CN101437999A - Desizing process - Google Patents

Abstract

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

Description

Desizing process

List of reference sequences

The application contains the sequence list of computer-reader form.Described computer-reader form is incorporated this paper into by reference.

Invention field

The present invention relates to during making particularly new fabric the destarch method of (sized) fabric of starching.

Background technology

Converted goods, for example cellulosic material makes it to become the material that is ready for use on the clothing manufacturing, relates to several steps: with described fiber spun yarn; Form (woven) of weaving or (knit) fabric of braiding from described yarn; Preparation subsequently, dyeing and arrangement (finishing) operation.Described preparatory technology, it can relate to destarch (for textiles (woven goods)), kiering (scouring) and bleaching, produces the fabric that is applicable to dyeing or arrangement.

In desizing process alkali alpha amylase is used as auxiliary ingredient (auxiliaries), to promote removing amyloid slurry (size), it is used as the protective layer on the yarn in weaving.Removing described pulp layer after the weaving fully to guaranteeing that the optimum in processing subsequently is important, in this following process usually with fabric kiering, bleaching, dyeing and/or printing.

The step that comprises mineral matter after the destarch step usually is worth and will does, to remove metal ion, for example Mn ²⁺, Fe ²⁺/ Fe ³⁺, Cu ²⁺Deng, if it is present on the fabric, may causes the inhomogeneous bleaching in procedure of processing after a while or even may form trachoma (pin-hole) in the BLEACHED FABRIC.Go mineral matter to follow acidic precipitation usually, and be usually directed to the interpolation of sour for example acetate or sulfuric acid.

Make new fabric particularly during, people's expectation can provide the improved destarch method of the fabric of starching that is used for.

Summary of the invention

What the present invention relates to is to provide the destarch method of the fabric of starching during making particularly new fabric.

First aspect of the present invention relates to during making fabric the destarch method of the fabric of starching, described fabric contains starch or starch derivatives, this method is included in the fabric of the described starching of incubation in the processing aqueous solution (aqueous treating solution) with pH scope between 1 and 5, and this processing aqueous solution comprises α-Dian Fenmei.

The present invention has been found that when implementing desizing process of the present invention, as defined in claims, does not need mineral matter.The described mineral matter that goes carries out in identical Treatment Solution in destarch sizing fabric and/or afterwards.Compare with the conventional method of going the mineral matter step with relating to alkaline destarch step, avoided the pH regulator step.Another advantage of the present invention is the saving/minimizing processing time, because destarch and go mineral matter to carry out simultaneously.Even cost will be saved/be reduced to described destarch and go mineral matter not finish dealing with one-step method promptly, side by side,, for example acid and be used to add the artificial of acid is because avoid in the alkaline destarch step of tradition and go pH regulator step between the mineral matter step.

In the context of the present invention, with term " fabric " (fabric) " and the interchangeable use of term " textiles (textile) "; the meaning contrasts with " old " washing fabric (used laundry fabric); coming of new, the finished fabric of preferred undyed fabric, clothing, fiber, yarn or other type.Can make fabric from fiber by weaving, braiding or on-woven operation.Weaving and braiding need yarn as input, and described bondedfibre fabric is the result's (can regard paper as on-woven) who connects fiber at random.

Textile fabric (woven fabric) is to pass through on the loom between warp thread with vertical weaving " filler/weft yarn (filling) " or the manufacturing of parallel yarn.The necessary starching of warp thread before the weaving is to lubricate and to protect them to avoid wearing and tearing when the weaving high speed is inserted weft yarn (filling yarn).Described weft yarn can pass warp thread and weave (plain weave (plain weave)) in the mode of " last-next one (over one-under the next) ", or weaves with the arrangement of " last-following two (over one-under two) " (twill weave) or any other countless versions.Dynamics, texture, the pattern not only type/quality with yarn are relevant, and also the type with weaving is relevant.Usually, clothes, shirt, trousers, coverlet class, towel, drapery etc. produce from textile fabric.

Braiding (knitting) is by chain yarn ring is connected together the formation fabric.With by two types yarn manufacturing and to have many " end " weaving (ends) opposite, braided fabric is from single continuous yarn production.The same with weaving, have many different modes that yarn ring (loop yarn) is compiled together, and the characteristic of final fabric depend on the type of yarn and braiding.Underwear, lint shirt, socks, sweater, undershirt etc. are from braided fabric production.

Bondedfibre fabric passes through the pieces of fabric that connection and/or chain fiber and fibril (filament) are made with method mechanical, heat, chemistry or the solvent mediation.The gained fabric can be the form with network structure, laminated material (laminates) or film.Representative instance be disposable infant diaper, towel, rag, operation with clothes, " environmental friendliness " (environmental friendly) formula fiber, filter medium, bedding, roof Material, two-dimensional fabric and many other.

According to the present invention, described method can be applied to any starching known in the described technical field fabric (weaving, braiding with on-woven).Described method is applied to the sizing fabric of coming of new, with the fabric with respect to used and/or pollution to be cleaned in the clothes washing process.Described in one embodiment fabric is the fiber manufacturing by natural and/or artificial source.Described in another embodiment fabric is the fiber manufacturing from animal origin.Especially, method of the present invention can be applied to fabric cellulose or that cellulose material is arranged, for example cotton, viscose (viscose), artificial fibre, ramie, sodolin, cellulose acetate, jean (denim), lyocell (Tencel for example ^TMMake by CourtauldsFibers), or its mixture, or any of these fiber and synthetic fiber (for example polyester, polyamide, acrylic acid, or polyurethanes, nylon, polyethylene terephthalate or PLA) together mixture, or with the mixture of other natural fabrics for example woollen (wool) and silk (silk), for example viscose/cotton mixture, the cotton mixture of lyocell/, viscose/woollen mixture, lyocell/ woolen cloth mixture, cotton/woollen mixture; Flax (sodolin (linen)), ramie (ramie) and other fabrics based on cellulose fibre, comprise the fabric of whole cellulose and other fibers mixture as woolen cloth, polyamide, acrylic acid and polyester fiber, for example, viscose/cotton/polyester mixture (blend), woolen cloth/cotton/polyester mixture, flax/cotton mixture etc.The fabric that described method can also be used to synthesize for example, is made up of 100% polyester, polyamide, nylon in fact respectively.The meaning of described term " woolen cloth " is the available animal hair product of any commerce, for example, from the woolen cloth of sheep, camel, rabbit, goat, lama, and the woolen cloth that is called as Merino wool (merino wool), Shetland wool (Shetland wool), cashmere wool/goat cashmere (cashmere wool), alpaca fibre (alpaca wool), mohair etc.The method of the invention can be used with woolen cloth or animal hair material, with the form of cloister fabric (top), fiber, yarn or weaving or braided fabric.

The α-Dian Fenmei consistent with the method for the invention can be any α-Dian Fenmei, but preferred bacterium or originated from fungus.Preferred α-Dian Fenmei is an acid alpha-amylase, for example the bacterial strain of the acid alpha-amylase of deriving from filamentous fungi, especially aspergillus (Aspergillus), Rhizomucor or Meripillus.

The meaning of described term " acid alpha-amylase " be 50 ℃ of temperature in pH in 1 to 7 scope, preferably 1 to 5, have the α-Dian Fenmei of optimum activity.

Described term " destarch " means understanding in a usual manner, that is, from fabric, for example the warp in textile fabric is degraded and/or the removal slurry formulations.

Described term " fabric that contains starch or starch derivatives " means the fabric that any form contains starch or starch derivatives, and is special in containing the textile fabric of cellulosic material preparation.Described fabric normally no dyeing and by manufacturings such as cotton, viscose, flax.The major part that is present in starch in the fabric or starch derivatives is slurry (size) normally, and it is used for being coated in yarn before weaving, normally on the warp (warp).

Described term " carbohydrate binding modules (carbohydrate-binding module; CBM) ", or common " sugar is in conjunction with territory (CBD) " of indication, be and polypeptid acid sequence poly-or that oligosaccharides (carbohydrate) preferentially combines, frequently, but and uniquely nonessential, combine with poly-or oligosaccharides water-fast (comprising crystallization) form.

Even without mentioning related with the method for the invention especially, should infer with " effective dose " and use described enzyme or preparation.The meaning of described term " effective dose " is that for example α-Dian Fenmei can provide the effect that expectation obtains, and promptly compares the amount of the described fabric of destarch with the fabric of handling without described enzyme.

Description of drawings

Fig. 1 is presented at the destarch effect of pH4.0 α-Dian Fenmei D for the Vlisco fabric

Fig. 2 is presented at the destarch effect of pH4.0 α-Dian Fenmei C for the Vlisco fabric

Detailed Description Of The Invention

The present invention is directly for being provided at the particularly destarch method of sizing fabric during the new fabric of manufacturing.

In preferred embodiments, follow the kiering step after the destarch step of the present invention, preferred enzymatic kiering step is preferably used for example pectase of kiering enzyme, for example, and transelminase, lipase, protease, or its combination; And blanching step, preferably include and use hydrogen peroxide and/or the bleaching of hydrogen peroxide product. Relevant kiering method is described in United States Patent (USP) numbering 5,578,489, United States Patent (USP) numbering 5,912,407 and United States Patent (USP) numbering 6,630,342 to some extent. Relevant method for bleaching is described in United States Patent (USP) numbering 5,851,233, United States Patent (USP) numbering 5,752,980 and United States Patent (USP) numbering 5,928,380 to some extent. The kiering of correlation combiner and method for bleaching are described in WO 2003/002810 (Novozymes) and WO 2003/002705 (Novozymes) respectively to some extent.

According to the present invention, fabric can side by side carry out destarch and remove mineral matter in the identical processing aqueous solution, or carries out continuously destarch and remove mineral matter in identical or two Treatment Solution of separating. In preferred embodiments, in identical Treatment Solution, carry out simultaneously described destarch and remove mineral matter. The method of the invention can use traditional starching/destarch instrument to finish, for example, and pad system, J-boxes, jets, jiggers etc. Usually, do not need extra process instrumentation.

According to while destarch of the present invention with to remove mineral matter be by having in the processing aqueous solution of pH scope between 1 and 5 the fabric of starching of incubation, the described processing aqueous solution comprises AMS. In preferred embodiments, the pH between incubation period makes scope between 1 and 4, especially between pH2 and 4.

Textile fabric is the common form of textile article. Weaving method needs " starching " warp to protect it to avoid wearing and tearing. Not modified or modify starch, polyvinyl alcohol (PVA), carboxymethyl cellulose (CMC), wax and acryloid cement, with and composition thereof be the example that is typically used as upper slurry formulation. Slurry formulation can be based on starch or based on the upper slurry formulation of starch derivatives on according to the present invention, but can also contain one or more non-starch or based on the upper slurry formulation of starch derivatives. Described upper slurry formulation is removed after weaving is processed usually, as the first step of preparation yarn fabric.

One or more other preparations comprise stabilizing agent, surfactant, wetting agent, dispersant, chelating agent and emulsifying agent, or its mixture, may reside in during the desizing process of the present invention. Described sizing fabric allows the sufficiently long period of incubation in water treatment solution, to realize the destarch of sizing fabric. Window of opportunity depends on type and the temperature of processing scheme, and can be changed to a few days from about 15 minutes, for example, and 48 hours. Method of the present invention is preferably carried out when from 5 to 90 ℃ of temperature ranges, particularly 20 to 90 ℃, depends on processing scheme.

Described processing scheme can be in batches or continuous fabric is contacted with the processing aqueous solution with open width (open width) or rope form (rope form) form.

Continuous operation can be used saturator, and the water treatment solution of the equivalent weight of about every fabric weight is applied to described fabric thus, then suspends case (heated dwell chamber) by heat, and chemical reaction carries out therein. Then clean link for the preparation of the fabric of next treatment step. For the pure white or good wettable and the generation dyeability that guarantee height, described destarch enzyme and other preparations must thoroughly be removed.

Operation in batches (batch process) can occur in a collection of (Treatment Solution), and fabric doubly contacts to the processing aqueous solution of its weight with for example about 8-15 thus. , will process the aqueous solution and drain after the period at incubation, rinse fabric, and start next treatment step. Discrete PB method (namely, pad-batch processes (pad-batch processes)) comprises saturated machine, the processing aqueous solution of the equivalent weight of about every fabric weight is applied to described fabric thus, (dwell period) follows by the blocking period, it may be one or a few days in the situation of CPB method (that is, cold pad-batch processes (cold pad-batch processes)). For example, the CPB method can in the pH scope between 1 and 5, be carried out between 20-40 ℃ 8-24 hour or more of a specified duration; Preferred pH scope is between about 1 and 4, especially between pH2 and 4. In addition, the PB method can be between about 1 and 5 in the pH scope, and preferably approximately 1-4, particularly 2-4 carried out 1-6 hour in 40-90 ℃.

Desizing process of the present invention can use the AMS of effective dose to carry out in one embodiment, preferred acidic AMS, and such as lactic acid or sulfuric acid etc. of acid.

Washing agent

In the context of the present invention, washing agent and surfactant are synonyms, and it can refer in particular to non-ionic surface active agent, anion surfactant, cationic surfactant, amphoteric surfactant, zwitterionic surfactant and semi-polarity surfactant, or its mixture.

Described surfactant is present in the composition of the present invention with 0.1% to 60% usually by weight.

Described surfactant preferably is deployed into the enzyme component compatibility that exists. In liquid or gel combination, described surfactant is most preferably prepared with this promotion or the mode of not cutting down at least any Enzymic stability in these compositions.

System preferably used according to the invention comprises surfactant, and it is one or more non-ionic and/or anionic surfactant described here.

The non-ionic surface active agent use that the polyethylene of alkyl phenol, polypropylene and polybutene oxide condensation product (condensates) are used for as surfactant system of the present invention is suitable, preferably polyethylene oxide condensation product.These compounds comprise the condensation product of the alkyl phenol with alkyl group, and described alkyl group contains has an appointment 6 to about 14 carbon atoms, and preferred about 8 to about 14 carbon atoms, with straight or branched with state alkylene oxide (alkylene oxide) combination.In the preferred embodiment kind, the amount that described alkylene oxide exists is equivalent to every mole of alkyl phenol about 2 to about 25 moles of alkylene oxides, more preferably from about 3 to about 15 moles.

Can draw together Igepal by this type of ionic surfactant pack that commercial sources obtains ^TMCO-630 is sold by GAF Corporation; And TRITON ^TMX-45, X-114, X-100 and X-102 are by Rohm ﹠amp; Haas Company sells.These surfactants are commonly referred to as alkyl phenolic alkoxy compound (for example alkylphenol ethoxylate).

The described condensation product of primary and secondary aliphatic alcohol has about 1 to about 25 moles of ethylene oxide (ethyleneoxide), and it is adapted at being used as in the non-ionic surfactant system non-ionic surface active agent.The alkyl chain of described aliphatic alcohol can be straight or branch, uncle or secondary, and contains usually and have an appointment 8 to about 22 carbon atoms.Preferably have and contain the 8 alcohols condensation products to the alkyl group of about 20 carbon atoms of having an appointment, more preferably from about 10 to about 18 carbon atoms, and every mole of alcohols of this condensation product has about 2 to about 10 moles of ethylene oxide.Exist every mole of alcohols about 2 to about 7 moles of ethylene oxide in the described condensation product, and 2 to 5 moles of ethylene oxide most preferably.The example of this type of non-ionic surface active agent that can be obtained by commercial sources comprises TERGITOL ^TM15-S-9 (the C that has 9 moles of ethylene oxide ₁₁-C ₁₅Linear alcohols condensation product), TERGITOL ^TM24-L-6NMW (the have a very limited distribution C that has 6 moles of ethylene oxide of (narrow) of molecular mass ₁₂-C ₁₄Primary alconol class condensation product), sell by Union Carbide Corporation; The NEODOL that sells by Shell Chemical Company ^TM45-9 (the C that has 9 moles of ethylene oxide ₁₄-C ₁₅Linear alcohols condensation product), NEODOL ^TM23-3 (the C that has 3.0 moles of ethylene oxide ₁₂-C ₁₃Linear alcohols condensation product), NEODOL ^TM45-7 (the C that has 7 moles of ethylene oxide ₁₄-C ₁₅Linear alcohols condensation product), NEODOL ^TM45-5 (the C that has 5 moles of ethylene oxide ₁₄-C ₁₅Linear alcohols condensation product), by The Procter; The KYRO that Gamble Company sells ^TMEOB (the C that has 9 moles of ethylene oxide ₁₃-C ₁₅The alcohols condensation product) and the Genapol LA 050 (C that has 5 moles of ethylene oxide that sells by Hoechst ₁₂-C ₁₄Linear alcohols condensation product).The preferable range of HLB is 8-11 in these products, and 8-10 most preferably.

Equally as surfactant system non-ionic surface active agent US 4 usefully, 565, disclosed alkyl polysaccharide class (alkylpolysaccharides) in 647, it has and contains from about 6 hydrophobic groupings to about 30 carbon atoms, preferably from about 10 to about 16 carbon atoms and glycan class, for example the glycan glycosides, contain from about 1.3 hydrophilic radicals to about 10 saccharide unit, preferably from about 1.3 to about 3, most preferably from about 1.3 to about 2.7.Can use any recuding sugars that contains 5 or 6 carbon atoms, for example, glucose, galactolipin and galactosyl part (galactosyl moieties) can replace glucosyl group part (glucosylmoieties) (alternatively, hydrophobic grouping is attached to positions such as 2-, 3-, 4-, thereby glucose or galactolipin with respect to glucoside or galactoside are provided).The sugar internal key can be, for example, position of additional carbohydrate unit and before between 2-, 3-, 4-and/or the 6-position of carbohydrate unit.

Preferred alkyl glycan class has general formula

R ²O(C _nH _2nO) _t(glycosyl) _x

R wherein ²Be selected from down group: alkyl, alkyl phenyl, hydroxyalkyl, hydroxyalkyl phenyl (hydroxyalkylphenyl) and its mixture, wherein alkyl group contains from about 10 to about 18, preferably from about 12 to about 14 carbon atoms; N is 2 or 3, preferred 2; T is from 0 to about 10, preferred 0; And 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.Described glycosyl is preferably derived from glucose.In order to prepare these compounds, at first form described alcohols or alkyl polyethoxy (alkylpolyethoxy) alcohols, and react with glucose or source of glucose subsequently, to form glucoside (being incorporated into the 1-position).Described additional glycosyl unit can then be bonded between 2-, 3-, 4-and/or the 6-position of its 1-position and described previous glycosyl unit, best preferred 2-position.

The described oxirane condensation product that has hydrophobic group that is synthesized by condensation propylene two pure and mild propylene oxides is suitable as additional non-ionic surfactant system equally.The hydrophobic part of these compounds will preferably have from about 1500 to about 1800 molecular weight, and will show water-insoluble.Partly trend towards increasing on the whole the water-soluble of molecule at the additional Polyethoxyolefin of this hydrophobic part, and the liquid property that keeps described product is about 50% a point of condensation product overall weight up to polyoxy ethyl content, and it is equivalent to condensation up to about 40 moles oxirane.The example of this compounds comprises that some can be by the PLURONIC of commercial sources acquisition ^TMSurfactant is sold by BASF.

Be suitable as the non-ionic surface active agent of non-ionic surfactant system equally from propylene oxide and ethylenediamine (ethylenediamine) reaction products therefrom and oxirane condensation product.The hydrophobic part of these products is made up of the product of ethylenediamine and excessive oxidation propylene, and has from about 2500 to about 3000 molecular weight usually.This hydrophobic part and ethylene oxide condensation to condensation product contains from about 40% to about 80% polyoxyethylated scope by weight, and has from about 5000 to about 11000 molecular weight.The example of this class non-ionic surface active agent comprises the TETRONIC that can obtain by commercial sources of some ^TM, sell by BASF.

The tween that is preferably used as surfactant system is the polyethylene oxide condensation compound of alkylphenol, have from about 1 condensation product, alkyl polysaccharide class and its mixture to the primary and secondary aliphatic alcohol of about 25 moles of ethylene oxide.The C that most preferably has from 3 to 15 ethyoxyl groups ₈-C ₁₄Alkyl phenol ethoxylate and C with from 2 to 10 ethyoxyl groups ₈-C ₁₈Alcohols ethoxylate (preferred average C ₁₀), with and composition thereof.

Particularly preferred non-ionic surface active agent is poly-hydroxy fatty acid acid amides (polyhydroxy fattyacid amide) surfactant, and its general formula is

R wherein ¹Be H, or R ¹Be C _1-4Alkyl, 2-ethoxy, 2-hydroxypropyl or its mixture, R ²Be C _5-31Alkyl, Z are the poly-hydroxy alkylene (polyhydroxyhydrocarbyl) with linear hydrocarbyl chain that at least 3 hydroxyls directly link to each other with this chain, or its alkoxy derivative.Preferably, R ¹Be methyl, R ²Be straight chain C _11-15Alkyl or C _16-18Alkyl or alkenyl chain such as coconut alkane (coconut alkyl) or its mixture, and Z is from reducing sugar for example glucose, fructose, maltose or lactose-derived with reductive amination reaction.

Special preferred anionic surfactant comprises alkyl alkoxylated suifate (surfate) surfactant.The example comprises that general formula is RO (A) _mThe water soluble salt of SO3M or acid, wherein R is unsubstituted C ₁₀-C ₂₄Alkyl or have C ₁₀-C ₂₄The hydroxyalkyl group of alkyl component, preferred C ₁₂-C ₂₀Alkyl or hydroxyalkyl, more preferably C ₁₂-C ₁₈Alkyl or hydroxyalkyl, A is ethyoxyl or propoxyl group unit, m is greater than 0, usually between about 0.5 and about 6, more preferably between about 0.5 and about 3, and M is H or CATION, and described CATION for example can be, metal cation (for example, sodium, potassium, lithium, calcium, magnesium etc.), ammonium or replacement ammonium (substituted ammonium) CATION.Consider alkyl ethoxyquin sulfate and the alkyl third sulfur oxide hydrochlorate herein.The instantiation that replaces ammonium cation comprises methyl, dimethyl, trimethyl ammonium CATION and quaternary ammonium cation such as tetramethyl-ammonium and dimethyl piperdinium CATION, and those from alkylamine such as ethamine, diethylamine, triethylamine and its mixture derive etc.Exemplary surfactants is C ₁₂-C ₁₈Alkyl gathers ethoxyquin (1.0) sulfate (C ₁₂-C ₁₈E (1.0) M), C ₁₂-C ₁₈Alkyl gathers ethoxyquin (2.25) sulfate (C ₁₂-C ₁₈M), and C (2.25) ₁₂-C ₁₈Alkyl gathers ethoxyquin (3.0) sulfate (C ₁₂-C ₁₈And C E (3.0) M), ₁₂-C ₁₈Alkyl gathers ethoxyquin (4.0) sulfate (C ₁₂-C ₁₈E (4.0) M), wherein M is fit to be selected from sodium and potassium.

The anion surfactant that is fit to use is an alkyl sulfonate surfactants, comprises C ₈-C ₂₀Carboxylic acid (that is) ol ester, aliphatic acid, it is according to " The Journal of the American Oil ChemistsSociety ", and 52 (1975), pp.323-329 is with the SO3 sulfonation of gaseous state.The parent material that is fit to can comprise the material that is derived from tallow, palm wet goods natural fat.

Preferred alkyl sulfonated ester surfactant comprises the following alkyl sulfonate surfactants of general structure:

R wherein ³Be C ₈-C ₂₀Alkyl, preferred alkyl, or its combination, R ⁴Be C ₁-C ₆Alkyl, preferred alkyl, or its combination, and M is CATION, itself and alkyl ester sulfonate form water soluble salt.The salt-forming cation that is fit to comprises metal such as sodium, potassium and lithium, and that replace or unsubstituted ammonium cation, as monoethanolamine, diethanol amine and triethanolamine.Preferred R ³Be C ₁₀-C ₁₆Alkyl, and R ⁴Be methyl, ethyl or isopropyl.Be preferably methyl ester sulfonate especially, wherein R ³Be C ₁₀-C ₁₆Alkyl.

Other anion surfactants that are fit to comprise alkyl sulfate surfactant, and it is ROSO for general formula ₃The water soluble salt of M or acid, the wherein preferred C of R _10-24Alkyl preferably has C ₁₀-C ₂₀The alkyl of alkyl component or hydroxyalkyl, more preferably C ₁₂-C ₁₈Alkyl or hydroxyalkyl, and M is H or CATION, for example, the alkalinous metal CATION (for example, sodium, potassium, lithium), ammonium or replace ammonium (for example, methyl-, dimethyl-and trimethyl ammonium CATION and quaternary ammonium cation such as tetramethyl-ammonium and dimethyl piperdinium CATION and the tetramethyl-ammonium of deriving from alkylamine such as ethamine, diethylamine, triethylamine and its mixture etc.).Usually, C ₁₂-C ₁₆The preferably lower wash temperature of alkyl chain (for example, about below 50 ℃) and for the preferred C of higher wash temperature (for example about more than 50 ℃) ₁₆-C ₁₈Alkyl chain.

Other anion surfactants that are used for washing (detersive) purpose comprise soap (soap) salt (comprise, for example, sodium, potassium, ammonium and substituted ammonium salt be for example single-two-and triethanolamine salt), C ₈-C ₂₂Uncle or secondary paraffin sulfonate, C ₈-C ₂₄Olefin(e) sulfonate, by the sulfonation polycarboxylic acids of sulfonation alkaline earth metal citrate thermal decomposition product preparation, for example,, C is described in 082,179 as BP specification NO.1 ₈-C ₂₄Alkyl polyglycol ether sulfate (comprise and reach 10 moles of ethylene oxide); Alkyl glycerol sulfonate (alkylglycerol sulfonates); the fatty acyl glycerol sulfonate; fat oil base glycerol sulfate (fatty oleylglycerol sulfates); alkylphenol ethene oxidation ether sulfate (alkyl phenol ethylene oxide ethersulfates); paraffin sulfonate (paraffin surfonate); alkylphosphonate; isethionate such as acyl-hydroxyethyl sulfonate; the N-acyl taurate; alkyl succinic acid (salt/ester) (alkyl succinamates) and sulfosuccinic acid (ester/salt) are (sulfosuccinates); sulfosuccinic acid monoether (particularly saturated and undersaturated C ₁₂-C ₁₈Monoesters) and sulfosuccinic acid diesters (particularly saturated and undersaturated C ₆-C ₁₂Diester), acyl sarcosinates/ester, alkyl polysaccharide sulfate/ester such as alkyl poly glucoside sulfate (nonionic non-sulfuric acid compound hereinafter described), chain primary alkyl sulfate and alkyl polyethoxy carboxylate such as those general formulas are RO (CH ₂CH ₂O) _k-CH ₂COO-M+'s, wherein R is C ₈-C ₂₂Alkyl, k are from 1 to 10 integer, and M is for forming the CATION of soluble-salt.Resin acid and hydrogenated resin acid are suitable equally, for example rosin, Foral, and be present in or derived from resin acid and the hydrogenated resin acid of tall oil (tall oil).

Special preferred alkyl benzene sulfonate/ester.Be preferably linearity (straight chain) alkylbenzenesulfonate/ester (LAS) especially, wherein alkyl group preferably comprises from 10 to 18 carbon atoms.

Further example is described in " Surface Active Agents and Detergents " (Vol.I and II bySchwartz, Perrry and Berch) to some extent.At US 3,929, in 678 (Column 23, line 58through Column 29, line 23, this is the part of list of references) multiple such surfactant is disclosed briefly also.

In the time of in being included in those compositions of the present invention, comprise usually by weight from about 1% to about 40%, preferably such anion surfactant of from about 3% to about 20%.

Composition of the present invention can also comprise cationic, both sexes, zwitterionic and the semi-polarity surfactant, and is different from the non-ionic and/or anion surfactant that those this paper have described.

Be applicable to that CATION spatter property surfactant in the present composition is to have those of long chain hydrocarbon groups group.The example of this cationic surfactant comprises ammonium (ammonium) surfactant, and for example alkyl trimethyl ammonium halide and those have the surfactant of following general formula:

[R ²(OR ³) _y][R ⁴(OR ³) _y] ₂R ⁵N+X-

R wherein ²For having about 8 alkyl or alkylbenzene methyl groups in the alkyl chain to about 18 carbon atoms; Each R ³Be selected from down group :-CH ₂CH ₂-, CH ₂CH (CH ₃)-,-CH ₂CH (CH ₂OH)-,-CH ₂CH ₂CH ₂-and its mixture; Each R ⁴Be selected from down group: C ₁-C ₄Alkyl, C ₁-C ₄Hydroxyalkyl, by connecting two R ⁴The benzyl ring structure that group forms ,-CH ₂CHOHCHOHCOR ⁶CHOHCH ₂OH, wherein R ⁶For having hexose or hexose polymer arbitrarily less than about 1000 molecular weight, and hydrogen, when y is not 0; R ⁵With R ⁴Identical or be alkyl chain, wherein the total number of carbon atoms or R ²Add R ⁵No more than about 18; Each y is from 0 to about 10, and the summation of y value is from 0 to about 15; And X is any compatibility (compatible) anion.

Special preferred cationic surfactant is the water-soluble quaternary ammonium compound that is used for this composition, and it has general formula:

R ₁R ₂R ₃R ₄N ⁺X ^- (i)

R wherein ₁Alkyl for C8-C16; R ₂, R ₃And R ₄Be independently C ₁-C ₄Alkyl, C ₁-C ₄Hydroxyalkyl, benzyl and-(C ₂H ₄₀) _xH, wherein x has from 2 to 5 numerical value; And X is an anion.R ₂, R ₃Or R ₄In should no more than one be benzyl.

Preferred R ₁Alkyl chain length is C ₁₂-C ₁₅, when alkyl group was the chain length mixture, this mixture was from coconut or palm nuclear adipose-derived especially, or was synthesized and synthetic deriving by alkene accumulation (olefin build up) or OXO alcohols.

R ₂R ₃And R ₄Preferred group be that methyl and hydroxyethyl groups and anion X can be selected from halide, dimethyl suflfate, acetate and phosphate ion.

The suitable example of general formula (i) quaternary ammonium compound is as used herein:

Coconut trimethyl ammonium chloride or bromide;

Coconut methyl dihydroxy ethyl ammonium chloride or bromide;

The decyl triethyl ammonium chloride;

Decyl dimethyl hydroxyethyl ammonium chloride or bromide;

C _12-15Dimethyl hydroxyethyl ammonium chloride or bromide;

Coconut dimethyl hydroxyethyl ammonium chloride or bromide;

Myristyl trimethyl ammonium Methylsulfate;

Lauryl dimethyl benzyl ammonium chloride or bromide;

Lauryl dimethyl (vinyl) ₄Ammonium chloride or bromide;

(compound of general formula (i) is R wherein for cholinester ₁For

Alkyl and R ₂R ₃R ₄Be methyl).

Dialkylimidazolium quinoline [compound of general formula (i)]

Other useful cationic surfactants of this paper are equally at US 4,228,044 and EP 000 224 in describe to some extent.

When being included in this moment, composition of the present invention comprises by weight usually from 0.2% to about 25%, preferably from about 1% to about 8% this cationic surfactant.

Amphoteric surfactant is equally applicable to composition of the present invention.These surfactants can be described as the aliphatic derivatives of secondary amine or tertiary amine widely, or wherein fatty root can be the heterocyclic secondary of straight or branched or the aliphatic derivatives of tertiary amine.Fat one of substituting group contains at least 8 carbon atoms, and usually from about 8 to about 18 carbon atoms, and at least one contains anionic water solubilization (water-solubilizing) group, for example, and carboxyl, sulfonate radical, sulfate radical.Examples of amphoteric surfactants is referring to US 3,929,678 (column 19, lines 18-35).

When being included in this moment, composition of the present invention comprises by weight usually from 0.2% to about 15%, preferably from about 1% to about 10% amphoteric surfactant.

Zwitterionic surfactant also is applicable to composition of the present invention.These surfactants can be described as the derivative of secondary amine or tertiary amine widely, the derivative of heterocyclic secondary or tertiary amine, or the derivative of quaternary amine, season or tertiary sulfonium compound.The example of zwitterionic compound is referring to US3,929,678 (column 19, line 38 through column 22, line 48).

When being included in this moment, composition of the present invention comprises by weight usually from 0.2% to about 15%, preferably from about 1% to about 10% zwitterionic surfactant.

Semi-polar nonionic surfactants is the Special Category of non-ionic surface active agent, it comprises water-soluble amine oxides, water soluble oxidized phosphine (phosphineoxide) and water-soluble sulfoxide, water-soluble amine oxides contain one from about 10 to the moieties of about 18 carbon atoms and 2 parts that are selected from down group: contain from about 1 alkyl group and hydroxyalkyl group to about 3 carbon atoms; The water soluble oxidized phosphine contain one from about 10 to the moieties of about 18 carbon atoms and 2 parts that are selected from down group: contain from about 1 alkyl group and hydroxyalkyl group to about 3 carbon atoms; Water-soluble sulfoxide contain one from about 10 to the moieties of about 18 carbon atoms and parts that are selected from down group: from about 1 alkyl and hydroxyalkyl part to about 3 carbon atoms.

Semi-polarity nonionic detergent (detergent) surfactant comprises the amine oxide surfactant with following general formula:

R wherein ³For containing from about 8 alkyl, hydroxyalkyl or alkyl phenyl group or its mixture to about 22 carbon atoms; R ⁴For containing from about 2 alkylidene or hydroxyl alkylidene group or its mixtures to about 3 carbon atoms; X is from 0 to about 3; And each R ⁵For containing from about 1 to the alkyl or the hydroxyalkyl group of about 3 carbon atoms or contain from about 1 poly(ethylene oxide) group to about 3 ethylene oxide groups.Described R ⁵Group can interconnect, for example, and by oxygen or nitrogen-atoms, to form ring structure.

These amine oxide surfactants comprise C especially ₁₀-C ₁₈Alkyl dimethyl amine oxide and C ₈-C ₁₂Alkoxyethyl dihydroxy ethyl amine oxide.

When being included in this moment, composition of the present invention comprises by weight usually from 0.2% to about 15%, preferably this type of semi-polar nonionic surfactants of from about 1% to about 10%.

Enzyme

α-Dian Fenmei

The α-Dian Fenmei of using in the method for the invention can be any α-Dian Fenmei, preferred source from bacterium or fungi.In preferred embodiments, described α-Dian Fenmei is an acid alpha-amylase, for example among the WO2005/003311 disclosed α-Dian Fenmei or hybridization α-Dian Fenmei, it is hereby incorporated by.

In preferred embodiments, described α-Dian Fenmei comprises WO 2005/003311 defined carbohydrate binding modules (CBM), preferred WO 2005/003311 defined 20 CBM of family.

The special CBMs that considers comprises and is selected from down group: the disclosed valley aspergillus of SEQ ID NO:2 (Aspergillus kawachii); The hot bacillus of the disclosed Huang of SEQ ID NO:5 (Bacillusflavothermus); The disclosed bacillus kind of SEQ ID NO:6; The disclosed Alcaliphilic Bacillus of SEQ ID NO:7; The disclosed Hormoconis resinae of SEQ ID NO:8; The disclosed Lentinula edodes of SEQ ID NO:9; The disclosed Neurospora crassa of SEQ ID NO:10 (Neurosporacrassa); The disclosed Talaromyces byssochlamydiodes of SEQ ID NO:11; The disclosed Geosmithia cylindrospora of SEQ ID NO:12; The disclosed Scorias spodiosa of SEQ ID NO:13; The disclosed Eupenicillium ludwigii of SEQID NO:14; The disclosed aspergillus japonicus of SEQ ID NO:15 (Aspergillus japonicus); The disclosed Penicillium cf.miczynskii of SEQ ID NO:16; The disclosed Mz1 mould of SEQID NO:17 kind (Mz1 Penicillium sp.); The disclosed Thysanospora sp. of SEQ ID NO:18; The disclosed Humicola grisea of SEQ ID NO:19 var.thermoidea; The disclosed aspergillus niger of SEQID NO:20; Or the disclosed Althea rolfsii of SEQ ID NO:21.

Fungiα -amylase

In preferred embodiments, fungal alpha-amylase is derived from yeast or filamentous fungi.Fungal alpha-amylase is an acid alpha-amylase in preferred embodiments.

Preferred α-Dian Fenmei comprises, for example, α-Dian Fenmei from the aspergillus bacterium acquisition, particularly from aspergillus niger, Aspergillus oryzae (A.oryzae) and aspergillus awamori (A.awamori), valley aspergillus, the for example disclosed acid alpha-amylase of SWISSPROT P56271, or description (embodiment 3) in more detail in WO 89/01969.Ripe acid alpha-amylase has the amino acid sequence shown in the 22-511 of SEQ ID NO:4, by the dna sequence encoding shown in the SEQ ID NO:3, or the amino acid sequence shown in the SEQ ID NO:38.Amino acid sequence shown in same preferred and SEQ ID NOS:4 or 38 has the α-Dian Fenmei sequence greater than 50% homogeneity respectively, for example greater than 60%, greater than 70%, greater than 80% or greater than 90%, greater than 95%, greater than 96%, greater than 97%, greater than 98%, or even greater than 99% homogeneity.

In a further preferred embodiment, described α-Dian Fenmei sequence is to be derived from Aspergillus oryzae acid starch enzyme.Amino acid sequence has the homogeneity greater than 50% shown in more preferably described α-Dian Fenmei sequence and the SEQ ID NOS:39, for example greater than 60%, greater than 70%, greater than 80% or greater than 90%, greater than 95%, greater than 96%, greater than 97%, greater than 98%, or even greater than 99% homogeneity.

In one embodiment, described α-Dian Fenmei is a disclosed valley aspergillus α-Dian Fenmei in SEQ ID NO:37, and it comprises that in the wild type form sugar shown in the SEQ ID NO:2 is in conjunction with territory (CBD).

In preferred embodiments, described α-Dian Fenmei is to have respectively with amino acid sequence shown in the SEQ ID NOS:43,44,46,47 homogeneity greater than 50% is arranged, for example greater than 60%, greater than 70%, greater than 80% or greater than 90%, greater than 95%, greater than 96%, greater than 97%, greater than 98%, or even greater than 99% homogeneity.

It is 1-3 that described α-Dian Fenmei may reside in concentration, the fabric of 000AFAU/kg, preferred 10-1, the fabric of 000AFAU/kg, in the fabric of special 100-500AFAU/kg or 1-3, the Treatment Solution of 000AFAU/L, preferred 10-1, the Treatment Solution of 000AFAU/L is in the Treatment Solution of special 100-500AFAU/L.

Bacterial

In embodiments, α-Dian Fenmei is a bacterial origin.In preferred embodiments, described bacterial is an acid alpha-amylase.

Described bacterial is preferably derived from Bacillus strain, bacillus licheniformis (Bacillus licheniformis) for example, bacillus amyloliquefaciens (Bacillus amyloliquefaciens), bacillus stearothermophilus (Bacillus stearothermophilus), bacillus subtilis (Bacillus subtilis) or other bacillus kinds NCIB 12289, NCIB 12512 (WO 95/26397), NCIB 12513 (WO95/26397), DSM 9375 (WO 95/26397), DSMZ 12648 (WO 00/60060), DSMZ12649 (WO 00/60060), KSM AP1378 (WO 97/00324), (EP 1 for KSM K36 or KSM K38,022,334).Bacillus kind α-Dian Fenmei among the preferred SEQ ID NOS.1 and 2 that is disclosed in WO 95/26393 respectively, be disclosed among the SEQ ID NO:2 of WO 00/60060 the AA560 α-Dian Fenmei (promptly, the SEO ID NO:40 of this paper), and by Tsukamoto etc. Biochemical and Biophysical Research Communications, 151 (1988), the disclosed #707 α-Dian Fenmei of pp.25-31.

In embodiments of the invention, described bacterial is SP722 α-Dian Fenmei or the described AA560 α-Dian Fenmei (the SEQ IDNO:40 of this paper) that is disclosed among the SEQ ID NO:2 of WO 95/26397.

In preferred embodiments, parent's α-Dian Fenmei have one or more down column positions or with the disappearance of following column position corresponding position: D183 and G184, preferred wherein said alpha-amylase variants further are included in the replacement (numbering with SEQ ID NO:40) of position N195F or its corresponding position.

In a further preferred embodiment, parent's α-Dian Fenmei has one or more following disappearances/replacement or with following disappearance/replacement is pairing: Delta (R81-G182), Delta (D183-G184), Delta (D183-G184)+N195F, R181Q+N445Q+K446N, Delta (D183-G184)+R181Q, Delta (D183-G184) and one or more following or corresponding replacement: R188K, N195F, R320K, R458K, wherein said especially variant has following sudden change: Δ (D183+G184)+R118K+N195F+R320K+R458K (utilizing SEQ ID NO:40 numbering).

In a further preferred embodiment, described α-Dian Fenmei is the AA560 α-Dian Fenmei that SEQ ID NO:40 shows, further comprise one or more following replacements: M9L, M202L, V214T, M323T, M382Y, E345R or described A560 α-Dian Fenmei have following whole replacement: M9L, M202L, V214T, M323T, M382Y or M9L, M202L, V214T, M323T and E345R.

α-Dian Fenmei product that can be obtained by commerce or the product that comprises α-Dian Fenmei comprise the product of selling with following trade name: NATALASE ^TM, STAINZYME ^TM(Novozymes A/S), Bioamylase-D (G), BIOAMYLASE ^TML (Biocon India Ltd.), KEMZYME ^TMAT 9000 (BiozymGes.m.b.H, Austria), PURASTAR ^TMST, PURASTAR ^TMHPAm1, PURAFECT ^TMOxAm, RAPIDASE ^TMTEC (Genencor Int.Inc, the U.S.), KAM (KAO, Japan)

Described α-Dian Fenmei may reside in concentration and is the Treatment Solution from about 0.05-150KNU/L, the Treatment Solution of preferred 1-100KNU/L, the Treatment Solution of special 2-20KNU/L or the fabric of 0.05-150KNU/Kg, the fabric of preferred 1-100KNU/Kg, the fabric of special 2-20KNU/kg

Heterozyme

In preferred embodiments, described α-Dian Fenmei can be for comprising the α-Dian Fenmei of sugar in conjunction with territory (CBD).This α-Dian Fenmei that has a CBD can be wild-type enzyme (referring to for example, top valley aspergillus) or the heterozyme (fusion) that will be further described below.The wild-type enzyme of heterozyme or the genetic modification that herein relates to comprises following kind, and described kind comprises and comprises the link to each other amino acid sequence (EC 3.2.1.1) of α-Dian Fenmei of (covalent bond) of the amino acid sequence of sugar in conjunction with territory (CBD).

The heterozyme that contains CBD, and preparation and their detailed description of purifying, be known in the art [referring to, for example WO 90/00609, WO 94/24158 and WO 95/16782 see Greenwood etc. again Biotechnology and Bioengineering 44(1994) DD.1295-1305].They can be, for example, by DNA construct is transformed in the host cell, this DNA construct comprise at least with the dna sequence dna of coding purpose enzyme by or the coding sugar that do not link to each other by joint in conjunction with the dna fragmentation in territory, cultivate transformed host cells again to express described fusion.Gained recombinant products (heterozyme)---is often referred to " fusion " in the art---and can be described by following general formula:

A-CBD-MR-X

In the general formula of back, A-CBD comprises at least N end or the C stub area of sugar in conjunction with the amino acid sequence in territory itself.MR is middle zone (described " joint "), and the serve as reasons amino acid residue sequence of polypeptide of dna sequence encoding of the enzyme (or other albumen) that coding will be connected with CBD of X.

Can there be (such A-CBD is CBD itself, that is, do not comprise those and constitute CBD amino acid residue in addition) in the A part, maybe can be the sequence (end as CBD itself extends the performance function) of one or more amino acid residues.Described joint (MR) can be key (bond) or comprise from the about 2 short linking groups to about 100 carbon atoms, special from about 2 to about 40 carbon atom.But MR is preferably from about 2 sequences to about 100 amino acid residues, more preferably from 2 to 40 amino acid residues, for example from 2 to the fifteen amino acid residue.

Described X part can be the N end or the C stub area of whole heterozymes.

By above apparent, it is terminal or wherein to be positioned at C end, the N of heterozyme at CBD described in the heterozyme of the type of being discussed.

Joint sequence

Described joint sequence can be any suitable joint sequence.In preferred embodiments, described joint sequence is from Athelia rolfsii glucoamylase, the aspergillus niger glucoamylase, valley aspergillus α-Dian Fenmei is derived, and for example is selected from down the joint sequence of group: aspergillus niger glucoamylase joint: TGGTTTTATPTGSGSVTSTSKTTATASKTSTSTSSTSA (SEQ ID NO:22), valley aspergillus α-Dian Fenmei joint: T T T T T T A A A T S T S K A T T S S S S S S A A A T T SS S (SEQ ID NO:23), Athelia rolfsii glucoamylase joint: G A T S P G G S S G S (SEQ ID NO:24), and PEPT joint: P E P T P E P T (SEQ ID NO:25).In a further preferred embodiment, described heterozyme has the catenation sequence different with following sequence: the amino acid sequence shown in SEQ IDNO:22, SEQ ID NO:23, SEQ ID NO:24 or the SEQ ID NO:25, no more than 10 positions of difference, no more than 9 positions, no more than 8 positions, no more than position, no more than 6 positions, no more than 5 positions, no more than 4 positions, no more than 3 positions, no more than 2 positions, or not even more than 1 position.

Sugar is in conjunction with the territory

Sugar is in conjunction with territory (CBD), or usually indication is, carbohydrate binding modules (CBM) be a polypeptid acid sequence, it preferentially combines with polysaccharide or oligosaccharides (carbohydrate), and is frequent---but unnecessary eliminating combines with their water-insoluble (comprising crystallization) form.

The CBD that derives from starch degrading enzyme is commonly called starch binding domain (SBD) or starch binding modules (SBM).SBD is for appearing at CBD in the specific amylolytic enzyme, for example at specific glucoamylase, or appears at enzyme for example in the cyclodextrin glycosyl transferase, or appears in the α-Dian Fenmei.Similarly, the CBD of other subclass will comprise, for example cellulose binding domain (from the CBD of decomposition of cellulose enzyme), chitin binding domain (appearing at the CBD in the chitinase usually), xylan is in conjunction with territory (being present in the CBD in the zytase usually), and mannosan is in conjunction with territory (appearing at the CBD in the mannase (mannanases) usually).

CBD is found to be by the regional large-scale polypeptide of forming of two or more polypeptid acid sequences or the major part of protein, special in the enzyme (hydrolase) of the generation hydrolysis that comprises catalytic domain usually, described catalytic domain contains the active site that is useful on substrate hydrolysis and is used to be attached to the sugar of the carbohydrate substrate of being discussed in conjunction with territory (CBD).Such enzyme can comprise catalytic domain and one, two or three CBD more than, and randomly further comprises one or more polypeptid acid sequence zones that CBD is connected with catalytic domain, and the zone of latter type is represented as " joint " usually.The example that produces the enzyme of hydrolysis comprises CBD, and---wherein a part is mentioned hereinbefore---is cellulase, zytase, mannase (mannanases), arabinofuranosidase, acetylesterase and chitinase.CBD is present in the algae equally, for example, with the polysaccharide of non-hydrolysis-type in conjunction with the albumen form in red algae Porphyra purpurea.

In the protein/polypeptide of CBD occurring (enzyme for example is generally the enzyme that produces hydrolysis), CBD can be positioned at N or C is terminal or in the position of inside.

The part that constitutes the polypeptide of CBD itself or protein (for example producing the enzyme of hydrolysis) is usually by more than about 30 and be less than about 250 amino acid residue and form.

Described " carbohydrate binding modules of family 20 " or CBM-20 module are defined as about 100 amino acid whose sequences in the context of the present invention, and (1997) carbohydrate binding modules of disclosed polypeptide in Biotechnol.Lett.19:1027-1031 Fig. 1 such as itself and Joergensen has 45% homology at least.CBM comprises 102 last amino acid of described polypeptide, i.e. 683 the subsequence (subsequence) from amino acid 582 to amino acid.The numbering of the glycoside hydrolysis enzyme family of using in this open source literature (Glycoside HydrolaseFamilies) is followed the theory in following: Coutinho, P.M.﹠amp; Henrissat, B. (1999) CAZy-Carbohydrate-Active Enzymes server are at URL: Http:// afmb.cnrs-mrs.fr/～cazy/CAZY/index.htmlOr Coutinho, P.M.﹠amp; Henrissat, B.1999; The modular structure of cellulases and other carbohydrate-active enzymes:an integrated database approach.In ＂ Genetics, Biochemistry and Ecology ofCellulose Degradation ＂, K.Ohmiya, K.Hayashi, K.Sakka, Y.Kobayashi, S.Karita and T.Kimura eds., Uni Publishers Co., Tokyo, pp.15-23 and Bourne, Y.﹠amp; Henrissat, B.2001; Glycoside hydrolases and glycosyltransferases:families andfunctional modules, Current Opinion in Structural Biology 11:593-600.

The example that comprises the enzyme of the CBD that is applicable to the context of the invention is α-Dian Fenmei, product maltose (maltogenic) α-Dian Fenmei, cellulase, zytase, mannase (mannanases), arabinofuranosidase, acetylesterase and chitinase.The purpose CBD relevant with the present invention further comprises from glucoamylase (EC 3.2.1.3) or the CBD that derives from CGTases (EC 2.4.1.19).

The CBD that is derived from fungi, bacterium or plant source will generally be applicable to context of the present invention.The CBD in preferred fungi source more preferably plants from aspergillus bacterium, bacillus kind, Klebsiella kind or rhizopus (Rhizopus).Relevant therewith, be applicable to that the technology of separating related gene is known in the art.

The CBD of the preferred carbohydrate binding modules of the present invention family 20.The CBD that is suitable for carbohydrate binding modules of the present invention family 20 can be derived from aspergillus awamori (SWISSPROT Q12537), valley aspergillus (SWISSPROT P23176), aspergillus niger (SWISSPROT P04064), the glucoamylase of Aspergillus oryzae (SWISSPROTP36914), valley aspergillus (EMBL:#AB008370), the α-Dian Fenmei of aspergillus nidulans (Aspergillusnidulans) (NCBI AAF17100.1), the beta amylase of Bacillus cercus (Bacillus cereus) (SWISSPROT P36924), or the CGTases of Bacillus circulans (Bacillus circulans) (SWISSPROT P43379).Preferably from the CBD of valley aspergillus (EMBL:#AB008370) α-Dian Fenmei, and have at least 50%, 60%, 70% with the CBD of valley aspergillus (EMBL:#AB008370) α-Dian Fenmei, 80% or even at least 90%, 95%, 96%, 97%, 98%, or the CBD of 99% homogeneity, promptly the amino acid sequence with SEQ ID NO:2 has at least 50%, 60%, 70%, 80% or even at least 90%, 95%, 96%, 97%, 98%, or the CBD of 99% homogeneity.The same preferred sugar of the present invention is in conjunction with the CBD of territory family 20, it has amino acid sequence shown in SEQ ID NO:5, SEQ ID NO:6 and the SEQ IDNO:7, and is disclosed in the amino acid sequence shown in the PCT application no.PCT/DK2004/000456 (or Danish Patent Application PA 200300949) with SEQ ID NO:1, SEQ ID NO:2 and SEQ IDNO:3 respectively.Further preferred CBD comprises the CBD from following listed glucoamylase: from Hormoconis sp. Hormoconis resinae. (Syn.Creosote mushroom or Amorphotheca resinae) for example, as from SWISSPROT:Q03045The CBD of (SEQ ID NO:8); From Lentinula sp. Lentinula edodes (shiitake mushroom) for example, as CBD from SPTREMBL:Q9P4C5 (SEQ ID NO:9); From Neurospora kind (Neurospora sp.) Neurospora crassa (Neurospora crassa) for example, as from SWISSPROT:P14804The CBD of (SEQ IDNO:10); From ankle joint bacterial classification (Talaromyces sp.) silk clothes mustiness ankle joint bacterium for example, as CBD from NN005220 (SEQ ID NO:11); From Geosmithia sp. Geosmithiacylindrospora for example, as CBD from NN48286 (SEQ ID NO:12); From Scorias sp. Scorias spongiosa for example, as CBD from NN007096 (SEQ ID NO:13); From penicillium kind (Eupenicillium sp.) Eupenicilliumludwigii for example, as CBD from NN005968 (SEQ ID NO:14); From aspergillus kind aspergillus japonicus for example, as CBD from NN001136 (SEQID NO:15); From mould kind (Penicillium sp.) Penicillium cf.miczynskii for example, as CBD from NN48691 (SEQ ID NO:16); From Mz1 mould kind (Mz1Penicillium sp.), as CBD from NN48690 (SEQ ID NO:17); From Thysanophora sp., as CBD from NN48711 (SEQ ID NO:18); And from humicola lanuginosa kind (Humicola sp.) Humicola grisea var.thermoidea for example, as CBD from SPTREMBL:Q12623 (SEQ ID NO:19).Most preferred CBD comprises from the aspergillus kind glucoamylase of aspergillus niger for example, as SEQ ID NO:20, and the Athelia sp. glucoamylase of Atheliarolfsii for example, as the CBD of SEQ ID NO:21.Same preferred any and above-mentioned CBD amino acid sequence has at least 50%, 60%, 70%, 80% or even the CBD of at least 90%, 95%, 96%, 97%, 98% or 99% homogeneity according to the present invention.

The CBD of further suitable carbohydrate binding modules family 20 can be at URL: Http:// afmb.cnrs-mrs.fr/～cazy/CAZY/index.htmlOn find.

In case the nucleotide sequence of coding substrate calmodulin binding domain CaM identified, no matter be form with cDNA or chromosomal DNA, can handle this nucleotide sequence with several different methods with that, the dna sequence dna of the enzyme that itself and coding paid close attention to is merged.Connect the dna fragmentation of the described sugared binding amino acid sequence of coding and the DNA of the enzyme that coding is paid close attention to afterwards, have or do not have joint.Can handle the DNA of gained connection to obtain expression with several different methods afterwards.

In embodiments, be included in α-Dian Fenmei in the crossbred be before the described α-Dian Fenmei of " α-Dian Fenmei " part.In preferred embodiments, described α-Dian Fenmei is an originated from fungus.In a more preferred embodiment, described α-Dian Fenmei is an acid alpha-amylase.

In preferred embodiments, described sugar is originated from fungus in conjunction with territory and/or joint sequence.Described sugar can be derived from α-Dian Fenmei in conjunction with the territory, but can for example, have the enzyme of glucoamylase activity from protein derived equally.

In embodiments, described α-Dian Fenmei is derived from aspergillus or Athelia bacterial strain.In embodiments, described α-Dian Fenmei is derived from Aspergillus oryzae or aspergillus niger.In a particularly embodiment, described α-Dian Fenmei is the disclosed Aspergillus oryzae acid alpha-amylase of SEQID NO:39.In a particularly embodiment, described joint sequence can be derived from the aspergillus bacterial strain, for example valley aspergillus α-Dian Fenmei (SEQ ID NO:23) or Athelia rolfsii glucoamylase (SEQ ID NO:24).In embodiments, described CBD is derived from aspergillus or Athelia bacterial strain.In a particularly embodiment, described CBD is the Athelia rolfsii glucoamylase shown in valley aspergillus α-Dian Fenmei shown in the SEQ ID NO:1 or the SEQ ID NO:21.

In the embodiment preferred, described heterozyme comprises the α-Dian Fenmei sequence of deriving from aspergillus niger acid alpha-amylase catalytic domain, the sequence of this catalytic domain is shown in SEQ ID NO:38, and/or the joint sequence of deriving of the Athelia rolfsii glucoamylase from shown in valley aspergillus α-Dian Fenmei or the SEQ ID NO:24 shown in the SEQ ID NO:23, and/or described CBD is from the valley aspergillus α-Dian Fenmei shown in the SEQ ID NO:2, aspergillus niger glucoamylase shown in Athelia rolfsii glucoamylase shown in the SEQ ID NO:21 or the SEQ ID NO:22 is derived.

In preferred embodiments, described heterozyme comprises it and has the CBD that sequence is shown in the valley aspergillus α-Dian Fenmei shown in aspergillus niger acid alpha-amylase catalytic domain, the valley aspergillus α-Dian Fenmei joint shown in the SEQ ID NO:23 and the SEQ ID NO:2 of SEQ ID NO:38.

In a particularly embodiment, described heterozyme is the maturing part of amino acid sequence shown in the following sequence: SEQ ID NO:28 (aspergillus niger acid alpha-amylase catalytic domain-valley aspergillus α-Dian Fenmei joint-aspergillus niger glucoamylase CBD), SEQ ID NO:30 (aspergillus niger acid alpha-amylase catalytic domain- Valley aspergillusα-Dian Fenmei joint-Athelia rolfsii glucoamylase CBD), or SEQ ID NO:32 (Aspergillus oryzae acid alpha-amylase catalytic domain-valley aspergillus α-Dian Fenmei joint-valley aspergillus α-Dian Fenmei CBD), or SEQ ID NO:34 (aspergillus niger acid alpha-amylase catalytic domain-Athelia rolfsii glucoamylase joint-Athelia rolfsii glucoamylase CBD), or SEQ ID NO:36 (Aspergillus oryzae acid alpha-amylase catalytic domain-Athelia rolfsii glucoamylase joint-Athelia rolfsii glucoamylase CBD), or described crossbred is made up of aspergillus niger acid alpha-amylase catalytic domain (being respectively SEQ ID NOS:4 or 38)-valley aspergillus glucoamylase joint (SEQ IDNO:23)-valley aspergillus glucoamylase CBD (SEQ ID NO:2), or heterozyme, it has with any aforementioned amino acid sequence and has at least 50%, 60%, 70%, 80% or even at least 90%, 95%, 96%, 97%, the amino acid sequence of 98% or 99% homogeneity.

In a further preferred embodiment, described heterozyme has the amino acid sequence that is different from amino acid sequence shown in the following sequence: SEQ ID NO:28 (aspergillus niger acid alpha-amylase catalytic domain-valley aspergillus α-Dian Fenmei joint-aspergillus niger glucose starch CBD), SEQ ID NO:30 (aspergillus niger acid alpha-amylase catalytic domain-valley aspergillus α-Dian Fenmei joint-Athelia rolfsii glucoamylase CBD), SEQ ID NO:32 (Aspergillus oryzae acid alpha-amylase catalytic domain-valley aspergillus α-Dian Fenmei joint-valley aspergillus α-Dian Fenmei CBD), SEQID NO:34 (aspergillus niger acid alpha-amylase catalytic domain-Athelia rolfsii glucoamylase joint-Atheliarolfsii glucoamylase CBD) or SEQ ID NO:36 (Aspergillus oryzae acid alpha-amylase catalytic domain-Atheliarolfsii glucoamylase joint-Athelia rolfsii glucoamylase CBD), or described crossbred is by no more than 10 sites, no more than 9 sites, no more than 8 sites, no more than 7 sites, no more than 6 sites, no more than 5 sites, no more than 4 sites, no more than 3 sites, no more than 2 sites, or even do not form more than the aspergillus niger acid alpha-amylase catalytic domain in 1 site (SEQ ID NOS:4 or 38 respectively)-valley aspergillus glucoamylase joint (SEQ ID NO:23)-valley aspergillus glucoamylase CBD (SEQ ID NO:2).

Preferred described heterozyme comprises and has and any SEQ of being shown in ID NO:5, SEQ ID NO:6, SEQ IDNO:7, SEQ ID NO:8, SEQ ID NO:9, SEQ ID NO:10, SEQ ID NO:11, SEQ IDNO:12, SEQ ID NO:13, SEQ ID NO:14, SEQ ID NO:15, SEQ ID NO:16, SEQID NO:17, SEQ ID NO:18, SEQ ID NO:19, the amino acid sequence of SEQ ID NO:20 or SEQ ID NO:21 has at least 50%, 60%, 70%, 80% or even at least 90%, 95%, 96%, 97%, the CBD sequence of 98% or 99% homogeneity.More preferably described heterozyme comprises the CBD sequence with the amino acid sequence that is shown in SEQ IDNO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:9, SEQ IDNO:10, SEQ ID NO:11, SEQ ID NO:12, SEQ ID NO:13, SEQ ID NO:14, SEQID NO:15, SEQ ID NO:16, SEQ ID NO:17, SEQ ID NO:18, SEQ ID NO:19, SEQ ID NO:20 or SEQ ID NO:21.Also in a further preferred embodiment, described CBD sequence is at no more than 10 amino acid positions, no more than 9 positions, no more than 8 positions, no more than 7 positions, no more than 6 positions, no more than 5 positions, no more than 4 positions, no more than 3 positions, no more than 2 positions or even do not have the amino acid sequence that is different from the amino acid sequence shown in the following sequence: SEQ ID NO:5 more than 1 position, SEQ ID NO:6, SEQID NO:7, SEQ ID NO:8, SEQ ID NO:9, SEQ ID NO:10, SEQ ID NO:11, SEQID NO:12, SEQ ID NO:13, SEQ ID NO:14, SEQ ID NO:15, SEQ ID NO:16, SEQ ID NO:17, SEQ ID NO:18, SEQ ID NO:19, SEQ ID NO:20 or SEQ IDNO:21.

In the most preferred embodiment, described heterozyme comprises the CBD that derives from Athelia rolfsii glucose starch, and for example the US patent 4,727, the glucoamylase of 026 disclosed Athelia rolfsiiAHU 9627.

The present invention is not to be in order to come limited field with specific embodiments disclosed herein this description and requirement because these embodiments be intended that illustrations as many aspects of the present invention.Any equal embodiment all falls within the scope of the present invention.。In fact, on the basis of foregoing, show here and the scheme described outside various modifications all be clearly for a person skilled in the art.This modification also within the scope of the appended claims.Under situation about clashing, the disclosure that comprises definition among the present invention will be started to control making usefulness.

The disclosure of the various lists of references of quoting here all is incorporated herein by reference.

Material and method

Enzyme

The disclosed wild type acid alpha-amylase of deriving of-acid alpha-amylase A:SEQ ID NO:38 from aspergillus niger.

-acid alpha-amylase B: by the hybridization acid alpha-amylase that aspergillus niger acid alpha-amylase catalytic domain (SEQ ID NO:38)-valley aspergillus glucoamylase joint (SEQ ID NO:23)-valley aspergillus glucoamylase CBD (SEQ ID NO:2) forms.

Hybridization α-Dian Fenmei shown in the-α-Dian Fenmei C:SEQ ID NO:44 comprises the catalytic domain (CD) from the Rhizomucorpusillus α-Dian Fenmei, and it has sugar from Athelia rolfsii glucoamylase in conjunction with territory (CBD).

The disclosed wild type Rhizomucor pusillus of-α-Dian Fenmei D:SEQ ID NO:43 α-Dian Fenmei.

In this manual enzyme classification numbering (EC number) indication with Recommendations (1992) of the Nomenclature Committee of the International Union of Biochemistry and Molecular Biology, Academic Press Inc, consistent in 1992.

Fabric

-from Vlisco BV, the Vlisco fabric of NL.

Buffer solution

Citrate buffer solution

10mM citrate buffer solution (pH4.0)

1.376g one hydration (monohydrate) citric acid and 1.015g two hydrations (dihydrate) natrium citricum are dissolved in the 1L deionized water.

Method

Measure homology/homogeneity

For the purposes of the present invention, homology (homology) degree is to judge as homogeneity (identity) degree between two amino acid, uses LASERGENE by the method (Higgins, 1989, CABIOS 5:151-153) of Clustal ^TMMEGALIGN ^TMSoftware (DNASTAR, Inc., Madison, WI), described software has homogeneity form and following multiple sequence reduced parameter (multiple alignmentparameters): breach point penalty (gap penalty) is 10, and the notch length point penalty is 10.The sequence reduced parameter is Ktuple=1, breach point penalty=3, window (windows)=5 and diago-nals=5 in pairs.

Acid alpha-amylase activity (AFAU mensuration)

When using according to the present invention, the activity of any acid alpha-amylase can be measured with AFAU (AcidFungal Alpha-amylase Units, fungal alpha-amylase unit), and it is with respect to the enzyme standard test.1 FAU is defined in the amount of the enzyme of the 5.260mg starch dry matter of per hour degrading under the following standard conditions.

The acid starch enzyme, and interior-α-Dian Fenmei (endo-alpha-amylase) (1,4-α-D-glucan-glucano-hydrolase, the E.C.3.2.1.1) α in hydrolyzed starch intramolecule zone-1, the 4-glycosidic bond is to form the dextrin and the oligosaccharides of different chain length.The formed color intensity of iodine is direct and starch concentration is proportional.Hydrolytic enzyme activities is measured in the minimizing (reduction) of using anti-phase colorimetric method (reverse colorimetry) to measure starch concentration under the particular analysis condition.

λ＝590nm

Indigo plant/purple (blue/violet) t=23 discolor second (decoloration)

Standard conditions/reaction condition:

Substrate: soluble starch, about 0.17g/L

Buffer solution: citric acid, about 0.03M

Iodine (I ₂): 0.03g/L

CaCl ₂： 1.85mM

pH： 2.50±0.05

Heated culture temperature: 40 ℃

Reaction time: 23 seconds

Wavelength 590nm

Enzyme concentration 0.025AFAU/mL

Enzyme working range 0.01-0.04AFAU/mL

The file EB-SM-0259.02/01 that has described this analysis method in more detail can ask for to Denmark Novozymes A/S request, so this document is hereby incorporated by.

Alpha-amylase activity (KNU)

Described amylolytic activity can be measured as substrate with potato starch.The method is based on the decomposition of enzyme to the potato starch of modified, and reaction mixes starch/enzyme solutions sample afterwards with iodine solution.Initial form black and blue color, but in the starch decomposable process, described blueness is thin out and become rufous gradually, this color can with the standard control of coloured glass.

1,000 Novo α-Dian Fenmei units (Kilo Novo alpha amylase Unit) (KNU) are defined under the standard conditions (promptly 37 ℃+/-0.05; 0.0003M Ca ²⁺And pH5.6), the amount of the enzyme of dextrinize 5260mg starch dry matter substrate Merck Amylum solubile.

The file EB-SM-0009.02/01 that has described this analysis method in more detail can ask for to Denmark Novozymes A/S request, so this document is hereby incorporated by.

Measure acid starch degrading activity (FAU)

One fungal alpha-amylase unit (Fungal Alpha-amylase Unit) (1 FAU) is defined in based under the following standard conditions, method with the mensuration α-Dian Fenmei of Novozymes standard, the amount of per hour decomposing the enzyme of 5.26g starch (Merck Amylum solubile Erg.B.6, lot number 9947275):

The substrate soluble starch

37 ℃ of temperature

pH 4.7

Reaction time 7-20 minute

Measure the detailed description of the Novozymes method of KNU and FAU and can ask to ask for standard method EB-SM-0009.02/01.

Destarch (Tegewa method)

Compare by the standard photographs with iodine staining fabric sample and a series of 1-9 scale (scale), wherein 1 is that dark blue dyeing and 9 is dye-free, visually measures starch starching residue.Described iodine staining solution is to be made by following, and dissolving 10g KI adds 0.635g I in 10ml water ₂In deionized water, make the solution that total amount is 1L with 200mL ethanol.Cutting was also soaked fabric sample 60 seconds in iodine solution, about 5 seconds of rinsing in deionized water again.After unnecessary water was extruded in sample, described fabric sample was by at least two professional's assessments.Provide average.Can be from Verband TEGEWA, Karlstrasse 21, Frankfurt a.M., Germany preparation method and standard scale.

Embodiment

Embodiment 1

With wild type acid alpha-amylase A destarch COTTON FABRIC

100% COTTON FABRIC (270g/m ²) from Sweden

Kungsfors AB.It adopts Cupper 3/1 structure (construction) to be manufactured in 2003.Described fabric contains 28 lines (thread)/cm warp thread and 14 lines/cm weft yarn.Described warp thread has Ne 11 and weft yarn has Ne 8.Two kinds of yarns are the open end (end) that shows.Dry pulp hydroscopicity pick up on warp thread is 8%.Described slurry mainly contains Kollotex 5, Solvitose XO, and the butter wax (beef tallow wax) that has emulsifying agent.Kollotex5 is low-viscosity potato starch ester.Solvitose XO is the starch ether that has the high viscosity of about 0.07DS.The fabric sample is cut to each 25g approximately.

For three kinds of buffer solutions are made in this research.PH of buffer 2 is made by following method: dissolving 11.53g 85% phosphoric acid in 4.5 liters of pure water to pH1.95, adds water to 5 liters with 5N HCl titration then.PH of buffer 3 is made by following method: dissolving 11.53g85% phosphoric acid in 4.5 liters of pure water to pH2.95, adds water to 10 liters with 5N NaOH titration then.PH of buffer 4 is made by following method: dissolving 6.005g 85% acetate in 4.5 liters of pure water to pH3.95, adds water to 10 liters with 5N HCl titration then.In each buffer solution, add 2g/l non-ionic surface active agent wetting agent.The final pH value of the mensuration of these three kinds of buffer solutions is respectively 2,2.99 and 3.98.

For each processing, get 1 liter of cushioning liquid.The wild type acid alpha-amylase A that adds specified rate (a given amount) is to buffer solution.Fabric sample soaked in solution 30 seconds, then by padding mangles (padder) (Werner Mathis) pad dyeing (padded) to reach 85% wet pickup.Described fabric sample is rolled-up and be sealed in the plastic bag.After a while in 50 ℃ of baking oven incubations 2 hours.Described fabric sample is washing in containing the padding of four little rinsing boxes-decatize scope (pad-steam range) (Werner Mathis).Water temperature in the rinsing box is respectively 95,95,90 and 90 ℃.

In air after the dried overnight, with the iodine solution described fabric sample that dyes.Visually will shown in dyeing fabric sample and 1-9 scale the TEGEWA standard photographs relatively, wherein 1 be that dark color and 9 is a dye-free.Therefore higher numerical value shows that starch is removed preferably.Described visual assessment is finished by at least three professionals, provides average T EGEWA value for each fabric sample.The results are shown in table 1.The fabric of handling through wild type acid alpha-amylase A demonstrates the TEGEWA value that is higher than the control fabric of handling without enzyme under whole three pH conditions, acid alpha-amylase A hydrolysis is described and has removed the starch slurry on the fabric.The enzymatic activity that strengthens causes higher TEGEWA that the starch removal of enhancing has been described.

Embodiment 2

With α-Dian Fenmei B destarch COTTON FABRIC

Fabric sample and buffer solution that preparation is identical with embodiment 1.α-Dian Fenmei B is different from the α-Dian Fenmei A among the embodiment 1, and α-Dian Fenmei B is fabricated and comprises wild type acid alpha-amylase A (embodiment 1) from aspergillus niger, and it is connected with CBD from valley aspergillus acid alpha-amylase.Used enzyme has the activity of 316AFAU/g in this research.Implement with embodiment 1 in identical processing and measurement.The described table 1 that the results are shown in.In the identical activity of all conditions, the acid alpha-amylase B that has CBD demonstrates the TEGEWA value that is higher than wild type acid alpha-amylase A, illustrates that the acid alpha-amylase B that has CBD has improved α-Dian Fenmei destarch performance.

Table 1

pH	The enzyme type	[enzyme] (AFAU/kg fabric)	TEGEWA value (on average)
				2	No enzyme acid alpha-amylase A acid alpha-amylase B (having CBD)	0 45.4 45.4	1.0 2.5 2.8
3	No enzyme acid alpha-amylase A acid alpha-amylase B (having CBD)	0 11.4 45.4 11.4 45.4	1.0 1.7 2.3 2.0 2.7
				4	No enzyme acid alpha-amylase A acid alpha-amylase B (having CBD)	0 11.4 45.4 11.4 45.4	1.0 1.7 2.5 1.8 3.7

Embodiment 3

The fabric sample that preparation is identical with embodiment 1.Prepare pH of buffer 3 by the following method: 11.53g 85% phosphoric acid is dissolved in 4.5 liters of pure water, to pH2.95, adds water to 5 liters then with 5N NaOH titration.Add 2g/L non-ionic surface active agent (wetting agent) afterwards in buffer solution, measuring described pH of buffer at 25 ℃ is 3.05.Use the enzyme identical with embodiment 1.

Implementing described destarch in Lab-o-mat (Werner Mathis) handles.In each beaker, add the 250mL buffer solution.Add the α-Dian Fenmei of specified quantitative.In each beaker, put into a fabric sample (25g).Seal described beaker and place Lab-o-mat.By the infrared ray heating system of equipping among the Lab-o-mat at 5 ℃ of/minute heating beakers to 50 ℃.In 30rpm, 50 ℃ of rotation beakers 45 minutes.After enzyme is handled, next in identical beaker, wash described fabric sample three times with water, respectively at 95,75 and 40 ℃.

After air drying spends the night, with embodiment 1 in identical method assess described sample.The results are shown in table 2.Can draw the conclusion identical with embodiment 1.Above 5 TEGEWA values finish for chamber equipment by experiment, illustrate that in the acid condition destarch be feasible method.

Metal ion residue on the same assessment fabric.At first cut described fabric by 1mm filter screen (sieve) with Thomas-Wiley mill.4.00 (+/-0.01) g fabric pulping (mash) is mixed with 80mL 1g/LEDTA solution.Described mixture incubation 15 hours in the oscillator (shaker) (new Brunswick Scientific Co.Inc, Series 25) of 70 ℃ and 200rpm.After cooling off about 30 minutes, that described mixture is centrifugal 10 minutes at 20 ℃ with 2500rpm.Collect the metal content analysis that supernatant is used for the Perkinelmer atomic absorption spectrophotometer.

Embodiment 4

Implement identical experiment with identical with embodiment 3 in essence method and set, except changing the α-Dian Fenmei described in the utilization embodiment 2 into.Carry out identical experiment condition and assessment.The results are shown in table 2.From then on sample can obtain with embodiment 2 in identical conclusion.Obtain the TEGEWA value more increase in this experiment, and peak is 5.8.

Table 2

N/a=does not measure

Embodiment 5

Use

Kungsfors AB (Sweden) with embodiment 1 in the fabric of same type, and cut described sample to every part of about 62g.Be equipped with pH of buffer 4 in order to the below legal system: 12.01g acetate is dissolved in 4.5 liters of pure water, to pH3.95, adds water to 10 liters then with 5N HCl titration.In buffer solution, add 2g/l non-ionic surface active agent (wetting agent).Described buffer solution final pH value is 3.99.

For each processing, use 1.2 liters of cushioning liquid.The α-Dian Fenmei that in buffer solution, adds specified quantitative.The fabric sample was soaked 30 seconds in described solution, then by padding mangles (Werner Mathis) pad dyeing to reach 90% fiber hydroscopicity.Described fabric sample is cut into the sample of two equidimensions.Two sample standard deviations are rolled-up and be sealed in the plastic bag.A bag is at 40 ℃ of baking oven incubations, and another is at room temperature (20 ℃).Two sample standard deviations were handled 16 hours.Described fabric sample is washing in containing the padding of four little rinsing boxes-decatize scope (Werner Mathis).Water temperature in the rinsing box is respectively 95,95,90 and 90 ℃.

Implement the assessment identical with embodiment 1.The results are shown in table 3.Starch slurry on the COTTON FABRIC is removed by α-Dian Fenmeishuixie and in the destarch process.At pH4, two kinds of amylase fermentoids all can be finished destarch at 20 ℃ and 40 ℃.

Table 3

Embodiment 6

Be used for α-Dian Fenmei D at pH4.0 destarch fabric

Inject deionized water in beaker, height is extremely apart from bottom 13cm.Adjust water temperature to 60 ℃.In each beaker, add 200ml 10mM citrate buffer solution (pH4.0) and place Lab-o-Mat.At 60 ℃ of described solution of heating.The α-Dian Fenmei D that in described buffer solution, adds different amounts.Final enzyme concentration is several, 200FAU/L of 50FAU/L, 100FAU and 400FAU/L respectively.The sample of two Vlisco fabrics is fixed with pair of forceps, and immersed in the dipping bath (impregnation bath) pad dyeing then 30 seconds.Repeat described dipping and extruding.Fiber hydroscopicity (wet pickup) is about 100%.

A sample is placed the double-layer plastic bag, squeeze out air and described sack is placed 60 ℃ of water-baths 2 hours.Another sample is placed the double-layer plastic bag, squeezes out air, and with sack room temperature preservation 24 hours.After reaching essential time, from water-bath, shift out described sample.Described sample is fixed in the tweezers, immersed hot rinsing solution (90 ℃) 30 seconds and extruding.Repeat described dipping and push twice.

Described fabric was immersed cold running water about 60 seconds, and squeeze the water out with hand.Described sample is placed on the shelf and in drying at room temperature.By residual content of starch on the described fabric of TEGEWA grade (rating) check.Assay is shown in Fig. 1.

Embodiment 7

α-Dian Fenmei C is in the destarch of pH4.0

Repeat embodiment 6 described destarch processes, except using α-Dian Fenmei C.Assay is shown in Fig. 2.

Sequence table

＜110〉Novozymes North America, Inc. (Novozymes North America, Inc.)

Liu，Jim

Salmon，Sonja

Wu，Guifang

＜120〉desizing process

<130>10692.204-WO

<160>47

<170>PatentIn version 3.3

<210>1

<211>396

<212>DNA

＜213〉valley aspergillus (Aspergillus kawachii)

<220>

<221>CDS

<222>(1)..(396)

＜223〉valley aspergillus CBD

<400>1

<210>2

<211>131

<212>PRT

＜213〉valley aspergillus

<400>2

<210>3

<211>1533

<212>DNA

＜213〉aspergillus niger (Aspergillus niger)

<220>

<221>CDS

<222>(1)..(1533)

＜223〉aspergillus niger acid alpha-amylase

<220>

＜221〉mat_ peptide

<222>(64)..(1533)

<400>3

<210>4

<211>511

<212>PRT

＜213〉aspergillus niger

<400>4

<210>5

<211>102

<212>PRT

＜213〉yellow hot bacillus (Bacillus flavothermus)

<220>

<221>DOMAIN

<222>(1)..(102)

<223>CBD

<400>5

<210>6

<211>99

<212>PRT

＜213〉bacillus kind

<220>

<221>DOMAIN

<222>(1)..(99)

<223>CBD

<400>6

<210>7

<211>102

<212>PRT

<213>Alcaliphilic Bacillus

<220>

<221>DOMAIN

<222>(1)..(102)

<223>CBD

<400>7

<210>8

<211>112

<212>PRT

<213>Hormoconis resinae

<220>

<221>DOMAIN

<222>(1)..(112)

<223>CBD

<400>8

<210>9

<211>95

<212>PRT

<213>Lentinula edodes

<220>

<221>DOMAIN

<222>(1)..(95)

<223>CBD

<400>9

<210>10

<211>107

<212>PRT

＜213〉Neurospora crassa (Neurospora crassa)

<220>

<221>DOMAIN

<222>(1)..(107)

<223>CBD

<400>10

<210>11

<211>115

<212>PRT

<213>Talaromyces byssochlamydioides

<220>

<221>DOMAIN

<222>(1)..(115)

<223>CBD

<400>11

<210>12

<211>115

<212>PRT

<213>Geosmithia cylindrospora：

<220>

<221>DOMAIN

<222>(1)..(115)

<223>CBD

<400>12

<210>13

<211>139

<212>PRT

<213>Scorias spongiosa

<220>

<221>DOMAIN

<222>(1)..(139)

<223>CBD

<400>13

<210>14

<211>126

<212>PRT

<213>Eupenicillium ludwigii

<220>

<221>DOMAIN

<222>(1)..(126)

<223>CBD

<400>14

<210>15

<211>116

<212>PRT

＜213〉aspergillus japonicus (Aspergillus japonicus)

<220>

<221>DOMAIN

<222>(1)..(116)

<223>CBD

<400>15

<210>16

<211>133

<212>PRT

<213>Penicillium cf.miczynskii

<220>

<221>DOMAIN

<222>(1)..(133)

<223>CBD

<400>16

<210>17

<211>116

<212>PRT

＜213〉Mz1 mould kind (Penicillium sp.)

<220>

<221>DOMAIN

<222>(1)..(116)

<223>CBD

<400>17

<210>18

<211>114

<212>PRT

<213>Thysanophora sp

<220>

<221>DOMAIN

<222>(1)..(114)

<223>CBD

<400>18

<210>19

<211>111

<212>PRT

＜213〉grey humicola lanuginosa (Humicola grisea) var.thermoidea

<220>

<221>DOMAIN

<222>(1)..(111)

<223>CBD

<400>19

<210>20

<211>108

<212>PRT

＜213〉aspergillus niger

<220>

<221>DOMAIN

<222>(1)..(108)

<223>CBD

<400>20

<210>21

<211>97

<212>PRT

<213>Althea rolfsii

<220>

<221>DOMAIN

<222>(1)..(97)

<223>CBD

<400>21

<210>22

<211>38

<212>PRT

＜213〉aspergillus niger

<220>

<221>DOMAIN

<222>(1)..(38)

＜223〉joint (linker)

<400>22

<210>23

<211>31

<212>PRT

＜213〉valley aspergillus

<220>

<221>DOMAIN

<222>(1)..(31)

＜223〉joint

<400>23

<210>24

<211>11

<212>PRT

<213>Athelia rolfsii

<220>

<221>DOMAIN

<222>(1)..(11)

＜223〉joint

<400>24

<210>25

<211>8

<212>PRT

＜213〉artificial

<220>

＜223〉artificial

<220>

<221>DOMAIN

<222>(1)..(8)

＜223〉joint

<400>25

<210>26

<211>498

<212>PRT

＜213〉Aspergillus oryzae

<220>

<221>SIGNAL

<222>(1)..(20)

<220>

＜221〉mat_ peptide

<222>(20)..(498)

<400>26

<210>27

<211>1860

<212>DNA

＜213〉artificial

<220>

＜223〉artificial

<220>

<221>CDS

<222>(1)..(1860)

＜223〉by crossbred that aspergillus niger acid alpha-amylase catalytic domain-valley aspergillus α-Dian Fenmei joint-aspergillus niger glucoamylase CBD forms

<400>27

<210>28

<211>619

<212>PRT

＜213〉artificial

<220>

＜223〉He Cheng construct

<400>28

<210>29

<211>1827

<212>DNA

＜213〉artificial

<220>

＜223〉artificial

<220>

<221>CDS

<222>(1)..(1827)

＜223〉contain aspergillus niger acid alpha-amylase catalytic domain-valley aspergillus α-Dian Fenmei joint-Athelia rolfsii glucoamylase CBD's

Crossbred

<400>29

<210>30

<211>608

<212>PRT

＜213〉artificial

<220>

＜223〉He Cheng construct

<400>30

<210>31

<211>1863

<212>DNA

＜213〉artificial

<220>

＜223〉artificial

<220>

<221>CDS

<222>(1)..(1863)

＜223〉by crossbred that Aspergillus oryzae catalytic domain-valley aspergillus α-Dian Fenmei joint-valley aspergillus α-Dian Fenmei CBD forms

<400>31

<210>32

<211>620

<212>PRT

＜213〉artificial

<220>

＜223〉He Cheng construct

<400>32

<210>33

<211>1767

<212>DNA

＜213〉artificial

<220>

＜223〉artificial

<220>

<221>CDS

<222>(1)..(1767)

＜223〉by crossbred that aspergillus niger acid alpha-amylase-A.rolfsii glucoamylase joint-A.rolfsii glucoamylase CBD forms

<400>33

<210>34

<211>588

<212>PRT

＜213〉artificial

<220>

＜223〉He Cheng construct

<400>34

<210>35

<211>1767

<212>DNA

＜213〉artificial

<220>

＜223〉artificial

<220>

<221>CDS

<222>(1)..(1767)

＜223〉by crossbred that Aspergillus oryzae α-Dian Fenmei-A.rolfsii glucoamylase joint-A.rolfsii glucoamylase joint is formed

<400>35

<210>36

<211>588

<212>PRT

＜213〉artificial

<220>

＜223〉He Cheng construct

<400>36

<210>37

<211>640

<212>PRT

＜213〉valley aspergillus

<220>

＜221〉mat_ peptide

<222>(22)..(640)

<400>37

<210>38

<211>505

<212>PRT

＜213〉aspergillus niger

<220>

＜221〉mat_ peptide

<222>(22)..(505)

<400>38

<210>39

<211>476

<212>PRT

＜213〉Aspergillus oryzae

<220>

＜221〉mat_ peptide

<222>(1)..(476)

<400>39

<210>40

<211>1455

<212>DNA

＜213〉bacillus kind

<220>

<221>CDS

<222>(1)..(1455)

<223>AA560

<220>

＜221〉mat_ peptide

<222>(1)..()

<400>40

<210>41

<211>485

<212>PRT

＜213〉bacillus kind

<400>41

<210>42

<211>1350

<212>DNA

<213>Rhizomucor pusillus

<220>

<221>CDS

<222>(1)..(1350)

<400>42

<210>43

<211>450

<212>PRT

<213>Rhizomucor pusillus

<400>43

<210>44

<211>558

<212>PRT

＜213〉artificial

<220>

＜223〉crossbred

<220>

<221>MISC_FEATURE

<222>(1)..(461)

＜223〉Rhizomucor pusillus α-Dian Fenmei

<220>

<221>MISC_FEATURE

<222>(462)..(558)

<223>CBD

<400>44

<210>45

<211>1398

<212>DNA

<213>Meripilus giganteus

<220>

<221>CDS

<222>(1)..(1398)

<400>45

<210>46

<211>466

<212>PRT

<213>Meripilus giganteus

<400>46

<210>47

<211>574

<212>PRT

<213>Meripilus giganteus

<220>

<221>MISC_FEATURE

<222>(1)..(477)

＜223〉Meripilus giganteus α-Dian Fenmei

<220>

<221>MISC_FEATURE

<222>(478)..(574)

<223>CBD

<400>47

Claims (31)

1. the destarch method of the fabric of starching during making fabric, described fabric contains starch or starch derivatives, this method is included in the fabric with described starching of incubation in the processing aqueous solution of pH scope between 1 to 5, and this processing aqueous solution comprises α-Dian Fenmei.

2. the process of claim 1 wherein that described pH is the scope between pH 1 to 4, particularly between pH 2 to 4.

3. claim 1 or 2 method, wherein said α-Dian Fenmei be belong to bacterial origin or originated from fungus, for example filamentous fungi source.

4. each method of claim 1-3, wherein said α-Dian Fenmei is an acid alpha-amylase.

5. claim 3 or 4 method, wherein said α-Dian Fenmei is to be derived from the aspergillus bacterial strain, preferred aspergillus niger, aspergillus awamori or Aspergillus oryzae; Or be derived from Rhizomucor (Rhizomucor) bacterial strain, preferred Rhizomucor pusillus; Or be derived from the Meripilus bacterial strain, preferred Meripilus giganteus bacterial strain.

6. the method for claim 5, wherein said aspergillus α-Dian Fenmei is a disclosed acid black aspergillus α-Dian Fenmei among the SEQ ID NO:38, or its variant.

7. the method for claim 5, wherein said Rhizomucor α-Dian Fenmei is a disclosed Rhizomucor pusillus α-Dian Fenmei among the SEQ ID NO:43, or its variant.

8. each method among the claim 1-7, wherein said α-Dian Fenmei, the preferred acidic α-Dian Fenmei is with 1-3, and the concentration of 000AFAU/kg fabric exists, preferred 10-1, the fabric of 000AFAU/kg, the particularly fabric of 100-500AFAU/kg or 1-3, the Treatment Solution of 000AFAU/L, preferred 10-1, the concentration of the Treatment Solution of 000AFAU/L Treatment Solution, particularly 100-500AFAU/L exists.

9. the method for claim 1-8, the α-Dian Fenmei of wherein said bacterium is to be derived from Bacillus strain, preferred source is from the bacterial classification bacterial strain of bacillus, the more preferably bacterial classification of lichens bacillus, bacillus amyloliquefaciens, bacillus stearothermophilus, bacillus subtilis or bacillus, for example bacillus bacterial classification NCIB 12289, NCIB 12512, NCIB 12513, DSM 9375, DSMZ 12648, DSMZ 12649, KSM AP1378, KSM K36 or KSM K38.

10. the method for claim 9, the α-Dian Fenmei of wherein said bacterium is as the disclosed AA560 α-Dian Fenmei of SEQ ID NO:40.

11. the method for claim 9 or 10, wherein said bacillus α-Dian Fenmei has one or more disappearances at position D183 and G184, and the variant of preferred wherein said bacillus α-Dian Fenmei further has replacement (using the numbering of SEQ ID NO:40) at position N195F.

12. the method for claim 9, wherein said bacillus α-Dian Fenmei has one or more disappearances at position D183 and G184, the variant of preferred wherein said bacillus α-Dian Fenmei further has one or more following replacements: R118K, N195F, R320K, R458K, particularly wherein said variant has following sudden change: Δ (D183+G184)+R118K+N195F+R320K+R458K (using the numbering of SEQ ID NO:40).

13. each method of claim 1-12, wherein said α-Dian Fenmei exists with the concentration of 0.05-150KNU/L Treatment Solution, preferably, with the 1-100KNU/L Treatment Solution, particularly 2-20KNU/L Treatment Solution or 0.05-150KNU/kg fabric, preferably, 1-100KNU/kg fabric, the particularly concentration of 2-20KNU/kg fabric exist.

14. each method of claim 1-13, wherein said α-Dian Fenmei is a heterozyme, and it comprises sugar in conjunction with the territory.

15. each method of claim 1-14, wherein said α-Dian Fenmei comprises starch binding domain originated from fungus or bacterial origin.

16. the method for claim 14 or 15, wherein said sugar is derived from the aspergillus bacterial strain in conjunction with territory (CBD), and preferred source is from bacterial classification bacterial strain, Athelia bacterial strain or the Talaromyces bacterial strain of aspergillus.

17. the method for claim 16, wherein said sugar is derived from valley aspergillus in conjunction with the amino acid sequence of territory (CBD), the CBD that for example has amino acid sequence shown in the SEQ ID NO:2, or described sugar is derived from aspergillus niger in conjunction with the amino acid sequence of territory (CBD), preferred described CBD with amino acid sequence shown in the SEQ ID:20, or be derived from Athelia sp., preferred source is from the sugar of the Athelia rolfsii amino acid sequence, particularly CBD amino acid sequence shown in the SEQ ID NO:21 in conjunction with territory (CBD).

18. the method for claim 15, the α-Dian Fenmei of the wherein said CBD of comprising comprises joint between α-Dian Fenmei and CBD or starch binding domain.

19. the method for claim 18, wherein said joint is derived from the Athelia bacterial strain, preferred Atheliarolfsii glucoamylase joint (SEQ ID NO:24), the aspergillus bacterial strain, preferred aspergillus niger glucoamylase joint (SEQ ID NO:22), or valley aspergillus α-Dian Fenmei joint (SEQ ID NO:23).

20. each method of claim 1-19, wherein said α-Dian Fenmei is the α-Dian Fenmei of the heterozygosis shown in the SEQ ID NO:44, it comprises the catalytic domain (CD) of Rhizomucor pusillus α-Dian Fenmei, and described Rhizomucor pusillus α-Dian Fenmei comprises the sugar of Athelia rolfsii glucoamylase in conjunction with territory (CBD).

21. each method of claim 1-20, wherein said method is that temperature is carried out in the scope from 5-90 ℃, particularly 20 to 90 ℃.

22. the method for claim 21, wherein said method are to carry out one suitable period between 20 to 40 ℃ in temperature, preferably between 8-24 hour.

23. the method for claim 21, wherein said method are carried out one suitable period between 40 to 90 ℃ of temperature the time, preferably between 1 to 6 hour.

24. each method of claim 1-23, wherein said method is carried out in the presence of surfactant, and preferred described surfactant exists with the concentration of 0.1-10g/L, preferably approximately 1g/L.

25. each method of claim 1-24, wherein said fabric is to be made by the fiber in natural or artificial source.

26. each method of claim 1-25, wherein said fabric is COTTON FABRIC, jean, sodolin, viscose, lyocell or cellulose acetate.

27. each method of claim 1-26, wherein said fabric is by the fiber manufacturing that is derived from animal, particularly silk or woollen.

28. each method of claim 1-27, wherein said fabric is to be made by the polyester fiber in natural or artificial source, for example polyethylene terephthalate or PLA.

29. each method of claim 1-28, wherein said fabric is made by nylon, fiber acrylic acid or polyurethanes.

30. each method of claim 1-29, wherein said fabric are to contain the fabric of polyester or the clothing of being made up of 100% polyester basically.

31. the method for claim 1-30, wherein said polyester textile is polyester mixture (blend), for example polyester and cellulosic mixture, comprise polyester and cotton mixture, polyester and woollen mixture, polyester and silk mixture, polyester and acrylic acid mixture, polyester and nylon mixture, polyester, nylon and polyurethane mixture, polyester and polyurethane mixture, artificial fibre (viscose), cellulose acetate and tencel.

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