HK1104329B - Simultaneous desizing and scouring process - Google Patents

Description

Method for simultaneous desizing and scouring

Sequence listing reference

The application contains information in the form of a sequence table, which information is attached to the application later and is also filed on a data carrier along with the application. The contents of the data carrier are hereby incorporated by reference in their entirety.

Technical Field

The invention relates to a method for simultaneously desizing and scouring sized fabrics (sized fabric). The invention also relates to compositions suitable for use in the methods of the invention.

Background

In the textile processing industry, alpha-amylases are traditionally used as an aid in desizing processes to facilitate the removal of starch-containing size as a protective coating on yarns (yarn) during the textile process. The complete removal of the size coating after weaving is essential to ensure optimal results in subsequent processes, wherein the fabric is typically scoured, bleached, dyed and/or printed. Enzymatic starch breakdown is preferred because it does not have any deleterious effect on the textile material. In order to reduce processing costs while increasing throughput, desizing processes are sometimes combined with scouring steps.

WO 95/21417 suggests the use of an oxidation stable alpha-amylase for simultaneous desizing and scouring of sized fabrics.

However, it would be desirable to provide further improved methods of simultaneous desizing and scouring.

Disclosure of Invention

The present invention aims to propose an improved simultaneous desizing and scouring process.

In a first aspect, the present invention relates to a process for simultaneously desizing and scouring of sized fabric containing starch or starch derivatives, which comprises treating the fabric with an alkaline alpha-amylase and an alkaline scouring enzyme.

In the context of the present invention, the term "fabric" includes garments (garment), fibers, yarns and other kinds of processed fabrics. Fabrics may be obtained from fibers by weaving, knitting or non-weaving operations. Weaving and knitting require yarn as input, while nonwovens are the result of random fiber attachment (paper can be considered a nonwoven).

Woven fabrics (woven fabrics) are woven by weaving "filling" or weft yarns (weft yarns) between warp yarns (warp yarns) extending in the longitudinal axial direction on a loom (loom). In order to lubricate and prevent abrasion when weft yarns are inserted at high speed during weaving, warp yarns must be sized before weaving. The weft yarns may be woven through the warp yarns in an "over-one-under-the-next" pattern (plain weave), or in an "over-one-under-two" pattern (bias weave) or in other endless arrangements. The strength (strength), texture (texture) and pattern (pattern) are not only related to the type/quality of the yarn, but also to the type of knitting. Generally, coats (stress), shirts, pants, sheets, towels, drapes (drapery), etc. are made of woven fabric.

Knitting forms fabrics by joining interlocking loops of yarn together. In contrast to woven fabrics which are woven from two types of yarns and contain a plurality of ends (ends), knitted fabrics are woven from a single continuous yarn. There are many different ways of knitting the loops of yarn, and the characteristics of the final fabric depend on both the type of yarn and the type of knitting. Underwear, sweaters (sweater), socks, sweaters (sweat shirt), and the like are made of knitted fabric.

Nonwovens are fabric sheets made by joining and/or interlocking fibers and threads by mechanical, thermal, chemical, or solvent-mediated methods. The resulting product may be in the form of a network, a sheet or a film. Specific examples are: disposable baby diapers, towels, wipes (wipes), surgical gowns (surgical gown), "environmentally friendly" fibers, filter media, bedding (padding), roofing materials, backings for bi-directional fabrics (backing for two-dimensional fabrics), and many other products.

The method according to the invention can be applied to any fabric (woven, knitted, or nonwoven) known in the art. In particular, the method of the invention is applicable to cellulose-containing or cellulosic fabrics, such as cotton, viscose (viscose), rayon (rayon), ramie (ramie), flax (linen), lyocell (for example: Tence1, manufactured by Courtaulds Fibers), or blends (blends) of any of these Fibers with synthetic Fibers (for example: polyester, polyamide, nylon) or other natural Fibers such as wool and silk, for example viscose/cotton blends, green/cotton blends, viscose/wool blends, green/wool blends, cotton/wool blends; and flax (flex), ramie and other cellulosic fiber-based fabrics, 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. This method can also be used for synthetic textiles, for example synthetic textiles consisting of substantially 100% polyester, polyamide, or nylon, respectively. The term "hair (wool)" refers to any commercially useful animal wool, such as wool made from sheep (sheet), camels, rabbits, goats, llamas (llama) and the well-known merinowool (merinowool), Shetland wool (Shetland wool), kefir wool (casemerie wool), alpaca wool (alpaca wool), mohair (mohair), and the like; also includes wool fiber and animal hair. The method of the present invention may be applied to wool fabrics or animal hair materials in the form of tops (tops), fibres, yarns, or woven or knitted fabrics.

The alkaline alpha-amylase used in the process of the invention may preferably be of bacterial origin, e.g. derived in particular from a strain of bacillus.

The alkaline scouring enzyme used in the process of the invention may be an enzyme selected from the group consisting of alkaline pectinase, cellulase, lipase, protease, or a mixture thereof.

According to the invention, the enzyme is "alkaline" when the optimum pH is above 7, preferably above 8, especially above 9, for example between pH7-11, such as between pH8-11, or between pH9-11 during simultaneous desizing and scouring.

The term "desizing" may be understood in the usual way, i.e. the degradation and/or removal of sizing agents (sizing agents) from warp yarns in a fabric, such as a woven fabric.

The term "scouring" is also understood in the usual way, i.e. the removal of non-cellulosic materials on the fabric, such as grease (grease), wax (wax), protein, hemicellulosic materials, pectin, ash, dirt and oil.

The term "simultaneous" means that desizing and scouring are performed in a single operation step. This has the advantage that washing, rinsing and other treatments, which are normally carried out between separate desizing and scouring steps, are no longer required. Thus, the water, energy requirements and the need for different equipment to be used in each treatment process is greatly reduced. According to a preferred embodiment, the process of the invention is carried out in a single bath (single bath). Scouring enzymes may be added prior to, simultaneously with, or after desizing enzymes.

The term "fabric comprising starch or starch derivatives" refers to any type of fabric, in particular a woven fabric made of cellulose-containing material, which contains starch or starch derivatives. These fabrics are typically made of cotton, viscose, linen and the like. The major part of the starch or starch derivative present on the fabric is usually the size applied to the yarn, usually warp, before weaving.

Even if no particular mention is made in connection with the method of the invention, it is understood that the enzyme or reagent is used in an "effective amount". The term "effective amount" refers to the amount of alkaline alpha-amylase and alkaline scouring enzyme that is capable of achieving the desired effect, i.e., desizing and scouring of the fabric, as compared to a fabric that has not been treated with the enzyme.

In a second aspect, the present invention relates to a composition suitable for use in a simultaneous desizing and scouring process comprising an alkaline alpha-amylase and an alkaline scouring enzyme.

Detailed Description

The invention aims to provide a method for synchronously desizing and scouring. According to the invention, scouring may be carried out while desizing the fabric. The method of the present invention can be performed using conventional sizing/desizing equipment such as pad systems, J-boxes, jet machines, jiggers, etc., without any additional processing equipment. This can be accomplished by simultaneous treatment of the fabric with a combination of alkaline alpha-amylase and alkaline scouring enzyme. The inventors have found that, in addition to the advantages obtained by carrying out simultaneous desizing and scouring (see above), also other advantages are obtained. Examples include one or more of the following: reduced enzyme usage, improved desizing, improved pectin removal, improved wettability (wetability), improved whiteness (whiteness), improved fabric handling, improved fabric smoothness, reduced pilling (pilling). The results of the experiments supporting the present invention are shown in table 1 following examples 1-13.

Woven fabrics are a popular form of fabric construction. The weaving process requires "sizing" of the warp yarns to protect them from abrasion. Starches (modified and unmodified), polyvinyl alcohol (PVA), carboxymethyl cellulose (CMC), waxes and acrylic binders, and mixtures thereof are examples of commonly used sizing agents. According to the invention, the sizing agent may be a starch-based or starch derivative-based sizing agent, but may also comprise one or more non-starch-based or starch derivative-based sizing agents. After weaving, the sizing agent must be removed as a first step in the preparation of the woven article.

Furthermore, textile fibers contain natural, non-cellulosic impurities that must be removed prior to subsequent processing steps, such as bleaching, dyeing, printing, and finishing (finishing). Scouring removes a large number of natural non-cellulosic impurities, including in particular the stratum corneum (cuticle), which consists mainly of wax, and the primary cell wall, which consists mainly of pectin, protein and xyloglucan (xyloglucan). In order to obtain high wettability, which is a measure for obtaining good dyeing, proper wax removal is necessary. Removal of the primary cell walls (especially pectin) improves wax removal and ensures more even dyeing, which in addition improves whiteness during the bleaching process. In addition, scouring can remove dirt (dirt), soil (soil) and residues introduced during manufacturing such as spinning (spinning), coning (coning) or sizing agents.

Method of the invention

According to the method of the invention, the sized fabric, in rope form or open width (open width), is contacted with a treatment liquid, such as a treatment solvent. In case the size (in addition to the starch-based or starch derivative-based sizing agent) contains polyvinyl alcohol or carboxymethylcellulose, the process of the invention is preferably carried out with hot water, surfactants and mild bases.

According to the invention, desizing and scouring are carried out simultaneously and under conventional textile desizing conditions.

Accordingly, in a first aspect, the present invention relates to a process for simultaneously desizing and scouring a sized fabric containing starch or starch derivatives, which process comprises treating the fabric with an alkaline alpha-amylase and an alkaline scouring enzyme.

According to the present invention, the sized fabric is treated with a combination of water, alkaline alpha-amylase and alkaline scouring enzyme (as described in further detail below), preferably in combination with one or more agents including stabilizers, surfactants, wetting agents, dispersants, chelating agents and emulsifiers and mixtures thereof. The sized fabric is allowed to remain in the treatment liquor for a sufficient period of "hold time" to complete desizing and scouring. The holding time depends on the type and temperature of the processing process (processing region) and may vary from 15 minutes to 2 hours, or in some cases days.

The treatment process may be a batch or continuous process of contacting a flat web or rope of fabric with a stream of treatment liquid.

Continuous operation typically uses a saturator (saturator) in which an approximately equal weight of treatment liquid per weight of fabric is applied to the fabric, followed by a heated dwell chamber (heated dwell chamber) in which the chemical reaction takes place. The washing section then prepares the fabric for the next processing step. To ensure high whiteness or better wettability, and resulting dyeability, desizing and scouring enzymes, as well as other agents, must be thoroughly removed.

In one embodiment, the process of the present invention is a continuous process carried out at a temperature of about 100 deg.C, such as 90-100 deg.C, and a pH of 7-11, for 5-30 minutes.

Batch processing is typically carried out in one treatment bath, i.e. a single bath, wherein the fabric is contacted with about 8-15 times its own weight of treatment liquid. After the reaction period, the treatment liquid is drained, the fabric rinsed and the next treatment step started. Discontinuous PB treatment (i.e. pad-batch process) involves a saturator in which approximately equal weight of treatment liquid per fabric weight is applied to the fabric, followed by a dwell period which in the case of CPB treatment (i.e. cold pad-batch process) may be one or several days. For example, the CPB treatment may be carried out at 20-40 deg.C at a pH of 7-11, preferably about 8-9.5, for 8-24 hours or more. In addition, the PB treatment may be carried out at 50-85 ℃ and pH7-11, preferably 8-9.5, for 1-6 hours.

In one embodiment, the combined desizing and scouring process of the present invention may be carried out using alkaline alpha-amylase and alkaline scouring enzyme and a strong base, such as sodium hydroxide, or related caustic agents, such as sodium carbonate, potassium hydroxide, or mixtures thereof, under conditions known in the art for desizing and scouring.

At present, commercial alpha-amylases for desizing, e.g. AQUAZYM^TM120L (Novozymes A/S, Denmark), the recommended concentration is in the range of about 180-. According to the invention, this concentration can be reduced.

In a preferred embodiment, the alkaline alpha-amylase is present in a concentration of 0.05-150KNU/L of treatment solution, preferably 1-100KNU/L of treatment solution, especially 2-20KNU/L of treatment solution or 0.05-150KNU/kg of fabric, preferably 1-100KNU/kg of fabric, especially 2-20KNU/kg of fabric.

In addition, commercial pectinases for scouring, such as SCOURZYME^TML (Novozymes A/S, Denmark), the recommended concentration is in the range of about 1500-1875APSU/L, which corresponds to about 1500-1875APSU/kg of fabric. According to the invention, this concentration can be reduced.

In a preferred embodiment, the pectinase is a pectate lyase present at a concentration in the range of 1-1,500APSU/kg fabric, preferably 10-1, 200APSU/kg fabric, especially 100-1,000APSU/kg fabric.

Detergent composition

Typically, an alkali-stable surfactant is added to the treatment process to enhance the dissolution (solvation) of the hydrophobic compound and/or to prevent its redeposition (redeposition) back onto the fabric. In the context of the present invention, detergent is synonymous with surfactant and may in particular be a nonionic, anionic, cationic, amphoteric, zwitterionic and semi-polar surfactant, or a mixture thereof.

Surfactants are typically present in the compositions of the present invention at a level of from 0.1 to 60% by weight.

The surfactant is preferably formulated to be compatible with the enzyme component of the composition. In liquid or gel compositions, the surfactant is most preferably formulated in a manner that promotes, or at least does not reduce, the stability of any enzyme in these compositions.

According to the present invention, preferred systems to be used comprise as surfactant one or more of the nonionic and/or anionic surfactants as described herein.

Polyethylene oxide, polypropylene oxide and polybutylene oxide polycondensates of alkyl phenols are suitable for use as the nonionic surfactant in the surfactant systems of the present invention, with polyethylene oxide polycondensates being preferred. These compounds include condensation polymers of alkyl phenols having an alkyl group in a straight or branched chain configuration containing from about 6 to about 14 carbon atoms, preferably from about 8 to about 14 carbon atoms, with an olefin oxide (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 alkylphenol.

Commercially available such nonionic surfactants include GAF corporation ioIgepal sold by n^TMCO-630; and are all Rohm&Triton sold by Haas Company^TMX-45, X-114, X-100 and X-102. These surfactants are commonly referred to as alkylphenol alkoxylates (e.g., alkylphenol ethoxylates).

The condensation products of primary and secondary aliphatic alcohols with from about 1 to about 25 moles of ethylene oxide are suitable for use as nonionic surfactants in nonionic surfactant systems. The alkyl chain of the aliphatic alcohol can be straight or branched, can be primary or secondary, and typically contains from about 8 to about 22 carbon atoms. Preferred are the polycondensation products of alcohols having an alkyl group containing from about 8 to about 20, preferably from about 10 to about 18 carbon atoms with from about 2 to about 10 moles of ethylene oxide per mole of alcohol. The polycondensation product has from about 2 to about 7 moles of ethylene oxide present, and most preferably from 2 to 5 moles of ethylene oxide per mole of alcohol. Examples of commercially available such nonionic surfactants include Tergitol, all sold by Union Carbide Corporation of the United states (Union Carbide Corporation)^TM 15-S-9(C₁₁-C₁₅Polycondensates of linear alcohols with 9 mol of ethylene oxide), Tergitol^TM 24-L-6NMW(C₁₂-C₁₄Polycondensation products of primary alcohols with 6 moles of ethylene oxide, which have a narrow molecular weight distribution); neodol sold by Shell chemical company^TM 45-9(C₁₄-C₁₅Polycondensates of linear alcohols with 9 mol of ethylene oxide), Neodol^TM 23-3(C₁₂-C₁₃Polycondensates of linear alcohols with 3.0 mol of ethylene oxide), Neodol^TM 45-7(C₁₄-C₁₅Polycondensates of linear alcohols with 7 mol of ethylene oxide), Neodol^TM 45-5(C₁₄-C₁₅Polycondensates of linear alcohols with 5 moles of ethylene oxide); from Procter, Inc&Gamble Company) sold Kyro^TM EOB(C₁₃-C₁₅Polycondensates of alcohols with 9 moles of ethylene oxide); and Genapol LA 050 (C) sold by Hoechst₁₂-C₁₄Polycondensates of alcohols with 5 moles of ethylene oxide). The preferred range of HLB in these products is from 8 to11, and most preferably from 8 to 10.

Also useful as nonionic surfactants in the surfactant system are alkyl polysaccharides (alkyl polysaccharaides) disclosed in U.S. patent No.4,565,647 having a hydrophobic group containing from about 6 to about 30, preferably from about 10 to about 16 carbon atoms and a polysaccharide such as a polyglycoside, a 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 (saccharoide units). Any reducing saccharide containing 5 or 6 carbon atoms such as glucose, galactose can be used, and the galactosyl moiety can be substituted with a glucosyl moiety (as opposed to a glucoside or galactoside, the hydrophobic group is optionally attached at the 2-, 3-, 4-, etc. position, thereby yielding glucose or galactose). The intersaccharide linkage may be located, for example, between one position of the additional saccharide unit and the 2-, 3-, 4-and/or 6-position on the preceding saccharide unit.

Preferred alkyl polyglycosides have the formula:

R²O(C_nH_2nO)_t(glycoside)_x

Wherein R is²Selected from the group consisting of alkyl, alkylphenyl, hydroxyalkyl, hydroxyalkylphenyl, and mixtures thereof, wherein the alkyl group contains from about 10 to about 18, preferably from about 12 to about 14, carbon atoms; n is 2 or 3, preferably 2; t is 0 to about 10, preferably 0; x is from about 1.3 to about 10, preferably from about 1.3 to about 3, and 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 first formed 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 2-, 3-, 4-and/or 6-position, preferably predominantly the 2-position, of the preceding glycosyl unit.

Ethylene oxide polycondensation products containing hydrophobic materials formed by the polycondensation of propylene oxide with propylene glycol are also suitable for use as the additional nonionic surfactant system. The hydrophobic moieties of these compoundsPreferably has a molecular weight of about 1500 to about 1800, and exhibits water insolubility. The addition of a polyoxyethylene moiety to this hydrophobic moiety is intended to increase the water solubility of the molecule as a whole and when the polyoxyethylene content is about 50% of the total weight of the polycondensation product, it is equivalent to polycondensation with up to about 40 moles of ethylene oxide until the liquid character of the product is maintained. Examples of such compounds include certain commercially available pluronics sold by BASF^TMA surfactant.

Also suitable for use as the nonionic surfactant in the nonionic surfactant system is the polycondensation product of ethylene oxide and the reaction product of propylene oxide and ethylenediamine. The hydrophobic portion of these compounds consists of the reaction product of ethylenediamine with excess propylene oxide and typically has a molecular weight of from about 2500 to about 3000. The hydrophobic moiety is condensed with ethylene oxide to an extent such that the condensation product contains from about 40% to about 80% by weight polyoxyethylene and has a molecular weight of from about 5000 to about 11000. Examples of such nonionic surfactants include certain commercially available Tetronic surfactants sold by BASF^TMA compound is provided.

Preferred for use as the nonionic surfactant in 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 ethylene oxide, alkyl polysaccharides, and mixtures thereof. Most preferred are C's containing 3 to 15 ethoxy groups₈-C₁₄Alkylphenol ethoxylates and C having 2-10 ethoxy groups₈-C₁₈(average C is preferred)₁₀) Alcohol ethoxylates, and mixtures thereof.

Highly preferred nonionic surfactants are polyhydroxy fatty acid amide surfactants of the formula:

wherein R is¹Is H, or R¹Is C_1-4A hydrocarbyl group, 2-hydroxyethyl group, 2-hydroxypropyl group, or mixtures thereof; r²Is C_5-31A hydrocarbyl group; and Z is a polyhydroxyhydrocarbyl or an alkoxy derivative thereof having a linear hydrocarbyl chain with at least 3 hydroxyl groups directly attached to the chain. Preferably, R¹Is methyl, R²Is straight chain (straight) C_11-15Alkyl or C_16-18An 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.

Highly preferred anionic surfactants include alkyl alkoxy sulfate surfactants. Examples thereof are of the formula RO (A)_mSO₃Water soluble salts or acids of M, wherein R is unsubstituted C₁₀-C₂₄Alkyl or having C₁₀-C₂₄Hydroxyalkyl of the alkyl component, preferably C₁₂-C₂₀Alkyl or hydroxyalkyl, more preferably C₁₂-C₁₈Alkyl or hydroxyalkyl; a is an ethoxy or propoxy unit; m is greater than 0, typically from about 0.5 to about 6, more preferably from about 0.5 to about 3; m is H or a cation which may be, for example, a metal cation (e.g., sodium, potassium, lithium, calcium, magnesium, etc.), ammonium, or a substituted ammonium cation. Also included herein are alkyl ethoxy sulfates as well as alkyl propoxy sulfates. Specific examples of substituted ammonium cations include methyl-, dimethyl, trimethyl-ammonium cations and quaternary ammonium cations such as tetramethyl-ammonium and dimethyl piperidinium (dimethyl piperidinium) cations and those derived from alkylamines such as ethylamine, diethylamine, triethylamine, mixtures thereof, and the like. A representative surfactant is C₁₂-C₁₈Alkyl polyethoxylate (1.0) sulfate (C)₁₂-C₁₈E(1.0)M)，C₁₂-C₁₈Alkyl polyethoxylate (2.25) sulfate (C)₁₂-C₁₈(2.25)M)，C₁₂-C₁₈Alkyl polyethoxylate (3.0) sulfate (C)₁₂-C₁₈E (3.0) M), and C₁₂-C₁₈Alkyl poly(s)Ethoxylate (4.0) sulfate (C)₁₂-C₁₈E (4.0) M), wherein M is conveniently selected from sodium and potassium.

Suitable anionic surfactants to be used are alkyl ester sulfonate surfactants comprising C₈-C₂₀Linear esters of carboxylic acids (i.e.fatty acids) with gaseous SO according to "The Journal of The American oil chemists Society, 52(1975), pp.323-329₃And (4) sulfonating. Suitable starting materials include natural fatty substances such as those derived from tallow (tallow), palm oil, etc.

Preferred alkyl ester sulfonate surfactants include alkyl ester sulfonate surfactants of the formula:

wherein R is³Is C₈-C₂₀A hydrocarbyl group, preferably an alkyl group, or a combination thereof; r⁴Is C₁-C₆A hydrocarbyl group, preferably an alkyl group, or a combination thereof; and M is a cation that 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, diethanolamine, and triethanolamine. Preferably, R³Is C₁₀-C₁₆Alkyl radical, R⁴Is methyl, ethyl or isopropyl. Especially preferred are the methyl ester sulfonates, wherein R³Is C₁₀-C₁₆An alkyl group.

Other suitable anionic surfactants include alkyl sulfate surfactants having the formula ROSO₃Water-soluble salts or acids of M, wherein R is preferably C₁₀-C₂₄A hydrocarbon group, preferably having C₁₀-C₂₀Alkyl or hydroxyalkyl of the alkyl moiety, more preferably C₁₂-C₁₈Alkyl or hydroxyalkyl, and M is H or a cation, e.g. an alkali metal cation (e.g. sodium, potassium, lithium), or ammoniumSalts or substituted ammonium (e.g., methyl-, dimethyl-, trimethylammonium cations and quaternary ammonium cations such as tetramethylammonium and dimethylpiperidinium cations, as well as quaternary ammonium cations derived from alkylammonium such as ethylamine, diethylamine, triethylamine, and mixtures thereof, and the like). Generally, lower wash temperatures (e.g., below about 50℃.) are preferred to C₁₂-C₁₆While higher washing temperatures (e.g., above about 50 ℃) are preferred to be C₁₆-C₁₈Alkyl chain of (2).

Other anionic surfactants useful for detersive purposes include soap (soap) salts (which include, for example, sodium, potassium, ammonium and substituted ammonium salts such as the monoethanol, diethanol and triethanol amine salts), C₈-C₂₂Salts of primary or secondary alkanesulfonic acids, C₈-C₂₄Olefin sulfonates, sulfonated polycarboxylic acids prepared by sulfonating the pyrolysis (pyrolyzed) product of alkaline earth metal citrates, as described in British patent 1,082, 179 Specification, C₈-C₂₄Alkyl polyglycol ether sulfates (which contain up to 10 moles of ethylene oxide); alkyl glycerol sulfonates, fatty acyl glycerol sulfonates, fatty oleoyl glycerol sulfates, alkylphenol oxyethane ether sulfates, paraffin sulfonates, alkyl phosphates, isethionates (isethionates) such as acyl isethionates, N-acyl taurates, alkyl succinamates (alkyl succinamates) and sulfosuccinates (sulfosuccinates), monoesters of sulfosuccinates (especially saturated and unsaturated C)₁₂-C₁₈Monoesters) and diesters of sulfosuccinic acid salts (especially saturated and unsaturated C)₆-C₁₂Diesters), acyl sarcosinates (acyl sarcosinates), alkyl polysaccharide sulfates such as alkyl polyglucoside sulfates (the nonionic nonsulfated compound is described below), branched primary alkyl sulfates, and alkyl polyethoxy carboxylates such as RO (CH)₂CH₂O)_k-CH₂COO-M + wherein R is C₈-C₂₂Alkyl, k is an integer from 1 to 10, and M is a cation which is a soluble salt. Resin acids (resin acids) and hydrogenated resin acids are also suitable, such as rosin (rosin), hydrogenated rosin, andresin acids present in or derived from tall oil (tall oil) and hydrogenated resin acids.

Very preferred is alkyl benzene sulfonate. Especially preferred are linear (straight-chain) alkylbenzene sulfonates (LAS) in which the alkyl group preferably contains from 10 to 18 carbon atoms.

Other examples are described in "Surface Active Agents and Detergents" (Vo1.I and II by Schwartz, Perrryand Berch). A number of such surfactants are also generally disclosed in U.S. patent No.3, 929, 678 (column 23, line 58 to column 29, line 23, incorporated herein by reference).

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 the anionic surfactant.

The compositions of the present invention may also comprise cationic, amphoteric, zwitterionic and semi-polar surfactants, as well as nonionic and/or anionic surfactants, in addition to those already described herein.

Cationic cleaning (detersive) surfactants suitable for use in the compositions of the present invention are those containing a long chain hydrocarbyl group. Examples of such cationic surfactants include ammonium surfactants such as alkyltrimethylammonium halides and those having the formula:

[R²(OR³)_y][R⁴(OR³)_y]₂R⁵N+X-

wherein R is²Is an alkyl group or an alkylphenyl group having from about 8 to about 18 carbon atoms in the alkyl chain; each R³Selected from: -CH₂CH₂-，-CH₂CH(CH₃)-，-CH₂CH(CH₂OH)-，-CH₂CH₂CH₂-, and mixtures thereof; each R⁴Is selected from C₁-C₄Alkyl radical, C₁-C₄Hydroxyalkyl by reacting two R⁴Benzyl ring structure formed by connecting groups, -CH₂CHOHCHOHCOR⁶CHOHCH₂OH, wherein R⁶Is any hexose or hexose polymer having a molecular weight below about 1000, and hydrogen (when y is not equal to 0); r⁵And R⁴Identical or are alkyl chains in which the total number of carbon atoms or R²Adding R⁵No more than about 18; each y is from about 0 to about 10, and the sum of the values of y is from 0 to about 15; and X is any suitable anion.

Highly preferred cationic surfactants are the water soluble quaternary ammonium compounds suitable for use in the present compositions having the formula:

R₁R₂R₃R₄N⁺X^-(i)

wherein R is₁Is C₈-C₁₆Alkyl radical, R₂、R₃And R₄Each of which is independently C₁-C₄Alkyl radical, C₁-C₄Hydroxyalkyl, benzyl and- (C)₂H₄₀)_xH, wherein X has a value of 2 to 5 and X is an anion. R₂、R₃Or R₄At most one of which is benzyl.

R₁Preferred alkyl chain lengths are C₁₂-C₁₅Especially where the alkyl group is a mixture of chain lengths from coconut or palm kernel fat, or is synthesized by olefin building (olefin built up) or OXO alcohol synthesis.

R₂、R₃And R₄Preferred groups of (a) are methyl and hydroxyethyl groups and the anion X may be selected from halide, methyl sulfate, acetate and phosphate ions.

Examples of quaternary ammonium compounds of formula (i) suitable for use in the present invention are:

coconut trimethyl ammonium chloride or coconut trimethyl ammonium bromide;

coconut methyl dihydroxyethyl ammonium chloride or coconut methyl dihydroxyethyl ammonium bromide;

decyl trimethyl ammonium chloride;

decyl dimethyl hydroxyethyl ammonium chloride or decyl dimethyl hydroxyethyl ammonium bromide;

C_12-15dimethylhydroxyethylammonium chloride or C_12-15Dimethyl hydroxyethyl ammonium bromide;

coconut dimethylhydroxyethylammonium chloride or coconut dimethylhydroxyethylammonium bromide;

tetradecyltrimethylammonium methylsulfate;

dodecyl dimethyl benzyl ammonium chloride or dodecyl dimethyl benzyl ammonium bromide;

dodecyl dimethyl (ethyleneoxy) 4 ammonium chloride or dodecyl dimethyl (ethyleneoxy)₄Ammonium bromide;

choline esters (compounds of formula (i) wherein R₁Is that

Alkyl, and R₂、R₃、R₄Is methyl).

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

Other cationic surfactants useful in the present invention are also described in U.S. Pat. No.4,228,044 and EP 000224.

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 the cationic surfactant.

Amphoteric 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 or tertiary amines in which the aliphatic radical can be straight or branched chain. One of these aliphatic substituents contains at least about 8 carbon atoms, usually from about 8 to about 18 carbon atoms, and at least one contains a water-solubilizing anionic group, e.g., carboxy, sulfonate, sulfate. See U.S. Pat. No.3,929,678 (column 19, lines 18-35), for examples of amphoteric surfactants.

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 the amphoteric surfactant.

Zwitterionic surfactants are also suitable for use in the compositions of the present invention. These surfactants can be broadly described as derivatives of secondary or tertiary amines, derivatives of heterocyclic secondary or 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 to column 22, line 48), for examples of zwitterionic surfactants.

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 the zwitterionic surfactant.

Semi-polar nonionic surfactants are a particular class of nonionic surfactants which include the water-soluble amine oxides (amine oxides) containing one alkyl moiety of from about 10 to about 18 carbon atoms and two moieties selected from the group consisting of alkyl and hydroxyalkyl radicals containing from about 1 to about 3 carbon atoms; water-soluble phosphine oxides containing one alkyl moiety of from about 10 to about 18 carbon atoms and two moieties selected from the group consisting of alkyl and hydroxyalkyl containing from about 1 to about 3 carbon atoms; and water-soluble sulfoxides (sulfoxides) containing one alkyl moiety of from about 10 to about 18 carbon atoms and one moiety selected from the group consisting of alkyl and hydroxyalkyl moieties containing from about 1 to about 3 carbon atoms.

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

wherein R is³Is an alkyl, hydroxyalkyl, or alkylbenzyl group containing from about 8 to about 22 carbon atoms or mixtures thereof; r⁴Is an alkylene (alkylene) or hydroxyalkylene group containing from about 2 to about 3 carbon atoms or mixtures thereof; x is 0 to about 3; and each R⁵Is 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. R⁵The groups may be linked to each other, for example, by oxygen or nitrogen atoms to form a cyclic structure.

These amine oxide surfactants include, in particular, those containing C₁₀-C₁₈Alkyl dimethyl amine oxide and C₈-C₁₂Alkoxyethyl dihydroxyethyl amine oxide.

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 the semi-polar nonionic surfactant.

Enzyme

Amylase

Any alkaline alpha-amylase may be used according to the invention. In the context of the present invention, an amylase is "alkaline" when the pH optimum is greater than 7, preferably greater than 8, especially greater than 9, under the conditions present during simultaneous desizing and scouring.

Suitable alpha-amylases include those of bacterial or fungal origin. Chemically or genetically modified mutants (variants) are included. Preferred alkaline alpha-amylases are derived from strains of Bacillus, e.g.Bacillus licheniformis, Bacillus amyloliquefaciens, Bacillus stearothermophilus, Bacillus subtilis or other Bacillus species, e.g.Bacillus NCIB 12289, NCIB 12512, NCIB 12513, DSM 9375, DSMZ No.12649, KSM AP1378(WO 97/00324), KSM K36 or KSM K38(EP1,022, 334). Preferred are the Bacillus alpha-amylases disclosed in WO 95/26397 as SEQ ID NOS.1 and 2, respectively (i.e., SEQ ID NO: 4 herein), the Bacillus alpha-amylases disclosed in WO 00/60060 as SEQ ID NO: 2 (i.e., SEQ ID NO: 6 herein), and #707 alpha-amylase, disclosed by Tsukamoto et al, Biochemical and Biophysical Research Communications, Vo1.15l, pp.25-31 (1988).

Commercially available alkaline alpha-amylase products or products containing alpha-amylase include those sold under the following trade names: NATALASE^TM，STAINZYME^TM(NovozymesA/S)，BIOAMYLASE-D(G)，BIOAMYLASE^TM L(Biocon India Ltd.)，KEMZYM^TMAT9000(Biozym Ges.m.b.H, Switzerland), PURASTAR^TM ST，PURASTAR^TM HPAmL，PURAFECT^TM OxAm，RAPIDASE^TMTEX (Genencor int. inc., usa), KAM (KAO, japan)

In a particular embodiment of the invention, the alkaline alpha-amylase is a polypeptide having the amino acid sequence of SEQ ID NO: 4, or an alpha-amylase having the amino acid sequence of SEQ ID NO: 6, or an alpha-amylase identical to the amino acid sequence of SEQ ID NO: 4 or SEQ ID NO: 6, preferably at least 70%, more preferably at least 80%, still more preferably at least 90%, such as at least 95%, at least 96%, at least 97%, at least 98% or at least 99% identity to a degree of at least 60%, preferably at least 70%, more preferably at least 80%, still more preferably at least 90%, such as at least 95%, at least 96%, at least 97%, at least 98% or at least 99%

For the purposes of the present invention, LASERGENE was used by the method Clusta1 (Higgins, 1989, CABIOS 5: 151-^TM MEGALIGN^TMThe software (DNASTAR, inc., Madison, WI) together with the identity table and the following multiple alignment parameters (multiple alignment parameter) were used to determine the degree of identity between two amino acid sequences: a Gap penalty (Gap penalty) of 10 and a Gap length penalty of 10. Pairwise alignment parameter (Pairwise)alignment parameter) is: ktule ═ 1, gap penalty ═ 3, window (windows) ═ 5, and diagonal (diago-nals) ═ 5]。

In a preferred embodiment, the parent alpha-amylase has one or more deletions at positions D183 and G184, preferably wherein said alpha-amylase variant further has a substitution at position N195F (numbered using SEQ ID NO: 4).

In another preferred embodiment, the parent alpha-amylase has one or more of 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: R118K, N195F, R320K, R458K, in particular wherein the variant has the following mutations: delta (D183+ G184) + R118K + N195F + R320K + R458K (numbering using SEQ ID NO: 6).

In another preferred embodiment, the alkaline alpha-amylase is SEQ ID NO: 6, further comprising one or more of the following substitutions: M9L, M202L, V214T, M323T, M382Y, E345R, or a560 α -amylase with all of the following substitutions: M9L, M202L, V214T, M323T, M382Y or M9L, M202L, V214T, M323T and E345R.

In an embodiment of the process of the invention, the alkaline alpha-amylase may preferably be present in a concentration of 0.05-150KNU/L treatment solution, preferably 1-100KNU/L treatment solution, especially 2-20KNU/L treatment solution or 0.05-150KNU/kg fabric, preferably 1-100KNU/kg fabric, especially 2-20KNU/kg fabric.

Alkaline scouring enzyme

Any alkaline scouring enzyme may be used according to the invention. The alkaline scouring enzyme may be an alkaline enzyme selected from pectinases, cellulases, lipases, proteases, xyloglucanases, cutinases and mixtures thereof. In the context of the present invention, a scouring enzyme is "alkaline" when the pH optimum is greater than 7, preferably greater than 8, especially greater than 9, under the conditions present during simultaneous desizing and scouring.

In a preferred embodiment, the alkaline pectinase is a pectate lyase, a polygalacturonase, or a polygalacturonate lyase.

Pectinase

The term "pectinase" is intended to include any alkaline pectinase. Pectinases are a group of enzymes that hydrolyze the glycosidic linkages in pectic substances, primarily poly-1.4-alpha-D-galacturonide and its derivatives (see Sakai et al, published in Advances in Applied Microbiology, Vo1.39, pp.213-294(1993), Pectin, peptide and protein: production, properties and applications), which are understood to include mature proteins or their precursor forms, or functional fragments thereof that have substantially the activity of the full-length enzyme. Furthermore, the term pectinase is intended to include homologues or analogues of these enzymes.

Preferably, the alkaline pectinase is an enzyme that catalyzes the random cleavage of alpha-1, 4-glycosidic bonds in pectic acid (also known as polygalacturonic acid) by trans-elimination (transalimination), such as the polygalacturonic acid lyase (EC 4.2.2.2) (PGL), also known as poly (1, 4-alpha-D-galacturonoside) lyase, also known as pectate lyase. Also preferred are pectinases that catalyze the random hydrolysis of alpha-1, 4-glycosidic bonds in pectic acids, such as the polygalacturonases (EC 3.2.1.15) (PG), also known as endo-PG. Also preferred are pectinases that catalyze the random cleavage of alpha-1, 4-glycosidic bonds in pectin, such as polymethylgalacturonase (EC 4.2.2.10) (PMGL), also known as Endo-PMGL, also known as poly (methoxygalacturonide) lyase, also known as pectin lyase. Other preferred pectinases are galactase (EC 3.2.1.89), arabinase (EC 3.2.1.99), pectinesterase (EC 3.1.1.11) and mannanase (EC 3.2.1.78).

The enzyme is preferably derived from a microorganism, preferably from a bacterium, archaea (archea) or a fungus, especially from a bacterium, e.g. a bacterium belonging to the genus bacillus, preferably a strain belonging to the genus bacillus alkalophilus, which may be selected from bacillus licheniformis and highly related bacillus species, wherein all species have at least 90% homology (identity) with bacillus licheniformis based on the 16S rDNA sequence. Specific examples of such species are: bacillus licheniformis, Bacillus alcalophilus, Bacillus pseudoalcalophilus, and Bacillus clarkii. A specific and highly preferred example is Bacillus licheniformis strain, ATCC 14580 (U.S. Pat. No.6,284,524). Other useful pectate lyases are from Bacillus agaradhaeerens, especially from the strain deposited as NCIMB 40482; and from Bacillus subtilis, Bacillus stearothermophilus, Bacillus pumilus (Bacillus pumilus), Bacillus koshii (Bacillus cohnii), Bacillus pseudoalkalophilus, Erwinia 9482, in particular the FERM BP-5994 strain and Paenibacillus polymyxa (Paenibacillus polymyxa).

Pectinases may be a component present in an enzyme system produced by a given microorganism, which enzyme system mostly contains several different pectinase components, including those identified above.

In addition, the pectinase may be a single ingredient, i.e., one ingredient that may be present in the enzyme system produced by a given microorganism is substantially free of other pectinases. The single component is typically a recombinant component, i.e., a component produced by cloning a DNA sequence encoding the single component, and then transforming a cell with the DNA sequence and expressing in a host. These useful recombinant enzymes, especially pectate lyase, pectin lyase and polygalacturonase, are described in detail in, for example, WO 99/27083 and WO 99/27084 (from Novozymes A/S), the entire contents of which are incorporated herein by reference. The host is preferably a heterologous host, but under certain conditions the host may also be a homologous host.

In a preferred embodiment, the pectate lyase used in the present invention is derived from the genus Bacillus, preferably Bacillus licheniformis, Bacillus alkalophilus, Bacillus pseudoalkalophilus and Bacillus clarkia, especially Bacillus licheniformis, ATCC 14580.

In a more preferred embodiment, the pectate lyase is the amino acid sequence of SEQ ID NO: 2, mature pectate lyase. This pectate lyase is also disclosed in U.S. Pat. No.6,284,524, which is incorporated herein by reference.

Pectinases, especially pectate lyases, may preferably be present at a concentration of 1-1,500APSU/kg fabric, preferably 10-1, 200APSU/kg fabric, especially 100-1,000APSU/kg fabric.

Commercially available alkaline pectate lyases include BIOPREP from Novozymes A/S, Denmark^TMAnd SCOURZYME^TM L。

Protease enzyme

Any protease suitable for use in alkaline solutions may be used. Suitable proteases include those of animal, vegetable and microbial origin. Preferably of microbial origin. Chemically or genetically modified mutants are included. The protease may be a serine protease, preferably an alkaline microbial protease or a trypsin-like (trypsin-like) protease. Examples of alkaline proteases are subtilisins (subtilisins), especially those from the genus Bacillus, preferably Bacillus lentus or Bacillus clausii, such as subtilisin Novo, subtilisin Carlsberg, subtilisin 309, subtilisin 147 and subtilisin 168 (described in WO 89/06279).

Preferred commercially available proteases include those sold under the following trade names: ALCALASE (alcohol calcium citrate)^TM，SAVINASE^TM16L Type Ex，PRIMASE^TM，DURAZYM^TMAnd ESPERASE^TM(Novozymes A/S, Denmark), those proteases sold under the following trade names: OPTICLEAN, produced by Genencor International Inc. (USA)^TM，OPTIMASE^TM，PROPARASE^TM，PURAFECT^TM，PURAPECT^TMMA and PURAPECT^TM OX，PURAFECT^TMOX-1 and PURAFECT^TM OX-2。

In an embodiment of the process of the invention, the protease may be present in a concentration of 0.001-10KNPU/L, preferably 0.1-1KNPU/L, especially about 0.3KNPU/L or 0.001-10KNPU/kg fabric, preferably 0.1-1KNPU/kg fabric, especially about 0.3KNPU/kg fabric.

Lipase enzyme

Any lipase suitable for use in alkaline solutions may be used. Suitable lipases include those of bacterial or fungal origin. Chemically or genetically modified mutants are included. Examples of useful lipases include the humicola lanuginosa lipase, as described in EP 258068 and EP 305216; rhizomucor miehei (Rhizomucor miehei) lipase, as described in EP 238023; candida (Candida) lipases, such as c.antarctica lipase, e.g. c.antarctica lipase a or B as described in EP 214761; pseudomonas lipases such as pseudomonas alcaligenes (p.alcaligenes) and pseudomonas pseudoalcaligenes (p.pseudoalcaligenes) lipases, as described in EP 218272; pseudomonas cepacia (p.cepacia) lipase as described in EP 331376; pseudomonas stutzeri (p.stutzeri) lipase, as disclosed in GB 1, 372,034; pseudomonas fluorescens (P.. fluoroscens) lipase, Bacillus lipases, such as Bacillus subtilis lipase (Dartois et al, biochemicala Biophysica Acta 1131, 253-260(1993)), Bacillus stearothermophilus lipase (JP 64/744992) and Bacillus pumilus lipase (WO 91/16422).

In addition, a number of cloned lipases may be useful, including the lipases Penicillium salmeterum (Penicillium camembertii) described by Yamaguchi et al, Gene 103, 61-67(1991), the lipases Geotrichum candidum (Geotrichum candidum) (Schimada, Y. et al, J.biochem., Vo1.106, pp.383-388(1989)), and various Rhizopus (Rhizopus) lipases such as Rhizopus delemar (R.delemar) lipase (Hass, M.J et al, Gene, Vo1.109, pp.117-113(1991)), Rhizopus niveus (R.niveus) lipase (Kugimiya et al, biosci.Biotech., biochem., Vo1.56, pp.716-1992 (R.biozazae) and Rhizopus oryzae (R.oryzae) lipases.

Particularly suitable are LIPASEs such as M1LIPASE^TM，LUMA FAST^TMAnd LIPOMAX^TM(Genencor Intemationa1Inc, USA), LIPOLASE^TMAnd LIPOLASE ULTRA^TMSP735(Novozymes A/S, Denmark), and LIPASE P "Amano" (Amano pharmaceutical Co. Ltd.).

In one embodiment of the process of the invention, the lipase may be present in a concentration of from 0.01 to 100LU/L of treatment solution, preferably from 1 to 10LU/L of treatment solution, especially from about 1LU/L of treatment solution or from 0.01 to 100LU/kg of fabric, preferably from 1 to 10LU/kg of fabric, especially about 1LU/kg of fabric.

Cellulase enzymes

In this context, the term "cellulase" or "cellulolytic enzyme" refers to an enzyme that catalyzes the degradation of cellulose to glucose, cellobiose, triose and other cellooligosaccharides (cellooligosaccharides). Cellulose is a polymer of glucose linked by β -1, 4-glycosidic bonds. Cellulose chains constitute numerous intra-and intermolecular hydrogen bonds, which lead to the formation of insoluble cellulose microfibrils (microfibrils). The hydrolysis of cellulose to glucose by microorganisms involves three main types of cellulases: endo-1, 4-beta-glucanases (EC 3.2.1.4), which randomly cleave the beta-1, 4-glycosidic bonds throughout the cellulose molecule; cellobiohydrolases (EC 3.2.1.91) (exoglucanases) which digest cellulose from non-reducing ends; and β -glucosidase (EC 3.2.1.21), which hydrolyzes cellobiose and low molecular weight cellodextrin (cellodextrin) to release glucose. Most cellulases consist of a Cellulose Binding Domain (CBD) and a catalytic domain (CAD) separated by a linker (linker) rich in proline and hydroxyl amino acid residues. In the present description and claims, the term "endoglucanase" refers to an enzyme having cellulolytic activity, in particular an endo-1, 4-beta-glucanase activity classified as EC 3.2.1.4 according to the enzyme nomenclature (1992) and capable of catalyzing the (endo) hydrolysis of 1, 4 bonds in cellulose, lichenin and cereal beta-D-glucans, including also 1, 3 bonds in beta-D-glucans. Any cellulase suitable for use in alkaline solutions may be used. Suitable cellulases include those of bacterial or fungal origin. Chemically or genetically modified mutants are included. Suitable cellulases are disclosed in U.S. patent No.4,435,307, which discloses fungal cellulases produced by Humicola insolens. Particularly suitable cellulases are the cellulases having color protection (colour care) benefits. Examples of such cellulases are the cellulases described in european patent application nos. 0495257, WO 91/17243 and WO 96/29397.

In a preferred embodiment, the alkaline cellulase is an alkaline endoglucanase, preferably a Humicola (Humicola) endoglucanase, in particular a Humicola insolens endoglucanase, more preferably an EG I or EG V endoglucanase from Humicola insolens DSM 1800, or a variant thereof, or a Thielavia (Thielavia) endoglucanase, preferably a Thielavia terrestris (Thielavia terrestris) endoglucanase or a variant thereof.

Commercially available cellulases include CELLUZYME produced by a strain of Humicola insolens^TMAnd DENIMAX^TM399S (Novozymes A/S), and KAC-500(B)^TM(Kao Corporation)。

In one embodiment of the process of the invention, the cellulase may be used in a concentration of from 0.001 to 10g enzyme protein per liter of treatment solution, preferably from 0.005 to 5g enzyme protein per liter of treatment solution, especially from 0.01 to 3g enzyme protein per liter of treatment solution or from 0.001 to 10g enzyme protein per kg fabric, preferably from 0.005 to 5g enzyme protein per kg fabric, especially from 0.01 to 3g enzyme protein per kg fabric. In one embodiment, the cellulase is used at a concentration of from 0.1 to1,000 ECU/g fabric, preferably from 0.5 to 200ECU/g fabric, especially from 1 to 500ECU/g fabric.

Cutinase

Cutinases are enzymes capable of degrading cutin, as described in Lin T S & Kolattukudy P E, J.Bacteriol.1978133(2)942-951, for example, cutinases differ from classical lipases in that no measurable activation is observed near the Critical Micelle Concentration (CMC) of tributyrin substrates. Also, cutinases are considered to belong to one class of serine esterases. The cutinase may also be a cutinase from humicola insolens as disclosed in WO 96/13580. The cutinase may be a variant, for example one or more of the variants disclosed in WO00/34450 and WO 01/92502, which are incorporated herein by reference.

Examples of cutinases are those from Humicola insolens (U.S. patent No.5, 827, 719); cutinases from Fusarium (Fusarium) strains, such as Fusarium culmorum (f.roseum culmorum), or in particular f.solani pisi (WO 90/09446, WO 94/14964, WO 94/03578). Cutinases may also be derived from a strain of the genus Rhizoctonia (Rhizoctonia), such as Rhizoctonia solani (r. solani), or from a strain of the genus Alternaria (Alternaria), such as a. brassicicola (WO 94/03578), or variants thereof, such as those described in WO00/34450 and WO 01/92502. The cutinase may also be of bacterial origin, such as a strain of Pseudomonas, preferably Pseudomonas mendocina as disclosed in WO 01/34899).

Cutinases may be added at a concentration of 0.001-25,000 micrograms of enzyme protein per gram of fabric, preferably 0.01-10,000 micrograms of enzyme protein per gram of fabric, especially 0.05-1,000 micrograms of enzyme protein per gram of fabric.

Xyloglucanase (Xvloglucanase)

Xyloglucanase is a xyloglucan specific enzyme that is capable of catalyzing the breakdown (solvabilization) of xyloglucan to xyloglucan oligosaccharides. Xyloglucanases are classified as EC 3.2.1.151 according to IUBMB enzyme nomenclature (2003). Pauly et al, Glycobiology 9(1999) p.93-100 disclose a xyloglucan-specific endo-beta-1, 4-glucanase from Aspergillus aculeatus (Aspergillus aculeatus). The xyloglucanase used according to the invention may be derived from a microorganism, such as a fungus or a bacterium. Examples of useful xyloglucanases are family 12 xyloglucan hydrolyzing endoglucanases, in particular family 12 xyloglucan hydrolyzing endoglucanases from e.g. aspergillus aculeatus as described in WO 94/14953. Another useful example is xyloglucanase produced by Trichoderma (Trichoderma), in particular EGIII. Xyloglucanases may also be derived from bacteria of the genus Bacillus, including Bacillus licheniformis, Bacillus agaradharens and Bacillus firmus. The xyloglucanase may also be an endoglucanase with xyloglucanase activity and low activity towards insoluble cellulose and high activity towards soluble cellulose, such as family 7 endoglucanases from e.g. Humicola insolens.

Xyloglucanase may be added at a concentration of 0.001-25,000 micrograms of enzyme protein per gram of fabric, preferably 0.01-10,000 micrograms of enzyme protein per gram of fabric, more preferably 0.05-1,000 micrograms of enzyme protein per gram of fabric, especially 0.5-500 micrograms of enzyme protein per gram of fabric.

Compositions of the invention

In a second aspect, the present invention relates to compositions suitable for use in the methods of the present invention. The composition may be a solid or liquid (aqueous) composition, and may be a concentrated composition or a ready-to-use composition.

Thus, in this aspect, the invention relates to a composition comprising an alkaline alpha-amylase and an alkaline scouring enzyme.

The enzymes comprised may preferably be those mentioned in the "enzymes" section above.

In a preferred embodiment, the alkaline alpha-amylase is derived from a strain of bacillus, preferably a strain derived from bacillus licheniformis, bacillus amyloliquefaciens, bacillus stearothermophilus, bacillus NCIB 12289, NCIB 12512, NCIB 12513 or DSM 9375, or DSMZ No.12649, KSM AP1378, or KSM K36 or KSM K38.

The Bacillus alpha-amylase may be a variant having one or more deletions at positions D183 and G184, respectively, and may further have a substitution at position N195F (numbered according to SEQ ID NO: 4). The bacillus alpha-amylase variant may also have one or more deletions at positions D183 and G184, and may also have one or more of the following substitutions: R118K, N195F, R320K, R458K (numbering according to SEQ ID NO: 6).

In particular, the bacillus variant may have a double deletion at positions D183 and G184 and further comprise the following substitutions: R118K + N195F + R320K + R458K (numbered with SEQ ID NO: 6).

The alkaline scouring enzyme is selected from: alkaline pectinase, cellulase, lipase, protease, cutinase, xyloglucanase, and mixtures thereof.

In a preferred embodiment, the alkaline pectinase is a pectate lyase, preferably a pectate lyase derived from a strain of bacillus, preferably a strain of bacillus licheniformis, bacillus alcalophilus, bacillus pseudoalcalophilus and bacillus clarkia, especially the species bacillus licheniformis, ATCC 14580.

Other formulations suitable for use in practicing the present method may be added separately or included in the compositions of the present invention. Examples of such formulations include stabilizers, surfactants, wetting agents, dispersants, chelating agents, and emulsifiers, and mixtures thereof.

Although alkaline alpha-amylase and alkaline scouring enzyme may be added as such, they are preferably formulated into a suitable composition. Thus, the enzyme may be used in the form of granules, preferably non-dusting granules, liquids, especially stable liquids, slurries (slury), or in a protected form. As disclosed in U.S. patent nos. 4,106, 991 and 4, 661, 452 (both to Novozymes a/S), dust-free granules can be produced and can optionally be coated by methods well known in the art.

Depending on the established method, the liquid enzyme preparation may be stabilized, for example, by adding polyols, such as propylene glycol, sugars or sugar alcohols or acetic acid. Other enzyme stabilizers are well known in the art. The protected enzymes may be prepared according to the method disclosed in EP 238216.

In principle, the composition of the invention comprising an alkaline alpha-amylase and a scouring enzyme may comprise any other agent to be used in the combined process of the invention. In a preferred embodiment, the composition of the invention comprises at least one further ingredient selected from the group consisting of stabilizers, surfactants, wetting agents, dispersants, chelating agents and emulsifiers. All of these other ingredients suitable for use in textiles are well known in the art.

Suitable surfactants include those mentioned in the "detergents" section above. The size is used to increase the wettability of the fibres so that a rapid and uniform desizing and scouring can be obtained. The emulsifier serves to emulsify the hydrophobic impurities present on the fabric. Dispersants are used to prevent the extracted impurities from redepositing on the fabric. The chelating agent is used to remove ions such as Ca, Mg and Fe that may have a negative effect on the process, and preferred examples include caustic soda (sodium hydroxide) and soda ash (sodium carbonate).

Use of the composition of the invention

In a third aspect, the present invention relates to the use of the composition of the invention in a simultaneous desizing and scouring process, preferably the process of the invention. In a preferred embodiment, the composition of the invention is used in the method of the invention.

Materials and methods

Alkaline alpha-amylase:

alkaline alpha-amylase SZIs as set forth in WO 00/60060 in SEQ ID NO: 2a variant alpha-amylase of the bacillus alpha-amylase backbone (backbone) disclosed. The amino acid sequence of the backbone has the following 6 amino acid deletions/substitutions:

D183^*+G184^*+R118K+N195F+R320K+R458K。

this variant is also disclosed in WO 01/66712. The alkaline alpha-amylase is prepared in batch 03AGE014-4, which is available on demand from Novozymes A/S.

Alkaline alpha-Amylase NLIs as set forth in WO 95/26397 in SEQ ID NO: 2A variant alpha-amylase of the bacillus alpha-amylase backbone disclosed in bacillus NCIB 12512 and having the amino acid sequence at D183^*+G184^*Double deletion of (a). The alkaline alpha-amylase is prepared in the batch APN00012, which is available from Novozymes A/S as required.

Pectate lyase SPIs as set forth in U.S. Pat. No.6,284, 524 SEQ ID NO: 2 discloses a bacillus licheniformis pectate lyase. This pectate lyase from Bacillus was prepared in the batch KND 01001. The enzyme may be obtained on demand from Novozymes A/S.

The method comprises the following steps:

alpha-Amylase Activity (KNU)

The amylolytic activity can be determined using potato starch as substrate. The method is based on the reaction of modified potato starch by enzymatic breakdown and then mixing a sample solution of starch/enzyme with an iodine solution. In contrast to standard colored glass standards, a deep blue color initially forms, but the blue color becomes lighter during starch breakdown and gradually becomes reddish brown.

One thousand Novo alpha-amylase units (KNU) are defined as Ca at standard conditions (i.e. +/-0.05; 0.0003M at 37 ℃.)²⁺(ii) a And pH 5.6), an amount of 5260mg dry starch substrate Merck Amylum soluble enzyme dextrinized.

Folder for describing this analysis method in more detailEB-SM-0009.02/01Available on demand from Novozymes a/S, denmark, which is incorporated herein by reference.

Viscosity measurement APSU

APSU unit: APSU unit measurement is a viscosity measurement method using the substrate polygalacturonic acid without the addition of calcium.

Substrate 5% polygalacturonic acid sodium salt (Sigma P-1879) was dissolved in 0.1M glycine buffer at pH 10. The 4m1 substrate was preincubated at 40 ℃ for 5 minutes, the enzyme was added (in a volume of 250. mu.L) and mixed in the mixer for 10 seconds at maximum speed, then incubated at 40 ℃ for 20 minutes. To obtain a standard curve, dilutions were measured twice with enzyme concentrations ranging from 5APSU/ml to above 100APSU/ml, with at least 4 concentrations between 10-60 APSU/ml.

The viscosity was determined using MIVI 600 from Sofraser corporation, 45700Villemandeur, france. The viscosity was measured in mV after 10 sec.

To calculate APSU units, standard enzyme dilutions as described above were used to obtain a standard curve. The GrafPad Prism program using a non-linear fit with monophasic exponential decay and plateau (plateau) was used for the calculation. The plateau and range are the mV obtained without addition of enzyme. The plateau was mV of greater than 100APSU/ml, and the viscosity half-decay value was found to be 12APSU units, and a standard error of 1.5APSU in both examples.

Lyase assay (at 235nm)

To determine β -elimination, an assay was performed to determine the increase in absorbance at 235nm using 0.1% polygalacturonic acid sodium salt (Sigma P-1879) dissolved in 0.1M glycine buffer at pH 10 as substrate. To calculate the catalytic rate, an increase in absorbance (absorbance) at 235 (5.2 units per minute) corresponds to1

Formation of μmol of unsaturated product (Nasuna and Starr, (1966) J.biol.chem., Vo1.241page5298-5306 (1966); and Bartleg, Wegener and Olsen, Microbiology, Vo1.141page873-881 (1995)).

Steady State conditions the absorbance was measured continuously at 235nm in a temperature-controlled cuvette vessel on an HP diode array spectrophotometer using a 0.5ml cuvette with a 1cm optical path. To reach steady state, a linear increase of at least 200 seconds is used for the rate calculation. Which is converted to micromoles per minute of product formed.

Lipase Activity (LU)

The cutinase activity was determined as the lipolytic activity using tributyrin as substrate. The method is based on the enzymatic hydrolysis of tributyrin and the consumption of base is recorded as a function of time.

One Lipase Unit (LU) is defined as the amount of enzyme that releases 1 micromole of titratable butyric acid per minute under standard conditions (i.e., at 30 ℃; pH 7.0; using Gum Arabic as an emulsifier and tributyrin as a substrate). Folder AF 95/5, which describes the analysis method in detail, is available on demand from Novozymes A/S, Denmark and is incorporated herein by reference.

Determination of cellulase Activity (ECU)

Cellulolytic activity can be measured by measuring the ability of an enzyme to reduce the viscosity of a carboxymethyl cellulose (CMC) solution, in-cut cellulose units (ECU).

The ECU assay quantifies the amount of catalytic activity present in a sample by determining the ability of the sample to reduce the viscosity of a carboxymethyl cellulose (CMC) solution. The test was carried out in a vibration viscometer (e.g., MIVI 3000 from Sofraser, france) at 40 ℃, pH 7.5, 0.1M phosphate buffer, for 30 minutes, using a relative enzyme standard that reduces the viscosity of CMC substrates (Hercules 7 LFD), at an enzyme concentration of approximately 0.15 ECU/ml. The primary standard was defined as 8200 ECU/g.

One ECU is the amount of enzyme that reduces the viscosity by half under the conditions described above.

The following non-limiting examples illustrate the invention.

Examples

Example 1

Desizing of cotton fabric in the absence of enzymes:

100% cotton woven fabric (270 g/m)²The fabric structure is Cupper 3/1. Warp yarn: 28 strands/cm, weft: 14 strands/cm) from Boras Wafveri Kungsfors, Sweden. The fabric has 8% starch based (starch based) size on the warp. A 0.25 m x o.5m fabric sample was cut and used. A25 mM buffer of pH9 was prepared with sodium tetraborate. Mixing O.5g/l surfactant BRIJ78 from Unichema and 0.5g/l surfactant from SASOLTDA-7 ethoxylate was added to the above buffer. The fabric samples were immersed in 1 liter of buffer solution containing surfactant for about 30 seconds and then padded by a pad-dyeing machine (Mathis) at about 50 ℃ to achieve a 90% fiber wet pick-up (wetpick up). The sample was quickly sealed in a plastic bag, which was incubated at 50 ℃ for 1 hour. After incubation, the fabric samples were rinsed in 90 ℃ water through four rinse boxes in a Mathis pad-steam oven (pad-steam range). The total time to pass through the rinse box was approximately 8 minutes. Prior to analysis, the fabric samples were air dried and then allowed to equilibrate in a conditioning chamber at 21 ℃ (70 ° F) and 65% relative humidity for at least 24 hours.

Desizing (Tegewa method)

The residue of the starch slurry was determined visually by comparing an iodine-dyed fabric sample with a standard set of photographs having a scale of 1-9 (scale) where 1 is dark blue and 9 is colorless. By dissolving 10g KI in 10ml water, 0.635gI was added₂A1 liter solution of 1 gram iodine and 200ml ethanol in deionized water was prepared to obtain an iodine staining solution. The fabric samples were cut and soaked in iodine solution for 60 seconds and then rinsed in deionized water for approximately 5 seconds. After excess water in the sample is squeezed out (press out), the fabric is worked on by at least two professionalsSamples were evaluated and the average given. Methods and standard scales are available from Verband TEGEWA, karlslasse 21, Frankfurt a.m., germany.

Pectin removal

The pectin residues on the fabric were quantitatively determined. The principle is as follows: ruthenium red (ruthenium red) binds to polyanionic compounds, such as unmethylated pectin. The pectin level on the fabric is proportional to the ruthenium red concentration on cotton fabric, which is also linearly proportional to the Kulberka-Munk function (i.e., K/S). The color reflectance (R) of the ruthenium red dyed fabric was measured at 540nm (Macbeth colorimeter, model 1# CE-7000) and automatically calculated as the K/S value by the following formula:

K/S＝(1-R)²/2R)。

the percent pectin removal was calculated by the following formula:

pectin removal percentage 1-% res pectin 1-100^*(K/S-K/S₀)/(K/S₁₀₀-K/S₀)

Wherein K/S₁₀₀From a fabric containing 100% pectin, typically a virgin untreated fabric; and K/S₀Then from fabrics with 0% residual pectin, typically heavily scoured and bleached fabrics. The staining solution was obtained by dissolving 0.2g/L Ruthenium red, 1.0g/L ammonium chloride, 2.5 ml/L28% ammonium hydroxide solution, 1.0g/L Silwet L-77 and 1.0g/L Tergito115-S-12 in distilled water to make a total of one liter of solution based on the information described in John H.Luft and article "Ruthenium red and Violet I.chemistry" 1971. The solution is prepared daily before use. For dyeing, 100mL of dye solution was used for 1 gram of fabric. The fabric samples were incubated in ruthenium red solution for 15 minutes at room temperature. The swatches were rinsed in a dyer (stainer) and then in distilled water (100ml/1g fabric) at 60 ℃ for 10 minutes. After drying, the color reflection coefficient (R) was determined.

Fabric wettability:

fabric wettability was determined according to AATCC test method 79-1995 using the drop test (drop test) experiment. A drop of water is allowed to drip from a fixed height (1 cm) onto the surface of the flat (taut) test specimen. The time required for the specular reflection (specific reflection) of the water droplets to disappear was measured and recorded as wetting time (wetting time).

Whiteness of fabrics

The desized fabric samples were passed through a bleach bath and padded to a wet pick up (wet up) of 90-100%. The bleach bath contained 10ml/L sodium silicate 40-42Be, 5 g/L40% EDTA, 16 ml/L50% w/v sodium hydroxide, and 16 ml/L50% hydrogen peroxide. The fabric swatches were incubated at 100 ℃ for 40 minutes and then rinsed in four rinse boxes at 95 ℃, 65 ℃ and 75 ℃, respectively. The total time to pass through the rinse box was approximately 8 minutes. After air drying and conditioning (conditioning), the whiteness of the fabric was measured according to AATCC test method 110-. CIE tristimulus (tristimulus) values were measured using a reflectance colorimeter (Macbeth colorimeter, Mode1# CE-7000) with a 300-700nm CIE illuminant D65 and a 196410 DEG observer. The whiteness degree is calculated from a formula based on CIE chromaticity coordinates (chromaticity coordinates).

The test results are shown in table 1.

Example 2

Desizing cotton fabric with alpha-amylase

The fabric and desizing process were essentially the same as in example 1, except that 10KNU/L of alpha-amylase SZ was also added to the surfactant containing buffer solution prior to desizing.

Starch size residues, pectin residues, fabric wettability and whiteness were evaluated in the same manner as in example 1. The test results are shown in table 1.

Example 3

Synchronous desizing and biological scouring of cotton fabrics

The fabric and desizing process was essentially the same as in example 2, except that 250APSU/L of pectate lyase SP was also added to the desizing solution prior to desizing.

Starch size residues, pectin residues, fabric wettability and whiteness were evaluated in the same manner as in example 1. The test results are shown in table 1.

Example 4

Synchronous desizing and biological scouring of cotton fabrics

The fabric and desizing process was essentially the same as in example 2, except that 750APSU/L of pectate lyase SP was also added to the desizing solution prior to desizing.

Starch size residues, pectin residues, fabric wettability and whiteness were evaluated in the same manner as in example 1. The test results are shown in table 1.

Example 5

Synchronous desizing and biological scouring of cotton fabrics

The fabric and the desizing process were essentially the same as in example 2, except that 1500APSU/L of pectate lyase SP was also added to the desizing solution prior to desizing.

Starch size residues, pectin residues, fabric wettability and whiteness were evaluated in the same manner as in example 1. The test results are shown in table 1.

Example 6

Desizing of cotton fabrics with alpha-amylase (NL)

The fabric and desizing process was essentially the same as in example 1, except that 10KNU/L of alpha-amylase NL was added to the surfactant containing buffer solution prior to desizing.

Starch size residues, pectin residues, fabric wettability and whiteness were evaluated in the same manner as in example 1. The test results are shown in table 1.

Example 7

Synchronous desizing and biological scouring of cotton fabrics

The fabric and desizing process were essentially the same as in example 6, except that 250APSU/L of pectate lyase SP was added to the desizing solution prior to desizing.

Starch size residues, pectin residues, fabric wettability and whiteness were evaluated in the same manner as in example 1. The test results are shown in table 1.

Example 8

Synchronous desizing and biological scouring of cotton fabrics

The fabric and desizing process were essentially the same as in example 6, except that 750APSU/L of pectate lyase SP was added to the desizing solution prior to desizing.

Starch size residues, pectin residues, fabric wettability and whiteness were evaluated in the same manner as in example 1. The test results are shown in table 1.

Example 9

Synchronous desizing and biological scouring of cotton fabrics

The fabric and desizing process were essentially the same as in example 6, except that 1500APSU/L of pectate lyase SP was added to the desizing solution prior to desizing.

Starch size residues, pectin residues, fabric wettability and whiteness were evaluated in the same manner as in example 1. The test results are shown in table 1.

Example 10

Desizing of cotton fabrics with alpha-amylase (SZ)

The fabric and desizing process were essentially the same as in example 1, except that 50KNU/L of alpha-amylase SZ was added to the surfactant containing buffer solution prior to desizing.

Starch size residues, pectin residues, fabric wettability and whiteness were evaluated in the same manner as in example 1. The test results are shown in table 1.

Example 11

Synchronous desizing and biological scouring of cotton fabrics

The fabric and desizing process was essentially the same as in example 10, except that 250APSU/L of pectate lyase SP was also added to the desizing solution prior to desizing.

Starch size residues, pectin residues, fabric wettability and whiteness were evaluated in the same manner as in example 1. The test results are shown in table 1.

Example 12

Synchronous desizing and biological scouring of cotton fabrics

The fabric and desizing process was essentially the same as in example 10, except that 750APSU/L of pectate lyase SP was also added to the desizing solution prior to desizing.

Starch size residues, pectin residues, fabric wettability and whiteness were evaluated in the same manner as in example 1. The test results are shown in table 1.

Example 13

Synchronous desizing and biological scouring of cotton fabrics

The fabric and desizing process were essentially the same as in example 10, except that 1500APSU/L of pectate lyase SP was also added to the desizing solution prior to desizing.

Starch size residues, pectin residues, fabric wettability and whiteness were evaluated in the same manner as in example 1. The test results are shown in table 1.

TABLE 1

Example #	Desizing (Tegewa)	Pectin removal Rate (%)	Soaking time (seconds)	Whiteness after bleaching CIE Ganz82
					1234567891O111213	3.04.754.54.54.754.254.54.54.O5.56.256.255.75	11.O13.928.545.852.314.725.742.149.517.431.945.250.9	6.413.67.33.42.38.86.44.94.46.64.O3.13.O	63.064.264.365.464.765.565.365.165.466.266.667.367.6

Sequence listing

<110> Novozymes North America Inc (Novozymes North America, Inc)

Novozymes corporation (Novozymes A/S)

<120> method for simultaneously desizing and scouring

<130>10657.204-WO

<160>6

<170>PatentIn version 3.3

<210>1

<211>1026

<212>DNA

<213> Bacillus licheniformis

<220>

<221>CDS

<222>(1)..(1026)

<400>1

atg aag aaa tta atc agc atc atc ttt atc ttt gta tta ggg gtt gtc 48

Met Lys Lys Leu Ile Ser Ile Ile Phe Ile Phe Val Leu Gly Val Val

1 5 10 15

ggg tca ttg aca gcg gcg gtt tcg gca gaa gca gct tct gcc tta aac 96

Gly Ser Leu Thr Ala Ala Val Ser Ala Glu Ala Ala Ser Ala Leu Asn

20 25 30

tcg ggc aaa gta aat ccg ctt gcc gac ttc agc tta aaa ggc ttt gcc 144

Ser Gly Lys Val Asn Pro Leu Ala Asp Phe Ser Leu Lys Gly Phe Ala

35 40 45

gca cta aac ggc gga aca acg ggc gga gaa ggc ggt cag acg gta acc 192

Ala Leu Asn Gly Gly Thr Thr Gly Gly Glu Gly Gly Gln Thr Val Thr

50 55 60

gta aca acg gga gat cag ctg att gcg gca tta aaa aat aag aat gca 240

Val Thr Thr Gly Asp Gln Leu Ile Ala Ala Leu Lys Asn Lys Asn Ala

65 70 75 80

aat acg cct tta aaa att tat gtc aac ggc acc att aca aca tca aat 288

Asn Thr Pro Leu Lys Ile Tyr Val Asn Gly Thr Ile Thr Thr Ser Asn

85 90 95

aca tcc gca tca aag att gac gtc aaa gac gtg tca aac gta tcg att 336

Thr Ser Ala Ser Lys Ile Asp Val Lys Asp Val Ser Asn Val Ser Ile

100 105 110

gtc gga tca ggg acc aaa ggg gaa ctc aaa ggg atc ggc atc aaa ata 384

Val Gly Ser Gly Thr Lys Gly Glu Leu Lys Gly Ile Gly Ile Lys Ile

115 120 125

tgg cgg gcc aac aac atc atc atc cgc aac ttg aaa att cac gag gtc 432

Trp Arg Ala Asn Asn Ile Ile Ile Arg Asn Leu Lys Ile His Glu Val

130 135 140

gcc tca ggc gat aaa gac gcg atc ggc att gaa ggc cct tct aaa aac 480

Ala Ser Gly Asp Lys Asp Ala Ile Gly Ile Glu Gly Pro Ser Lys Asn

145 150 155 160

att tgg gtt gat cat aat gag ctt tac cac agc ctg aac gtt gac aaa 528

Ile Trp Val Asp His Asn Glu Leu Tyr His Ser Leu Asn Val Asp Lys

165 170 175

gat tac tat gac gga tta ttt gac gtc aaa aga gat gcg gaa tat att 576

Asp Tyr Tyr Asp Gly Leu Phe Asp Val Lys Arg Asp Ala Glu Tyr Ile

180 185 190

aca ttc tct tgg aac tat gtg cac gat gga tgg aaa tca atg ctg atg 624

Thr Phe Ser Trp Asn Tyr Val His Asp Gly Trp Lys Ser Met Leu Met

195 200 205

ggt tca tcg gac agc gat aat tac aac agg acg att aca ttc cat cat 672

Gly Ser Ser Asp Ser Asp Asn Tyr Asn Arg Thr Ile Thr Phe His His

210 215 220

aac tgg ttt gag aat ctg aat tcg cgt gtg ccg tca ttc cgt ttc gga 720

Asn Trp Phe Glu Asn Leu Asn Ser Arg Val Pro Ser Phe Arg Phe Gly

225 230 235 240

gaa ggc cat att tac aac aac tat ttc aat aaa atc atc gac agc gga 768

Glu Gly His Ile Tyr Asn Asn Tyr Phe Asn Lys Ile Ile Asp Ser Gly

245 250 255

att aat tcg agg atg ggc gcg cgc atc aga att gag aac aac ctc ttt 816

Ile Asn Ser Arg Met Gly Ala Arg Ile Arg Ile Glu Asn Asn Leu Phe

260 265 270

gaa aac gcc aaa gat ccg att gtc tct tgg tac agc agt tca ccg ggc 864

Glu Asn Ala Lys Asp Pro Ile Val Ser Trp Tyr Ser Ser Ser Pro Gly

275 280 285

tat tgg cat gta tcc aac aac aaa ttt gta aac tct agg ggc agt atg 912

Tyr Trp His Val Ser Asn Asn Lys Phe Val Asn Ser Arg Gly Ser Met

290 295 300

ccg act acc tct act aca acc tat aat ccg cca tac agc tac tca ctc 960

Pro Thr Thr Ser Thr Thr Thr Tyr Asn Pro Pro Tyr Ser Tyr Ser Leu

305 310 315 320

gac aat gtc gac aat gta aaa tca atc gtc aag caa aat gcc gga gtc 1008

Asp Asn Val Asp Asn Val Lys Ser Ile Val Lys Gln Asn Ala Gly Val

325 330 335

ggc aaa atc aat cca taa 1026

Gly Lys Ile Asn Pro

340

<210>2

<211>341

<212>PRT

<213> Bacillus licheniformis

<400>2

Met Lys Lys Leu Ile Ser Ile Ile Phe Ile Phe Val Leu Gly Val Val

1 5 10 15

Gly Ser Leu Thr Ala Ala Val Ser Ala Glu Ala Ala Ser Ala Leu Asn

20 25 30

Ser Gly Lys Val Asn Pro Leu Ala Asp Phe Ser Leu Lys Gly Phe Ala

35 40 45

Ala Leu Asn Gly Gly Thr Thr Gly Gly Glu Gly Gly Gln Thr Val Thr

50 55 60

Val Thr Thr Gly Asp Gln Leu Ile Ala Ala Leu Lys Asn Lys Asn Ala

65 70 75 80

Asn Thr Pro Leu Lys Ile Tyr Val Asn Gly Thr Ile Thr Thr Ser Asn

85 90 95

Thr Ser Ala Ser Lys Ile Asp Val Lys Asp Val Ser Asn Val Ser Ile

100 105 110

Val Gly Ser Gly Thr Lys Gly Glu Leu Lys Gly Ile Gly Ile Lys Ile

115 120 125

Trp Arg Ala Asn Asn Ile Ile Ile Arg Asn Leu Lys Ile His Glu Val

130 135 140

Ala Ser Gly Asp Lys Asp Ala Ile Gly Ile Glu Gly Pro Ser Lys Asn

145 150 155 160

Ile Trp Val Asp His Asn Glu Leu Tyr His Ser Leu Asn Val Asp Lys

165 170 175

Asp Tyr Tyr Asp Gly Leu Phe Asp Val Lys Arg Asp Ala Glu Tyr Ile

180 185 190

Thr Phe Ser Trp Asn Tyr Val His Asp Gly Trp Lys Ser Met Leu Met

195 200 205

Gly Ser Ser Asp Ser Asp Ash Tyr Asn Arg Thr Ile Thr Phe His His

210 215 220

Asn Trp Phe Glu Asn Leu Asn Ser Arg Val Pro Ser Phe Arg Phe Gly

225 230 235 240

Glu Gly His Ile Tyr Asn Asn Tyr Phe Asn Lys Ile Ile Asp Ser Gly

245 250 255

Ile Asn Ser Arg Met Gly Ala Arg Ile Arg Ile Glu Asn Asn Leu Phe

260 265 270

Glu Asn Ala Lys Asp Pro Ile Val Ser Trp Tyr Ser Ser Ser Pro Gly

275 280 285

Tyr Trp His Val Ser Asn Asn Lys Phe Val Asn Ser Arg Gly Ser Met

290 295 300

Pro Thr Thr Ser Thr Thr Thr Tyr Asn Pro Pro Tyr Ser Tyr Ser Leu

305 310 315 320

Asp Asn Val Asp Asn Val Lys Ser Ile Val Lys Gln Asn Ala Gly Val

325 330 335

Gly Lys Ile Asn Pro

340

<210>3

<211>1455

<212>DNA

<213> Bacillus

<220>

<221> mat _ peptide

<222>(1)..()

<223>SP722

<220>

<221>CDS

<222>(1)..(1455)

<400>3

cat cat aat ggg aca aat ggg acg atg atg caa tac ttt gaa tgg cac 48

His His Asn Gly Thr Asn Gly Thr Met Met Gln Tyr Phe Glu Trp His

1 5 10 15

ttg cct aat gat ggg aat cac tgg aat aga tta aga gat gat gct agt 96

Leu Pro Asn Asp Gly Asn His Trp Asn Arg Leu Arg Asp Asp Ala Ser

20 25 30

aat cta aga aat aga ggt ata acc gct att tgg att ccg cct gcc tgg 144

Asn Leu Arg Asn Arg Gly Ile Thr Ala Ile Trp Ile Pro Pro Ala Trp

35 40 45

aaa ggg act tcg caa aat gat gtg ggg tat gga gcc tat gat ctt tat 192

Lys Gly Thr Ser Gln Asn Asp Val Gly Tyr Gly Ala Tyr Asp Leu Tyr

50 55 60

gat tta ggg gaa ttt aat caa aag ggg acg gtt cgt act aag tat ggg 240

Asp Leu Gly Glu Phe Asn Gln Lys Gly Thr Val Arg Thr Lys Tyr Gly

65 70 75 80

aca cgt agt caa ttg gag tct gcc atc cat gct tta aag aat aat ggc 288

Thr Arg Ser Gln Leu Glu Ser Ala Ile His Ala Leu Lys Asn Asn Gly

85 90 95

gtt caa gtt tat ggg gat gta gtg atg aac cat aaa gga gga gct gat 336

Val Gln Val Tyr Gly Asp Val Val Met Asn His Lys Gly Gly Ala Asp

100 105 110

gct aca gaa aac gtt ctt gct gtc gag gtg aat cca aat aac cgg aat 384

Ala Thr Glu Asn Val Leu Ala Val Glu Val Asn Pro Asn Asn Arg Asn

115 120 125

caa gaa ata tct ggg gac tac aca att gag gct tgg act aag ttt gat 432

Gln Glu Ile Ser Gly Asp Tyr Thr Ile Glu Ala Trp Thr Lys Phe Asp

130 135 140

ttt cca ggg agg ggt aat aca tac tca gac ttt aaa tgg cgt tgg tat 480

Phe Pro Gly Arg Gly Asn Thr Tyr Ser Asp Phe Lys Trp Arg Trp Tyr

145 150 155 160

cat ttc gat ggt gta gat tgg gat caa tca cga caa ttc caa aat cgt 528

His Phe Asp Gly Val Asp Trp Asp Gln Ser Arg Gln Phe Gln Asn Arg

165 170 175

atc tac aaa ttc cga ggt gat ggt aag gca tgg gat tgg gaa gta gat 576

Ile Tyr Lys Phe Arg Gly Asp Gly Lys Ala Trp Asp Trp Glu Val Asp

180 185 190

tcg gaa aat gga aat tat gat tat tta atg tat gca gat gta gat atg 624

Ser Glu Asn Gly Asn Tyr Asp Tyr Leu Met Tyr Ala Asp Val Asp Met

195 200 205

gat cat ccg gag gta gta aat gag ctt aga aga tgg gga gaa tgg tat 672

Asp His Pro Glu Val Val Asn Glu Leu Arg Arg Trp Gly Glu Trp Tyr

210 215 220

aca aat aca tta aat ctt gat gga ttt agg atc gat gcg gtg aag cat 720

Thr Asn Thr Leu Asn Leu Asp Gly Phe Arg Ile Asp Ala Val Lys His

225 230 235 240

att aaa tat agc ttt aca cgt gat tgg ttg acc cat gta aga aac gca 768

Ile Lys Tyr Ser Phe Thr Arg Asp Trp Leu Thr His Val Arg Asn Ala

245 250 255

acg gga aaa gaa atg ttt gct gtt gct gaa ttt tgg aaa aat gat tta 816

Thr Gly Lys Glu Met Phe Ala Val Ala Glu Phe Trp Lys Asn Asp Leu

260 265 270

ggt gcc ttg gag aac tat tta aat aaa aca aac tgg aat cat tct gtc 864

Gly Ala Leu Glu Asn Tyr Leu Asn Lys Thr Asn Trp Asn His Ser Val

275 280 285

ttt gat gtc ccc ctt cat tat aat ctt tat aac gcg tca aat agt gga 912

Phe Asp Val Pro Leu His Tyr Asn Leu Tyr Asn Ala Ser Asn Ser Gly

290 295 300

ggc aac tat gac atg gca aaa ctt ctt aat gga acg gtt gtt caa aag 960

Gly Asn Tyr Asp Met Ala Lys Leu Leu Asn Gly Thr Val Val Gln Lys

305 310 315 320

cat cca atg cat gcc gta act ttt gtg gat aat cac gat tct caa cct 1008

His Pro Met His Ala Val Thr Phe Val Asp Asn His Asp Ser Gln Pro

325 330 335

ggg gaa tca tta gaa tca ttt gta caa gaa tgg ttt aag cca ctt gct 1056

Gly Glu Ser Leu Glu Ser Phe Val Gln Glu Trp Phe Lys Pro Leu Ala

340 345 350

tat gcg ctt att tta aca aga gaa caa ggc tat ccc tct gtc ttc tat 1104

Tyr Ala Leu Ile Leu Thr Arg Glu Gln Gly Tyr Pro Ser Val Phe Tyr

355 360 365

ggt gac tac tat gga att cca aca cat agt gtc cca gca atg aaa gcc 1152

Gly Asp Tyr Tyr Gly Ile Pro Thr His Ser Val Pro Ala Met Lys Ala

370 375 380

aag att gat cca atc tta gag gcg cgt caa aat ttt gca tat gga aca 1200

Lys Ile Asp Pro Ile Leu Glu Ala Arg Gln Asn Phe Ala Tyr Gly Thr

385 390 395 400

caa cat gat tat ttt gac cat cat aat ata atc gga tgg aca cgt gaa 1248

Gln His Asp Tyr Phe Asp His His Asn Ile Ile Gly Trp Thr Arg Glu

405 410 415

gga aat acc acg cat ccc aat tca gga ctt gcg act atc atg tcg gat 1296

Gly Asn Thr Thr His Pro Asn Ser Gly Leu Ala Thr Ile Met Ser Asp

420 425 430

ggg cca ggg gga gag aaa tgg atg tac gta ggg caa aat aaa gca ggt 1344

Gly Pro Gly Gly Glu Lys Trp Met Tyr Val Gly Gln Asn Lys Ala Gly

435 440 445

caa gtt tgg cat gac ata act gga aat aaa cca gga aca gtt acg atc 1392

Gln Val Trp His Asp Ile Thr Gly Asn Lys Pro Gly Thr Val Thr Ile

450 455 460

aat gca gat gga tgg gct aat ttt tca gta aat gga gga tct gtt tcc 1440

Asn Ala Asp Gly Trp Ala Asn Phe Ser Val Asn Gly Gly Ser Val Ser

465 470 475 480

att tgg gtg aaa cga 1455

Ile Trp Val Lys Arg

485

<210>4

<211>485

<212>PRT

<213> Bacillus

<400>4

His His Asn Gly Thr Asn Gly Thr Met Met Gln Tyr Phe Glu Trp His

1 5 10 15

Leu Pro Asn Asp Gly Asn His Trp Asn Arg Leu Arg Asp Asp Ala Ser

20 25 30

Asn Leu Arg Asn Arg Gly Ile Thr Ala Ile Trp Ile Pro Pro Ala Trp

35 40 45

Lys Gly Thr Ser Gln Asn Asp Val Gly Tyr Gly Ala Tyr Asp Leu Tyr

50 55 60

Asp Leu Gly Glu Phe Asn Gln Lys Gly Thr Val Arg Thr Lys Tyr Gly

65 70 75 80

Thr Arg Ser Gln Leu Glu Ser Ala Ile His Ala Leu Lys Asn Asn Gly

85 90 95

Val Gln Val Tyr Gly Asp Val Val Met Asn His Lys Gly Gly Ala Asp

100 105 110

Ala Thr Glu Asn Val Leu Ala Val Glu Val Asn Pro Asn Asn Arg Asn

115 120 125

Gln Glu Ile Ser Gly Asp Tyr Thr Ile Glu Ala Trp Thr Lys Phe Asp

130 135 140

Phe Pro Gly Arg Gly Asn Thr Tyr Ser Asp Phe Lys Trp Arg Trp Tyr

145 150 155 160

Hi s Phe Asp Gly Val Asp Trp Asp Gln Ser Arg Gln Phe Gln Asn Arg

165 170 175

Ile Tyr Lys Phe Arg Gly Asp Gly Lys Ala Trp Asp Trp Glu Val Asp

180 185 190

Ser Glu Asn Gly Asn Tyr Asp Tyr Leu Met Tyr Ala Asp Val Asp Met

195 200 205

Asp His Pro Glu Val Val Asn Glu Leu Arg Arg Trp Gly Glu Trp Tyr

210 215 220

Thr Asn Thr Leu Asn Leu Asp Gly Phe Arg Ile Asp Ala Val Lys His

225 230 235 240

Ile Lys Tyr Ser Phe Thr Arg Asp Trp Leu Thr His Val Arg Asn Ala

245 250 255

Thr Gly Lys Glu Met Phe Ala Val Ala Glu Phe Trp Lys Asn Asp Leu

260 265 270

Gly Ala Leu Glu Asn Tyr Leu Asn Lys Thr Asn Trp Asn His Ser Val

275 280 285

Phe Asp Val Pro Leu His Tyr Asn Leu Tyr Asn Ala Ser Asn Ser Gly

290 295 300

Gly Asn Tyr Asp Met Ala Lys Leu Leu Asn Gly Thr Val Val Gln Lys

305 310 315 320

His Pro Met His Ala Val Thr Phe Val Asp Asn His Asp Ser Gln Pro

325 330 335

Gly Glu Ser Leu Glu Ser Phe Val Gln Glu Trp Phe Lys Pro Leu Ala

340 345 350

Tyr Ala Leu Ile Leu Thr Arg Glu Gln Gly Tyr Pro Ser Val Phe Tyr

355 360 365

Gly Asp Tyr Tyr Gly Ile Pro Thr His Ser Val Pro Ala Met Lys Ala

370 375 380

Lys Ile Asp Pro Ile Leu Glu Ala Arg Gln Asn Phe Ala Tyr Gly Thr

385 390 395 400

Gln His Asp Tyr Phe Asp Hi s His Asn Ile Ile Gly Trp Thr Arg Glu

405 410 415

Gly Asn Thr Thr His Pro Asn Ser Gly Leu Ala Thr Ile Met Ser Asp

420 425 430

Gly Pro Gly Gly Glu Lys Trp Met Tyr Val Gly Gln Asn Lys Ala Gly

435 440 445

Gln Val Trp His Asp Ile Thr Gly Asn Lys Pro Gly Thr Val Thr Ile

450 455 460

Asn Ala Asp Gly Trp Ala Asn Phe Ser Val Asn Gly Gly Ser Val Ser

465 470 475 480

Ile Trp Val Lys Arg

485

<210>5

<211>1455

<212>DNA

<213> Bacillus

<220>

<221>CDS

<222>(1)..(1455)

<223>AA560

<220>

<221> mat _ peptide

<222>(1)..()

<400>5

cac cat aat ggt acg aac ggc aca atg atg cag tac ttt gaa tgg tat 48

His His Asn Gly Thr Asn Gly Thr Met Met Gln Tyr Phe Glu Trp Tyr

1 5 10 15

cta cca aat gac gga aac cat tgg aat aga tta agg tct gat gca agt 96

Leu Pro Asn Asp Gly Asn His Trp Asn Arg Leu Arg Ser Asp Ala Ser

20 25 30

aac cta aaa gat aaa ggg atc tca gcg gtt tgg att cct cct gca tgg 144

Asn Leu Lys Asp Lys Gly Ile Ser Ala Val Trp Ile Pro Pro Ala Trp

35 40 45

aag ggt gcc tct caa aat gat gtg ggg tat ggt gct tat gat ctg tat 192

Lys Gly Ala Ser Gln Asn Asp Val Gly Tyr Gly Ala Tyr Asp Leu Tyr

50 55 60

gat tta gga gaa ttc aat caa aaa gga acc att cgt aca aaa tat gga 240

Asp Leu Gly Glu Phe Asn Gln Lys Gly Thr Ile Arg Thr Lys Tyr Gly

65 70 75 80

acg cgc aat cag tta caa gct gca gtt aac gcc ttg aaa agt aat gga 288

Thr Arg Asn Gln Leu Gln Ala Ala Val Asn Ala Leu Lys Ser Asn Gly

85 90 95

att caa gtg tat ggc gat gtt gta atg aat cat aaa ggg gga gca gac 336

Ile Gln Val Tyr Gly Asp Val Val Met Asn His Lys Gly Gly Ala Asp

100 105 110

gct acc gaa atg gtt agg gca gtt gaa gta aac ccg aat aat aga aat 384

Ala Thr Glu Met Val Arg Ala Val Glu Val Asn Pro Asn Asn Arg Asn

115 120 125

caa gaa gtg tcc ggt gaa tat aca att gag gct tgg aca aag ttt gac 432

Gln Glu Val Ser Gly Glu Tyr Thr Ile Glu Ala Trp Thr Lys Phe Asp

130 135 140

ttt cca gga cga ggt aat act cat tca aac ttc aaa tgg aga tgg tat 480

Phe Pro Gly Arg Gly Asn Thr His Ser Asn Phe Lys Trp Arg Trp Tyr

145 150 155 160

cac ttt gat gga gta gat tgg gat cag tca cgt aag ctg aac aat cga 528

His Phe Asp Gly Val Asp Trp Asp Gln Ser Arg Lys Leu Asn Asn Arg

165 170 175

att tat aaa ttt aga ggt gat gga aaa ggg tgg gat tgg gaa gtc gat 576

Ile Tyr Lys Phe Arg Gly Asp Gly Lys Gly Trp Asp Trp Glu Val Asp

180 185 190

aca gaa aac ggt aac tat gat tac cta atg tat gca gat att gac atg 624

Thr Glu Asn Gly Asn Tyr Asp Tyr Leu Met Tyr Ala Asp Ile Asp Met

195 200 205

gat cac cca gag gta gtg aat gag cta aga aat tgg ggt gtt tgg tat 672

Asp His Pro Glu Val Val Asn Glu Leu Arg Asn Trp Gly Val Trp Tyr

210 215 220

acg aat aca tta ggc ctt gat ggt ttt aga ata gat gca gta aaa cat 720

Thr Asn Thr Leu Gly Leu Asp Gly Phe Arg Ile Asp Ala Val Lys His

225 230 235 240

ata aaa tac agc ttt act cgt gat tgg att aat cat gtt aga agt gca 768

Ile Lys Tyr Ser Phe Thr Arg Asp Trp Ile Asn His Val Arg Ser Ala

245 250 255

act ggc aaa aat atg ttt gcg gtt gcg gaa ttt tgg aaa aat gat tta 816

Thr Gly Lys Asn Met Phe Ala Val Ala Glu Phe Trp Lys Asn Asp Leu

260 265 270

ggt gct att gaa aac tat tta aac aaa aca aac tgg aac cat tca gtc 864

Gly Ala Ile Glu Asn Tyr Leu Asn Lys Thr Asn Trp Asn His Ser Val

275 280 285

ttt gat gtt ccg ctg cac tat aac ctc tat aat gct tca aaa agc gga 912

Phe Asp Val Pro Leu His Tyr Asn Leu Tyr Asn Ala Ser Lys Ser Gly

290 295 300

ggg aat tat gat atg agg caa ata ttt aat ggt aca gtc gtg caa aga 960

Gly Asn Tyr Asp Met Arg Gln Ile Phe Asn Gly Thr Val Val Gln Arg

305 310 315 320

cat cca atg cat gct gtt aca ttt gtt gat aat cat gat tcg caa cct 1008

His Pro Met His Ala Val Thr Phe Val Asp Asn His Asp Ser Gln Pro

325 330 335

gaa gaa gct tta gag tct ttt gtt gaa gaa tgg ttc aaa cca tta gcg 1056

Glu Glu Ala Leu Glu Ser Phe Val Glu Glu Trp Phe Lys Pro Leu Ala

340 345 350

tat gct ttg aca tta aca cgt gaa caa ggc tac cct tct gta ttt tat 1104

Tyr Ala Leu Thr Leu Thr Arg Glu Gln Gly Tyr Pro Ser Val Phe Tyr

355 360 365

gga gat tat tat ggc att cca acg cat ggt gta cca gcg atg aaa tcg 1152

Gly Asp Tyr Tyr Gly Ile Pro Thr His Gly Val Pro Ala Met Lys Ser

370 375 380

aaa att gac ccg att cta gaa gcg cgt caa aag tat gca tat gga aga 1200

Lys Ile Asp Pro Ile Leu Glu Ala Arg Gln Lys Tyr Ala Tyr Gly Arg

385 390 395 400

caa aat gac tac tta gac cat cat aat atc atc ggt tgg aca cgt gaa 1248

Gln Asn Asp Tyr Leu Asp His His Asn Ile Ile Gly Trp Thr Arg Glu

405 410 415

ggg aat aca gca cac ccc aac tcc ggt tta gct act atc atg tcc gat 1296

Gly Asn Thr Ala His Pro Asn Ser Gly Leu Ala Thr Ile Met Ser Asp

420 425 430

ggg gca gga gga aat aag tgg atg ttt gtt ggg cgt aat aaa gct ggt 1344

Gly Ala Gly Gly Asn Lys Trp Met Phe Val Gly Arg Asn Lys Ala Gly

435 440 445

caa gtt tgg acc gat atc act gga aat cgt gca ggt act gtt acg att 1392

Gln Val Trp Thr Asp Ile Thr Gly Asn Arg Ala Gly Thr Val Thr Ile

450 455 460

aat gct gat gga tgg ggt aat ttt tct gta aat gga gga tca gtt tct 1440

Asn Ala Asp Gly Trp Gly Asn Phe Ser Val Asn Gly Gly Ser Val Ser

465 470 475 480

att tgg gta aac aaa 1455

Ile Trp Val Asn Lys

485

<210>6

<211>485

<212>PRT

<213> Bacillus

<400>6

His His Asn Gly Thr Asn Gly Thr Met Met Gln Tyr Phe Glu Trp Tyr

1 5 10 15

Leu Pro Asn Asp Gly Asn His Trp Asn Arg Leu Arg Ser Asp Ala Ser

20 25 30

Asn Leu Lys Asp Lys Gly Ile Ser Ala Val Trp Ile Pro Pro Ala Trp

35 40 45

Lys Gly Ala Ser Gln Asn Asp Val Gly Tyr Gly Ala Tyr Asp Leu Tyr

50 55 60

Asp Leu Gly Glu Phe Asn Gln Lys Gly Thr Ile Arg Thr Lys Tyr Gly

65 70 75 80

Thr Arg Asn Gln Leu Gln Ala Ala Val Asn Ala Leu Lys Ser Asn Gly

85 90 95

Ile Gln Val Tyr Gly Asp Val Val Met Asn His Lys Gly Gly Ala Asp

100 105 110

Ala Thr Glu Met Val Arg Ala Val Glu Val Asn Pro Asn Asn Arg Asn

115 120 125

Gln Glu Val Ser Gly Glu Tyr Thr Ile Glu Ala Trp Thr Lys Phe Asp

130 135 140

Phe Pro Gly Arg Gly Asn Thr His Ser Asn Phe Lys Trp Arg Trp Tyr

145 150 155 160

His Phe Asp Gly Val Asp Trp Asp Gln Ser Arg Lys Leu Asn Asn Arg

165 170 175

Ile Tyr Lys Phe Arg Gly Asp Gly Lys Gly Trp Asp Trp Glu Val Asp

180 185 190

Thr Glu Asn Gly Asn Tyr Asp Tyr Leu Met Tyr Ala Asp Ile Asp Met

195 200 205

Asp His Pro Glu Val Val Asn Glu Leu Arg Asn Trp Gly Val Trp Tyr

210 215 220

Thr Asn Thr Leu Gly Leu Asp Gly Phe Arg Ile Asp Ala Val Lys His

225 230 235 240

Ile Lys Tyr Ser Phe Thr Arg Asp Trp Ile Asn His Val Arg Ser Ala

245 250 255

Thr Gly Lys Asn Met Phe Ala Val Ala Glu Phe Trp Lys Asn Asp Leu

260 265 270

Gly Ala Ile Glu Asn Tyr Leu Asn Lys Thr Asn Trp Asn His Ser Val

275 280 285

Phe Asp Val Pro Leu His Tyr Asn Leu Tyr Asn Ala Ser Lys Ser Gly

290 295 300

Gly Asn Tyr Asp Met Arg Gln Ile Phe Asn Gly Thr Val Val Gln Arg

305 310 315 320

His Pro Met His Ala Val Thr Phe Val Asp Asn His Asp Ser Gln Pro

325 330 335

Glu Glu Ala Leu Glu Ser Phe Val Glu Glu Trp Phe Lys Pro Leu Ala

340 345 350

Tyr Ala Leu ThrLeu Thr Arg Glu Gln Gly Tyr Pro Ser Val Phe Tyr

355 360 365

Gly Asp Tyr Tyr Gly Ile Pro Thr His Gly Val Pro Ala Met Lys Ser

370 375 380

Lys Ile Asp Pro Ile Leu Glu Ala Arg Gln Lys Tyr Ala Tyr Gly Arg

385 390 395 400

Gln Asn Asp Tyr Leu Asp His His Asn Ile Ile Gly Trp Thr Arg Glu

405 410 415

Gly Asn Thr Ala His Pro Asn Ser Gly Leu Ala Thr Ile Met Ser Asp

420 425 430

Gly Ala Gly Gly Asn Lys Trp Met Phe Val Gly Arg Asn Lys Ala Gly

435 440 445

Gln Val Trp Thr Asp Ile Thr Gly Asn Arg Ala Gly Thr Val Thr Ile

450 455 460

Asn Ala Asp Gly Trp Gly Asn Phe Ser Val Asn Gly Gly Ser Val Ser

465 470 475 480

Ile Trp Val Asn Lys

485

Claims (30)

1. A method for simultaneously desizing and scouring a sized fabric containing starch or starch derivatives, which method comprises treating said fabric with an alkaline alpha-amylase and an alkaline pectinase at a pH above 7 and a temperature between 30 ℃ and 115 ℃.

2. The method of claim 1, wherein the alkaline pectinase is a pectate lyase.

3. The method of claim 2, wherein the pectate lyase is derived from a strain of Bacillus.

4. The method of claim 3, wherein the strain is from Bacillus licheniformis, Bacillus alkalophilus, Bacillus pseudoalkalophilus, and Bacillus clarkia.

5. The method of claim 2, wherein the pectate lyase is the amino acid sequence of SEQ ID NO: 2, mature pectate lyase.

6. The method of claim 1, wherein the alkaline alpha-amylase is derived from a Bacillus species.

7. The method of claim 6, wherein the alkaline alpha-amylase is SEQ ID NO: 4 or SEQ ID NO: 6.

8. The method of claim 7, wherein the Bacillus alpha-amylase is produced using the polypeptide of SEQ ID NO: numbering 4, one or more deletions at position D183+ G184.

9. The method of claim 7, wherein the Bacillus alpha-amylase is produced using the polypeptide of SEQ ID NO: numbering 6, there are one or more deletions at positions D183 and G184.

10. The method of any one of claims 1-9, wherein the alkaline alpha-amylase is present at a concentration of 0.05-150KNU/L treatment solution.

11. The method of any one of claims 1-9, wherein the pectinase is a pectate lyase present at a concentration of 1-1,500APSU/kg fabric.

12. The process of any one of claims 1-9, wherein the process is carried out at a pH of 7-11.

13. The process of claim 12, wherein the process is carried out at a pH of 8 to 10.

14. The process of any one of claims 1 to 9, wherein the process is carried out at a temperature of from 30 ℃ to 60 ℃ or from 50 ℃ to 110 ℃.

15. The process of any one of claims 1 to 9, wherein the process is carried out in the presence of a surfactant, the surfactant being present at a concentration of 0.1 to 10 g/L.

16. The method of any one of claims 1-9, wherein the fabric is a cellulosic fabric.

17. The method of any one of claims 1-9, wherein the fabric is silk or wool.

18. The method of any of claims 1-9, wherein the fabric is a polyester-containing fabric, or a garment consisting essentially of 100% polyester.

19. The method of claim 18, wherein said polyester fabric is a polyester blend.

20. The method of any one of claims 1-9, wherein the method is performed in a single bath.

21. A composition comprising alkaline alpha-amylase and alkaline pectinase.

22. The composition of claim 21, wherein the alkaline alpha-amylase is derived from a strain of bacillus species.

23. The composition of claim 21 or 22, wherein the alkaline pectinase is pectate lyase.

24. The composition of claim 23, wherein the pectate lyase is a pectate lyase derived from a strain of bacillus.

25. The composition of claim 24, wherein the strain is a strain from Bacillus licheniformis, Bacillus alkalophilus, Bacillus pseudoalkalophilus, and Bacillus clarkia.

26. The composition of claim 22, wherein the bacillus alpha-amylase is modified in the presence of a protease in a nucleic acid sequence as set forth in SEQ ID NO: numbering 4, there are one or more deletions at positions D183 and G184.

27. The composition of claim 22, wherein the bacillus alpha-amylase is modified in the presence of a protease in a nucleic acid sequence as set forth in SEQ ID NO: in the case of numbering 6, there are one or more deletions at the positions D183 and G184.

28. The composition of any one of claims 22-27, wherein the composition further comprises a stabilizer, a surfactant, a wetting agent, a dispersant, a chelating agent, and an emulsifier, or a mixture thereof.

29. Use of the composition of any one of claims 22-27 for simultaneous desizing and scouring.

30. The use of claim 29, wherein the composition is used in the method of any one of claims 1-20.