Cys Leu Ile Thr Asp Trp Asp Asn Leu Thr Met145 150 155 160Val Gln Asp Cys Trp Glu Gly Asp Thr Ile Val Ser Leu Pro Asp Leu 165 170 175Asn Thr Thr Glu Thr Ala Val Arg Thr Ile Trp Tyr Asp Trp Val Ala 180 185 190Asp Leu Val Ser Asn Tyr Ser Val Asp Gly Leu Arg Ile Asp Ser Val 195 200 205Leu Glu Val Glu Pro Asp Phe Phe Pro Gly Tyr Gln Glu Ala Ala Gly 210 215 220Val Tyr Cys Val Gly Glu Val Asp Asn Gly Asn Pro Ala Leu Asp Cys225 230 235 240Pro Tyr Gln Lys Val Leu Asp Gly Val Leu Asn Tyr Pro Ile Tyr Trp 245 250 255Gln Leu Leu Tyr Ala Phe Glu Ser Ser Ser Gly Ser Ile Ser Asn Leu 260 265 270Tyr Asn Met Ile Lys Ser Val Ala Ser Asp Cys Ser Asp Pro Thr Leu 275 280 285Leu Gly Asn Phe Ile Glu Asn His Asp Asn Pro Arg Phe Ala Ser Tyr 290 295 300Thr Ser Asp Tyr Ser Gln Ala Lys Asn Val Leu Ser Tyr Ile Phe Leu305 310 315 320Ser Asp Gly Ile Pro Ile Val Tyr Ala Gly Glu Glu Gln His Tyr Ser 325 330 335Gly Gly Lys Val Pro Tyr Asn Arg Glu Ala Thr Trp Leu Ser Gly Tyr 340 345 350Asp Thr Ser Ala Glu Leu Tyr Thr Trp Ile Ala Thr Thr Asn Ala Ile 355 360 365Arg Lys Leu Ala Ile Ser Ala Asp Ser Ala Tyr Ile Thr Tyr Ala Asn 370 375 380Asp Ala Phe Tyr Thr Asp Ser Asn Thr Ile Ala Met Arg Lys Gly Thr385 390 395 400Ser Gly Ser Gln Val Ile Thr Val Leu Ser Asn Lys Gly Ser Ser Gly 405 410 415Ser Ser Tyr Thr Leu Thr Leu Ser Gly Ser Gly Tyr Thr Ser Gly Thr 420 425 430Lys Leu Ile Glu Ala Tyr Thr Cys Thr Ser Val Thr Val Asp Ser Ser 435 440 445Gly Asp Ile Pro Val Pro Met Ala Ser Gly Leu Pro Arg Val Leu Leu 450 455 460Pro Ala Ser Val Val Asp Ser Ser Ser Leu Cys Gly Gly Ser Gly Arg465 470 475 480Gly Ala Thr Ser Pro Gly Gly Ser Ser Gly Ser Val Glu Val Thr Phe 485 490 495Asp Val Tyr Ala Thr Thr Val Tyr Gly Gln Asn Ile Tyr Ile Thr Gly 500 505 510Asp Val Ser Glu Leu Gly Asn Trp Thr Pro Ala Asn Gly Val Ala Leu 515 520 525Ser Ser Ala Asn Tyr Pro Thr Trp Ser Ala Thr Ile Ala Leu Pro Ala 530 535 540Asp Thr Thr Ile Gln Tyr Lys Tyr Val Asn Ile Asp Gly Ser Thr Val545 550 555 560Ile Trp Glu Asp Ala Ile Ser Asn Arg Glu Ile Thr Thr Pro Ala Ser 565 570 575Gly Thr Tyr Thr Glu Lys Asp Thr Trp Asp Glu Ser 580 585351767DNAArtificial sequenceSynthetic construct 35gca acg cct gcg gac tgg cga tcg caa tcc att tat ttc ctt ctc acg 48Ala Thr Pro Ala Asp Trp Arg Ser Gln Ser Ile Tyr Phe Leu Leu Thr1 5 10 15gat cga ttt gca agg acg gat ggg tcg acg act gcg act tgt aat act 96Asp Arg Phe Ala Arg Thr Asp Gly Ser Thr Thr Ala Thr Cys Asn Thr 20 25 30gcg gat cag aaa tac tgt ggt gga aca tgg cag ggc atc atc gac aag 144Ala Asp Gln Lys Tyr Cys Gly Gly Thr Trp Gln Gly Ile Ile Asp Lys 35 40 45ttg gac tat atc cag gga atg ggc ttc aca gcc atc tgg atc acc ccc 192Leu Asp Tyr Ile Gln Gly Met Gly Phe Thr Ala Ile Trp Ile Thr Pro 50 55 60gtt aca gcc cag ctg ccc cag acc acc gca tat gga gat gcc tac cat 240Val Thr Ala Gln Leu Pro Gln Thr Thr Ala Tyr Gly Asp Ala Tyr His65 70 75 80ggc tac tgg cag cag gat ata tac tct ctg aac gaa aac tac ggc act 288Gly Tyr Trp Gln Gln Asp Ile Tyr Ser Leu Asn Glu Asn Tyr Gly Thr 85 90 95gca gat gac ttg aag gcg ctc tct tcg gcc ctt cat gag agg ggg atg 336Ala Asp Asp Leu Lys Ala Leu Ser Ser Ala Leu His Glu Arg Gly Met 100 105 110tat ctt atg gtc gat gtg gtt gct aac cat atg ggc tat gat gga gcg 384Tyr Leu Met Val Asp Val Val Ala Asn His Met Gly Tyr Asp Gly Ala 115 120 125ggt agc tca gtc gat tac agt gtg ttt aaa ccg ttc agt tcc caa gac 432Gly Ser Ser Val Asp Tyr Ser Val Phe Lys Pro Phe Ser Ser Gln Asp 130 135 140tac ttc cac ccg ttc tgt ttc att caa aac tat gaa gat cag act cag 480Tyr Phe His Pro Phe Cys Phe Ile Gln Asn Tyr Glu Asp Gln Thr Gln145 150 155 160gtt gag gat tgc tgg cta gga gat aac act gtc tcc ttg cct gat ctc 528Val Glu Asp Cys Trp Leu Gly Asp Asn Thr Val Ser Leu Pro Asp Leu 165 170 175gat acc acc aag gat gtg gtc aag aat gaa tgg tac gac tgg gtg gga 576Asp Thr Thr Lys Asp Val Val Lys Asn Glu Trp Tyr Asp Trp Val Gly 180 185 190tca ttg gta tcg aac tac tcc att gac ggc ctc cgt atc gac aca gta 624Ser Leu Val Ser Asn Tyr Ser Ile Asp Gly Leu Arg Ile Asp Thr Val 195 200 205aaa cac gtc cag aag gac ttc tgg ccc ggg tac aac aaa gcc gca ggc 672Lys His Val Gln Lys Asp Phe Trp Pro Gly Tyr Asn Lys Ala Ala Gly 210 215 220gtg tac tgt atc ggc gag gtg ctc gac ggt gat ccg gcc tac act tgt 720Val Tyr Cys Ile Gly Glu Val Leu Asp Gly Asp Pro Ala Tyr Thr Cys225 230 235 240ccc tac cag aac gtc atg gac ggc gta ctg aac tat ccc att tac tat 768Pro Tyr Gln Asn Val Met Asp Gly Val Leu Asn Tyr Pro Ile Tyr Tyr 245 250 255cca ctc ctc aac gcc ttc aag tca acc tcc ggc agc atg gac gac ctc 816Pro Leu Leu Asn Ala Phe Lys Ser Thr Ser Gly Ser Met Asp Asp Leu 260 265 270tac aac atg atc aac acc gtc aaa tcc gac tgt cca gac tca aca ctc 864Tyr Asn Met Ile Asn Thr Val Lys Ser Asp Cys Pro Asp Ser Thr Leu 275 280 285ctg ggc aca ttc gtc gag aac cac gac aac cca cgg ttc gct tct tac 912Leu Gly Thr Phe Val Glu Asn His Asp Asn Pro Arg Phe Ala Ser Tyr 290 295 300acc aac gac ata gcc ctc gcc aag aac gtc gca gca ttc atc atc ctc 960Thr Asn Asp Ile Ala Leu Ala Lys Asn Val Ala Ala Phe Ile Ile Leu305 310 315 320aac gac gga atc ccc atc atc tac gcc ggc caa gaa cag cac tac gcc 1008Asn Asp Gly Ile Pro Ile Ile Tyr Ala Gly Gln Glu Gln His Tyr Ala 325 330 335ggc gga aac gac ccc gcg aac cgc gaa gca acc tgg ctc tcg ggc tac 1056Gly Gly Asn Asp Pro Ala Asn Arg Glu Ala Thr Trp Leu Ser Gly Tyr 340 345 350ccg acc gac agc gag ctg tac aag tta att gcc tcc gcg aac gca atc 1104Pro Thr Asp Ser Glu Leu Tyr Lys Leu Ile Ala Ser Ala Asn Ala Ile 355 360 365cgg aac tat gcc att agc aaa gat aca gga ttc gtg acc tac aag aac 1152Arg Asn Tyr Ala Ile Ser Lys Asp Thr Gly Phe Val Thr Tyr Lys Asn 370 375 380tgg ccc atc tac aaa gac gac aca acg atc gcc atg cgc aag ggc aca 1200Trp Pro Ile Tyr Lys Asp Asp Thr Thr Ile Ala Met Arg Lys Gly Thr385 390 395 400gat ggg tcg cag atc gtg act atc ttg tcc aac aag ggt gct tcg ggt 1248Asp Gly Ser Gln Ile Val Thr Ile Leu Ser Asn Lys Gly Ala Ser Gly 405 410 415gat tcg tat acc ctc tcc ttg agt ggt gcg ggt tac aca gcc ggc cag 1296Asp Ser Tyr Thr Leu Ser Leu Ser Gly Ala Gly Tyr Thr Ala Gly Gln 420 425 430caa ttg acg gag gtc att ggc tgc acg acc gtg acg gtt ggt tcg gat 1344Gln Leu Thr Glu Val Ile Gly Cys Thr Thr Val Thr Val Gly Ser Asp 435 440 445gga aat gtg cct gtt cct atg gca ggt ggg cta cct agg gta ttg tat 1392Gly Asn Val Pro Val Pro Met Ala Gly Gly Leu Pro Arg Val Leu Tyr 450 455 460ccg act gag aag ttg gca ggt agc aag atc tgt agt agc tcg gga aga 1440Pro Thr Glu Lys Leu Ala Gly Ser Lys Ile Cys Ser Ser Ser Gly Arg465 470 475 480ggt gct aca agc ccg ggt ggc tcc tcg ggt agt gtc gag gtc act ttc 1488Gly Ala Thr Ser Pro Gly Gly Ser Ser Gly Ser Val Glu Val Thr Phe 485 490 495gac gtt tac gct acc aca gta tat ggc cag aac atc tat atc acc ggt 1536Asp Val Tyr Ala Thr Thr Val Tyr Gly Gln Asn Ile Tyr Ile Thr Gly 500 505 510gat gtg agt gag ctc ggc aac tgg aca ccc gcc aat ggt gtt gca ctc 1584Asp Val Ser Glu Leu Gly Asn Trp Thr Pro Ala Asn Gly Val Ala Leu 515 520 525tct tct gct aac tac ccc acc tgg agt gcc acg atc gct ctc ccc gct 1632Ser Ser Ala Asn Tyr Pro Thr Trp Ser Ala Thr Ile Ala Leu Pro Ala 530 535 540gac acg aca atc cag tac aag tat gtc aac att gac ggc agc acc gtc 1680Asp Thr Thr Ile Gln Tyr Lys Tyr Val Asn Ile Asp Gly Ser Thr Val545 550 555 560atc tgg gag gat gct atc agc aat cgc gag atc acg acg ccc gcc agc 1728Ile Trp Glu Asp Ala Ile Ser Asn Arg Glu Ile Thr Thr Pro Ala Ser 565 570 575ggc aca tac acc gaa aaa gac act tgg gat gaa tct tag 1767Gly Thr Tyr Thr Glu Lys Asp Thr Trp Asp Glu Ser 580 58536588PRTArtificial sequenceSynthetic Construct 36Ala Thr Pro Ala Asp Trp Arg Ser Gln Ser Ile Tyr Phe Leu Leu Thr1 5 10 15Asp Arg Phe Ala Arg Thr Asp Gly Ser Thr Thr Ala Thr Cys Asn Thr 20 25 30Ala Asp Gln Lys Tyr Cys Gly Gly Thr Trp Gln Gly Ile Ile Asp Lys 35 40 45Leu Asp Tyr Ile Gln Gly Met Gly Phe Thr Ala Ile Trp Ile Thr Pro 50 55 60Val Thr Ala Gln Leu Pro Gln Thr Thr Ala Tyr Gly Asp Ala Tyr His65 70 75 80Gly Tyr Trp Gln Gln Asp Ile Tyr Ser Leu Asn Glu Asn Tyr Gly Thr 85 90 95Ala Asp Asp Leu Lys Ala Leu Ser Ser Ala Leu His Glu Arg Gly Met 100 105 110Tyr Leu Met Val Asp Val Val Ala Asn His Met Gly Tyr Asp Gly Ala 115 120 125Gly Ser Ser Val Asp Tyr Ser Val Phe Lys Pro Phe Ser Ser Gln Asp 130 135 140Tyr Phe His Pro Phe Cys Phe Ile Gln Asn Tyr Glu Asp Gln Thr Gln145 150 155 160Val Glu Asp Cys Trp Leu Gly Asp Asn Thr Val Ser Leu Pro Asp Leu 165 170 175Asp Thr Thr Lys Asp Val Val Lys Asn Glu Trp Tyr Asp Trp Val Gly 180 185 190Ser Leu Val Ser Asn Tyr Ser Ile Asp Gly Leu Arg Ile Asp Thr Val 195 200 205Lys His Val Gln Lys Asp Phe Trp Pro Gly Tyr Asn Lys Ala Ala Gly 210 215 220Val Tyr Cys Ile Gly Glu Val Leu Asp Gly Asp Pro Ala Tyr Thr Cys225 230 235 240Pro Tyr Gln Asn Val Met Asp Gly Val Leu Asn Tyr Pro Ile Tyr Tyr 245 250 255Pro Leu Leu Asn Ala Phe Lys Ser Thr Ser Gly Ser Met Asp Asp Leu 260 265 270Tyr Asn Met Ile Asn Thr Val Lys Ser Asp Cys Pro Asp Ser Thr Leu 275 280 285Leu Gly Thr Phe Val Glu Asn His Asp Asn Pro Arg Phe Ala Ser Tyr 290 295 300Thr Asn Asp Ile Ala Leu Ala Lys Asn Val Ala Ala Phe Ile Ile Leu305 310 315 320Asn Asp Gly Ile Pro Ile Ile Tyr Ala Gly Gln Glu Gln His Tyr Ala 325 330 335Gly Gly Asn Asp Pro Ala Asn Arg Glu Ala Thr Trp Leu Ser Gly Tyr 340 345 350Pro Thr Asp Ser Glu Leu Tyr Lys Leu Ile Ala Ser Ala Asn Ala Ile 355 360 365Arg Asn Tyr Ala Ile Ser Lys Asp Thr Gly Phe Val Thr Tyr Lys Asn 370 375 380Trp Pro Ile Tyr Lys Asp Asp Thr Thr Ile Ala Met Arg Lys Gly Thr385 390 395 400Asp Gly Ser Gln Ile Val Thr Ile Leu Ser Asn Lys Gly Ala Ser Gly 405 410 415Asp Ser Tyr Thr Leu Ser Leu Ser Gly Ala Gly Tyr Thr Ala Gly Gln 420 425 430Gln Leu Thr Glu Val Ile Gly Cys Thr Thr Val Thr Val Gly Ser Asp 435 440 445Gly Asn Val Pro Val Pro Met Ala Gly Gly Leu Pro Arg Val Leu Tyr 450 455 460Pro Thr Glu Lys Leu Ala Gly Ser Lys Ile Cys Ser Ser Ser Gly Arg465 470 475 480Gly Ala Thr Ser Pro Gly Gly Ser Ser Gly Ser Val Glu Val Thr Phe 485 490 495Asp Val Tyr Ala Thr Thr Val Tyr Gly Gln Asn Ile Tyr Ile Thr Gly 500 505 510Asp Val Ser Glu Leu Gly Asn Trp Thr Pro Ala Asn Gly Val Ala Leu 515 520 525Ser Ser Ala Asn Tyr Pro Thr Trp Ser Ala Thr Ile Ala Leu Pro Ala 530 535 540Asp Thr Thr Ile Gln Tyr Lys Tyr Val Asn Ile Asp Gly Ser Thr Val545 550 555 560Ile Trp Glu Asp Ala Ile Ser Asn Arg Glu Ile Thr Thr Pro Ala Ser 565 570 575Gly Thr Tyr Thr Glu Lys Asp Thr Trp Asp Glu Ser 580 58537640PRTAspergillus kawachiimat_peptide(22)..(640) 37Met Arg Val Ser Thr Ser Ser Ile Ala Leu Ala Val Ser Leu Phe Gly -20 -15 -10Lys Leu Ala Leu Gly Leu Ser Ala Ala Glu Trp Arg Thr Gln Ser Ile-5 -1 1 5 10Tyr Phe Leu Leu Thr Asp Arg Phe Gly Arg Thr Asp Asn Ser Thr Thr 15 20 25Ala Thr Cys Asn Thr Gly

Asp Gln Ile Tyr Cys Gly Gly Ser Trp Gln 30 35 40Gly Ile Ile Asn His Leu Asp Tyr Ile Gln Gly Met Gly Phe Thr Ala 45 50 55Ile Trp Ile Ser Pro Ile Thr Glu Gln Leu Pro Gln Asp Thr Ser Asp60 65 70 75Gly Glu Ala Tyr His Gly Tyr Trp Gln Gln Lys Ile Tyr Tyr Val Asn 80 85 90Ser Asn Phe Gly Thr Ala Asp Asp Leu Lys Ser Leu Ser Asp Ala Leu 95 100 105His Ala Arg Gly Met Tyr Leu Met Val Asp Val Val Pro Asn His Met 110 115 120Gly Tyr Ala Gly Asn Gly Asn Asp Val Asp Tyr Ser Val Phe Asp Pro 125 130 135Phe Asp Ser Ser Ser Tyr Phe His Pro Tyr Cys Leu Ile Thr Asp Trp140 145 150 155Asp Asn Leu Thr Met Val Gln Asp Cys Trp Glu Gly Asp Thr Ile Val 160 165 170Ser Leu Pro Asp Leu Asn Thr Thr Glu Thr Ala Val Arg Thr Ile Trp 175 180 185Tyr Asp Trp Val Ala Asp Leu Val Ser Asn Tyr Ser Val Asp Gly Leu 190 195 200Arg Ile Asp Ser Val Glu Glu Val Glu Pro Asp Phe Phe Pro Gly Tyr 205 210 215Gln Glu Ala Ala Gly Val Tyr Cys Val Gly Glu Val Asp Asn Gly Asn220 225 230 235Pro Ala Leu Asp Cys Pro Tyr Gln Lys Tyr Leu Asp Gly Val Leu Asn 240 245 250Tyr Pro Ile Tyr Trp Gln Leu Leu Tyr Ala Phe Glu Ser Ser Ser Gly 255 260 265Ser Ile Ser Asn Leu Tyr Asn Met Ile Lys Ser Val Ala Ser Asp Cys 270 275 280Ser Asp Pro Thr Leu Leu Gly Asn Phe Ile Glu Asn His Asp Asn Pro 285 290 295Arg Phe Ala Ser Tyr Thr Ser Asp Tyr Ser Gln Ala Lys Asn Val Leu300 305 310 315Ser Tyr Ile Phe Leu Ser Asp Gly Ile Pro Ile Val Tyr Ala Gly Glu 320 325 330Glu Gln His Tyr Ser Gly Gly Asp Val Pro Tyr Asn Arg Glu Ala Thr 335 340 345Trp Leu Ser Gly Tyr Asp Thr Ser Ala Glu Leu Tyr Thr Trp Ile Ala 350 355 360Thr Thr Asn Ala Ile Arg Lys Leu Ala Ile Ser Ala Asp Ser Asp Tyr 365 370 375Ile Thr Tyr Lys Asn Asp Pro Ile Tyr Thr Asp Ser Asn Thr Ile Ala380 385 390 395Met Arg Lys Gly Thr Ser Gly Ser Gln Ile Ile Thr Val Leu Ser Asn 400 405 410Lys Gly Ser Ser Gly Ser Ser Tyr Thr Leu Thr Leu Ser Gly Ser Gly 415 420 425Tyr Thr Ser Gly Thr Lys Leu Ile Glu Ala Tyr Thr Cys Thr Ser Val 430 435 440Thr Val Asp Ser Asn Gly Asp Ile Pro Val Pro Met Ala Ser Gly Leu 445 450 455Pro Arg Val Leu Leu Pro Ala Ser Val Val Asp Ser Ser Ser Leu Cys460 465 470 475Gly Gly Ser Gly Asn Thr Thr Thr Thr Thr Thr Ala Ala Thr Ser Thr 480 485 490Ser Lys Ala Thr Thr Ser Ser Ser Ser Ser Ser Ala Ala Ala Thr Thr 495 500 505Ser Ser Ser Cys Thr Ala Thr Ser Thr Thr Leu Pro Ile Thr Phe Glu 510 515 520Glu Leu Val Thr Thr Thr Tyr Gly Glu Glu Val Tyr Leu Ser Gly Ser 525 530 535Ile Ser Gln Leu Gly Glu Trp His Thr Ser Asp Ala Val Lys Leu Ser540 545 550 555Ala Asp Asp Tyr Thr Ser Ser Asn Pro Glu Trp Ser Val Thr Val Ser 560 565 570Leu Pro Val Gly Thr Thr Phe Glu Tyr Lys Phe Ile Lys Val Asp Glu 575 580 585Gly Gly Ser Val Thr Trp Glu Ser Asp Pro Asn Arg Glu Tyr Thr Val 590 595 600Pro Glu Cys Gly Ser Gly Ser Gly Glu Thr Val Val Asp Thr Trp Arg 605 610 61538505PRTAspergillus nigermat_peptide(22)..(505) 38Met Arg Leu Ser Thr Ser Ser Leu Phe Leu Ser Val Ser Leu Leu Gly -20 -15 -10Lys Leu Ala Leu Gly Leu Ser Ala Ala Glu Trp Arg Thr Gln Ser Ile-5 -1 1 5 10Tyr Phe Leu Leu Thr Asp Arg Phe Gly Arg Thr Asp Asn Ser Thr Thr 15 20 25Ala Thr Cys Asp Thr Gly Asp Gln Ile Tyr Cys Gly Gly Ser Trp Gln 30 35 40Gly Ile Ile Asn His Leu Asp Tyr Ile Gln Gly Met Gly Phe Thr Ala 45 50 55Ile Trp Ile Ser Pro Ile Thr Glu Gln Leu Pro Gln Asp Thr Ala Asp60 65 70 75Gly Glu Ala Tyr His Gly Tyr Trp Gln Gln Lys Ile Tyr Asp Val Asn 80 85 90 Ser Asn Phe Gly Thr Ala Asp Asp Leu Lys Ser Leu Ser Asp Ala Leu 95 100 105His Ala Arg Gly Met Tyr Leu Met Val Asp Val Val Pro Asn His Met 110 115 120Gly Tyr Ala Gly Asn Gly Asn Asp Val Asp Tyr Ser Val Phe Asp Pro 125 130 135Phe Asp Ser Ser Ser Tyr Phe His Pro Tyr Cys Leu Ile Thr Asp Trp140 145 150 155Asp Asn Leu Thr Met Val Gln Asp Cys Trp Glu Gly Asp Thr Ile Val 160 165 170Ser Leu Pro Asp Leu Asn Thr Thr Glu Thr Ala Val Arg Thr Ile Trp 175 180 185Tyr Asp Trp Val Ala Asp Leu Val Ser Asn Tyr Ser Val Asp Gly Leu 190 195 200Arg Ile Asp Ser Val Leu Glu Val Glu Pro Asp Phe Phe Pro Gly Tyr 205 210 215Gln Glu Ala Ala Gly Val Tyr Cys Val Gly Glu Val Asp Asn Gly Asn220 225 230 235Pro Ala Leu Asp Cys Pro Tyr Gln Lys Val Leu Asp Gly Val Leu Asn 240 245 250Tyr Pro Ile Tyr Trp Gln Leu Leu Tyr Ala Phe Glu Ser Ser Ser Gly 255 260 265Ser Ile Ser Asn Leu Tyr Asn Met Ile Lys Ser Val Ala Ser Asp Cys 270 275 280Ser Asp Pro Thr Leu Leu Gly Asn Phe Ile Glu Asn His Asp Asn Pro 285 290 295Arg Phe Ala Ser Tyr Thr Ser Asp Tyr Ser Gln Ala Lys Asn Val Leu300 305 310 315Ser Tyr Ile Phe Leu Ser Asp Gly Ile Pro Ile Val Tyr Ala Gly Glu 320 325 330Glu Gln His Tyr Ser Gly Gly Lys Val Pro Tyr Asn Arg Glu Ala Thr 335 340 345Trp Leu Ser Gly Tyr Asp Thr Ser Ala Glu Leu Tyr Thr Trp Ile Ala 350 355 360Thr Thr Asn Ala Ile Arg Lys Leu Ala Ile Ser Ala Asp Ser Ala Tyr 365 370 375Ile Thr Tyr Ala Asn Asp Ala Phe Tyr Thr Asp Ser Asn Thr Ile Ala380 385 390 395Met Arg Lys Gly Thr Ser Gly Ser Gln Val Ile Thr Val Leu Ser Asn 400 405 410Lys Gly Ser Ser Gly Ser Ser Tyr Thr Leu Thr Leu Ser Gly Ser Gly 415 420 425Tyr Thr Ser Gly Thr Lys Leu Ile Glu Ala Tyr Thr Cys Thr Ser Val 430 435 440Thr Val Asp Ser Ser Gly Asp Ile Pro Val Pro Met Ala Ser Gly Leu 445 450 455Pro Arg Val Leu Leu Pro Ala Ser Val Val Asp Ser Ser Ser Leu Cys460 465 470 475Gly Gly Ser Gly Arg Leu Tyr Val Glu 48039476PRTAspergillus oryzaemat_peptide(1)..(476) 39Ala Thr Pro Ala Asp Trp Arg Ser Gln Ser Ile Tyr Phe Leu Leu Thr1 5 10 15Asp Arg Phe Ala Arg Thr Asp Gly Ser Thr Thr Ala Thr Cys Asn Thr 20 25 30Ala Asp Gln Lys Tyr Cys Gly Gly Thr Trp Gln Gly Ile Ile Asp Lys 35 40 45Leu Asp Tyr Ile Gln Gly Met Gly Phe Thr Ala Ile Trp Ile Thr Pro 50 55 60Val Thr Ala Gln Leu Pro Gln Thr Thr Ala Tyr Gly Asp Ala Tyr His65 70 75 80Gly Tyr Trp Gln Gln Asp Ile Tyr Ser Leu Asn Glu Asn Tyr Gly Thr 85 90 95Ala Asp Asp Leu Lys Ala Leu Ser Ser Ala Leu His Glu Arg Gly Met 100 105 110Tyr Leu Met Val Asp Val Val Ala Asn His Met Gly Tyr Asp Gly Ala 115 120 125Gly Ser Ser Val Asp Tyr Ser Val Phe Lys Pro Phe Ser Ser Gln Asp 130 135 140Tyr Phe His Pro Phe Cys Phe Ile Gln Asn Tyr Glu Asp Gln Thr Gln145 150 155 160Val Glu Asp Cys Trp Leu Gly Asp Asn Thr Val Ser Leu Pro Asp Leu 165 170 175Asp Thr Thr Lys Asp Val Val Lys Asn Glu Trp Tyr Asp Trp Val Gly 180 185 190Ser Leu Val Ser Asn Tyr Ser Ile Asp Gly Leu Arg Ile Asp Thr Val 195 200 205Lys His Val Gln Lys Asp Phe Trp Pro Gly Tyr Asn Lys Ala Ala Gly 210 215 220Val Tyr Cys Ile Gly Glu Val Leu Asp Gly Asp Pro Ala Tyr Thr Cys225 230 235 240Pro Tyr Gln Asn Val Met Asp Gly Val Leu Asn Tyr Pro Ile Tyr Tyr 245 250 255Pro Leu Leu Asn Ala Phe Lys Ser Thr Ser Gly Ser Met Asp Asp Leu 260 265 270Tyr Asn Met Ile Asn Thr Val Lys Ser Asp Cys Pro Asp Ser Thr Leu 275 280 285Leu Gly Thr Phe Val Glu Asn His Asp Asn Pro Arg Phe Ala Ser Tyr 290 295 300Thr Asn Asp Ile Ala Leu Ala Lys Asn Val Ala Ala Phe Ile Ile Leu305 310 315 320Asn Asp Gly Ile Pro Ile Ile Tyr Ala Gly Gln Glu Gln His Tyr Ala 325 330 335Gly Gly Asn Asp Pro Ala Asn Arg Glu Ala Thr Trp Leu Ser Gly Tyr 340 345 350Pro Thr Asp Ser Glu Leu Tyr Lys Leu Ile Ala Ser Ala Asn Ala Ile 355 360 365Arg Asn Tyr Ala Ile Ser Lys Asp Thr Gly Phe Val Thr Tyr Lys Asn 370 375 380Trp Pro Ile Tyr Lys Asp Asp Thr Thr Ile Ala Met Arg Lys Gly Thr385 390 395 400Asp Gly Ser Gln Ile Val Thr Ile Leu Ser Asn Lys Gly Ala Ser Gly 405 410 415Asp Ser Tyr Thr Leu Ser Leu Ser Gly Ala Gly Tyr Thr Ala Gly Gln 420 425 430Gln Leu Thr Glu Val Ile Gly Cys Thr Thr Val Thr Val Gly Ser Asp 435 440 445Gly Asn Val Pro Val Pro Met Ala Gly Gly Leu Pro Arg Val Leu Tyr 450 455 460Pro Thr Glu Lys Leu Ala Gly Ser Lys Ile Cys Ser465 470 475401455DNABacillus speciesCDS(1)..(1455)AA560 40cac cat aat ggt acg aac ggc aca atg atg cag tac ttt gaa tgg tat 48His His Asn Gly Thr Asn Gly Thr Met Met Gln Tyr Phe Glu Trp Tyr1 5 10 15cta cca aat gac gga aac cat tgg aat aga tta agg tct gat gca agt 96Leu Pro Asn Asp Gly Asn His Trp Asn Arg Leu Arg Ser Asp Ala Ser 20 25 30aac cta aaa gat aaa ggg atc tca gcg gtt tgg att cct cct gca tgg 144Asn Leu Lys Asp Lys Gly Ile Ser Ala Val Trp Ile Pro Pro Ala Trp 35 40 45aag ggt gcc tct caa aat gat gtg ggg tat ggt gct tat gat ctg tat 192Lys Gly Ala Ser Gln Asn Asp Val Gly Tyr Gly Ala Tyr Asp Leu Tyr 50 55 60gat tta gga gaa ttc aat caa aaa gga acc att cgt aca aaa tat gga 240Asp Leu Gly Glu Phe Asn Gln Lys Gly Thr Ile Arg Thr Lys Tyr Gly65 70 75 80acg cgc aat cag tta caa gct gca gtt aac gcc ttg aaa agt aat gga 288Thr Arg Asn Gln Leu Gln Ala Ala Val Asn Ala Leu Lys Ser Asn Gly 85 90 95att caa gtg tat ggc gat gtt gta atg aat cat aaa ggg gga gca gac 336Ile Gln Val Tyr Gly Asp Val Val Met Asn His Lys Gly Gly Ala Asp 100 105 110gct acc gaa atg gtt agg gca gtt gaa gta aac ccg aat aat aga aat 384Ala Thr Glu Met Val Arg Ala Val Glu Val Asn Pro Asn Asn Arg Asn 115 120 125caa gaa gtg tcc ggt gaa tat aca att gag gct tgg aca aag ttt gac 432Gln Glu Val Ser Gly Glu Tyr Thr Ile Glu Ala Trp Thr Lys Phe Asp 130 135 140ttt cca gga cga ggt aat act cat tca aac ttc aaa tgg aga tgg tat 480Phe Pro Gly Arg Gly Asn Thr His Ser Asn Phe Lys Trp Arg Trp Tyr145 150 155 160cac ttt gat gga gta gat tgg gat cag tca cgt aag ctg aac aat cga 528His Phe Asp Gly Val Asp Trp Asp Gln Ser Arg Lys Leu Asn Asn Arg 165 170 175att tat aaa ttt aga ggt gat gga aaa ggg tgg gat tgg gaa gtc gat 576Ile Tyr Lys Phe Arg Gly Asp Gly Lys Gly Trp Asp Trp Glu Val Asp 180 185 190aca gaa aac ggt aac tat gat tac cta atg tat gca gat att gac atg 624Thr Glu Asn Gly Asn Tyr Asp Tyr Leu Met Tyr Ala Asp Ile Asp Met 195 200 205gat cac cca gag gta gtg aat gag cta aga aat tgg ggt gtt tgg tat 672Asp His Pro Glu Val Val Asn Glu Leu Arg Asn Trp Gly Val Trp Tyr 210 215 220acg aat aca tta ggc ctt gat ggt ttt aga ata gat gca gta aaa cat 720Thr Asn Thr Leu Gly Leu Asp Gly Phe Arg Ile Asp Ala Val Lys His225 230 235 240ata aaa tac agc ttt act cgt gat tgg att aat cat gtt aga agt gca 768Ile Lys Tyr Ser Phe Thr Arg Asp Trp Ile Asn His Val Arg Ser Ala 245 250 255act ggc aaa aat atg ttt gcg gtt gcg gaa ttt tgg aaa aat gat tta 816Thr Gly Lys Asn Met Phe Ala Val Ala Glu Phe Trp Lys Asn Asp Leu 260 265 270ggt gct att gaa aac tat tta aac aaa aca aac tgg aac cat tca gtc 864Gly Ala Ile Glu Asn Tyr Leu Asn Lys Thr Asn Trp Asn His Ser Val 275 280 285ttt gat gtt ccg ctg cac tat aac ctc tat aat gct tca aaa agc gga 912Phe Asp Val Pro Leu His Tyr Asn Leu Tyr Asn Ala Ser Lys Ser Gly 290 295 300ggg aat tat gat atg agg caa ata ttt aat ggt aca gtc gtg caa aga 960Gly Asn Tyr Asp Met Arg Gln Ile Phe Asn Gly Thr Val Val Gln Arg305 310 315 320cat cca atg cat gct gtt aca ttt gtt gat aat cat gat tcg caa cct 1008His Pro Met His Ala Val Thr Phe Val Asp Asn His Asp Ser Gln Pro 325 330 335gaa gaa gct tta gag tct ttt gtt gaa gaa tgg ttc aaa cca tta gcg 1056Glu Glu Ala Leu Glu Ser Phe Val Glu Glu Trp Phe Lys Pro Leu Ala 340 345 350tat gct ttg aca tta aca cgt gaa caa ggc tac cct tct gta ttt tat 1104Tyr Ala Leu Thr Leu Thr Arg Glu Gln Gly Tyr Pro Ser Val Phe Tyr 355 360 365gga gat tat tat ggc att cca acg cat ggt gta cca gcg atg aaa tcg 1152Gly Asp Tyr Tyr Gly Ile Pro Thr His Gly Val Pro Ala Met Lys Ser 370 375 380aaa att gac ccg att cta gaa gcg cgt caa aag tat gca tat gga aga 1200Lys Ile Asp Pro Ile Leu Glu Ala Arg Gln Lys Tyr Ala Tyr Gly Arg385 390 395 400caa aat gac tac tta gac cat cat aat atc atc ggt tgg aca cgt gaa 1248Gln Asn Asp Tyr Leu Asp His His Asn Ile Ile Gly Trp Thr Arg Glu 405 410 415ggg aat aca gca cac ccc aac tcc ggt tta gct act atc atg tcc gat 1296Gly Asn Thr Ala His Pro Asn Ser Gly Leu Ala Thr Ile Met Ser Asp 420 425 430ggg gca gga gga aat aag tgg atg ttt gtt ggg cgt aat aaa gct ggt 1344Gly Ala Gly Gly Asn Lys Trp Met Phe Val Gly Arg Asn Lys Ala Gly 435 440 445caa gtt tgg acc gat atc act gga aat cgt gca ggt act gtt acg att 1392Gln Val Trp Thr Asp Ile Thr Gly Asn Arg Ala Gly Thr Val Thr Ile 450 455 460aat gct gat gga tgg ggt aat ttt tct gta aat gga gga tca gtt tct 1440Asn Ala Asp Gly Trp Gly Asn Phe Ser Val Asn Gly Gly Ser Val Ser465 470 475 480att tgg gta aac aaa 1455Ile Trp Val Asn Lys 48541485PRTBacillus species 41His His Asn Gly Thr Asn Gly Thr Met Met Gln Tyr Phe Glu Trp Tyr1 5 10 15Leu Pro Asn Asp Gly Asn His Trp Asn Arg Leu Arg Ser Asp Ala Ser 20 25 30Asn Leu Lys Asp Lys Gly Ile Ser Ala Val Trp Ile Pro Pro Ala Trp 35 40 45Lys Gly Ala Ser Gln Asn Asp Val Gly Tyr Gly Ala Tyr Asp Leu Tyr 50 55 60Asp Leu Gly Glu Phe Asn Gln Lys Gly Thr Ile Arg Thr Lys Tyr Gly65 70 75 80Thr Arg Asn Gln Leu Gln Ala Ala Val Asn Ala Leu Lys Ser Asn Gly 85 90 95Ile Gln Val Tyr Gly Asp Val Val Met Asn His

Lys Gly Gly Ala Asp 100 105 110Ala Thr Glu Met Val Arg Ala Val Glu Val Asn Pro Asn Asn Arg Asn 115 120 125Gln Glu Val Ser Gly Glu Tyr Thr Ile Glu Ala Trp Thr Lys Phe Asp 130 135 140Phe Pro Gly Arg Gly Asn Thr His Ser Asn Phe Lys Trp Arg Trp Tyr145 150 155 160His Phe Asp Gly Val Asp Trp Asp Gln Ser Arg Lys Leu Asn Asn Arg 165 170 175Ile Tyr Lys Phe Arg Gly Asp Gly Lys Gly Trp Asp Trp Glu Val Asp 180 185 190Thr Glu Asn Gly Asn Tyr Asp Tyr Leu Met Tyr Ala Asp Ile Asp Met 195 200 205Asp His Pro Glu Val Val Asn Glu Leu Arg Asn Trp Gly Val Trp Tyr 210 215 220Thr Asn Thr Leu Gly Leu Asp Gly Phe Arg Ile Asp Ala Val Lys His225 230 235 240Ile Lys Tyr Ser Phe Thr Arg Asp Trp Ile Asn His Val Arg Ser Ala 245 250 255Thr Gly Lys Asn Met Phe Ala Val Ala Glu Phe Trp Lys Asn Asp Leu 260 265 270Gly Ala Ile Glu Asn Tyr Leu Asn Lys Thr Asn Trp Asn His Ser Val 275 280 285Phe Asp Val Pro Leu His Tyr Asn Leu Tyr Asn Ala Ser Lys Ser Gly 290 295 300Gly Asn Tyr Asp Met Arg Gln Ile Phe Asn Gly Thr Val Val Gln Arg305 310 315 320His Pro Met His Ala Val Thr Phe Val Asp Asn His Asp Ser Gln Pro 325 330 335Glu Glu Ala Leu Glu Ser Phe Val Glu Glu Trp Phe Lys Pro Leu Ala 340 345 350Tyr Ala Leu Thr Leu Thr Arg Glu Gln Gly Tyr Pro Ser Val Phe Tyr 355 360 365Gly Asp Tyr Tyr Gly Ile Pro Thr His Gly Val Pro Ala Met Lys Ser 370 375 380Lys Ile Asp Pro Ile Leu Glu Ala Arg Gln Lys Tyr Ala Tyr Gly Arg385 390 395 400Gln Asn Asp Tyr Leu Asp His His Asn Ile Ile Gly Trp Thr Arg Glu 405 410 415Gly Asn Thr Ala His Pro Asn Ser Gly Leu Ala Thr Ile Met Ser Asp 420 425 430Gly Ala Gly Gly Asn Lys Trp Met Phe Val Gly Arg Asn Lys Ala Gly 435 440 445Gln Val Trp Thr Asp Ile Thr Gly Asn Arg Ala Gly Thr Val Thr Ile 450 455 460Asn Ala Asp Gly Trp Gly Asn Phe Ser Val Asn Gly Gly Ser Val Ser465 470 475 480Ile Trp Val Asn Lys 485421350DNARhizomucor pusillusCDS(1)..(1350) 42agc cct ttg ccc caa cag cag cga tat ggc aaa aga gca act tcg gat 48Ser Pro Leu Pro Gln Gln Gln Arg Tyr Gly Lys Arg Ala Thr Ser Asp1 5 10 15gac tgg aaa ggc aag gcc att tat cag ctg ctt aca gat cga ttt ggc 96Asp Trp Lys Gly Lys Ala Ile Tyr Gln Leu Leu Thr Asp Arg Phe Gly 20 25 30cgc gcc gat gac tca aca agc aac tgc tct aat tta tcc aac tac tgt 144Arg Ala Asp Asp Ser Thr Ser Asn Cys Ser Asn Leu Ser Asn Tyr Cys 35 40 45ggt ggt acc tac gaa ggc att acg aag cat ctt gac tac att tcc ggt 192Gly Gly Thr Tyr Glu Gly Ile Thr Lys His Leu Asp Tyr Ile Ser Gly 50 55 60atg ggc ttt gat gct atc tgg ata tcg cca att ccc aag aac tcg gat 240Met Gly Phe Asp Ala Ile Trp Ile Ser Pro Ile Pro Lys Asn Ser Asp65 70 75 80gga ggc tac cac ggc tac tgg gct aca gat ttc tac caa cta aac agc 288Gly Gly Tyr His Gly Tyr Trp Ala Thr Asp Phe Tyr Gln Leu Asn Ser 85 90 95aac ttt ggt gat gaa tcc cag ctc aaa gcg ctc atc cag gct gcc cat 336Asn Phe Gly Asp Glu Ser Gln Leu Lys Ala Leu Ile Gln Ala Ala His 100 105 110gaa cgt gac atg tat gtt atg ctt gat gtc gta gcc aat cat gca ggt 384Glu Arg Asp Met Tyr Val Met Leu Asp Val Val Ala Asn His Ala Gly 115 120 125ccc acc agc aat ggc tac tcg ggt tac aca ttc ggc gat gca agt tta 432Pro Thr Ser Asn Gly Tyr Ser Gly Tyr Thr Phe Gly Asp Ala Ser Leu 130 135 140tat cat cct aaa tgc acc ata gat tac aat gat cag acg tct att gag 480Tyr His Pro Lys Cys Thr Ile Asp Tyr Asn Asp Gln Thr Ser Ile Glu145 150 155 160caa tgc tgg gtt gct gac gag ttg cct gat att gac act gaa aat tct 528Gln Cys Trp Val Ala Asp Glu Leu Pro Asp Ile Asp Thr Glu Asn Ser 165 170 175gac aac gtg gcc att ctc aac gac atc gtc tcc ggc tgg gtg ggt aac 576Asp Asn Val Ala Ile Leu Asn Asp Ile Val Ser Gly Trp Val Gly Asn 180 185 190tat agc ttt gac ggc atc cgc att gat act gtc aag cat att cgc aag 624Tyr Ser Phe Asp Gly Ile Arg Ile Asp Thr Val Lys His Ile Arg Lys 195 200 205gac ttt tgg aca ggc tac gca gaa gct gcc ggc gta ttc gca act gga 672Asp Phe Trp Thr Gly Tyr Ala Glu Ala Ala Gly Val Phe Ala Thr Gly 210 215 220gag gtc ttc aat ggt gat ccg gcc tac gtt gga cct tat caa aag tac 720Glu Val Phe Asn Gly Asp Pro Ala Tyr Val Gly Pro Tyr Gln Lys Tyr225 230 235 240ctg cca tct ctc atc aat tac cca atg tat tac gct ttg aac gac gtc 768Leu Pro Ser Leu Ile Asn Tyr Pro Met Tyr Tyr Ala Leu Asn Asp Val 245 250 255ttt gta tcc aaa agc aaa gga ttc agc cgc atc agc gaa atg cta gga 816Phe Val Ser Lys Ser Lys Gly Phe Ser Arg Ile Ser Glu Met Leu Gly 260 265 270tca aat cgc aat gcg ttt gag gat acc agc gta ctt aca acg ttt gta 864Ser Asn Arg Asn Ala Phe Glu Asp Thr Ser Val Leu Thr Thr Phe Val 275 280 285gac aac cat gac aat ccg cgc ttc ttg aac agt caa agc gac aag gct 912Asp Asn His Asp Asn Pro Arg Phe Leu Asn Ser Gln Ser Asp Lys Ala 290 295 300ctc ttc aag aac gct ctc aca tac gta ctg cta ggt gaa ggc atc cca 960Leu Phe Lys Asn Ala Leu Thr Tyr Val Leu Leu Gly Glu Gly Ile Pro305 310 315 320att gtg tat tat ggt tct gag caa ggt ttc agc gga gga gcg gat cct 1008Ile Val Tyr Tyr Gly Ser Glu Gln Gly Phe Ser Gly Gly Ala Asp Pro 325 330 335gct aac cgt gaa gtg ctg tgg acc acc aat tat gat aca tcc agc gat 1056Ala Asn Arg Glu Val Leu Trp Thr Thr Asn Tyr Asp Thr Ser Ser Asp 340 345 350ctc tac caa ttt atc aag aca gtc aac agt gtc cgc atg aaa agc aac 1104Leu Tyr Gln Phe Ile Lys Thr Val Asn Ser Val Arg Met Lys Ser Asn 355 360 365aag gcc gtc tac atg gat att tat gtt ggc gac aat gct tac gcc ttc 1152Lys Ala Val Tyr Met Asp Ile Tyr Val Gly Asp Asn Ala Tyr Ala Phe 370 375 380aag cac ggc gat gct ttg gtt gtt ctc aat aac tat gga tca ggt tcc 1200Lys His Gly Asp Ala Leu Val Val Leu Asn Asn Tyr Gly Ser Gly Ser385 390 395 400aca aac caa gtc agc ttc agc gtt agt ggc aag ttc gat agc ggc gca 1248Thr Asn Gln Val Ser Phe Ser Val Ser Gly Lys Phe Asp Ser Gly Ala 405 410 415agc ctc atg gat att gtc agt aac att acc acc acg gtg tcc tcg gat 1296Ser Leu Met Asp Ile Val Ser Asn Ile Thr Thr Thr Val Ser Ser Asp 420 425 430gga aca gtc act ttc aac ctt aaa gat gga ctt ccg gct atc ttc acc 1344Gly Thr Val Thr Phe Asn Leu Lys Asp Gly Leu Pro Ala Ile Phe Thr 435 440 445tct gct 1350Ser Ala45043450PRTRhizomucor pusillus 43Ser Pro Leu Pro Gln Gln Gln Arg Tyr Gly Lys Arg Ala Thr Ser Asp1 5 10 15Asp Trp Lys Gly Lys Ala Ile Tyr Gln Leu Leu Thr Asp Arg Phe Gly 20 25 30Arg Ala Asp Asp Ser Thr Ser Asn Cys Ser Asn Leu Ser Asn Tyr Cys 35 40 45Gly Gly Thr Tyr Glu Gly Ile Thr Lys His Leu Asp Tyr Ile Ser Gly 50 55 60Met Gly Phe Asp Ala Ile Trp Ile Ser Pro Ile Pro Lys Asn Ser Asp65 70 75 80Gly Gly Tyr His Gly Tyr Trp Ala Thr Asp Phe Tyr Gln Leu Asn Ser 85 90 95Asn Phe Gly Asp Glu Ser Gln Leu Lys Ala Leu Ile Gln Ala Ala His 100 105 110Glu Arg Asp Met Tyr Val Met Leu Asp Val Val Ala Asn His Ala Gly 115 120 125Pro Thr Ser Asn Gly Tyr Ser Gly Tyr Thr Phe Gly Asp Ala Ser Leu 130 135 140Tyr His Pro Lys Cys Thr Ile Asp Tyr Asn Asp Gln Thr Ser Ile Glu145 150 155 160Gln Cys Trp Val Ala Asp Glu Leu Pro Asp Ile Asp Thr Glu Asn Ser 165 170 175Asp Asn Val Ala Ile Leu Asn Asp Ile Val Ser Gly Trp Val Gly Asn 180 185 190Tyr Ser Phe Asp Gly Ile Arg Ile Asp Thr Val Lys His Ile Arg Lys 195 200 205Asp Phe Trp Thr Gly Tyr Ala Glu Ala Ala Gly Val Phe Ala Thr Gly 210 215 220Glu Val Phe Asn Gly Asp Pro Ala Tyr Val Gly Pro Tyr Gln Lys Tyr225 230 235 240Leu Pro Ser Leu Ile Asn Tyr Pro Met Tyr Tyr Ala Leu Asn Asp Val 245 250 255Phe Val Ser Lys Ser Lys Gly Phe Ser Arg Ile Ser Glu Met Leu Gly 260 265 270Ser Asn Arg Asn Ala Phe Glu Asp Thr Ser Val Leu Thr Thr Phe Val 275 280 285Asp Asn His Asp Asn Pro Arg Phe Leu Asn Ser Gln Ser Asp Lys Ala 290 295 300Leu Phe Lys Asn Ala Leu Thr Tyr Val Leu Leu Gly Glu Gly Ile Pro305 310 315 320Ile Val Tyr Tyr Gly Ser Glu Gln Gly Phe Ser Gly Gly Ala Asp Pro 325 330 335Ala Asn Arg Glu Val Leu Trp Thr Thr Asn Tyr Asp Thr Ser Ser Asp 340 345 350Leu Tyr Gln Phe Ile Lys Thr Val Asn Ser Val Arg Met Lys Ser Asn 355 360 365Lys Ala Val Tyr Met Asp Ile Tyr Val Gly Asp Asn Ala Tyr Ala Phe 370 375 380Lys His Gly Asp Ala Leu Val Val Leu Asn Asn Tyr Gly Ser Gly Ser385 390 395 400Thr Asn Gln Val Ser Phe Ser Val Ser Gly Lys Phe Asp Ser Gly Ala 405 410 415Ser Leu Met Asp Ile Val Ser Asn Ile Thr Thr Thr Val Ser Ser Asp 420 425 430Gly Thr Val Thr Phe Asn Leu Lys Asp Gly Leu Pro Ala Ile Phe Thr 435 440 445Ser Ala 45044558PRTArtificial sequenceSynthetic construct 44Ser Pro Leu Pro Gln Gln Gln Arg Tyr Gly Lys Arg Ala Thr Ser Asp1 5 10 15Asp Trp Lys Ser Lys Ala Ile Tyr Gln Leu Leu Thr Asp Arg Phe Gly 20 25 30Arg Ala Asp Asp Ser Thr Ser Asn Cys Ser Asn Leu Ser Asn Tyr Cys 35 40 45Gly Gly Thr Tyr Glu Gly Ile Thr Lys His Leu Asp Tyr Ile Ser Gly 50 55 60Met Gly Phe Asp Ala Ile Trp Ile Ser Pro Ile Pro Lys Asn Ser Asp65 70 75 80Gly Gly Tyr His Gly Tyr Trp Ala Thr Asp Phe Tyr Gln Leu Asn Ser 85 90 95Asn Phe Gly Asp Glu Ser Gln Leu Lys Ala Leu Ile Gln Ala Ala His 100 105 110Glu Arg Asp Met Tyr Val Met Leu Asp Val Val Ala Asn His Ala Gly 115 120 125Pro Thr Ser Asn Gly Tyr Ser Gly Tyr Thr Phe Gly Asp Ala Ser Leu 130 135 140Tyr His Pro Lys Cys Thr Ile Asp Tyr Asn Asp Gln Thr Ser Ile Glu145 150 155 160Gln Cys Trp Val Ala Asp Glu Leu Pro Asp Ile Asp Thr Glu Asn Ser 165 170 175Asp Asn Val Ala Ile Leu Asn Asp Ile Val Ser Gly Trp Val Gly Asn 180 185 190Tyr Ser Phe Asp Gly Ile Arg Ile Asp Thr Val Lys His Ile Arg Lys 195 200 205Asp Phe Trp Thr Gly Tyr Ala Glu Ala Ala Gly Val Phe Ala Thr Gly 210 215 220Glu Val Phe Asn Gly Asp Pro Ala Tyr Val Gly Pro Tyr Gln Lys Tyr225 230 235 240Leu Pro Ser Leu Ile Asn Tyr Pro Met Tyr Tyr Ala Leu Asn Asp Val 245 250 255Phe Val Ser Lys Ser Lys Gly Phe Ser Arg Ile Ser Glu Met Leu Gly 260 265 270Ser Asn Arg Asn Ala Phe Glu Asp Thr Ser Val Leu Thr Thr Phe Val 275 280 285Asp Asn His Asp Asn Pro Arg Phe Leu Asn Ser Gln Ser Asp Lys Ala 290 295 300Leu Phe Lys Asn Ala Leu Thr Tyr Val Leu Leu Gly Glu Gly Ile Pro305 310 315 320Ile Val Tyr Tyr Gly Ser Glu Gln Gly Phe Ser Gly Gly Ala Asp Pro 325 330 335Ala Asn Arg Glu Val Leu Trp Thr Thr Asn Tyr Asp Thr Ser Ser Asp 340 345 350Leu Tyr Gln Phe Ile Lys Thr Val Asn Ser Val Arg Met Lys Ser Asn 355 360 365Lys Ala Val Tyr Met Asp Ile Tyr Val Gly Asp Asn Ala Tyr Ala Phe 370 375 380Lys His Gly Asp Ala Leu Val Val Leu Asn Asn Tyr Gly Ser Gly Ser385 390 395 400Thr Asn Gln Val Ser Phe Ser Val Ser Gly Lys Phe Asp Ser Gly Ala 405 410 415Ser Leu Met Asp Ile Val Ser Asn Ile Thr Thr Thr Val Ser Ser Asp 420 425 430Gly Thr Val Thr Phe Asn Leu Lys Asp Gly Leu Pro Ala Ile Phe Thr 435 440 445Ser Ala Gly Ala Thr Ser Pro Gly Gly Ser Ser Gly Ser Val Glu Val 450 455 460Thr Phe Asp Val Tyr Ala Thr Thr Val Tyr Gly Gln Asn Ile Tyr Ile465 470 475 480Thr Gly Asp Val Ser Glu Leu Gly Asn Trp Thr Pro Ala Asn Gly Val 485 490 495Ala Leu Ser Ser Ala Asn Tyr Pro Thr Trp Ser Ala Thr Ile Ala Leu 500 505 510Pro Ala Asp Thr Thr Ile Gln Tyr Lys Tyr Val Asn Ile Asp Gly Ser 515 520 525Thr Val Ile Trp Glu Asp Ala Ile Ser Asn Arg Glu Ile Thr Thr Pro 530 535 540Ala Ser Gly Thr Tyr Thr Glu Lys Asp Thr Trp Asp Glu Ser545 550 555451398DNAMeripilus giganteusCDS(1)..(1398) 45cgc cct act gtc ttt gac gcc ggc gcg gac gca cac tcg ctg cat gcc 48Arg Pro Thr Val Phe Asp Ala Gly Ala Asp Ala His Ser Leu His Ala1 5 10 15cgg gcc ccc tcc ggc agc aag gat gtc atc atc cag atg ttt gag tgg 96Arg Ala Pro Ser Gly Ser Lys Asp Val Ile Ile Gln Met Phe Glu Trp 20 25 30aac tgg gac agc gtc gct gcc gag tgc act aac ttc atc ggc ccc gcc 144Asn Trp Asp Ser Val Ala Ala Glu Cys Thr Asn Phe Ile Gly Pro Ala 35 40 45ggg tac ggc ttc gtg caa gtg agc ccg ccc cag gag acc atc cag ggc 192Gly Tyr Gly Phe Val Gln Val Ser Pro Pro Gln Glu Thr Ile Gln Gly 50 55 60gcg cag tgg tgg acc gac tac cag ccg gtg tcg tac acg ctc act ggg 240Ala Gln Trp Trp Thr Asp Tyr Gln Pro Val Ser Tyr Thr Leu Thr Gly65 70 75 80aag cgg ggc gac cgc tcc cag ttt gcg aac atg att act acg tgc cac 288Lys Arg Gly Asp Arg Ser Gln Phe Ala Asn Met Ile Thr Thr Cys His 85 90 95gcc gcg ggc gtc ggc gtg atc gtt gac acc atc tgg aac cac atg gcg 336Ala Ala Gly Val Gly Val Ile Val Asp Thr Ile Trp Asn His Met Ala 100 105 110ggc gtc gac tcc ggc acg ggt acc gcc ggc tcg tcc ttc acg cac tac 384Gly Val Asp Ser Gly Thr Gly Thr Ala Gly Ser Ser Phe Thr His Tyr 115 120 125aac tac ccc ggc atc tac caa aac cag gac ttt cac cac tgc ggc ctc 432Asn Tyr Pro Gly Ile Tyr Gln Asn Gln Asp Phe His His Cys Gly Leu 130 135 140gag ccg ggc gat gac atc gtc aac tac gac aac gcg gtt gag gtc cag 480Glu Pro Gly Asp Asp Ile Val Asn Tyr Asp Asn Ala Val Glu Val Gln145 150 155 160acc tgc gag ctt gtc aac ctc gct gac ctc gcc acc gac acg gag tat 528Thr Cys Glu Leu Val Asn Leu Ala Asp Leu Ala Thr Asp Thr Glu Tyr 165 170 175gtg cgc ggt cgc ctt gcc cag tac gga aac gac ctg ctc tcg ctc ggt 576Val Arg Gly Arg Leu Ala Gln Tyr Gly Asn Asp Leu Leu Ser Leu Gly 180 185 190gcc gat ggc ctg cgt ctt gac gct tcc aaa cac att cct gtg ggc gac 624Ala Asp Gly Leu Arg Leu Asp Ala Ser Lys His Ile Pro Val Gly Asp 195 200

205atc gcg aac atc ctg tct cgc ctc agt cgc tct gtc tac atc acc cag 672Ile Ala Asn Ile Leu Ser Arg Leu Ser Arg Ser Val Tyr Ile Thr Gln 210 215 220gaa gtc atc ttt ggg gcc ggc gag ccc atc acg ccg aac cag tac acc 720Glu Val Ile Phe Gly Ala Gly Glu Pro Ile Thr Pro Asn Gln Tyr Thr225 230 235 240ggg aac ggc gac gtt cag gag ttc cgc tac acc tct gcg cta aag gac 768Gly Asn Gly Asp Val Gln Glu Phe Arg Tyr Thr Ser Ala Leu Lys Asp 245 250 255gcc ttc ttg agc tcg ggc ata tcc aac ctg cag gac ttc gaa aac cgt 816Ala Phe Leu Ser Ser Gly Ile Ser Asn Leu Gln Asp Phe Glu Asn Arg 260 265 270gga tgg gta cct ggc tcg ggc gcc aac gtg ttc gtc gtc aac cat gac 864Gly Trp Val Pro Gly Ser Gly Ala Asn Val Phe Val Val Asn His Asp 275 280 285acc gag cgg aac ggc gcg tcg ctg aac aac aac tcg cct tcg aac acc 912Thr Glu Arg Asn Gly Ala Ser Leu Asn Asn Asn Ser Pro Ser Asn Thr 290 295 300tac gtc acc gcg acg atc ttc tcg ctc gca cac ccg tac ggc acg ccc 960Tyr Val Thr Ala Thr Ile Phe Ser Leu Ala His Pro Tyr Gly Thr Pro305 310 315 320acg atc ctc tcc tcg tat gat ggc ttc acg aac acc gac gcc ggt gcg 1008Thr Ile Leu Ser Ser Tyr Asp Gly Phe Thr Asn Thr Asp Ala Gly Ala 325 330 335ccg aac aac aac gtc ggc aca tgc tcg acc agc ggt ggt gcg aac ggg 1056Pro Asn Asn Asn Val Gly Thr Cys Ser Thr Ser Gly Gly Ala Asn Gly 340 345 350tgg ctc tgc cag cac cgc tgg acc gcg atc gcc ggc atg gtc ggc ttc 1104Trp Leu Cys Gln His Arg Trp Thr Ala Ile Ala Gly Met Val Gly Phe 355 360 365cgc aac aac gtc ggc agc gct gca ctc aac aac tgg cag gcc ccg cag 1152Arg Asn Asn Val Gly Ser Ala Ala Leu Asn Asn Trp Gln Ala Pro Gln 370 375 380tcg cag cag att gcg ttc ggt cgc ggc gca ctt ggc ttc gtc gcg atc 1200Ser Gln Gln Ile Ala Phe Gly Arg Gly Ala Leu Gly Phe Val Ala Ile385 390 395 400aac aac gcc gac tcg gcc tgg tct acg acg ttc acc act tcc ctc ccc 1248Asn Asn Ala Asp Ser Ala Trp Ser Thr Thr Phe Thr Thr Ser Leu Pro 405 410 415gat ggt tcc tac tgc gat gtc atc agc ggc aag gcc tcc ggc agt agc 1296Asp Gly Ser Tyr Cys Asp Val Ile Ser Gly Lys Ala Ser Gly Ser Ser 420 425 430tgc acc ggt tct tcg ttc acc gtc tcc ggc ggg aag ctg acc gcc acg 1344Cys Thr Gly Ser Ser Phe Thr Val Ser Gly Gly Lys Leu Thr Ala Thr 435 440 445gtg ccg gcg cgt agc gcc atc gcc gtg cac acc ggt cag aaa ggt tct 1392Val Pro Ala Arg Ser Ala Ile Ala Val His Thr Gly Gln Lys Gly Ser 450 455 460ggt ggt 1398Gly Gly46546466PRTMeripilus giganteus 46Arg Pro Thr Val Phe Asp Ala Gly Ala Asp Ala His Ser Leu His Ala1 5 10 15Arg Ala Pro Ser Gly Ser Lys Asp Val Ile Ile Gln Met Phe Glu Trp 20 25 30Asn Trp Asp Ser Val Ala Ala Glu Cys Thr Asn Phe Ile Gly Pro Ala 35 40 45Gly Tyr Gly Phe Val Gln Val Ser Pro Pro Gln Glu Thr Ile Gln Gly 50 55 60Ala Gln Trp Trp Thr Asp Tyr Gln Pro Val Ser Tyr Thr Leu Thr Gly65 70 75 80Lys Arg Gly Asp Arg Ser Gln Phe Ala Asn Met Ile Thr Thr Cys His 85 90 95Ala Ala Gly Val Gly Val Ile Val Asp Thr Ile Trp Asn His Met Ala 100 105 110Gly Val Asp Ser Gly Thr Gly Thr Ala Gly Ser Ser Phe Thr His Tyr 115 120 125Asn Tyr Pro Gly Ile Tyr Gln Asn Gln Asp Phe His His Cys Gly Leu 130 135 140Glu Pro Gly Asp Asp Ile Val Asn Tyr Asp Asn Ala Val Glu Val Gln145 150 155 160Thr Cys Glu Leu Val Asn Leu Ala Asp Leu Ala Thr Asp Thr Glu Tyr 165 170 175Val Arg Gly Arg Leu Ala Gln Tyr Gly Asn Asp Leu Leu Ser Leu Gly 180 185 190Ala Asp Gly Leu Arg Leu Asp Ala Ser Lys His Ile Pro Val Gly Asp 195 200 205Ile Ala Asn Ile Leu Ser Arg Leu Ser Arg Ser Val Tyr Ile Thr Gln 210 215 220Glu Val Ile Phe Gly Ala Gly Glu Pro Ile Thr Pro Asn Gln Tyr Thr225 230 235 240Gly Asn Gly Asp Val Gln Glu Phe Arg Tyr Thr Ser Ala Leu Lys Asp 245 250 255Ala Phe Leu Ser Ser Gly Ile Ser Asn Leu Gln Asp Phe Glu Asn Arg 260 265 270Gly Trp Val Pro Gly Ser Gly Ala Asn Val Phe Val Val Asn His Asp 275 280 285Thr Glu Arg Asn Gly Ala Ser Leu Asn Asn Asn Ser Pro Ser Asn Thr 290 295 300Tyr Val Thr Ala Thr Ile Phe Ser Leu Ala His Pro Tyr Gly Thr Pro305 310 315 320Thr Ile Leu Ser Ser Tyr Asp Gly Phe Thr Asn Thr Asp Ala Gly Ala 325 330 335Pro Asn Asn Asn Val Gly Thr Cys Ser Thr Ser Gly Gly Ala Asn Gly 340 345 350Trp Leu Cys Gln His Arg Trp Thr Ala Ile Ala Gly Met Val Gly Phe 355 360 365Arg Asn Asn Val Gly Ser Ala Ala Leu Asn Asn Trp Gln Ala Pro Gln 370 375 380Ser Gln Gln Ile Ala Phe Gly Arg Gly Ala Leu Gly Phe Val Ala Ile385 390 395 400Asn Asn Ala Asp Ser Ala Trp Ser Thr Thr Phe Thr Thr Ser Leu Pro 405 410 415Asp Gly Ser Tyr Cys Asp Val Ile Ser Gly Lys Ala Ser Gly Ser Ser 420 425 430Cys Thr Gly Ser Ser Phe Thr Val Ser Gly Gly Lys Leu Thr Ala Thr 435 440 445Val Pro Ala Arg Ser Ala Ile Ala Val His Thr Gly Gln Lys Gly Ser 450 455 460Gly Gly46547574PRTMeripilus giganteusMISC_FEATURE(1)..(477)Meripilus giganteus alpha-amylase 47Arg Pro Thr Val Phe Asp Ala Gly Ala Asp Ala His Ser Leu His Ala1 5 10 15Arg Ala Pro Ser Gly Ser Lys Asp Val Ile Ile Gln Met Phe Glu Trp 20 25 30Asn Trp Asp Ser Val Ala Ala Glu Cys Thr Asn Phe Ile Gly Pro Ala 35 40 45Gly Tyr Gly Phe Val Gln Val Ser Pro Pro Gln Glu Thr Ile Gln Gly 50 55 60Ala Gln Trp Trp Thr Asp Tyr Gln Pro Val Ser Tyr Thr Leu Thr Gly65 70 75 80Lys Arg Gly Asp Arg Ser Gln Phe Ala Asn Met Ile Thr Thr Cys His 85 90 95Ala Ala Gly Val Gly Val Ile Val Asp Thr Ile Trp Asn His Met Ala 100 105 110Gly Val Asp Ser Gly Thr Gly Thr Ala Gly Ser Ser Phe Thr His Tyr 115 120 125Asn Tyr Pro Gly Ile Tyr Gln Asn Gln Asp Phe His His Cys Gly Leu 130 135 140Glu Pro Gly Asp Asp Ile Val Asn Tyr Asp Asn Ala Val Glu Val Gln145 150 155 160Thr Cys Glu Leu Val Asn Leu Ala Asp Leu Ala Thr Asp Thr Glu Tyr 165 170 175Val Arg Gly Arg Leu Ala Gln Tyr Gly Asn Asp Leu Leu Ser Leu Gly 180 185 190Ala Asp Gly Leu Arg Leu Asp Ala Ser Lys His Ile Pro Val Gly Asp 195 200 205Ile Ala Asn Ile Leu Ser Arg Leu Ser Arg Ser Val Tyr Ile Thr Gln 210 215 220Glu Val Ile Phe Gly Ala Gly Glu Pro Ile Thr Pro Asn Gln Tyr Thr225 230 235 240Gly Asn Gly Asp Val Gln Glu Phe Arg Tyr Thr Ser Ala Leu Lys Asp 245 250 255Ala Phe Leu Ser Ser Gly Ile Ser Asn Leu Gln Asp Phe Glu Asn Arg 260 265 270Gly Trp Val Pro Gly Ser Gly Ala Asn Val Phe Val Val Asn His Asp 275 280 285Thr Glu Arg Asn Gly Ala Ser Leu Asn Asn Asn Ser Pro Ser Asn Thr 290 295 300Tyr Val Thr Ala Thr Ile Phe Ser Leu Ala His Pro Tyr Gly Thr Pro305 310 315 320Thr Ile Leu Ser Ser Tyr Asp Gly Phe Thr Asn Thr Asp Ala Gly Ala 325 330 335Pro Asn Asn Asn Val Gly Thr Cys Ser Thr Ser Gly Gly Ala Asn Gly 340 345 350Trp Leu Cys Gln His Arg Trp Thr Ala Ile Ala Gly Met Val Gly Phe 355 360 365Arg Asn Asn Val Gly Ser Ala Ala Leu Asn Asn Trp Gln Ala Pro Gln 370 375 380Ser Gln Gln Ile Ala Phe Gly Arg Gly Ala Leu Gly Phe Val Ala Ile385 390 395 400Asn Asn Ala Asp Ser Ala Trp Ser Thr Thr Phe Thr Thr Ser Leu Pro 405 410 415Asp Gly Ser Tyr Cys Asp Val Ile Ser Gly Lys Ala Ser Gly Ser Ser 420 425 430Cys Thr Gly Ser Ser Phe Thr Val Ser Gly Gly Lys Leu Thr Ala Thr 435 440 445Val Pro Ala Arg Ser Ala Ile Ala Val His Thr Gly Gln Lys Gly Ser 450 455 460Gly Gly Gly Ala Thr Ser Pro Gly Gly Ser Ser Gly Ser Val Glu Val465 470 475 480Thr Phe Asp Val Tyr Ala Thr Thr Val Tyr Gly Gln Asn Ile Tyr Ile 485 490 495Thr Gly Asp Val Ser Glu Leu Gly Asn Trp Thr Pro Ala Asn Gly Val 500 505 510Ala Leu Ser Ser Ala Asn Tyr Pro Thr Trp Ser Ala Thr Ile Ala Leu 515 520 525Pro Ala Asp Thr Thr Ile Gln Tyr Lys Tyr Val Asn Ile Asp Gly Ser 530 535 540Thr Val Ile Trp Glu Asp Ala Ile Ser Asn Arg Glu Ile Thr Thr Pro545 550 555 560Ala Ser Gly Thr Tyr Thr Glu Lys Asp Thr Trp Asp Glu Ser 565 570

Claims

1-31. (canceled)

32. A process for desizing a sized fabric containing starch or starch derivatives during manufacture of a fabric, which process comprises incubating said sized fabric in an aqueous treating solution having a pH in the range between 1 and 5 which aqueous treating solution comprises an alpha-amylase.

33. The process of claim 32, wherein the pH is in the range between 1 and 4.

34. The process of claim 32, wherein the alpha-amylase is of bacterial or fungal.

35. The process of claim 32, wherein the alpha-amylase is an acid alpha-amylase.

36. The process of claim 35, wherein the alpha-amylase is a derived from a strain of Aspergillus, Rhizomucor, or Meripilus.

37. The process of claim 36, wherein the Aspergillus alpha-amylase is the Aspergillus niger alpha-amylase disclosed in SEQ ID NO: 38, or a variant thereof.

38. The process of claim 36, wherein the Rhizomucor alpha-amylase is the Rhizomucor pusillus alpha-amylase disclosed in SEQ ID NO: 43, or a variant thereof.

39. The process of claim 32, wherein the alpha-amylase is present in a concentration of 1-3,000 AFAU/kg fabric or 1-3,000 AFAU/L treating solution.

40. The process of claim 35, wherein the bacterial alpha-amylase is derived from a strain of the genus Bacillus.

41. The process of claim 40, wherein the bacterial alpha-amylase is the alpha-amylase disclosed as SEQ ID NO: 40.

42. The process of claim 40, wherein the Bacillus alpha-amylase has one or more deletions in position D183 and G184 (using the SEQ ID NO: 40 numbering).

43. The process of claim 32, wherein the alpha-amylase is a hybrid enzyme comprising a carbohydrate-binding domain (CBD).

44. The process of claim 32, wherein the alpha-amylase comprises a starch-binding domain of fungal or bacterial origin.

45. The process of claim 43, wherein the carbohydrate-binding domain (CBD) is derived from a strain of Aspergillus, Athelia, or Talaromyces.

46. The process of claim 43, wherein the amino acid sequence of the carbohydrate-binding domain (CBD) is derived from Aspergillus kawachii, Aspergillus niger, or Athelia sp.

47. The process of claim 43, wherein the alpha-amylase comprising a CBD comprises a linker between the alpha-amylase and CBD or starch-binding domain.

48. The process of claim 47, wherein the linker is derived from is derived from a strain of Athelia or Aspergillus.

49. The process of claim 32, wherein the alpha-amylase is the hybrid alpha-amylase shown in SEQ ID NO: 44 comprising a catalytic domain (CD) from Rhizomucor pusillus alpha-amylase having a carbohydrate-binding domain (CBD) from the Athelia rolfsii glucoamylase.

50. The process of claim 32, wherein the process is carried out at a temperature in the range from 5-90.degree. C.

51. The process of claim 32, wherein the process is carried out in the presence of a surfactant, preferably the surfactant is present in a concentration of 0.1-10 g/L.