CN1276824A

CN1276824A - Detergent compositions comprising mannanase and cationic surfactant

Info

Publication number: CN1276824A
Application number: CN98810231A
Authority: CN
Inventors: J·L·P·贝蒂奥尔
Original assignee: Procter and Gamble Co
Current assignee: Procter and Gamble Co
Priority date: 1997-08-14
Filing date: 1998-06-10
Publication date: 2000-12-13
Also published as: DE69826294T2; ES2268780T3; CA2301168A1; DE69835214D1; DK1009795T3; MXPA00001567A; DE69837850D1; CZ2000502A3; WO1999009126A1; ATE332958T1; PT1009795E; EP1036151A1; HUP0003670A3; AU8065198A; EP0896998A1; JP4090688B2; BR9811195A; MXPA00001613A; AU7832798A; AU8064198A

Abstract

The present invention relates to laundry detergent and/or fabric tending and protecting compositions, comprising a saccharide gum degrading enzyme and positive ion superficial active reagent providing excellent pliability and cleaning performance.

Description

The detergent composition that contains mannase and cats product

Invention field

The present invention relates to contain the washing composition and/or the Fabrid care composition of mannase (mannanase) and cats product.

Background of invention

The performance of Betengent product is judged by many factors, comprises that soil removability and the degradation production of avoiding dirt or dirt deposit to the ability on the goods that are in the suds again.Therefore, present detergent composition comprises the complex combination of the active ingredient that has satisfied some special requirement.Specifically, present detergent formulation generally includes tensio-active agent and detergent enzyme so that washing and fabric nursing effect to be provided.

Cats product is well known in the prior artly to remove greasy spot preferably to provide, for example makeup and food stain effect.

The foods and cosmetics stain has been represented most of stain relevant with the human consumer, contains foodstuff additive usually, for example thickeners/stabilizers.In fact, hydro-colloid glue and emulsifying agent are usually as foodstuff additive.Polysaccharide (long chain polymer) or their derivative that one group of industry of term " glue " expression is used, their hydrations in heat or cold water form viscous soln, dispersion liquid or gel.Glue is categorized into natural and modification.Natural gum comprises Seaweed Extract, plant milk extract, the glue that is obtained by seed or root and the glue that is obtained by microbial fermentation.(semi-synthetic) glue of modification comprises Mierocrystalline cellulose and starch derivative and some synthetical glue, for example rudimentary methoxypectin, propanediol alginate and carboxyl methyl and hydroxypropyl guar gum (glue, complete works of 4th version 12 volumes of chemical technology, the 842-862 page or leaf, J.Baird, Kelco division of Merck).Also referring to Food science, carbohydrate chemistry (Eagan Press-1997), R.L.Whistlerand J.N.BeMiller, the 4th chapter, the direct food additive in 63-89 page or leaf and the processing fruits, P.Laslo, biological principle and application, the 1st volume, II chapter, 313-325 page or leaf (1996) Technomie publishing.The part of these glue, for example guar gum (E412), carob bean gum (E410) are widely used in food applications (Gums in ECT4th Ed. the 12nd volume, 842-862 page or leaf, J.Baird, Kelco division ofMerck) separately or in combination.

The guar gum that uses in these foods and cosmetics spots is obtained by the seed endosperm of bean Cyamopsistetragonoloba.The guar gum (being also referred to as melon ear sugar) that is extracted by dicotyledonous seed is by 1-4, b-D-pyran-mannose glycosylation unit skeleton is formed, in seasonings and frozen product and makeup, be used as thickening material (H.-D.Belitz. food chemistry, the 243rd page, English edition the 2nd edition, Springer-verlag, 1987, ISBN 0-387-15043-9 (US)) ﹠amp; (Food science, carbohydrate chemistry, R.L.Wilstler, eagan press, 1977, ISBN 0-913250-92-9) ﹠amp; (industrial glue, the 2nd edition, the R.L.Whistler308 page or leaf, the science communication, 1973, ISBN, 0-12-74-6252-x).Carob bean gum (being also referred to as tragon or St Jon ' s bread) also is used for foodstuffs industry, is extracted by the evergreen seed in the mediterranean region plantation.The difference of the structure of carob bean gum and guar gum only is fewer purpose D-galactosyl side chain, has identical 1-4, b-D-pyran-mannose glycosylation skeleton.In legume-seeds, water-soluble gala mannosans is main storage carbohydrate, in some cases, accounts for 20% of gross dry weight amount at the most.Polygalactomannan has α-semi-lactosi of the 0-6 that is connected in mannose residue, but on the 0-2 of mannose residue and 0-3 also acyl group change into various degree.

Yet people still need to prepare washing composition and/or Fabrid care composition, and it provides outstanding softness and scourability, especially to makeup and food stain.This purpose can contain the washing composition of mannase and cats product and/or Fabrid care composition satisfies by preparation.

We are surprised to find owing to remove the cats product of greasy spot and the synergy of the mannase of the residual water colloid glue of degrading, and said composition provides outstanding soft and washing.This tensio-active agent-enzyme mixed system provides outstanding softness and washing effect, especially for makeup and food stain.

We also find by adding other tensio-active agent, especially anion surfactant, preferred alkyl vitriol and/or sulfonate and/or nonionogenic tenside, for example has the C8-C20 chain length, preferred C12-C16 and 2-9, the alkyl ethoxy nonionogenic tenside and/or the alkyl chain length of the degree of ethoxylation of preferred 3-7 are C8-C20, and the alkyl methyl glucamide tensio-active agent of preferred C12-C18 has improved the performance of washing composition of the present invention and/or Fabrid care composition.

Oneself discerns mannase in some bacillus biologies.The practical environmental microorganism of Talbot etc. for example, 56 volumes, No.11,3505-3510 page or leaf (1990) have been described a kind of 'beta '-mannase of the dimeric forms that is obtained by bacstearothermophilus, and it has the molecular weight of 162kDa and the best pH of 5.5-7.5.World's microorganism biological technical journal of Mendoza etc., the 10th volume, no.5, it is 38kDa that 551-555 (1994) has described the MW that is obtained by subtilis, optimum activity is 4.8 'beta '-mannase at pH5.0/55 ℃ and pl.J0304706 disclose by having of obtaining of bacillus with the MW of gel filteration determining be 37+/-3kDa, best pH is that 8-10 and pl are the 'beta '-mannase of 5.3-5.4.J63056289 has described alkalescence, the heat-staple 'beta '-mannase of preparation, and its hydrolysis is the β of mannosans-1 for example, and 4-D-mannopyranose glycosidic bond produces manna oligosaccharide.J63036774 relates to micro-organisms bacillus FERM P-8856, and it produces 'beta '-mannase and beta-Mannosidase under alkaline pH.A kind of mannase and their preparation method who is used for the purifying of bleached pulp from bacillus amyloliquefaciens discloses at WO97/11164.WO91/18974 described a kind of under limit pH and temperature active hemicellulase, for example dextranase, zytase or mannase and their preparation method.WO94/25576 has described the active enzyme of demonstration mannase that is obtained by Aspergillus aculeatusCBS101.43, and it can be used for various uses, and wherein the degraded and the modification of plant or alga cells wall material are desirable.WO93/24622 discloses the isolating mannase that is used for the bleaching lignin cellulose pulp by Trichoderma reesie.

Yet people do not recognize that yet the collaborative combination in washing composition and/or Fabrid care composition of mannase and cats product is to provide outstanding softness and scourability so far.

Summary of the invention

The present invention relates to contain the washing composition and/or the Fabrid care composition of mannase and cats product, these compositions provide outstanding softness and scourability.

The detailed description of invention

Cats product is knownly to remove greasy spot to provide in detergent applications, especially for example makeup and food stain.Known modern makeup of people and food compositions contain increasing additive, for example are used as the hydro-colloid glue of thickening material.Mannosans, guar gum and angle beans be used for many makeup and food compositions (referring to Industrial Gum, the 2nd edition, R.L.Whistler, p308, Academic Press, 1973, ISBN, 0-12-74-6252-x).It is found that these hydro-colloid glue have very high avidity to cellulosic material.

In addition, it is found that the spot that contains hydro-colloid glue is difficult to remove, even representing surface-active agent, for example under the help of cats product.Although do not plan to be limited to theory, we believe that it is because the greasy spot on the hydro-colloid glue bond fabric that cats product is removed the reduction of the efficient of the makeup that contain hydro-colloid glue and food stain.We are surprised to find and are used in combination mannase with cats product and produce effective washing effect.In fact, we find that cats product removes greasy spot, mannosans enzyme liberating residual water colloid glue.Use this cats product-enzyme mixed system that outstanding scourability is provided, especially for makeup and food stain.

In addition, we find that also this blended tensio-active agent-enzyme system provides good soft character.

Mannase

The basal component of washing composition of the present invention and/or Fabrid care composition is a mannase.

Comprise in the present invention be following three kinds of mannosans lytic enzyme: EC3.2.1.25: beta-Mannosidase, EC3.2.1.78: interior-1, the 4-beta-Mannosidase, hereinafter referred to as " mannase " and EC3.2.1.100:1, the biological Glycosylase (IUPAC classification-enzyme nomenclature, 1992 ISBN 0-12-227165-3 Academic Press) of 4-β-sweet dew.

Detergent for washing clothes of the present invention and/or Fabrid care composition more preferably contain the β-1 that is called mannase, 4-mannosidase (E.C.3.2.1.78).The mannase that term " mannase " or " polygalactomannan enzyme " expression define officially according to prior art, it is called mannosans interior-1, the 4-beta-Mannosidase, and be called 'beta '-mannase and interior-1 in addition, the 4-mannase, and catalysis is in mannosans, polygalactomannan, glucomannan and galactoglucomannan 1, the reaction of the random hydrolysis of 4-β-D-seminose glycosidic bond.

Mannase (EC 3.2.1.78) especially constitutes one group of polysaccharidase, its mannosans of degrading, expression can be ruptured and be contained the enzyme of the unitary polysaccharide chain of seminose, promptly can break at the enzyme of the glycosidic link in mannosans, polygalactomannan, glucomannan and the galactoglucomannan.Mannosans is to contain by β-1, the polysaccharide of the skeleton that the seminose that 4-connects is formed; Glucomannan is to contain more or less to change β-1, the polysaccharide of the seminose that 4-connects and the skeleton of glucose; Polygalactomannan and galactoglucomannan are to have α-1, the mannosans and the polygalactomannan of the galactose side that 6-connects.These compounds can be by ethanoylization.

The degraded of polygalactomannan and galactoglucomannan becomes easy by removing galactose side wholly or in part.In addition, the degraded of the mannosans of ethanoylization, glucomannan, polygalactomannan galactoglucomannan becomes easy by deacetylationization wholly or in part.Ethanoyl can be removed by alkali or mannosans acetylase.Can further degrade to discharge free maltose by the oligopolymer that the mixture of mannase or mannase and alpha-galactosidase and/or mannosans acetylase discharges from beta-Mannosidase and/or beta-glucosidase enzyme.

Oneself discerns mannase in some bacillus biologies.The Appl.Environ.Microbiol. of Talbot etc. for example, 56 volumes, No.11,3505-3510 page or leaf (1990) have described a kind of 'beta '-mannase of the dimeric forms that is obtained by bacstearothermophilus, and it has the molecular weight of 162kDa and the best pH of 5.5-7.5.The World J.MicobioBoitech. of Mendoza etc., the 10th volume, No.5, it is 38kDa that 551-555 (1994) has described the MW that is obtained by subtilis, optimum activity is 4.8 'beta '-mannase at pH5.0/55 ℃ and pl.It is 373kDa that JP-0304706 discloses the MW that has with gel filteration determining that is obtained by Bacillaceae, and best pH is that 8-10 and pl are the 'beta '-mannase of 5.3-5.4.JP-63056289 has described alkalescence, the heat-staple 'beta '-mannase of preparation, and its hydrolysis is the β of mannosans-1 for example, and 4-D-mannopyranose glycosidic bond produces manna oligosaccharide.JP-63036774 relates to bacillus micro-organism FERM P-8856, and it produces 'beta '-mannase and beta-Mannosidase under alkaline pH.JP-08051975 discloses the alkaline ' beta '-mannase that is obtained by close alkali bacillus AM-001.A kind of pure mannase that is used for bleached pulp and paper and their preparation method from bacillus amyloliquefaciens is open at WO97/11164.WO91/18974 described a kind of under limit pH and temperature active hemicellulase, for example dextranase, zytase or mannase.WO94/25576 has described the active enzyme of demonstration mannase that is obtained by Aspergillus aculeatus CBS 101.43, and it can be used for degraded and improved plant or alga cells wall material.WO93/24622 discloses the isolating mannase that is used for the bleaching lignin cellulose pulp by Trichoderma reseei.The hemicellulase of the hemicellulose that contains mannosans of can degrading in WO91/18974, describe and the pure mannase that obtains by bacillus amyloliquefaciens open in WO97/11164.

This mannase is the alkali mannanase as giving a definition especially, most preferably the mannase that is obtained by bacterial origin.Washing composition of the present invention and/or Fabrid care composition especially contain alkali mannanase, and it is selected from by bacterial strain Bacillus agaradherens and/or bacillus subtilis strain 168, the mannase that gene yght obtains.

Term " alkali mannanase " is meant and is included in 7-12, has at least 10% of its maximum activity in the certain pH scope of preferred 7.5-10.5, preferably at least 25%, and the more preferably enzyme of at least 40% enzymic activity.

Washing composition of the present invention and/or Fabrid care composition most preferably contain the alkali mannanase that is obtained by Bacillusagaradherens, and described mannase is:

I) by Bacillus agaradherens, the polypeptide of NCIMB40482 preparation, or

Ii) contain just like the polypeptide of the aminoacid sequence shown in the position 32-343 of SEQ ID NO:2 or

Iii) at i) and ii) in the analogue of polypeptide of definition, it at least 70% with described polypeptide of the same race, or obtain by described polypeptide, or be immune response with polyclonal antibody that the described polypeptide of relative pure form produces by displacement, disappearance or additional one or more amino acid.

The present invention also comprises having the active isolated polypeptide of mannase, and it is selected from:

(a) coding has the polynucleotide molecule of mannosans peptide activity, its contain just like among the SEQ IDNO:1 by the nucleotide sequence shown in the Nucleotide 97-Nucleotide 1029;

(b) kind isoplassont (a);

(c) polynucleotide molecule, its coding have the mannosans enzymic activity, with SEQ ID NO:2 in by the identical polypeptide of the aminoacid sequence at least 70% of amino-acid residue 32-amino-acid residue 343;

(d) with (a) and (b) or (c) complementary molecule; With

(e) with (a) and (b), (c) or degenerate nucleotide sequence (d).

The plasmid pSJ1678 that contains the polynucleotide molecule (dna sequence dna) of the mannase of the present invention of encoding is converted into coli strain, it by the inventor according to budapest treaty about being the microbial preservation unit (Germany of patent purpose and program international recognition, D-38124) preserve, on May 18th, 1998 was deposited deposit numbers DSM12180.

Second kind of most preferred enzyme is the mannase that is obtained by bacillus subtilis strain 168, this mannase:

I) by the encoding part coding of the analogue of dna sequence dna shown in the SEQ ID No.5 or described sequence and/or

Ii) contain the aminoacid sequence shown in the SEQ ID NO:6 polypeptide or

Iii) ii) in the analogue of polypeptide of definition, it is at least 70% of the same race with described polypeptide, or is obtained by described polypeptide by displacement, disappearance or additional one or more amino acid, or the polyclonal antibody reaction of the pure form that produces with relative described polypeptide.

(a) polynucleotide molecule of coded polypeptide, described polypeptide has the mannosans enzymic activity, and contains just like the nucleotide sequence shown in the SEQ ID NO:5;

(b) kind isoplassont (a);

(c) polynucleotide molecule, its coding has a mannosans enzymic activity, the polypeptide identical with the aminoacid sequence at least 70% of SEQ ID NO:6;

(d) with (a) and (b) or (c) complementary molecule; With

(e) with (a) and (b), (c) or degeneration (d) (degeneracy) nucleotide sequence.

Definition

Before further going through the present invention, at first be defined as follows term:

Polypeptide or protein that term " positive isoplassont (ortholog) " (or " kind isoplassont ") expression is obtained by a kind, this kind has homology with the similar polypeptide or the protein of not acquisition of the same race.

Polypeptide or protein that term " secondary isoplassont (paralog) " expression is obtained by specific kind, this kind has homology with not homopolypeptide or protein by acquisition mutually of the same race.

Term " expression vector " expression straight chain or Circular DNA molecular structure, it contains the fragment of the interested polypeptide of encoding, and it is operatively connected in the additional clip that provides it to transcribe.This additional clip can comprise promotor and terminator sequence, but comprises that optionally one or more duplicate origin, one or more selection markers, enhanser, poly A addition signal etc.Expression vector is obtained by plasmid or viral DNA usually, or can contain two kinds of elements simultaneously.Expression vector of the present invention can be any expression vector, and it can carry out the recombinant DNA process easily, and the host cell that carrier is introduced is depended in the selection of carrier usually.Therefore, carrier can be an autonomously replicationg vector, and promptly carrier exists as the extrachromosome entity, and duplicating with chromosome duplication of it is irrelevant, for example plasmid.In addition, carrier can be a kind of carrier, and when it introduced host cell, it was integrated in the host cell gene group, duplicates with the karyomit(e) of being integrated.

According to the standard definition definition of prior art, recombinant expression of proteins is usually by using above-mentioned expression vector to carry out to be used for the term " recombinant expressed " relevant with polypeptide or protein expression of the present invention or " the ground expression of recombinating ".

When being used for polynucleotide molecule, term " isolating " expression polynucleotide are by taking out in its natural gene environment, thereby do not have other irrelevant or unwanted encoding sequence, are the forms that is suitable for the engineered protein production system.This isolating molecule is by the isolating material of their natural surroundings, comprises cDNA and genomic clone.Isolated DNA molecule of the present invention does not have and its normal associating other gene, but can comprise 5 ' and 3 ' non-translational region of natural generation, for example promotor and terminator.The identification in zone of associating is obvious (referring to for example, Dynan and Tijan, Nature316:774-78,1985) for those skilled in the art.

In addition, term " isolating polynucleotide " also can be described as " clone's polynucleotide ".When being used for protein/polypeptide, term " isolating " is illustrated in the protein of finding under the condition of non-its natural surroundings.In a preferred form, isolating protein does not have other protein on substantially, especially other homologous protein (i.e. " impurity of the same race " (seeing below)).Preferably with greater than 40% purity form, the purity form more preferably greater than 60% provides protein.More preferably, preferably with high-purity form, promptly by SDS-PAGE measure greater than 80% purity, more preferably greater than 95% purity with most preferably provide protein greater than 99% purity.

In addition, term " isolating protein/polypeptide " also can be described as " protein/polypeptide of purifying ".

Any impurity (other polypeptide outside the polypeptide for example of the present invention) represented in term " impurity of the same race ", and it is by the wherein original allogenic cell generation that obtains polypeptide of the present invention.

Term when being used for this paper specified microorganisms source " by ... obtaining " expression is by particular source or the polynucleotide and/or the polypeptide that are obtained by the cell that has wherein inserted from the gene in this source.

Term when being used for dna fragmentation " be operatively connected " expression arrange fragment make they be its required purposes is consistent moves, for example transcribe from promotor, encoded fragment proceeds to terminator.

Term " polynucleotide " expression is read the deoxynucleotide of 3 ' end or the strand or the dichain polymer of Nucleotide by 5 '.Polynucleotide comprise RNA and DNA, can be separated by natural origin, external synthetic or by natural and synthetic molecules in conjunction with preparation.

The polynucleotide molecule with complementary base sequence and opposed orientation is compared in term " complementation of polynucleotide molecule " expression with reference sequences.For example sequence 5 ' ATGCACGGG3 ' and 5 ' CCCGTGCAT3 ' complementation.

A kind of nucleotide sequence that comprises one or more degenerate codons (comparing) of term " nucleotide sequence of degeneration " expression with the reference polynucleotide molecule of coded polypeptide.The codon of degeneracy contains the different triplets of Nucleotide, but the identical amino-acid residue of encoding (be GAU and GAC triplet all encode Asp).

Term " promotor " expression contains the part of the gene of dna sequence dna, it provide the combination of RNA polymerase and transcribe initial.Promoter sequence is common, but does not always find in 5 ' non-coding region of gene.

Term " secretory signal sequence " expression dna sequence dna, its coded polypeptide (" secrete polypeptide ") as the component of big polypeptide, instructs big polypeptide by the Secretory Pathway of synthetic cell therein.When changing by Secretory Pathway, big polypeptide ruptures usually to remove the secretion peptide.

How to use sequence of the present invention to obtain other correlated series:

The sequence information that relates to the polynucleotide sequence of the mannase of the present invention of encoding disclosed by the invention can be used as instrument to identify other mannase of the same race.For example, polymerase chain reaction (PCR) can be used for amplifying coding by various microbial sources, the sequence of other mannase of the same race that especially different bacillus obtain.

The method that is used for activity test

Having the active polypeptide of the present invention of mannase can be according to standard test methods test mannosans enzymic activity well known in the prior art, for example be applied in the 4mm diametric hole of on the offset plate that contains 0.2%AZCL polygalactomannan (carob bean gum), getting by the solution that will be tested, promptly by Megazyme company (network address: http://www.megazyme.com/purchase/index.html). be used for inscribe-1, the substrate of 4-β-D-mannosans enzyme test as CatNo.1-AZGMA with what per 3 gram US$110.00 obtained.

Polynucleotide:

Isolating polynucleotide of the present invention under medium at least constraint condition, will hybridize in SEQ ID NO1 or with the similar size area of its complementary sequence.

Polynucleotide of the present invention are in medium at least constraint condition as described below, but preferably especially hybridize in the denatured double stranded dna probe of the full sequence shown in the position 97-1029 that contains SEQ ID NO:1 under high constraint condition or have any probe at least about the subsequence of the SEQ ID NO:1 of 100 base pair length.The suitable experiment condition that is used to measure hybridization under the medium or high constraint between nucleotide probe and DNA of the same race or the RNA sequence is included in 5 * SSC (sodium chloride/sodium citrate, Sambrook etc., 1989) pre-soaking contains the strainer of dna fragmentation or RNA to hybridize 10 minutes in, at 5 * SSC, 5 * Denhardt ' s solution (Sambrook etc., 1989), salmon sperm DNA (the Sambrook etc. of 0.5%SDS and 100 μ g/ml sex change supersound process, 1989) in the solution with the strainer prehybridization, containing the random primer (Feinberg that concentration is 10ng/ml subsequently, A.P. with Vogelstein B. (1983) Anal.Biochem.132:6-13), the same solution of 32P-dCTP mark (being higher than 1 * 109cpm/ μ g than living) probe was hybridized 12 hours down at about 45 ℃.Strainer is subsequently at least 60 ℃ (medium constraints), more preferably at least 65 ℃ (medium/high constraint), also preferably under at least 70 ℃ (high constraints) and most preferably at least 75 ℃ (very high constraint) at 2 * SSC, washed twice among the 0.5%SDS.

The molecule of oligonucleotide probe hybridization detects with the x-radiographic film under these conditions.

As mentioned above, isolating polynucleotide of the present invention comprise DNA and RNA, the method that is used for DNA isolation and RNA is well known in the prior art, and interested DNA and RNA encoding gene can be by currently known methods clones of the prior art in gene pool or DNA library.

The polynucleotide with the active polypeptide of mannase of the present invention of encoding are identified and are separated by for example hybridization or PCR subsequently.

The present invention also provides pairing polypeptide and the polynucleotide that obtained by different bacterium bacterial strain (positive isoplassont or secondary isoplassont).What cherish a special interest is by Gram-positive parent alkali bacterial strain, comprises the mannosans enzyme polypeptide that the kind of genus bacillus obtains.

Kind isoplassont with the active polypeptide of mannase of the present invention can be with information provided by the invention and component and in conjunction with conventional clone technology clone.The chromosomal DNA clone that dna sequence dna for example of the present invention can use the cell type by marking protein to obtain.The suitable source of DNA can be determined by surveying the RNA trace by the probe with sequences Design disclosed herein, chromosomal DNA by positive clone prepares the library subsequently, the dna sequence dna of the present invention that coding has the active polypeptide of mannase can separate by the whole bag of tricks subsequently, for example by surveying with the probe of disclosed sequences Design in specification sheets of the present invention and claims or based on one or more groups degeneration probe of disclosed sequence.(Mullis US4683202), uses the primer clone according to sequences Design disclosed herein for also available polymerase chain reaction of dna sequence dna of the present invention or PCR.In other method, the DNA library can be used for transforming or transfection host cell, the expression of interested DNA can be by with respect to by B.agaradherens, and NCIMB 40482 be the clone's and as Materials and Methods and expressing described in the embodiment 1 and antibody (mono-clonal or polyclone) that the mannase of purifying produces or by the activity test detection relevant with having the active polypeptide of mannase.

The mannase encoding part of being cloned among the intestinal bacteria DSM 12180 dna sequence dna among the plasmid pSJ1678 that exists and/or similar DNA sequence of the present invention can be by the bacterial strain of the bacterium kind Bacillus agaradherens of the enzyme that produces the mannosans degrading activity, preferred strain NCIMB40482, or other or associated biomolecule described herein clone.

In addition, similar sequence can be according to the dna sequence dna that can be obtained by the plasmid of existence among the intestinal bacteria DSM 12180 (it is considered to identical with appended SEQ ID NO:1), for example its subsequence constitutes, and/or by introducing the nucleotide subsitution formation, described displacement does not produce other aminoacid sequence by the mannase of dna sequence encoding, but it is selected corresponding to the codon that produces the required host living beings of enzyme, or by introducing the nucleotide subsitution formation, described displacement can produce different aminoacids sequence (being the mutation of mannosans degrading enzyme of the present invention).

Polypeptide:

The sequence of the amino acid nos.32-343 of SEQ ID NO:2 is ripe mannosans enzyme sequence.

The present invention also provides the mannosans enzyme polypeptide, and the polypeptide of it and SEQ ID NO:2 and its kind isoplassont (secondary isoplassont or positive isoplassont) is of the same race basically.Term " of the same race basically " expression that is used for this paper has 70%, preferably at least 80%, more preferably at least 85%, most preferably at least 90% with the identical polypeptide of sequence shown in the amino acid nos.32-343 of SEQ IDNO:2 or its positive isoplassont or secondary isoplassont.This polypeptide more preferably at least 95%, most preferably 98% or the amino acid nos.32-343 of higher and SEQ ID NO:2 or its positive isoplassont or secondary isoplassont shown in sequence identical.Sequence identity percentage ratio is measured by ordinary method, for example uses computer program well known in the prior art, for example the GAP that provides in the GCG routine package (Wisconsin routine package procedure manual, the 8th edition, in August, 1994, genetics computer set, 575 Science Drive, Madison, Wisconsin, USA 53711), as Needleman, S.B.and Wunsch, C.D. (1970), molecular biology magazine, 48, disclosed among the 443-453, classify this paper reference as.GAP carries out peptide sequence relatively with following setting: GAP produces and hinders 3.0, and GAP extends obstruction 0.1.

The sequence identity of peptide molecule is measured by similar approach, and GAP carries out dna sequence dna relatively with following setting: GAP produces and hinders 5.0, and GAP extends obstruction 0.3.

Zymin of the present invention is preferably by microorganism, preferably obtain by bacterium, archea or fungi, especially by bacterium, for example belong to bacillus, the bacterium of preferred close alkali Bacillus strain obtains, it can be selected from kind of Bacillus agaradherens and very relevant genus bacillus kind, and wherein all plant Bacillus agaradherens at least 95% preferred and based on the 16S rDNA sequence of arranging, and more preferably at least 98% is of the same race.Basically protein of the same race and polypeptide it is characterized by have one or more amino-acid substitutions, disappearance or additional.These change secondary properties preferably, i.e. conservative amino acid replacement (referring to table 2) and not obviously influence protein or polypeptide is folding or active other displacement; A small amount of disappearance, about 30 amino acid of 1-usually; Extend N-end methionine residues for example, the little joint peptide of about at the most 20-25 residue or help the little extension (affinity tag) of purifying, polyhistidine bundle for example, a-protein (Nilsson etc., EMBO J.4:1075,1985 with small amount of N or C-end; Nilsson etc., Methods Enzymol.198:3,1991).Referring to Ford etc., Protein Expression and Purification 2:95-107,1991, classify this paper reference as.The affinity tag of dna encoding obtains (PharmaciaBiotech for example, Piscataway, NJ by commercial supplier; New England Biolabs, Beverly, MA).Yet even above-mentioned change secondary properties preferably, but this change also can be big character, and for example nearly 300 amino acid or more big polypeptide extend as N-or C-end and be fused to mannosans enzyme polypeptide of the present invention.

Table 1

Conservative amino acid replacement

Alkalescence: arginine, Methionin, Histidine

Acid: L-glutamic acid, aspartic acid

Polarity: glutaminate, aspartate

Hydrophobic: leucine, Isoleucine, Xie Ansuan

Fragrance: phenylalanine, tryptophane, tyrosine

On a small quantity: glycine, L-Ala, Serine, Threonine, methionine(Met)

Except 20 kinds of standard amino acids, non-standard amino acid (for example 4-oxyproline, 6-N-methyllysine, the acid of 2-aminoisobutyric base, isovaline and a-methyl Serine) can be used for replacing the amino-acid residue of polypeptide of the present invention.The non-conserved amino acid that limits the number, do not use gene-code amino acids coding and the replaceable amino-acid residue of alpha-non-natural amino acid." alpha-non-natural amino acid " is being modified behind the protein synthesis and/or has the chemical structure different with standard amino acid in its side chain.But alpha-non-natural amino acid chemosynthesis, or preferred commercial obtaining, it comprises pipecolic acid, thiazolidine carboxylic acid, dehydroproline, 3-and 4-methylproline and 3,3-dimethyl proline(Pro).

Primary amino acid in mannosans enzyme polypeptide of the present invention can be definite according to method well known in the prior art, for example site-directed mutagenesis or alanine scanning mutagenesis (Cunningham and Wells, Science 244:1081-1085,1989).In one technology of back, introduce single alanine mutation on each residue in molecule, the amino-acid residue of the biological activity (being the mannosans enzymic activity) of the mutant molecule that test obtains to determine molecular activity is played a crucial role.Also referring to Hilton etc., J.Biol.Chem.271:4699-4708,1966.The reactive site of enzyme or other biological interaction also can be measured by the physical analysis of structure, as by following technology, as nucleus magnetic resonance, crystallography, electron diffraction or photoaffinity labeling, combine mensuration with the amino acid whose sudden change of contact part of supposition.Referring to for example de Vos etc., Science 255:306-312,1992; Smith etc., J.Mol.Biol.224:899-904,1992; Wlodaver etc., FEBS Lett.309:59-64,1992.The identity of primary amino acid also can be inferred by the analysis of the isoplassont with polypeptide relevant with polypeptide of the present invention.

Repeatedly amino-acid substitution can use mutagenesis, reorganization and/or confusion to prepare and test by the currently known methods of relevant scanning technique subsequently, for example by Reidhaar-Olson and Sauer (Science241:53-57.1988) .Bowie and Sauer (Proc.Natl.Acad.Sci.USA 86:2152-2156,1989), described in WO 95/17413 or the WO95/22625.In brief, these authors disclose following method, the two or more positions of randomization simultaneously in polypeptide, or the different sudden change of reorganization/confusion (WO95/17413, WO95/22625), the selection official can polypeptide subsequently, and the polypeptide of sequencing mutagenesis is to be determined at admissible metathetical spectrum on each position then.Spendable other method comprises that phage shows (Lowman etc. for example, Biochem.30:10832-10837,1991; Ladner etc., US5223409; Huse WO92/06204) and fixed regional mutagenesis (Derbyshire etc., Gene 46:145,1986; Ner etc., DNA 7:127,1988).

Above-mentioned mutagenesis/chaotic method can combine with high-throughput, automatic scanning method to detect the activity of polypeptide that clone, mutagenesis in host cell.The mutagenized dna molecule of coding active polypeptide can be reclaimed by host cell, carries out serializing rapidly with modern comfort.These methods can rapid test single amino acids residue in interested polypeptide importance, and can be used for the polypeptide of unknown structure.Use aforesaid method, those skilled in the art can identify and/or residue 32-343 each peptide species of the same race basically of preparation and SEQ ID NO:2 the mannosans enzymic activity of maintenance wild-type protein.

The protein preparation:

Protein of the present invention and polypeptide comprise overall length protein, its fragment and fused protein, can prepare in genetically engineered host cell according to routine techniques.Proper host cell is those cell types, and its available foreign DNA transforms or transfection, grows in substratum, comprises bacterium, fungal cell and the senior eukaryotic cell of cultivating.The preferred bacterium cell, especially the culturing cell of Gram-positive biology, especially the preferred gram-positive cell that obtains by bacillus, for example by subtilis, bacillus lentus, bacillus brevis, bacstearothermophilus, Alkaliphilic bacillus, bacillus amyloliquefaciens, Bacillus coagulans, Bacillus circulans, bacillus lautus, bacillus thuringiensis, Bacillus licheniformis and Bacillus agaradherens, the cell that obtains of Bacillus agarad herens especially.

The technology that is used for handling clone's dna molecular and introduces foreign DNAs at various host cells is by Sambrook etc., molecular cloning: laboratory manual, 2nd, laboratory, bay of cold spring, Cold Spring Harbor, NY, 1989; Ausubel etc., (eds.), the common protocol John of molecular biology Wiley and Sons, Inc., NY, 1987; " subtilis and other gram positive bacterium ", Sonensheim etc., 1993, U.S. microbiology association, Washington D.C. is open, classifies this paper reference as.Usually the dna sequence dna of coding mannase of the present invention is operated and is connected in because it expresses other required gene element, is usually included in transcripting promoter and terminator in the expression vector.This carrier also contains one or more selectable signs and one or more copy source usually, though person of skill in the art will appreciate that in some system, selectable sign can provide on isolated vectors, and duplicating of foreign DNA can provide by being incorporated in the host cell gene group.The selection of promotor, terminator, selectable sign, carrier and other element is those skilled in the art's a daily design transaction, and many these elements are described in the literature and can be obtained by commercial supplier.

For indicating polypeptide to enter the Secretory Pathway of host cell, in expression vector, provide secretory signal sequence (being also referred to as leader sequence, preceding former sequence or presequence).The excretory signal sequence can be peptide sequence or can be obtained or de novo synthesis by another excretory protein.Various suitable secretory signal sequences are known in the prior art, referring to " subtilis and other gram-bacteria ", Sonensheim etc., 1993, U.S. microbiology association, WashingtonD.C.; And Cutting, S.M. (eds) " the molecular biosciences method of bacillus ", John Wiley and Sons, 1990 have further described the suitable secretory signal sequence that is particularly useful for the genus bacillus secretory host cell.Secretory signal sequence is to be connected in dna sequence dna in the correct reading frame, secretory signal sequence is positioned 5 ' usually to the interested dna sequence dna of encoding, although some signal sequence can be positioned other position of interested dna sequence dna (referring to for example Welch etc., US5037743; Holland etc., US5143830).

The host cell of conversion or transfection is cultivated in the substratum medium that contains other required component of nutrient substance and selected host cell growth according to conventional methods.Various suitable medium comprise that the substratum and the complex medium of principal component are known in the prior art, generally include carbon source, nitrogenous source, primary amino acid, VITAMIN and mineral substance.Substratum also can comprise component as required, as somatomedin or serum.Growth medium will pass through usually, medicament selection for example, cell according to the DNA that contains the external source adding is selected, or selects according to the defective in the basic nutrition element, but described defective is by the selection marker complementation of transfection that have in expression vector or common to host cell.

Protein separation:

When the recombinant polypeptide of expressing was secreted, polypeptide can be by purifying in the growth medium.The host cell of expressing preferably reclaimed (for example by centrifugal) from substratum before peptide purification.

When the recombinant polypeptide of expressing is not during by secretory host cell, host cell preferably ruptures, and during polypeptide was released into moisture " extract ", this was the fs of this purification technique.Expression host cell preferably reclaimed (for example by centrifugal) in by substratum before cell fracture.

Cell fission can be undertaken by routine techniques, for example by N,O-Diacetylmuramidase digestion or by high pressure to the molecule application of force.Referring to (Robert K.Scobes, " protein purification ", Springer-Verlag), have further described this cell fission technology by the 2nd edition.

No matter whether the recombinant polypeptide (or chimeric polyeptides) of expressing is secreted its available classification and/or conventional purification process and medium purification.

Ammonium sulfate precipitation and acid or chaotropic agent extract the classification that can be used for sample.Purification step for example can comprise hydroxyapatite, size exclusion, FPLC and anti-phase high performance liquid chromatography.Suitable cation exchange medium comprises deutero-dextrin, agarose, Mierocrystalline cellulose, polyacrylamide, special silicon oxide etc.PEI, DEAE, QAE and Q derivative are preferred, and especially preferred DEAE Fast-FlowSepha rose (Pharmacia, Piscataway, NJ).Chromatographic media for example comprises with phenyl, butyl or octyl group deutero-medium, Phenyl-Sepharose FF (Pharmacia) for example, Toyopearl butyl 650 (Toso Haas, Montgomeryville, PA), Octyl-Sepharose (Pharmacia) etc.; Or polyacrylic resin, for example Amberchrom CG71 (Toso Haas) etc.Suitable solid carrier comprises glass bead, silica-based resin, celluosic resin, the thin pearl of agarose, the thin pearl of Sepharose, the thin pearl of polystyrene, cross-linked polyacrylamide resin etc., and they are undissolved under its working conditions.These carriers can be used the reactive group modification, can partly connect protein by amino, carboxyl, sulfydryl, hydroxyl and/or carbohydrate like this.The example of coupling chemical process comprises cyanogen bromide-activated, N-hydroxy-succinamide activation, epoxide activation, sulfydryl activation, hydrazides activation and is used for the carboxyl and the aminoderivative of carbonization two inferior acid amides coupling chemistry.These and other solid dielectric is well known in the prior art and widely used, can be obtained by commercial supplier.

The selection of concrete grammar is the approach design problem, partly by selected carrier character decision.Referring to for example affinity chromatography: principle and method.Pharmacopeia LKB biotechnology, Uppsala, Sweden, 1988.

Polypeptide of the present invention or its fragment also can prepare by chemosynthesis, and polypeptide of the present invention can be monomer or many bodies, glycosylation or nonglycosylated; Pegylated or non-pegylated; With can comprise or not comprise initial methionine(Met) amino-acid residue.

Based on sequence information disclosed herein, but clones coding mannase of the present invention and comprise the dna sequence dna shown in the SEQ ID No1, at least from the overall length dna sequence dna of the dna sequence dna of 97-position, position 1029.

The clone is undertaken by standard technique well known in the prior art, for example

By Bacillus strain, bacterial strain B.agaradherens especially, NCIMB 40482 preparation genomic libraries;

Inoculate this library at suitable substrate plate upper flat plate;

The clone who contains polynucleotide sequence of the present invention by use based on the standard hybridization technique identification of the probe of SEQ ID No1; Or

Use is cloned by described Bacillus agaradherens NCIMB 40482 genomic libraries identification by the inverse PCR strategy based on the primer of the sequence information of SEQ ID No1.Referring to (" the PCR practice is crossed the threshold " information publishing company, Oxford England) such as M.J.MCPherson inverse PCR is described in further detail.

According to sequence information disclosed herein (SEQ ID No1, SEQ ID No2), by similar strategy, use is by the genomic library of related microorganisms, especially by other bacterial strain of kind of genus bacillus, for example the polynucleotide sequence of the same race of the genomic library separation coding mannase of the same race of the present invention of the close alkali kind generation of genus bacillus is daily work for those skilled in the art.

In addition, encoding the DNA of mannosans of the present invention or galactomannan degradation enzyme can be according to known method easily by suitable source, for example any above-mentioned biology is by using the synthetic oligonucleotide probing pin clone based on the dna sequence dna preparation that is obtained by the plasmid that exists among the intestinal bacteria DSM 12180.

Therefore, polynucleotide molecule of the present invention can be separated by intestinal bacteria DSM 12180, and wherein the plasmid that obtains by for example above-mentioned clone is deposited.Equally, the present invention relates to the isolating pure basically biological culture thing of bacterial strain intestinal bacteria DSM12180.In the present invention, term " zymin " is meant the conventional enzymic fermentation product that the microorganism by single kind obtains, and separable and purifying, said preparation contain many different enzymic activitys usually; Or the mixture of single component enzyme, preferred enzyme by using conventional recombinant technology to obtain by bacterium or fungi kind, this enzyme oneself by fermentation with may separate respectively and purifying, it can be obtained by different sorts, preferred fungi or bacterial species; Or the tunning of microorganism, this microorganism is as the host cell of express recombinant mannase, but this microorganism produces other enzyme simultaneously, for example pectin degrading enzyme, proteolytic enzyme or cellulase, be the tunning of the natural generation of this microorganism, promptly by the conventional enzyme complex that produces of the microorganism of corresponding natural generation.

The method for preparing zymin of the present invention, this method comprises culturing micro-organisms, for example can produce the wild type strain of mannase and reclaim enzyme by culture under the generation condition that enzyme allowed.Cultivation can use conventional fermentation technique to carry out, and for example at the vibration flask or have in the fermentation container of stirring and cultivate to guarantee the abundant ventilation of growth medium, induces mannase to produce.Growth medium can contain conventional N-source, peptone for example, the conventional C-source of yeast extract or casamino acids, reduction amount, for example glucose or sucrose, and inductor, for example guar gum or carob bean gum.Recovery can use routine techniques to carry out, for example by centrifugal or filtering separation biological substance and supernatant liquor, if interested enzyme is intercellular, reclaim supernatant liquor or cell fission, perhaps described in EP406314 or described in WO97/15660, be further purified subsequently by crystallization.

The immunological cross-reaction activity:

Being used to measure the active polyclonal antibody of immunological cross-reaction can be by using pure mannase preparation.More particularly, the antiserum(antisera) of mannase of the present invention relatively can be according to N.Axelsen etc. at " quantitatively immunoelectrophoresis handbook ", Blackwell scientific publication thing, 1973, the 23rd chapter or A.Johnstone and R.Thorpe, " immunochemistry is put into practice ", Blackwell

The scientific publication thing, 1982, the method described in (27-31 page or leaf more specifically) produces by immune rabbit (or other rodents).The immunoglobulin (Ig) of purifying can by dialysis and ion exchange chromatography, for example obtain in DEAE-Sephadex subsequently by antiserum(antisera) by for example salt precipitation (ammonium sulfate).Proteinic immunochemistry sign can adopt Outcherlony double diffusion analysis, and (O.Ouchterlony is at " experiment immunization learns to do volume " (D.M.Weir, Ed) Blackwell scientific publication thing, 1967, the 655-706 page or leaf), by crossed immunoelectrophoresis (above-mentioned N.Axelsen etc., the 3rd and 4 chapters) or by rocket (rocket) immunoelectrophoresis (Axelsen etc., the 2nd chapter) undertaken.

The useful example that produces the bacterium of enzyme of the present invention or zymin is a gram positive bacterium, preferably obtain by genus bacillus/Bacterium lacticum subphylum, preferably from the bacterial strain of kind of genus bacillus, the more preferably bacterial strain of Bacillus agaradherens, especially bacterial strain Bacillus agaradherens, NCIMB40482.

The present invention includes the isolating mannase with above-mentioned character, it does not have impurity of the same race and uses conventional recombinant technology to produce.

The mensuration of mannosans enzymatic activity (ManU)

Colorimetric test: substrate: at 0.1M glycin damping fluid, the 0.2%AZCL-polygalactomannan (Megazyme, Australia) that obtains by carob bean gum among the pH10.0.Test is stirred and temperature is controlled on 40 ℃ the hot mixing tank to manage among the 1.5ml at Eppendorf Micro and carries out having.Cultivated the 0.750ml substrate 20 minutes with the 0.05ml enzyme, by stopping in centrifugal 4 minutes with 15000rpm.In the 1cm cuvette, under 600nm, measure the color of supernatant liquor.A ManU (mannosans unit of enzyme) obtains 0.24 (absolute value) in 1cm.

Obtain BACILLUS AGARADHERENS mannase NCIMB40482

Bacterial strain

Bacillus agaradherens NCIMB40482 contains the mannase of DNA sequences encoding.

Coli strain: prepare intestinal bacteria SJ2 cell (Diderichsen, B., Wedsted, U., Hede gaard, L., Jensen, B.R., Sjoholm, C. (1990) clone aldB, the extracellular enzyme that its coding for alpha-E.C. 4.1.1.4, bacillus brevis obtain, J.Bacteriol., 172,4315-4321), as mentioned above, use the Gene Pulser that obtains by BIO-RAD ^TMThe electroporation device transforms by electroporation as described in supplier.

This bacterial strain of subtilis PL2306. is to have division apr and npr gene subtilis DN 1885 (Diderichsen, B., Wedsted, U., Hedegaard, L., Jensen, B.R., Sjoholm, C. (1990) clone aldB, the extracellular enzyme that its coding for alpha-E.C. 4.1.1.4, bacillus brevis obtain, J.Bacteriol., 172,4315-4321), it divides in the transcription unit of known subtilis cellulose enzyme gene, produces the negative cell of cellulase.Divide basically as Eds.A.L.Sonenshein, J.A.Hoch and RichardLosick (1993), " subtilis and other gram-positive cell ", U.S. microorganism association carries out described in the 618th page.

As Yasbin, R.E., Wilson, G.A. and Young, F.E. (1975) " conversion in the lysogenic strain of subtilis and transfection: the proof of selecting to introduce prophage in competent cell ", preparation competent cell described in the J.Bacteriol.12l:296-304 also transforms.

Plasmid

PSJ1678 (in WO94/19454, describe in detail, classify this paper reference as).

PMOL944: this plasmid is mainly to contain subtilis, can make the element of plasmid breeding in the kalamycin resistance gene, and also has strong promoter and by the pUB110 derivative of the signal peptide of the amyL gene clone of Bacillus licheniformis ATCC14580.Signal peptide contains the Sacll position, makes its DNA of the protein maturation part that merges of clones coding and signal peptide easily.This causes preceding protein expression, and it points to the outside of cell.

Constitute plasmid by the conventional genetic engineering technique that is summarized as follows.

The formation of pMOL944:

PUB110 plasmid (McKenzie, T. etc., 1986, " plasmid " 15:93-103) with single restriction enzyme Ncil digestion, by coding plasmid pDN1981 (P.L.Jorgensen etc., 1990, " gene ", 96,37-41 page or leaf) the PCR fragment that amyL promotor is amplified digests with Ncil, inserts among the pUB110 of Ncil digestion and obtains plasmid pSJ2624.

Employed two kinds of PCR primers have following sequence:

#LWN5494 5’-GTCGCCGGGGCGGCCGCTATCAATTGGTAACTGTATC?TCAGC-3’

#LWN5495 5’-GTCGCCCGGGAGCTCTGATCAGGTACCAAGCTFGTCGA CCTGCAGAATGAGGCAGCAAGAAGAT-3’

Primer #LWN5495 inserts the Notl position of plasmid.

Plasmid pSJ2624 is subsequently with Sacl and Notl digestion, and by new PCR fragment Sacl and Notl digestion that the amyL promotor of coding pDN1981 is amplified, the pSJ2624 that this dna fragmentation is inserted Sacl-Notl digestion obtains plasmid pSJ2670.

This clones an amyL promotor that replaces using identical promoters but in the opposite direction clone, and two kinds of primers that are used for the PCR amplification have following sequence:

#LWN5938 5’-GTCGGCGGCCGCTGATCACGTACCAAGCTTGTCGACCTGCAGAAGAGGCAGCAAGAAGAT-3’

#LWN5939?5’-GTCGGAGCTCTATCAATTGGTAACTGTATCTCAGC-3’

Plasmid pSJ2670 is with restriction enzyme Pstl and Bcll digestion, and the PCR fragment of being amplified by the cloned dna sequence of coding alkali starch enzyme SP722 (open in WO95/26397, as to classify this paper reference as) obtains plasmid pMOL944 with Pstl and Bcll digestion and insertion.Two kinds of primers that are used for the PCR amplification have following sequence:

#LWN7864?5’-AACAGCTGATCACGACTGATCTTTTAGCTTGGCAC-3’

#LWN7901?5’-AACTGCAGCCGCGGCACATCATAATGGGACAAATGGG-3’

Primer #LWN7901 inserts in the Sacll position of plasmid.

Clone from the mannase gene of Bacillus agaradherens

The genomic dna preparation:

In liquid medium, breed described in bacterial strain Bacillus agaradherensNCIMB40482 such as the WO/01532.After 30 ℃ and 300rpm cultivate 16 hours, harvested cell, by (Pit chef such as Pitcher, D.G., Saunders, N.A.Owen, R.J. (1989) " with guanidine thiocyanate rapid extraction bacterial genomes DNA ", Lett.Appl.Microbiol., 8,151-156) the method isolation of genomic DNA of Miao Shuing.

The genomic library structure:

Genomic dna partly digests with restriction enzyme Sau3A, carries out the size classification by electrophoretic method on 0.7% sepharose.Size is separated (Dretzen by electrophoretic method in the fragment between the 2-7kb on the DEAE-cellulose paper, G.Bellard, M., Sassone-Corsi, P., Chambon, P. (1981) " reliable method of recovery dna fragmentation from agarose and acrylamide gel " Anal.biochem., 112,295-298).

The separated DNA fragment is connected in the pSJ1678 plasmid DNA of BamHI digestion, connects mixture and be used for transformed into escherichia coli SJ2.

The identification of just cloning:

Gou Zao e. coli dna library is scanned on the LB agar plate that contains 0.2%AZCL-polygalactomannan (Megazyme) and 9 μ g/ml paraxin and 37 ℃ of overnight incubation as mentioned above.Express the active clone of mannase and show blue look diffusion orifice ring, separate by Qiagen plasmid spin preps in the meat soup of 1ml incubated overnight (cell is containing among the TY of 9 μ g/ml paraxin and cultivates down at 37 ℃, and with the 250rpm vibration) by one of these clones plasmid DNA that obtains.

This clone (MB525) further characterizes with the segmental dna sequence dna mensuration of clone's Sau3ADNA, the terminal cyclic sequence metering equipment of dna sequence dna use Tag deoxidation (Perkin-Elmer, USA) terminator of fluorescent agent sign and (primerwalking) carry out by " primer walking " as the suitable oligonucleotide of primer.

The analysis of sequence data is according to (1984) " nucleic acids research " such as Devereux, and 12, to carry out described in the 387-395, the sequence of coding mannase shows that in SEQ ID No1 the deutero-protein sequence shows in SEQ ID No.2.

The subclone of mannase and expression in subtilis:

The mannase of dna sequence dna of the present invention of encoding is the PCR that uses the PCR primer sets be made up of following two kinds of oligonucleotide to amplify:

Mannase, on, Sacll

5’-CATTCTGCAGCCGCGGCAGCAAGTACAGGCTTTTATGTTGATGG-3’

Mannase, down, Notl

5’-GACGACGTACAAGCGGCCGCGCTATTTCCCTAACATGATGATATTT?TCG-3’

Underlined is agretope Sacll and Notll.

Be used as template by the instruction according to manufacturers in the PCR reaction of using Amplitaq archaeal dna polymerase (Perkin Elmer) of the isolating chromosomal DNA of B.agaradherensNCIMB40482 as mentioned above.PCR be reflected at the various dNTP that contain 200 μ M, 2.5 units the AmpliTaq polysaccharase (Perkin-Elmer, Cetus, USA) and PCR damping fluid (the 10mM Tris-HCl of the various primers of 100pmol, pH8.3,50mM KCl, 1.5mM magnesium chloride, 0.01% (w/v) gelatin) middle foundation.

(Landgraf Germany) carries out with the DNA thermo cycler in the PCR reaction.Cultivated 1 minute at 94 ℃, use subsequently, carry out 30 PCR circulations 60 ℃ of renaturation 1 minute with 72 ℃ of Recycle design of extending 2 minutes 94 ℃ of sex change 30 seconds.(NuSieve amplifies the separatory such as 5 μ l of product with the electrophoretic method analysis in FMC) at 0.7% sepharose.The outward appearance of dna fragmentation size 1.4kb is represented the suitable amplification of gene fragment.

The segmental subclone of PCR

(Qiagen, USA) purifying, the DNA of purifying be at 50 μ l 10mM Tris-HCl, wash-out among the pH8.5 with the QlAquickPCR purifier apparatus according to the instruction of manufacturers for separatory such as 45 μ l of the PCR product of above-mentioned generation.

The PCR fragment of 5 μ g pMOL944 and 25 μ l purifying digests with Sacll and Notl, agarose (SeaPlaque GTG at 0.8% low gel temperature, FMC) electrophoresis in the gel, the relevant fragment of excision from gel, according to the instruction of manufacturers QlAquick gel extraction equipment (Qiagen, USA) purifying.Isolating pcr dna fragment is connected in the pMOL944 of Sacll-Notl digestion and purifying subsequently.Connect that (Boeh ringer Mannheim Germany) spends the night at 16 ℃ with the T4DNA ligase enzyme of the various dna fragmentations of 0.5 μ g, 1U and T4 ligase enzyme damping fluid.

Connect mixture and be used for transformed competence colibacillus subtilis PL2306, cell transformed is carried out plating on LBPG-10 μ g/ml kantlex flat board.37 ℃ cultivate 18 hours after, on flat board, can see the clone, by by the some clones of the isolating plasmid DNA analysis of incubated overnight meat soup.

A kind of this just is being cloned in rules (restreak) several times again in the above-mentioned used agar plate, this clone is called MB594.37 ℃ of grow overnight, the instruction that was used for the subtilis process for preparing plasmid in second day according to manufacturers uses the 1ml cell from the cell separation quality grain with QiaprepSpin Plasmid Miniprep Kit #27106 to clone MB594 at TY-10 μ g/ml kantlex.This DNA is through dna sequence dnaization and show its maturing part corresponding to mannase, the position 94-1404 of promptly appended SEQ ID NO:3.The deutero-mature protein shows in SEQ ID NO:4.It shows by 3 ' end of the mannase of the sequence encoding of SEQ ID NO:1 changes into the sequence shown in the SEQ ID NO:3 owing to be used for the low primer design of PCR.The aminoacid sequence that obtains is shown among the SEQ ID NO:4, and it shows that the C of SEQ ID NO:2 holds (SHHVREIGVQFSAADNS SGQTALYVDNVTLR) to change into the D end (IIMLGK) of SEQ ID NO:4.

Substratum:

TY (as Ausubel, F.M. etc. (eds.) " molecular biology common protocol book " .JobnWiley and Sons is described in 1995).

LB agar (as Ausubel, F.M. etc. (eds.) " molecular biology common protocol book " .JohnWiley and Sons is described in 1995).

LBPG is with 0.5% glucose and 0.05M potassiumphosphate, the LB agar (as mentioned above) that pH7.0 replenishes

The BPX substratum is described in EP0506780 (WO91/09129).

Expression, purifying and the sign of the mannase that obtains by Bacillus agaradherens

The clone MB594 that obtains described in above-mentioned material and method was containing in 25 * 200mlBPX substratum of 10 μ g/ml kantlex under 37 ℃ and 300rpm growth 5 days in the two baffle plates vibration of 500ml flask.

Collect the vibration flask nutrient solution (lot number #9813) of 6500ml clone MB594, pH regulator to 5.5.Adding 146ml cationoid reagent (C521) and 292ml reagents for anion (A130) flocculate in whipping process.By centrifugal 20 minute separating the material that flocculate at 6 ℃ with 9000rpm with Sorval RC 3B sedimentator, supernatant liquor finally cuts 10kDa with filtron and concentrates with Whatman glass filter GF/D and C clarification.With sodium hydroxide the 750ml enriched material is adjusted to pH7.5, settled solution is used for anion-exchange chromatography, uses by 50mmolTris pH7.5 equilibrated 900ml Q-Sepharose post.With sodium-chlor gradient elution mannosans enzymic activity binding substances.

Pure enzyme obtains single band in SDS-PAGE, molecular weight is 38kDa, the aminoacid sequence of mannase, and promptly Fan Yi dna sequence dna is shown among the SEQ ID No.2.

Kinetic constant is measured:

Substrate: carob bean gum and go back raw sugar analysis (PHBAH).

The carob bean gum (G-0753) that obtains by Sigma.

Kinetic determination carried out 20 minutes in the pH10 cultivation at 40 ℃ with the carob bean gum of different concns

The Kcat:467 per second

Km：0.08g/l

MW：38kDa

Pl (iso-electric point): 4.2

The temperature optimum value of mannase is 60 ℃.

The pH activity curve shows that maximum activity is between pH8-10.

It is 77 ℃ that the DSC differential scanning calorimeter obtains at the fusing point of pH7.5 in the Tris damping fluid, shows that this enzyme is very heat-staple.

Use the 0.2%AZCL-polygalactomannan that obtains by carob bean gum as substrate with as mentioned above 40 ℃ of compatible abilities demonstrations of washing composition and the outstanding consistency of conventional liq washing composition and the consistency good of cultivating mensuration down with conventional powder detergent.

The acquisition of subtilis mannase 168

Sign and purifying subtilis 'beta '-mannase are as follows:

With the genomic isoplassont of known bacillus beta-mannase gene sequence (Mendoza etc., Biochemicaet Bio physica Acta 1243:552-554,1995) search subtilis.The coding region that its product still belongs to unknown ydhT shows that 58% is similar to known genus bacillus 'beta '-mannase.Following oligonucleotide is used to amplify the sequence of maturing part of the 'beta '-mannase of coding supposition: 5 '-GCT CAA TTG GCG CAT ACT GTG TCG CCT TGT-3 ' and 5 '-GAC GGA TCC CGG ATT CAC TCA ACG ATT GGC G-3 '.The total genome that is obtained by bacillus subtilis strain 1A95 uses above-mentioned primer to amplify the ripe zone of ydhT as template.(Foster City CA) carries out PCR for Perkin Elmer, Applier Biosystems with the AMPLITAQ archaeal dna polymerase with GENE-AMP PCR Kit.At first, carry out 25 following programs of round-robin subsequently:,, extended 2 minutes at 72 ℃ 55 ℃ of renaturation 2 minutes 95 ℃ of fusings 1 minute 95 ℃ of fusings 5 minutes.In the end after the circulation, be reflected at 72 ℃ and keep 10 minutes to finish extension, PCR product QlAquick PCR purifier apparatus (Qiagen, Chatsworth, CA) purifying.

Insert as follows among the expression vector pPG1524 (the previous description) by the ripe zone of ydhT that bacillus subtilis strain 1A95 amplifies.The 1028bp fragment of amplifying uses EcoR I and BamH I to digest with Mfe I and BamH I digestion, expression vector pPG1527.Restriction product QlAquickPCR purifier apparatus (Qiagen, Chatsworth, CA) purifying.Two fragments connect (13 hours, 16 ℃) with the T4DNA ligase enzyme, are used for transformed competence colibacillus coli strain DH5-α.For preparation DNA cultivates the amicillin resistance clone.DNA characterizes with restriction analysis subsequently.Plasmid pPG3200 contains the ripe zone of ydhT gene, and plasmid pPG3200 is used for transformed competence colibacillus bacillus subtilis strain PG632 (Saunders etc., 1992) subsequently.

Select 7 kalamycin resistance subtilis clones and 1 PG632 contrast clone, growth in 20ml 20/20/5 substratum (20g/l tryptone, 20g/l yeast extract and 5g/l sodium-chlor) that replenishes with 1ml25%maltrin, 120 μ l 10mM Manganous chloride tetrahydrates and 20 μ l 50mg/ml kantlex.Be cloned in 250ml with in the baffle plate flask of 250rpm vibration 37 ℃ of following grow overnight with marking protein.Cell was with 14000rpm spin 15 minutes, each 1 μ l supernatant liquor dilutes with the 50mM sodium acetate (pH6.0) of 99 μ l, with interior-1, (Megazyme Ireland) analyzes 4-'beta '-mannase β-Mannazyme Tabs 1 this diluent of μ l according to the instruction of manufacturers.Measure light absorption ratio at Beckman DU640 spectrophotometer at 590nm, clone 7 shows 1.67 the highest light absorption ratio, and PG632 does not show light absorption ratio to impinging upon 590nm.

(Novex, San Diego analyze to confirm the desired protein size of 38kDa by SDS-PAGE in Ca) supernatant liquor in the 10-20%Tris-glycine gels.Specimen preparation is as follows, 500 μ l samples of ydhT clone 7 and PG632 supernatant liquor precipitate with 55.5 μ l100% trichoroacetic acid(TCA)s (Sigma), with the washing of 100 μ l5% trichoroacetic acid(TCA)s, be suspended in again in the 50 μ lTris-glycine SDS sample damping fluids (Novex), seethed with excitement 5 minutes.1 μ l per sample (p.s.) in gel under 30mA electrophoresis 90 minutes, ydhT clone 7 is observed big protein belt at 38kDa.

10 liters of fermentations of subtilis ydhT clone 7 are carried out in B.Braun Biostat C fermentation container, and fermentation condition is as follows, cell in being similar to 20/20/5 rich substratum 37 ℃ of growths 18 hours down.When fermentation test finishes, remove cell, with tangential flow filtration system concentrated supernatant to 1 liter, the ultimate yield of the 'beta '-mannase in concentrated supernatant is determined as 3g/l.

Be performed as follows by fermented supernatant fluid purifying 'beta '-mannase: 4 ℃ with 500ml supernatant liquor centrifugal 10 minutes at 10000rpm, the centrifugal supernatant liquor cuts off film (Spectrum) dialysed overnight by Spectrapor 12000-14000mol.wt. at 4 ℃ in two 4 liters 10mM potassiumphosphates (pH7.2) conversion container.The supernatant liquor of dialysis 4 ℃ under 10000rpm centrifugal 10 minutes.200mlQ Sepharose flow fast (Pharmacia) ion exchange column with 1 liter of 10mM potassiumphosphate (pH7.2) 20 ℃ of following balances, load 300ml supernatant liquor in post.Collect two and flow through cut 210ml (Sample A) and 175ml (sample B), two cuts are as above analyzed, and just sample dilutes with 199 μ l 50mM sodium acetates (pH6.0), and they show the light absorption ratio of .38 and .52 respectively.The 2ul per sample (p.s.) add 8 μ lTris-glycine SDS sample damping fluids (Novex, CA) in, seethed with excitement 5 minutes.The sample that generates 10-20% the Tris-glycine gels (Novex, Ca) under 30mA electrophoresis 90 minutes, in per sample (p.s.), have main light belt corresponding to 38kDa, account for more than 95% of gross protein.Instruction according to manufacturers is carried out BCA protein test (Pierce) as standard to two kinds of samples with bovine serum albumin, and Sample A and sample B contain 1.3mg/ml and 1.6mg/ml 'beta '-mannase respectively.Proteinic evaluation is confirmed by ionspray mass spectrum and n terminal amino acid sequential analysis.

The 'beta '-mannase sample of purifying is following to be used to characterize enzymic activity.Above-mentioned inscribe-1,4-'beta '-mannase β-Mannazyme Tabs (Megazyme Ireland) are used in all tests.Activity at pH scope 3.0-9.0 is carried out in 50mM Citrate trianion phosphate buffered saline buffer, for the determination of activity of pH9.5, adopts 50mM CAPSO (Sigma), and for the pH10.0-11.0 scope, then adopts the 50mMCAPS damping fluid.The best pH of subtilis 'beta '-mannase is pH6.0-6.5.The temperature activity curve carries out in 50mM Citrate trianion phosphoric acid salt (pH6.5), and this enzyme shows optimum activity at 40-45 ℃.Be lower than 15 ℃ and when being higher than 80 ℃ the subtilis 'beta '-mannase keep significantly active.According to the guidance of manufacturers, with inscribe-1,4-'beta '-mannase β-Mannazyme Tabs (Megazyme Ireland) measure for β-1, the ratio work of 4-polygalactomannan is 160 000 μ mol/min.mg 'beta '-mannases.The Nucleotide of subtilis 'beta '-mannase and aminoacid sequence are shown in SEQ ID NO5 and 6.

Mannase is preferably to press composition weight meter 0.0001%-2%, more preferably 0.0005%-0.1%, and most preferably the content of the pure enzyme of 0.001%-0.02% adds in the composition of the present invention.

Enzyme of the present invention except the enzyme nuclear that contains catalytic domain, also contains cellulose binding domain (CBD), and the cellulose binding domain of enzyme and enzyme nuclear (catalytic activity territory) are operatively connected.The integral part that cellulose binding domain (CBD) can be used as codase exists, or can introduce enzyme from the CBD in other source, so form the enzyme hybrid.In this article, term " cellulose binding domain " can be regarded as by Peter Tomme etc., " cellulose binding domain: classification and character ", at " enzyme liberating of insoluble carbohydrate ", John N.Saddler and Michael H.Penner (Eds), ACS collection of thesis series, No.618, in 1996 define.This definition will be divided into 10 families (I-X) above 120 kinds of cellulose binding domains, illustrate that CBD is present in various enzymes, for example among cellulase, zytase, mannase, arabinofuranosidase, acetyl esterase and the chitinase.CBD for example finds among the red algae Porphyra purpurea as non-Polysaccharides conjugated protein also in algae, referring to the document of quoting as proof more than Tomme etc.Yet most of CBD are from cellulase and zytase, and CBD is at proteinic N and C end or in the centre.The enzyme hybrid is known in the prior art, for example referring to WO90/00609 and WO95/16782, can prepare to express fusion gene by DNA structure being transformed into host cell and growth host cell, described DNA structure contain at least one by or be not connected in the dna fragmentation of coding cellulose binding domain of the dna sequence dna of coding mannase by joint.The enzyme hybrid can be described by following formula:

CBD-MR-X wherein CBD is N-end or a C-end regions corresponding to the aminoacid sequence of cellulose binding domain at least; MR is the region intermediate connection portion, and can be key, or about 100 carbon atoms of preferably about 2-, the more preferably short connection chain of 2-40 carbon atom; Or about 100 amino acid of preferably about 2-, more preferably 2-40 amino acid; With X be the N-end or the C-end regions of enzyme of the present invention.

Above-mentioned enzyme can be any suitable source, for example plant, animal, bacterium, fungi and yeast source.The source can also be have a liking for temperature or have a liking for limiting condition (extremophilic) (have a liking for cold, cold nutrition, thermophilic, have a liking for pressure, have a liking for alkali, have a liking for acid, have a liking for halogen etc.).Pure or the impure form of these enzymes can both be used.At present, in common reality through protein/genetic engineering technique modification agriotype enzyme with their effectiveness of performance in washing composition of the present invention and/or Fabrid care composition of optimizing.For example but design variable is to increase the consistency of the component that runs into usually in enzyme and the said composition.In addition, but design variable makes best pH, SYNTHETIC OPTICAL WHITNER or sequestrant stability, the catalytic activity etc. of enzyme variants can adapt to specific washing uses.

Especially should notice that under the situation of bleach stability amino acid should note surface charge to the susceptibility of oxidation with for the compatible ability of tensio-active agent.The iso-electric point of this kind of enzyme can be passed through some charged amino acid whose displacement modification, for example increases iso-electric point and can help to improve consistency with anion surfactant.The stability of enzyme can further be improved by producing for example additional salt bridge, and the reinforced metal binding site is to increase sequestrant stability.

Cats product

Second kind of basal component of the present invention is cats product.The preferred cation detergent surfactant is the material that contains a long chain hydrocarbon groups, comprises ammonium surfactant and their ethoxylated derivative.

The preferred embodiment of this cats product comprises ammonium surfactant, for example alkyl trimethyl ammonium halogenide and have the tensio-active agent of following formula:

[R ²(OR ³) _y] [R ⁴(OR ³) _y] ₂R ⁵N ⁺X ^-R wherein ²Be alkyl or the alkyl benzyl that in alkyl chain, contains about 18 carbon atoms of the 8-that has an appointment, each R ³Be selected from-CH ₂CH ₂-,-CH ₂CH (CH ₃)-,-CH ₂CH (CH ₂OH)-,-CH ₂CH ₂CH ₂-and their mixture; Each R ⁴Be to be selected from C ₁-C ₄Alkyl, C ₁-C ₄Hydroxyalkyl, by connecting two R ⁴The benzyl rings structure that group forms ,-CH ₂CHOH-CHOHCOR ⁶CHOHCH ₂OH, wherein R ⁶Be that any hexose or molecular weight are lower than about 1000 hexose polymkeric substance and when y is not 0, are hydrogen; R ⁵With R ⁴Identical or alkyl chain, wherein R ²Add R ⁵The total number of carbon atoms be no more than about 18; Each y is that 0-is about 10, and the summation of y value is 0-about 15; With X be any compatible negatively charged ion.

Be applicable to that quaternary ammonium surfactant of the present invention has as shown in the formula (I):

Formula I wherein R1 is a short-chain alkyl (C6-C10) or as shown in the formula the alkyl amido alkyl of (II):

Formula II

Y is 2-4, and is preferred 3,

Wherein R2 is H or C1-C3 alkyl,

Wherein x is 0-4, preferred 0-2, and most preferably 0,

Wherein R3, R4 and R5 are identical or different, can be the alkoxylated alkyl of short-chain alkyl (C1-C3) or following formula III,

X wherein ^-Be counter ion, preferred halogenide, for example muriate or methylsulfate,

Formula III

R6 is C ₁-C ₄With z be 1 or 2.

Preferred quaternary ammonium surfactant is the compound by formula I definition, wherein

R ₁Be C ₈, C ₁₀Or their mixture, x=0,

R ₃, R ₄=CH ₃And R ₅=CH ₂CH ₂OH.

Preferred cats product is the water-soluble quaternary ammonium compound with following formula that is used for the present composition:

R ₁R ₂R ₃R ₄N ⁺X ^-(i) R wherein ₁Be C ₈-C ₁₆Alkyl, each R ₂, R ₃And R ₄Be respectively C ₁-C ₄Alkyl, C ₁-C ₄Hydroxyalkyl, benzyl and-(C ₂H ₄O) _xH, wherein x is 2-5, X is a negatively charged ion.Be no more than one R ₂, R ₃Or R ₄It should be benzyl.

R ₁The preferred alkyl chain length be C ₁₂-C ₁₅, especially wherein alkyl is by Oleum Cocois or palm nuclear fat mixture that obtain or or chain length that OXO pure synthetic method obtain synthetic by alkene.R ₂, R ₃And R ₄Preferred group be methyl and hydroxyethyl, negatively charged ion X can be selected from halogenide, methylsulfate, acetate moiety and phosphate anion.

The example that is used for suitable formula (i) quaternary ammonium compound of the present invention comprises:

Oleum Cocois trimethyl ammonium muriate or bromide;

Oleum Cocois methyl dihydroxy ethyl ammonium muriate or bromide;

Decyl triethyl ammonium muriate;

Decyl dimethyl hydroxyl ethyl ammonium muriate or bromide;

C _12-15Dimethyl hydroxyl ethyl ammonium muriate or bromide;

Oleum Cocois dimethyl hydroxyl ethyl ammonium muriate or bromide;

Tetradecyl trimethyl ammonium Methylsulfate;

Dodecyl dimethyl hexadecyldimethyl benzyl ammonium muriate or bromide;

Dodecyl dimethyl (oxyethane) ₄Ammonium muriate or bromide;

Cholinesterase (formula (i) compound, wherein R ₁Be CH ₂-CH ₂-O-C (O)-C _12-14Alkyl and R ₂, R ₃And R ₄Be methyl);

Dialkylimidazolium quinoline [formula (i) compound].

Be used for US4228044 and EP000224 that other cats product of the present invention also is described in the Cambre of promulgation on October 14th, 1980.

Typical cats product/fabric sofetening component comprises the soft active substance of water-insoluble quaternary ammonium fabric or their corresponding amine precursors, and the most frequently used is ammonium chloride or the methylsulfuric acid ammonium that contains two chain alkyl chains.

Wherein preferred monoalkyl cationic softening agent comprises:

1) stearyl benzyl dimethyl ammonium chloride;

2) Tallow, beef base trimethyl ammonium chloride;

3) hydrogenated tallow base trimethyl ammonium chloride;

4) C12-14 alkyl hydroxy ethyl alkyl dimethyl ammonium chloride;

5) C12-18 alkyl dihydroxy ethyl ammonio methacrylate.

Being used for preferred cats product of the present invention is C12-14 and C8-C11 alkyl hydroxy ethyl alkyl dimethyl ammonium chloride.

Same suitable be wherein dialkyl cationic softening agent, comprising:

1) two Tallow, beef alkyl dimethyl ammonium chlorides (DTDMAC);

2) dihydro Tallow, beef alkyl dimethyl ammonium chloride;

3) dihydro Tallow, beef dimethyl methyl ammonium sulfate;

4) VARISOFT TA100;

5) two oil base alkyl dimethyl ammonium chlorides;

6) two palmityl hydroxyethyl ammonio methacrylates;

7) two (stearoyl-oxy ethyl) alkyl dimethyl ammonium chlorides (DSOEDMAC);

8) two (Tallow, beef oxygen base ethyl) alkyl dimethyl ammonium chloride;

9) two Tallow, beef base imidazoles Methylsulfates;

10) 1-(2-Tallow, beef amido ethyl)-2-Tallow, beef base imidazoles Methylsulfate.

If comprise, it is about 25% that washing composition of the present invention and/or Fabrid care composition contain by weight 0.2%-usually, preferably about 1%-about 8% this cats product.

Detergent component

Washing composition of the present invention and/or Fabrid care composition must contain at least a other detergent component.The specific nature of these additional components and their add-on will be according to the physical form of composition and the character decisions of employed washing operation.

Washing composition of the present invention and/or Fabrid care composition preferably also contain the detergent component that is selected from other tensio-active agent, especially anion surfactant, preferred alkyl vitriol and/or sulfonate and/or nonionogenic tenside, for example have the C8-C20 chain length, preferred C12-C16, with degree of ethoxylation be 2-9, alkyl ethoxylated nonionogenic tenside and/or the alkyl chain length of preferred 3-7 are C8-C20, the alkyl methyl glucamide tensio-active agent of preferred C12-C18.

Washing composition of the present invention and/or Fabrid care composition can be liquid, paste, gel, piece, sheet, sprays, bubble end, powder or particle.Particulate composition can also be " densification " form, and liquid composition can be " concentrating " form.

In preferred embodiments, the present invention relates to contain the laundry detergent composition (embodiment 1-5) of mannase and cats product.In second embodiment, the present invention relates to dishwashing detergent or household detergent composition (embodiment 6-8).

Composition of the present invention can be mixed with for example hand dishwashing composition, craft and machine laundry cleaning composition, the composition that it comprises laundry additive composition and is applicable to the immersion and/or the pretreated composition of dirty fabric and is applicable to family's hard-surface cleaning operation.

When being formulated as the composition that is used for the manual dishwashing method, composition of the present invention preferably contains tensio-active agent and is preferably selected from other detergent compound of organic polyhydroxyl compound, suds booster, II family metal ion, solvent, solubilizing agent and additional enzyme.

When being formulated as the composition of the clothes washing method that is used to machine-wash, composition of the present invention preferably contains tensio-active agent and washing-aid compound and additional one or more detergent component simultaneously, and it is preferably selected from organic polymer, SYNTHETIC OPTICAL WHITNER, additional enzyme, suds suppressor, dispersion agent, lime soap dispersing agent, soil-suspending agent and anti-redeposition agent and corrosion inhibitor.Laundry composition also can contain the softening agent as additional detergent components.When being formulated as laundry detergent composition, the said composition that contains mannase and cats product can provide fabric washing, decontamination, softness and color outward appearance.

Composition of the present invention also can be used as the detergent additives product of solid or liquid form.This additive product is used for replenishing or strengthening the performance of conventional detergent composition, and can add in any stage of washing process.

As required, the density of laundry detergent composition of the present invention is to measure the 400-1200 grams per liter down at 20 ℃, preferred 500-950 grams per liter composition.

" densification " form of the present composition can preferably reflect with density, for composition, influenced by the amount of mineral filler salt; Mineral filler salt is the conventional component of the detergent composition of powder type; In conventional detergent composition, filling salt exists with fundamental quantity, is generally by general composition weight meter 17-35%.

In compact composition, filling salt preferably is no more than 10% to be no more than total composition 15% by composition weight meter, is most preferably not exceeding 5% amount existence.Mineral filler salt in the present composition for example is meant the vitriol and the muriate of basic metal and alkaline-earth metal.Preferred filling salt is a sodium sulfate.

Liquid washing agent of the present invention and/or Fabrid care composition also can be " conc forms ", in this case, compare with the conventional liq washing composition, and liquid detergent composition of the present invention will contain the water of low amount.The water content of concentrated liquid detergent is generally by washing composition and/or Fabrid care composition weight and is lower than 40%, more preferably less than 30%, most preferably is lower than 20%.

Be used for suitable detergent compound of the present invention and be selected from compound as described below.

Surfactant system

Except cats product, washing composition of the present invention and/or Fabrid care composition can contain other surfactant system usually, and wherein tensio-active agent can be selected from other cats product, nonionic and/or negatively charged ion and/or both sexes and/or zwitter-ion and/or semi-polarity tensio-active agent.Washing composition of the present invention and/or Fabrid care composition preferably also contain other tensio-active agent, especially anion surfactant, preferred alkyl vitriol and/or sulfonate and/or nonionogenic tenside, for example have the C8-C20 chain length, preferred C12-C16, with degree of ethoxylation be 2-9, alkyl ethoxylated nonionogenic tenside and/or the alkyl chain length of preferred 3-7 are C8-C20, the alkyl methyl glucamide tensio-active agent of preferred C12-C18.We are surprised to find said composition scourability preferably are provided.

Other tensio-active agent exists with the amount of 0.1%-60% by weight usually.In washing composition of the present invention and/or Fabrid care composition, more preferably add-on is 1%-35% by weight, most preferably 1%-30% by weight.

Tensio-active agent preferably be mixed with composition in the enzyme component compatibility that exists.In liquid or gelatinous composition, tensio-active agent most preferably is mixed with it is promoted, or does not destroy the stability of any enzyme in these compositions at least.

The polyoxyethylene of alkylphenol, polyoxypropylene and polyoxy croton condensation thing are suitable as the nonionogenic tenside of surfactant system of the present invention, wherein preferred polyoxyethylene condenses.These compounds comprise having and contain about 14 carbon atoms of about 6-, the alkylphenol of the alkyl of the straight or branched configuration of about 14 carbon atoms of preferably about 8-and the condensation product of alkylene oxide.In preferred embodiments, oxyethane equals about 25 moles of about 2-with every mole of alkylphenol, and more preferably from about the amount of about 15 moles of ethylene oxide of 3-exists.Commercial available such ionic surfactant pack is drawn together the Igepal that is sold by GAF company ^TMCO-630 is by Rohm ﹠amp; The Triton that Haas company sells ^TMX-45, X-114, X-100 and X-102.These tensio-active agents are commonly referred to alkylphenol alcoxylates (for example alkyl phenol ethoxylate).

The condensation product of the oxyethane that primary and secondary fatty alcohol and about 1-are about 25 moles is suitable as the nonionogenic tenside of nonionic surfactant system of the present invention.The alkyl chain of fatty alcohol can be a straight or branched, and uncle or secondary contains about 22 carbon atoms of the 8-that has an appointment usually.Preferably have and contain about 20 carbon atoms of about 8-, more preferably contain the alcohol of alkyl of about 18 carbon atoms of about 10-and the condensation product of about 10 moles of ethylene oxide of the about 2-of every mol of alcohol.Every mol of alcohol exists about 7 moles of ethylene oxide of about 2-, most preferably 2-5 moles of ethylene oxide in described condensation product.The example of commercial available such nonionogenic tenside comprises Tergitol ^TM15-S-9 (C ₁₁-C ₁₅The condensation product of straight chain alcohol and 9 moles of ethylene oxide), Tergitol ^TM24-L-6NMW (C with narrow molecular weight distributions ₁₂-C ₁₄The condensation product of primary alconol and 6 moles of ethylene oxide), sell by Union Carbide Corp; Neodol ^TM45-9 (C ₁₄-C ₁₅The condensation product of straight chain alcohol and 9 moles of ethylene oxide), Neodol ^TM23-3 (C ₁₂-C ₁₃The condensation product of straight chain alcohol and 3.0 moles of ethylene oxide), Neodol ^TM45-7 (C ₁₄-C ₁₅The condensation product of straight chain alcohol and 7 moles of ethylene oxide), Neodol ^TM45-5 (C ₁₄-C ₁₅The condensation product of straight chain alcohol and 5 moles of ethylene oxide), by the sale of shell chemical company, Kyro ^TMEOB (C ₁₃-C ₁₅The condensation product of alcohol and 9 moles of ethylene oxide), by Procter ﹠amp; Gamble sells; With Genapol LA 030 or 050 (C ₁₂-C ₁₄The condensation product of alcohol and 3 or 5 moles of ethylene oxide), sell by Hoechst.The preferred HLB scope of these products is 8-11, most preferably 8-10.

The nonionogenic tenside that also can be used as surfactant system of the present invention is a disclosed alkyl polysaccharide in the US4565647 of the Llenado of promulgation on January 21st, 1986, it contains about 30 carbon atoms of the 6-that has an appointment, the hydrophobic grouping and the polysaccharide of preferred about 16 carbon atoms of about 10-, polysaccharide glycosides for example, hydrophilic radical contains the 1.3-that has an appointment about 10, preferred about 1.3-is about 3, most preferably from about about 2.7 sugar units of 1.3-.Can use any recuding sugars that contains 5 or 6 carbon atoms, glucose for example, semi-lactosi and galactosyl can be used for substituting glucosyl (hydrophobic group optionally is connected positions such as 2-, 3-, 4-, thereby obtains glucose or semi-lactosi with respect to glucoside or galactoside).In sugared key can be between 2-, 3-, 4-and/or the 6-position of position of the sugar unit that for example adds and previous sugar unit.

Preferred APG has following formula:

R ²O (C _nH _2nO) _t(glycosyl) _xR wherein ²Be selected from alkyl, alkyl phenyl, hydroxyalkyl, hydroxyalkyl phenyl and their mixture, wherein to contain the 10-that has an appointment about 18 for alkyl, about 14 carbon atoms of preferably about 12-; N is 2 or 3, preferred 2; T is that 0-is about 10, and is preferred 0, and x is that about 1.3-is about 10, and preferably about 1.3-is about 3, and most preferably from about 1.3-about 2.7.Glycosyl is preferably obtained by glucose.For preparing these compounds, at first form the many ethoxy alcohols of alcohol or alkyl, form glucoside (being connected the 1-position) with glucose or source of glucose reaction subsequently.Other glycosyl can be connected between its 1-position and previous sugar unit 2-, 3-, 4-and/or the 6-position, preferably mainly in the 2-position subsequently.

Oxyethane also is suitable as additional nonionic surfactant system of the present invention with the condensation product of the hydrophobic grouping that forms by condensed epoxy propane and propylene glycol.The hydrophobic part of these compounds preferably has about 1500 to about 1800 molecular weight, and shows the water insoluble.Add the water-soluble that polyoxyethylene group will increase the branch subpopulation in this hydrophobic part, the fluid characteristics of product is retained to about 50% this point that polyoxyethylene content is the condensation product gross weight, and this is equivalent to be condensed to many about 40 moles oxyethane.Such examples for compounds comprises some commercially available Plurafac ^TMLF404 and Pluronic ^TMTensio-active agent is sold by BASF.

The nonionogenic tenside that is suitable as nonionic surfactant system of the present invention equally is an oxyethane and the condensation product of the product that obtains by propylene oxide and reacting ethylenediamine.The hydrophobic grouping of these products is made up of the reaction product of quadrol and excessive propylene oxide, 2500 to about 3000 the molecular weight of having an appointment usually.This hydrophobic grouping and ethylene oxide condensation are to making condensation product contain the polyoxyethylene of about by weight 40%-about 80% and having the degree of the molecular weight of about 5000-about 11000.The example of such nonionogenic tenside comprises some commercially available Tetronic ^TMCompound is sold by BASF.

The nonionogenic tenside that is preferably used as surfactant system of the present invention is the polyethylene oxide condensation compound of alkylphenol, condensation product, alkyl polysaccharide and their mixture of primary and secondary fatty alcohol and about 25 moles of ethylene oxide of about 1-.The C that most preferably contains the 3-15 oxyethyl group ₈-C ₁₄Alkyl phenol ethoxylate and the C that contains 2-10 oxyethyl group ₈-C ₁₈Alcohol ethoxylate (preferred average C ₁₀) and their mixture.

Preferred nonionogenic tenside is the following formula polyhydroxy fatty acid amide surfactant:

R ²-C (O)-N (R ¹)-Z is R wherein ¹Be H, or R ¹Be C _1-4Alkyl, 2-hydroxyethyl, 2-hydroxypropyl or their mixture, R ²Be C _5-31Alkyl and Z have at least 3 polyhydroxy alkyls with the straight-chain alkyl chain of the direct-connected hydroxyl of chain, or their alkoxy derivative.Preferred R ¹Be methyl, R ²It is straight chain C _11-15Alkyl or C _16-18Alkyl or alkenyl, for example Oleum Cocois alkyl or their mixture and Z by in reductive amination process by reducing sugar, for example glucose, fructose, maltose, lactose obtain.

Spendable suitable anion surfactant is linear alkylbenzene sulfonate, alkyl sulfonate surfactants, and it comprises C ₈-C ₂₀The linear ester of carboxylic acid (being lipid acid), this ester are according to " american petroleum chemistry meeting will ", the method gaseous sulfur trioxide sulfonation in 52 (1975) the 323-329 pages or leaves.Proper raw material will comprise the natural fat material that is obtained by Tallow, beef, palm wet goods.

The preferred alkyl sulfonate surfactants that is particularly useful for laundry applications comprises the alkyl sulfonate surfactants that structural formula is following:

R ³-CH (SO ₃M)-C (O)-OR ⁴R wherein ³Be C ₈-C ₂₀Alkyl, preferred alkyl or their mixture, R ⁴Be C ₁-C ₆Alkyl, preferred alkyl or their mixture, M is a positively charged ion, it and alkyl ester sulfonic acid form water-soluble salt.Suitable salt-forming cation comprises metal, for example sodium, potassium and lithium and replacement or unsubstituted ammonium cation, for example monoethanolamine, diethanolamine and trolamine.Preferred R ³Be C ₁₀-C ₁₆Alkyl, and R ⁴Be methyl, ethyl or sec.-propyl.Especially preferred is R wherein ³Be C ₁₀-C ₁₆The methyl ester sulfonate of alkyl.

Other suitable anion surfactant comprises alkyl sulfate surfactant, and it is formula ROSO ₃The water-soluble salt of M or acid, wherein R C preferably ₁₀-C ₂₄Alkyl preferably contains C ₁₀-C ₂₀The alkyl of moieties or hydroxyalkyl, more preferably C ₁₂-C ₁₈Alkyl or hydroxyalkyl, M is H or positively charged ion, for example the ammonium of alkali metal cation (for example sodium, potassium, lithium) or ammonium or replacement (as methyl-, dimethyl-and trimethylammonium-ammonium cation and quaternary ammonium cation, for example tetramethyl-ammonium and lupetidine positively charged ion and, for example quaternary ammonium cation that obtains of ethamine, diethylamine, triethylamine and their mixture etc.) by alkylamine.For low wash temperature (for example being lower than about 50 ℃), usually preferred C ₁₂-C ₁₆Alkyl chain, for the preferred C of higher wash temperature (for example being higher than about 50 ℃) _16-18Alkyl chain.

Other anion surfactant that is applicable to the washing purposes also can be included in washing composition of the present invention and/or the Fabrid care composition.They can comprise soap salt (comprise, the ammonium salt of sodium, potassium, ammonium and replacement for example, for example single-, two-and triethanolamine salt), C ₈-C ₂₂Uncle or secondary paraffin sulfonate, C ₈-C ₂₄Sulfonation poly carboxylic acid, the C of alkene sulfonate, the split product preparation of passing through the sulfonation alkaline earth metal citrate in GB1082179 for example, described ₈-C ₂₄Alkyl polyglycol ether sulfate (containing 10 moles of ethylene oxide at the most); Alkyl glycerol sulfonate, fatty acyl group glycerol sulfonate, fatty oil acylglycerol vitriol, alkylphenol oxyethane ether sulfate, paraffin sulfonate, alkylphosphonic, isethionate; for example, acyl isethinate, N-acyl taurine salt, amber alkyl amide salts and sulfosuccinate, sulfosuccinate monoesters (especially saturated and unsaturated C ₁₂-C ₁₈Monoesters) and sulfosuccinate diester (especially saturated and unsaturated C ₆-C ₁₂Diester), the vitriol of acyl sarcosinate, alkyl polysaccharide, alkyl polyglucoside vitriol (the not Sulfated compound of nonionic described below) for example, the many ethoxy carboxylates of chain primary alkyl sulfate and alkyl, for example formula RO (CH ₂CH ₂O) _k-CH ₂COO ^-M ⁺, wherein, R is C ₈-C ₂₂Alkyl, k are the 1-10 integers, and M is water miscible salt-forming cation.Resinous acid and hydrogenated resin acid also are suitable, for example rosin, staybelite and be present in Yatall MA or resinous acid therefrom and hydrogenated resin acid.

Other example is described in " tensio-active agent and washing composition " (I and II volume, Schwartz, Perry and Berch).Also general description is at the US3929678 of the Laughlin of on December 30th, 1975 promulgation etc. for various these class tensio-active agents, and the 23rd hurdle the 58th walks in the 29th hurdle the 23rd row (classifying this paper reference as).

If comprise, it is about 40% that detergent for washing clothes of the present invention and/or Fabrid care composition contain about by weight 1%-usually, preferably about 3%-about 20% this analog anion surfactants.

The ten minutes preferred anionic surfactants tensio-active agent that comprises its alkyl alkoxy sulfate tensio-active agent is formula RO (A) _mSO ₃The water-soluble salt of M or acid, wherein R is unsubstituted C ₁₀-C ₂₄Alkyl or contain C ₁₀-C ₂₄The hydroxyalkyl of moieties, preferred C ₁₂-C ₂₀Alkyl or hydroxyalkyl, more preferably C ₁₂-C ₁₈Alkyl or hydroxyalkyl, A are oxyethyl group or propoxy-unit, and m is greater than zero, usually between about 0.5 to about 6, more preferably from about between 0.5 to about 3, M is H or positively charged ion, and it can be the ammonium cation of metallic cation (for example sodium, potassium, lithium, calcium, magnesium etc.), ammonium or replacement.The present invention thinks over alkyl ethoxy sulfate and alkyl propoxy-vitriol.The specific examples of the ammonium cation that replaces comprise methyl-, dimethyl-, trimethylammonium-ammonium cation and quaternary ammonium cation, for example tetramethyl-ammonium and lupetidine positively charged ion and by alkylamine, for example ethamine, diethylamine, triethylamine, the positively charged ion that their mixture obtains etc.The tensio-active agent of illustrative is C ₁₂-C ₁₈The many ethoxylations of alkyl (1.0) vitriol (C ₁₂-C ₁₈E (1.0) M), C ₁₂-C ₁₈The many ethoxylations of alkyl (2.25) vitriol (C ₁₂-C ₁₈E (2.25) M), C ₁₂-C ₁₈The many ethoxylations of alkyl (3.0) vitriol (C ₁₂-C ₁₈E (3.0) M) and C ₁₂-C ₁₈The many ethoxylations of alkyl (4.0) vitriol (C ₁₂-C ₁₈E (4.0) M), wherein M is selected from sodium and potassium usually.

Washing composition of the present invention and/or Fabrid care composition also can contain other positively charged ions, both sexes, zwitter-ion and semi-polarity tensio-active agent and nonionic except that above having described and/or anion surfactant.

Biodegradable quaternary ammonium compound exists as two long alkyl chain ammonium chloride and the Methylsulfate surrogate that tradition is used.This quaternary ammonium compound contains by functional group, long-chain alkane (alkene) base of carboxyl fracture for example, and described material and the fabric sofetening composition that contains them are at many publications, and be for example open among EP-A-0040562 and the EP-A-0239910.

Quaternary ammonium compound of the present invention and amine parent have as shown in the formula (I) or (II):

Wherein Q be selected from-O-C (O)-,-C (O)-O-,-O-C (O)-O-, NR ⁴-C (O)-,-C (O)-NR ⁴-;

R ¹Be (CH ₂) _n-Q-T ²Or T ³

R ²Be (CH ₂) _m-Q-T ⁴Or T ⁵Or R ³

R ³Be C ₁-C ₄Alkyl or C ₁-C ₄Hydroxyalkyl or H;

R ⁴Be H or C ₁-C ₄Alkyl or C ₁-C ₄Hydroxyalkyl;

T ¹, T ², T ³, T ⁴And T ⁵Be respectively C ₁₁-C ₂₂Alkyl or alkenyl;

N and m are the integers of 1-4; With

X ^-It is the compatible negatively charged ion of softening agent.The anionic unrestricted example compatible with softening agent comprises chlorion or methylsulfate.

Alkyl or alkenyl chain T ¹, T ², T ³, T ⁴And T ⁵Must contain at least 11 carbon atoms, preferably at least 16 carbon atoms.Chain can be a straight or branched.Tallow, beef is the convenience and the cheap source of chain alkyl and thiazolinyl material.T wherein ¹, T ², T ³, T ⁴And T ⁵The compound of representing the long-chain substance mixture of common Tallow, beef is especially preferred.

The specific examples that is applicable to the quaternary ammonium compound of aqueous fabric soft compound of the present invention comprises:

1) N, N-two (Tallow, beef base oxygen base ethyl)-N, N-alkyl dimethyl ammonium chloride;

2) N, N-two (Tallow, beef base oxygen base ethyl) N-methyl, N-(2-hydroxyethyl) methylsulfuric acid ammonium;

3) N, N-two (2-Tallow, beef base oxygen base-2-oxygen base ethyl)-N, N-alkyl dimethyl ammonium chloride;

4) N, N-two (2-Tallow, beef base oxygen base-ethyl carbonyl-oxygen base ethyl)-N, N-alkyl dimethyl ammonium chloride;

5) N-(2-Tallow, beef base oxygen base-2-ethyl)-N-(2-Tallow, beef base oxygen base-2-oxygen base ethyl)-N, the N-alkyl dimethyl ammonium chloride;

6) N, N, N-three (Tallow, beef base-oxygen base-ethyl)-N-ammonio methacrylate;

7) N-(2-Tallow, beef base-oxygen base-2-oxygen base ethyl)-N-(Tallow, beef base)-N, the N-alkyl dimethyl ammonium chloride; With

8) 1,2-two Tallow, beef base oxygen base-3-trimethyl ammonium propane chloride;

Mixture with any above-mentioned substance.

Amphoterics also is applicable in washing composition of the present invention and/or the Fabrid care composition.These tensio-active agents can broadly be described as the aliphatic derivatives of the second month in a season or tertiary amine or heterocycle is secondary and the aliphatic derivatives of tertiary amine, and wherein aliphatic group can be a straight or branched.One of aliphatic series substituting group contains at least about 8 carbon atoms, and about 18 carbon atoms of about usually 8-and at least one aliphatic substituting group contain the anionic water solubilizing group, for example carboxyl, sulfonate radical, sulfate radical.Referring to the US3929678 at the Laughlin of on December 30th, 1975 promulgation etc., the amphoterics of the capable illustrated of the 19th hurdle 18-35.

If comprise, it is about 15% that washing composition of the present invention and/or Fabrid care composition contain by weight 0.2%-usually, preferably about 1%-about 10% this amphoterics.

Zwitterionics also is applicable in washing composition and/or the Fabrid care composition.These tensio-active agents can broadly be described as the derivative of derivative, the heterocycle second month in a season and the tertiary amine of secondary and tertiary amine, or the derivative of quaternary ammonium, quaternary phosphine or uncle's sulfonium compound.Referring to zwitterionics in the 38th row-22 hurdles, US 3929678, the 19 hurdles 48 row illustrated of the Laughlin of on December 30th, 1975 promulgation etc.

If comprise, it is about 15% that washing composition of the present invention and/or Fabrid care composition contain by weight 0.2%-usually, preferably about 1%-about 10% this zwitterionics.

Semi-polar nonionic surfactants is the nonionogenic tenside of particular variety, and it comprises that the alkyl that contains about 18 carbon atoms of about 10-and two are selected from the water-soluble amine oxides of the group of the alkyl that contains about 3 carbon atoms of the 1-that has an appointment and hydroxyalkyl; Contain the water soluble oxidized phosphine that the alkyl of about 18 carbon atoms of about 10-and two are selected from the group of the alkyl that contains about 3 carbon atoms of the 1-that has an appointment and hydroxyalkyl; Be selected from the water-soluble sulfoxide of the group of the alkyl that contains about 3 carbon atoms of the 1-that has an appointment and hydroxyalkyl with the alkyl that contains about 18 carbon atoms of about 10-and one.

Semi-polarity nonionic detergent tensio-active agent comprises the amine oxide surfactant with following formula:

R wherein ³Be alkyl, hydroxyalkyl or alkyl phenyl or its mixture that contains about 22 carbon atoms of the 8-that has an appointment; R ⁴Be alkylidene group or hydroxy alkylidene or their mixture that contains about 3 carbon atoms of the 2-that has an appointment; X is 0-about 3; Each R ⁵It is the polyoxyethylene group that contains the alkyl or the hydroxyalkyl of about 3 carbon atoms of the 1-that has an appointment or contain about 3 ethylene oxide groups of the 1-that has an appointment.R ⁵Group can for example be interconnected to form ring structure by oxygen or nitrogen-atoms.

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

If comprise, it is about 15% that washing composition of the present invention and/or Fabrid care composition contain by weight 0.2%-usually, this semi-polar nonionic surfactants of preferably about 1%-about 10%.

Washing composition of the present invention and/or Fabrid care composition also can contain the cosurfactant that is selected from uncle or tertiary amine.Be used for suitable primary amine of the present invention and comprise formula R ₁NH ₂Amine, R wherein ₁Be C ₆-C ₁₂, preferred C ₆-C ₁₀Alkyl chain or R ₄X (CH ₂) _n, X is-O-,-C (O) NH-or-NH-, R ₄Be C ₆-C ₁₂Alkyl chain, n are 1 to 5, preferred 3.R ₁Alkyl chain can be a straight or branched, can preferably be less than 5 ethylene oxide moieties and be interrupted by 12 at the most.

The preferred amines of following formula of the present invention is positive alkylamine.Be used for suitable amine of the present invention and be selected from 1-hexylamine, 1-octylame, 1-decyl amine and lauryl amine.Other preferred primary amine is C ₈-C ₁₀Oxygen base propylamine, octyloxy propylamine, 2-ethyl hexyl oxy propylamine, lauryl amido propylamine and amido propylamine.

Be used for suitable tertiary amine of the present invention and comprise having formula R ₁R ₂R ₃The tertiary amine of N, wherein R ₁And R ₂Be C ₁-C ₈Alkyl chain or R ₃Be C ₆-C ₁₂, preferred C ₆-C ₁₀Alkyl chain or R ₃Be R ₄X (CH ₂) _n, and X is-O-,-C (O) NH-or-NH-, R ₄Be C ₄-C ₁₂, n is 1 to 5, preferably 2-3.R ₅Be H or C ₁-C ₂Alkyl and x are 1-6.

R ₃And R ₄It can be straight or branched; R ₃Alkyl chain can preferably be less than 5 ethylene oxide moieties and be interrupted by 12 at the most.

Preferred tertiary amine is R ₁R ₂R ₃N, wherein R ₁Be C ₆-C ₁₂Alkyl chain, R ₂And R ₃Be C ₁-C ₃Alkyl or R wherein ₅Be H or methyl and x=1-2.

The amidoamines of following formula equally preferably:

R wherein ₁Be C ₆-C ₁₂Alkyl; N is 2-4, and preferred n is 3; R ₂And R ₃Be C ₁-C ₄

The most preferred amine of the present invention comprises 1-octylame, 1-hexylamine, 1-decyl amine, 1-lauryl amine, C ₈-C ₁₀Oxygen base propylamine, N-Oleum Cocois 1,3 diaminopropanes, Oleum Cocois alkyl dimethyl amine, lauryl dimethyl amine, lauryl two (hydroxyethyl) amine, Oleum Cocois alkyl two (hydroxyethyl) amine, 2 moles of propenoxylated lauryl amines, 2 moles of propenoxylated octylames, lauryl amido propyl-dimethyl amine, C ₈-C ₁₀Amido propyl-dimethyl amine and C ₁₀The amido propyl-dimethyl amine.

The most preferred amine that is used for the present composition is 1-hexylamine, 1-octylame, 1-decyl amine, 1-lauryl amine.Especially suitable is oleyl amine, lauryl amido propylamine and the Oleum Cocois amido propylamine of dodecyl dimethyl amine and two hydroxyethyl Oleum Cocois alkylamines and 7 ethoxylations.

SYNTHETIC OPTICAL WHITNER

Washing composition of the present invention and/or Fabrid care composition also can comprise SYNTHETIC OPTICAL WHITNER, hydrogen peroxide for example, and particle size is PB1, PB4 and the percarbonate of 400-800 micron.These bleaching components can comprise one or more oxygen bleaching agents and, according to selected SYNTHETIC OPTICAL WHITNER, one or more bleach activators.If present, the oxygen bleaching compound exists with the content of about 1%-about 25% usually.

Being used for SYNTHETIC OPTICAL WHITNER of the present invention can be any SYNTHETIC OPTICAL WHITNER that is used for washing composition and/or Fabrid care composition, comprises oxygen bleaching agent and other SYNTHETIC OPTICAL WHITNER well known in the prior art.Be applicable to that SYNTHETIC OPTICAL WHITNER of the present invention is activation or non-activated SYNTHETIC OPTICAL WHITNER.

Spendable a kind of oxygen bleaching agent comprises percarboxylic acids SYNTHETIC OPTICAL WHITNER and its salt.The suitable example of this class SYNTHETIC OPTICAL WHITNER comprises monoperphthalic acid magnesium hexahydrate, metachloroperbenzoic acid magnesium salts, 4-amino in the ninth of the ten Heavenly Stems-4-oxygen Perbutyric Acid and diperoxy dodecanedioic acid.This SYNTHETIC OPTICAL WHITNER is open in US4483781, US patent application 740446, EP0133354 and US4412934.Preferred SYNTHETIC OPTICAL WHITNER also is included in 6-amino in the ninth of the ten Heavenly Stems-6-oxygen of describing among the US4634551 and crosses oxy hexanoic acid.

Operable another kind of SYNTHETIC OPTICAL WHITNER comprises halogen bleaching agent.The example of hypohalite SYNTHETIC OPTICAL WHITNER for example comprises TCCA (Trichloroisocyanuric acid) and dichloroisocyanuric acid sodium and potassium and N-chlorine and N-bromoalkane sulphonamide.This material is usually in the weight 0.5-10% by final product, and preferably 1-5% adds by weight.

The hydrogen peroxide releasing agent can with bleach activator; for example tetra acetyl ethylene diamine (TAED), nonanoly acyloxy benzene sulfonate (NOBS; in US4412934, describe), 3; 5-trimethyl hexanol oxygen base benzene sulfonate (ISONOBS; in EP120591, describe) or penta-acetyl glucose (PAG) or N-nonanoyl-6-aminocaprolc acid sulfophenylate (NACA-OBS; in WO 94/28106, describe) be used in combination; they are crossed hydrolysis to form the peracid as active albic material, improved bleaching effect.Same suitable bleach activator is the acylations citrate, and is for example open in the european patent application of not examining 91870207.7, at Procter ﹠amp; The asymmetric acyclic inferior lactam bleach promoting agent of disclosed following formula among the patent application US № 60/022786 (application on July 30th, 1996) that does not examine of Gamble and the № 60/028122 (application on October 15th, 1996):

R wherein ₁Be C ₇-C ₁₃Saturated or the unsaturated alkyl of straight or branched, R ₂Be C ₁-C ₈Straight or branched saturated or unsaturated alkyl and R ₃Be C ₁-C ₄Saturated or the unsaturated alkyl of straight or branched.

Be used for the useful SYNTHETIC OPTICAL WHITNER of detergent composition of the present invention, the bleach system that comprises peroxy acid and contain bleach activator and peroxy bleaching compound is described in application USSN08/136626, PCT/US95/07823, WO95/27772, WO95/27773, WO95/27774 and WO95/27775 that we examine.

Hydrogen peroxide can also exist by adding enzyme system (being enzyme and its substrate), it can washing and/or rinse cycle begins or process in produce hydrogen peroxide, this enzyme system is open in the EP patent application 91202655.6 of application on October 9th, 1991.

The containing metal catalyzer that is used for whitener composition comprises cobalt-containing catalyst, five amine cobaltous acetate (III) salt and contain Mg catalyst, for example compound of in EPA549271, EPA549272, EPA458397, US5246621, EPA458398, US5194416 and US5114611, describing for example.The bleaching composition that contain peralcohol, contains magnesium bleach catalysts and sequestrant is described in patent application № 94870206.3.

SYNTHETIC OPTICAL WHITNER except that oxygen bleaching agent also is known in the prior art, can be used for the present invention.A kind of non-oxygen bleaching agent that cherishes a special interest comprises the photoactivation SYNTHETIC OPTICAL WHITNER, for example sulfonated zinc and/or aluminium phthalocyanine.These materials are precipitable on the dirt-carrying body in washing process.Under rayed and in the presence of oxygen, for example by clothes being hung in the daylight when dry, sulfonation zinc phthalocyanine phthalocyanine is activated, and the dirt-carrying body is bleached subsequently.Preferred zinc phthalocyanine and photoactivation bleaching process are described in US4033718.Washing composition and/or Fabrid care composition will contain about 1.25% sulfonation zinc phthalocyanine phthalocyanine of about by weight 0.025%-usually.

Builder system

Washing composition of the present invention and/or Fabrid care composition can further contain builder system.The builder system of any routine is applicable to the present invention, it comprises silico-aluminate material, silicate, multi-carboxylate, alkyl or alkenyl succsinic acid and lipid acid, material, for example edetate, two inferior second triamine pentamethylene acetates, metal ion chelation agent, for example aminopolyphosphonic acid salt, especially ethylenediamine tetramethylene phosphonic acid and two inferior second triamine pentamethylene phosphonic acids.Phosphate builders also can be used among the present invention.

Suitable washing assistant can be a mineral ion exchange material, normally inorganic hydrated aluminosilicate material, more particularly hydration synthetic zeolite, for example hydrated zeolite A, X, B, HS or MAP.

Other suitable inorganic builders material is a layered silicate, for example SKS-6 (Hoechst).SKS-6 is by water glass (Na ₂Si ₂O ₅) crystalline layered silicate formed.

The suitable multi-carboxylate of containing a carboxyl is included in disclosed lactic acid, oxyacetic acid and their ether derivant in belgian patent 831368,821369 and 821370.The multi-carboxylate of containing two carboxyls comprise succsinic acid, oxysuccinic acid, (ethylenedioxy) oxalic acid, toxilic acid, diglycollic acid, tartrate, tartronic acid and fumaric acid water-soluble salt and DE2446686 and 2446687 and US3935257 in the ether carboxylate described and the sulfinyl carboxylate salt of in belgian patent 840623, describing.The multi-carboxylate of containing three carboxyls especially comprises water-soluble citrate, itaconate and citraconate and succinate derivative, the carboxyl methoxy succinate of for example in GB1379241, describing, the newborn oxygen base succinate of in Holland's application 7205873, describing and the oxygen multi-carboxylate material of in GB1387447, describing, 2-oxygen-1 for example, 1,3-tricarballylic acid salt.

The multi-carboxylate of containing four carboxyls is included in disclosed oxygen di-succinate, 1,1,2 among the GB1261829,2-ethane tetracarboxylic acid hydrochlorate, 1,1,3,3-propane tetracarboxylic acid salt and 1,1,2,3-propane tetracarboxylic acid salt.Contain the substituent multi-carboxylate of sulfo group be included in GB1398421 and 1398422 and US3936448 in disclosed sulfo-succinic acid salt derivative and the sulfonation cracking Citrate trianion in GB1082179, described, and it is open in GB1439000 to contain the substituent multi-carboxylate of phosphono.

Alicyclic ring and heterocycle multi-carboxylate comprise pentamethylene-suitable, suitable, suitable-tetracarboxylic acid hydrochlorate, cyclopentadiene pentacarboxylic acid salt, 2,3,4,5-tetrahydrofuran (THF)-suitable, suitable, suitable-tetracarboxylic acid hydrochlorate, 2,5-tetrahydrofuran (THF)-suitable-dicarboxylate, 2,2,5,5-tetrahydrofuran (THF) tetracarboxylic acid hydrochlorate, 1,2,3,4,5,6-hexane hexacarboxylic acid salt and polyhydroxy-alcohol, for example carboxymethyl derivant of Sorbitol Powder, mannitol and Xylitol.Aromatic polycarboxylic acids salt is included in disclosed mellic acid among the GB1425343, pyromellitic acid and phthalic acid derivative.

Wherein, preferred multi-carboxylate is that per molecule contains the hydroxycarboxylate of three carboxyls, more particularly Citrate trianion at the most.

Be used for preferred builder system of the present invention and comprise water-soluble silicon aluminate washing assistant, for example zeolite A or layered silicate (SKS-6) and water-soluble carboxylate sequestrant, for example mixture of citric acid.Other preferred builder system comprises water-soluble silicon aluminate washing assistant, for example zeolite A and water-soluble carboxylate sequestrant, for example mixture of citric acid.The preferred builder system that is used for liquid washing agent of the present invention and/or Fabrid care composition comprises soap and multi-carboxylate.

But component part is used for other builder material of the builder system of particulate composition and comprises inorganic substance, for example alkaline carbonate, supercarbonate, silicate and organic substance, for example organic phosphonate, amino polyalkylene phosphonate and aminopolycanboxylic acid's salt.

Other suitable water-soluble organic salt is all or co-polymeric acids or their salt, and wherein poly carboxylic acid contains and is no more than at least two carboxyls that two carbon atoms are separated from each other.Such polymkeric substance is open in GB-A-1596756.The example of this salt is the multipolymer of the polyacrylate of MW2000-5000 and they and maleic anhydride, and this multipolymer has 20000-70000, especially about 40000 molecular weight.

Detergent builders salt is usually pressing composition weight meter 5%-80%, preferred 10%-70%, and the most common be the amount existence of 30%-60% by weight.

Conventional detergent enzyme

Except mannase, washing composition and/or Fabrid care composition also can contain one or more enzymes, and it provides scourability, fabric nursing and/or hygiene effect.

Described enzyme comprises the enzyme that is selected from cellulase, hemicellulase, peroxidase, proteolytic enzyme, glucoamylase, amylase, zytase, lipase, phosphide enzyme, esterase, at, polygalacturonase, M-Zyme, reductase enzyme, oxydase, phenol oxidase, lipoxygenase, lignoenzyme, Starch debranching enzyme, tannase, pentosanase, malic enzyme, beta-glucanase, arabinofuranosidase/xylosidase, hyaluronic enzyme, chondroitinase, laccase or their mixture.

Preferably combination is to contain enzyme commonly used, as the washing composition and/or the Fabrid care composition of proteolytic enzyme, amylase, lipase, at and/or cellulase and one or more plant cell-wall degrading enzymes bonded mixtures.

Suitable proteolytic enzyme is by the subtilysin of the special bacterial strain of subtilis and Bacillus licheniformis (subtilysin BPN and BPN ').A kind of suitable proteolytic enzyme is obtained by the special bacterial strain that the whole pH scope at 8-12 has the bacillus of maximum activity, and it is by NovoIndustries A/S (Denmark) exploitation, hereinafter referred to as " Novo ", and sells with ESPERASE .The preparation of this kind of enzyme and similar enzyme is described in the GB1243784 of Novo.Other suitable proteolytic enzyme comprises ALCALASE , DURAZYM  and SAVINASE  and the MAXATASE  that is obtained by Gist-Brocades, MAXACAL , PROPERASE  and the MAXAPEM  (Maxacal of protein engineering) that is obtained by Novo.Protease also comprises the modified bacteria serine protease, the proteolytic enzyme of description in the european patent application 87303761.8 (especially 17,24 and 98 pages) of on April 28th, 1987 application for example, it is referred to herein as " proteolytic enzyme B " and the proteolytic enzyme in the EP199404 of disclosed Venegas on the 29th October in 1986, it relates to the bacterial serine protease of modification, and it is referred to herein as " protease A ".Suitable is the proteolytic enzyme that is referred to herein as " proteolytic enzyme C ", it is the mutation of the alkaline serine protease that obtained by genus bacillus, wherein replace arginine with Methionin at 27, replace Xie Ansuan at 104 with tyrosine, replace aspartic acid and replace Threonine with L-Ala with Serine at 274 at 123.Proteolytic enzyme C is disclosed EP90915958.4 in 16 days Mays in 1991, corresponding to describing among the WO91/06637.The mutation of gene modification, especially proteolytic enzyme C is also included among the present invention.

The preferred protease that is called " proteolytic enzyme D " is the carbonylic hydrolase mutation that has at the undiscovered aminoacid sequence of occurring in nature, be called as the name of the C.Ghosh of WO95/10591 and on October 13rd, 1994 application etc. described in the patent application of " bleaching composition that contains proteolytic enzyme " (US series number № 08/322677), it by in above-mentioned carbonylic hydrolase, be equivalent to+replace the multiple amino acids residue with different aminoacids on 76 positions, and preferred also in conjunction with replacing numbering+99 that are equivalent to be selected from according to the bacillus amyloliquefaciens subtilysin, + 101, + 103, + 104, + 107, + 123, + 27, + 105, + 109, + 126, + 128, + 135, + 156, + 166, + 195, + 197, + 204, + 206, + 210, + 216, + 217, + 218, + 222, + 260, + 265 and/or+one or more amino-acid residue positions of 274 are obtained by the precursor carbonylic hydrolase.Same suitable be carbonylic hydrolase mutation at the proteolytic enzyme described in the WO95/10591, it have by on being equivalent to of pre-enzyme+210 positions with various radical amino acid replacements and in conjunction with one or more following residues :+33, + 62, + 67, + 76, + 100, + 101, + 103, + 104, + 107, + 128, + 129, + 130, + 132, + 135, + 156, + 158, + 164, + 166, + 167, + 170, + 209, + 215, + 217, + 218 and+222 aminoacid sequences that obtain, wherein Bian Hao position is equivalent to the bacillus amyloliquefaciens subtilysin of natural generation or is equivalent at other carbonylic hydrolase or subtilysin, for example the suitable amino-acid residue in the bacillus lentus subtilysin (the not careful patent application US series number 60/048550 of application on June 4th, 1997).

Being equally applicable to of the present invention is the proteolytic enzyme of describing in EP251446 and WO91/06637, the proteolytic enzyme BLAP  that describes in WO91/02792 and their mutation described in WO95/23221.

Also can be referring to the high pH proteolytic enzyme that obtains by bacillus NCIMB40338 described in the WO93/18140A of Novo.The enzyme-containing detergent that comprises proteolytic enzyme, one or more other enzymes and a kind of reversible protease inhibitors has been described among the WO92/03529A of Novo.If desired, can be as Procter ﹠amp; The WO95/07791 of Gamble is described to obtain having the absorptivity of reduction and the water-disintegrable proteolytic enzyme of improvement.A kind of recombinant trypsin shape proteolytic enzyme that is applicable to washing composition of the present invention has been described among the WO94/25583 of Novo.Other suitable proteolytic enzyme is described in the EP516200 of Unilever.

Protease to be pressing composition weight meter 0.0001%-2%, preferred 0.001%-0.2%, and more preferably the content of the pure enzyme of 0.005%-0.1% adds in washing composition of the present invention and/or the Fabrid care composition.

Be used for cellulase of the present invention and comprise bacterium or fungal cellulase.They preferably have the pH optimum value between the 5-12 and surpass the ratio work of 50CEVU (Mierocrystalline cellulose viscosity unit).Suitable cellulase is described in US4435307, the J61078384 of Barbesgoard etc. and WO96/02653, and they disclose the fungal cellulase that is produced by Humicola insolens, Trichoderma, Thielavia and Sporotrichum respectively.EP739982 has described by the new isolating cellulase of genus bacillus kind.Suitable cellulase also is disclosed among GB-A-2075028, GB-A-2095275, DE-OS-2247832 and the WO95/26398.

The example of this cellulase is the bacterial strain by Humicola insolens (Humicola grisea var.thermoi dea), especially the cellulase of Humicola strain DSM 1800 generations.

Other suitable cellulase is that the molecular weight that has that is obtained by Humicola insolens is about 50kDa, iso-electric point 5.5 and contain 415 amino acid whose cellulases; With the plain enzymic activity of the display fibers that obtains by Humicolainsolens DSM 1800 ^-The 43kD endoglucanase; Preferred endoglucanase component has disclosed aminoacid sequence among the PCT patent application WO91/17243.It is same that what be fit to is the EGIII cellulase of describing in the WO94/21801 of disclosed Genencor on the 29th September in 1994 that is obtained by Trichoderma longibrachiatum.Especially suitable cellulase is the cellulase with color nursing efficacy.The example of this cellulase is the cellulase of describing in the EP patent application № 91202879.2 (Novo) of application on November 6th, 1991.Carezyme and Celluzyme (NovoNordisk A/S) are particularly useful.Also referring to WO91/17244 and WO91/21801.The plain enzyme of other suitable fibers with fabric nursing and/or scourability is described in WO96/34092, WO96/17994 and WO95/24471.

Described other cellulase adds in washing composition and/or the Fabrid care composition in the content by washing composition and/or the pure enzyme of Fabrid care composition weight 0.0001%-2% usually.

Peroxidase and oxygen source, for example percarbonate, perborate, persulphate, hydrogen peroxide etc. and be used in combination as the phenolic aldehyde substrate of SYNTHETIC OPTICAL WHITNER synergy molecule.They are as " solution bleaching ", promptly avoid the dyestuff removed by the dirt-carrying body in washing operation or pigment to be transferred in other dirt-carrying body in washing soln.Peroxidase is known in the prior art, and it comprises, for example horseradish peroxidase, lignoenzyme and halogenation peroxidase, for example chlorine and bromo peroxidase.The detergent composition that contains peroxidase is open in the EP № 96870013.8 of the European patent application EP 91202882.6 of for example PCT International Application No. WO 89/099813, WO89/09813 and application on November 6th, 1991 and application on February 20th, 1996.Same suitable is laccase.

Synergistic agent exists with the content by general composition weight meter 0.1%-5% usually.The thiodiphenylamine that preferred synergistic agent is replacement and phenoxazine, lysivane propionic acid (PPT), 10-ethyl thiodiphenylamine-4-carboxylic acid (EPC), 10-phenoxazine propionic acid (POP) and 10-first base phenoxazine (in WO94/12621, describing) and the cloves acid esters (the alkyl cloves acid esters that C3-C5 replaces) and the phenol that replace.SPC-D or Sodium peroxoborate are preferred hydrogen peroxide cources.

Above-mentioned peroxidase adds in washing composition and/or the Fabrid care composition in the content by washing composition and/or the pure enzyme of Fabrid care composition weight 0.0001%-2% usually.

Other preferred enzyme that can be included in washing composition of the present invention and/or the Fabrid care composition comprises lipase, the suitable lipase that is used for the washing composition purposes comprises the microorganism of Rhodopseudomonas, the lipase that Situ Ci Shi aeruginosa atcc 19.154 disclosed in GB1372034 obtains.Suitable lipase comprises the lipase that shows positive immunological cross-reaction with lipase antibody, and it is produced by microorganism Pseudomonas fluorescens IAM 1057.This lipase can be obtained by Amano PharmaceuticalCo.Ltd. (Nagoya, Japan), and commodity are called lipase P " Amano ", hereinafter referred to as " Amano-P ".Other suitable commercial lipases comprises Amano-CES, from the Chromobacterviscosum lipase of the Chro mobacter viscosumvar.lipolyticum NRRLB 3673 of Toyo Jozo Co. (Tagata, Japan) for example; The Chromobacter viscosum lipase that obtains by U.S.Biochemical Corp. (US) and Disoynth (Holland); And the lipase that obtains by the gladiolus pseudomonas.Especially suitable lipase is lipase, for example M1 Lipase ^RAnd Lipomax ^R(Gist-Broxades) and Lipolase ^RWith Lipolase Ultra ^RWhen (Novo), they are found in and are used in combination with composition of the present invention is very effective.Same suitable be the lipolytic enzyme of in WO94/03578, the WO95/35381 of EP258068, the WO92/05249 of Novo Nordisk and WO95/22615 and Unilever and WO96/00292, describing.

That same suitable is at [EC 3.1.1.50], and it is considered to the lipase of particular variety, nominal lipase, but do not need interface activation.In washing composition and/or Fabrid care composition, add at for example WO-A-88/09367 (Genencor); Describe among WO90/09446 (PlantGenentic System) and WO94/14963 and the WO94/14964 (Unilever).

Lipase and/or at add in washing composition and/or the Fabrid care composition in the content by washing composition and/or the pure enzyme of Fabrid care composition weight 0.0001%-2% usually.

Can comprise that amylase (α and/or β) is to remove the spot of carbohydrate-based.On February 3rd, 1994, the WO94/02597 of disclosed Novo Nordisk A/S described diastatic washing composition of adding mutation and/or Fabrid care composition.Same WO95/10603 referring to disclosed NovoNordisk A/S on the 20th in April nineteen ninety-five.Other amylase that becomes known for detergent composition comprises α-and beta-amylase.α-Dian Fenmei is known in the prior art, is included in the amylase of describing among US5003257, EP252666, WO91/00353, FR2676456, EP285123, EP525610, EP368341 and the GB1296839 (Novo).Other suitable amylase is the amylase and as the amylase mutation that has additional modification in the intermediate parent that is obtained by Novo Nordisk A/S described among the disclosed WO95/10603 in April, 95 of the improved stability described among the disclosed WO96/05295 in 18 days Augusts in 1994 of Genencor disclosed WO94/18314 and on February 22nd, 1996.Same suitable amylase is described in EP277216, WO95/26397 and WO96/23873 (by Novo Nordisk).

The example of commercial α-Dian Fenmei product is Purafect Ox Am  and Termamyl , BAN , Fungamyl  and the Duramyl  that is obtained by Genencor, obtains by Novo Nordisk A/S Denmark.WO95/26397 has described other suitable amylase: α-Dian Fenmei, it is characterized in that, by the test determination of Phadebas  alpha-amylase activity, have under its pH in 25 ℃-55 ℃ temperature range and at 8-10 than the ratio of Termamyl  high at least 25% ratio alive and live.Suitable is the above-mentioned diastatic mutation of describing in WO96/23873 (Novo Nordisk).In WO95/35382, describe having other amylolytic enzyme that improves character aspect activity level and thermostability and the greater activity horizontal integration.

Amylolytic enzyme to be pressing composition weight meter 0.0001%-2%, preferred 0.00018%-0.06%, and more preferably the content of the pure enzyme of 0.00024%-0.048% adds in washing composition of the present invention and/or the Fabrid care composition.

Above-mentioned enzyme can be any suitable source, plant, animal, bacterium, fungi and yeast source for example, source can also be have a liking for temperature or have a liking for limiting condition (have a liking for cold, cold nutrition, thermophilic, have a liking for pressure, have a liking for alkali, have a liking for acid, have a liking for halogen etc.).Pure or the impure form of these enzymes can both be used.At present, in common reality through the enzyme of protein/genetic engineering technique modification agriotype with their effectiveness of performance in washing composition of the present invention and/or Fabrid care composition of optimizing.For example but design variable is to increase the consistency of the component that runs into usually in enzyme and the said composition.In addition, but design variable makes best pH, SYNTHETIC OPTICAL WHITNER or sequestrant stability, the catalytic activity etc. of enzyme variants can adapt to specific washing uses.

Especially should notice that under the situation of bleach stability amino acid should note surface charge to the susceptibility of oxidation with for the compatible ability of tensio-active agent.The iso-electric point of this kind of enzyme can be passed through some charged amino acid whose displacement modification, for example increases iso-electric point and can help to improve consistency with anion surfactant.The stability of enzyme can further be improved by producing for example additional salt bridge, and forces the calcium binding site to increase sequestrant stability.Especially must note cellulase because most of cellulase have separately in conjunction with territory (CBS), the character of this kind of enzyme can change by the modification in these territories.

Described enzyme adds in washing composition of the present invention and/or the Fabrid care composition in the content by washing composition and/or the pure enzyme of Fabrid care composition weight 0.0001%-2%.Enzyme can be used as independent component (containing a kind of bead, particle, stabilising liq of enzyme etc.) or adds as two or more mixture (for example agglomerated particle).

Other suitable detergent component that can add is the oxydasis scavenging agent, and it is described not examining in the european patent application 92870018.6 of application on January 21st, 1992, and the example of this oxydasis scavenging agent is ethoxylation four ethylidene polyamine.

The WO9307263A of Genencor International and WO9307260A, the US3553139 of the McCarty of the WO8908694A of Novo and promulgation on January 5th, 1971 etc. has also described the scope of enzyme material and the method in their adding synthesis of detergent compositions.The US4507219 of the US4101457 of the Place of promulgation on July 18th, 1978 etc. and the Hughes of promulgation on March 26th, 1985 further discloses enzyme.The US4261868 of the Hora of on April 14th, 1981 promulgation etc. has disclosed the proenzyme material that is used for the liquid scrubbing prescription and they and has added method in this prescription.The enzyme that is used for washing composition can in all sorts of ways and be stablized.The US3600319 of the Gedge of on August 17th, 1971 promulgation etc. and October in 1986 Venegas on the 29th EP199405 and EP200586 the enzyme stabilization technique is disclosed and enumerates.The enzyme stabilising system is also for example described in US3519570.The WO9401532A of Novo has described the useful bacillus AC13 that can access proteolytic enzyme, zytase and cellulase.

Color nursing and fabric nursing effect

Also can comprise the technology that color nursing efficacy type is provided, the example of these technology is to be used for the metal catalyst that color keeps, and this metal catalyst is described in the european patent application of not examining 928700181.2.Dye-fixing agent, be used for crease-resistant and improve polyolefine dispersion agent, the spices of water adsorptive power and be used for that color nursing is handled and the polymkeric substance (PCT/US97/16546) of the amido functional group that spices is firm is other example of color nursing/fabric nursing technology, and examine description among the patent application № 96870140.9 in application on November 7th, 1996.

Fabric softener also can add in washing composition of the present invention and/or the Fabrid care composition.These materials can be inorganic or organic types.Inorganic softening agent is the terre verte of for example describing in GB-A-1400898 and US5019292.The organic fabric softening agent comprises the water-insoluble tertiary amine as describing among GB-A1514276 and the EP-B0011340, the mixture of they and single C12-C14 quaternary salt in EP-B-0026527 and EP-B-0026528 open and as EP-B0242919 in disclosed two long-chain acid amides.Other useful organic constituent of fabric softener system is included in disclosed polyphosphazene polymer oxyethane material in EP-A0299575 and 0313146.

The consumption of terre verte is usually at 2%-20% by weight, and more preferably in the scope of 5%-15%, this material mixes in the rest part that component adds prescription as doing.The organic fabric softening agent, for example water-insoluble tertiary amine or two long-chain acid amides materials are with 0.5%-5% by weight, usually the consumption of 1%-3% adds, and polyphosphazene polymer oxyethane material and water-soluble cationic material be with 0.1%-2% by weight, and the consumption of 0.15%-1.5% adds usually.These materials add the spraying drying part of composition usually, though they can be more easily as doing mixed particle adding or being sprayed at as melt liquid on other solid ingredient of composition in some cases.

Sequestrant

Washing composition of the present invention and/or Fabrid care composition are optional will to contain one or more iron and/or manganese sequestrant in addition.This sequestrant can be selected from aminocarboxylic acid ester, amido phosphonate, and aromatic chelator that polyfunctional group replaces and composition thereof hereinafter will define them.Although do not want to be limited by theory, we think that the effect of these materials is in part because they remove the specific competence of iron and mn ion by forming water soluble chelate compound from washings.

The aminocarboxylate of the sequestrant of useful as selective comprises edetate, the N-hydroxyethyl-ethylenediamine triacetate, nitrilotriacetic acid(NTA) salt, ethylenediamine tetrapropionic acid(EDTP) salt, triethylenetetraaminehexaacetic acid salt, diethylentriamine pentacetate and ethanol Diglycocol, their basic metal, ammonium and substituted ammonium salt and composition thereof.

When allowing to use the total phosphorus of minimum content at least in detergent composition, amino phosphonates do also is adapted at being used as in the present composition sequestrant, and it comprises that ethylenediamine tetraacetic (methylene phosphonic acid salt) is as DEQUEST.These amino phosphonates do preferably do not comprise alkyl or the alkylidene group more than about 6 carbon atoms.

The aromatic chelator that polyfunctional group replaces also is suitable for the present composition.US3812044 referring to the Connor of on May 21st, 1974 promulgation etc.This compounds of preferred sour attitude is the dihydroxyl disulfobenzene, as 1, and 2-dihydroxyl-3,5-disulfobenzene.

Being used for preferred biodegradable cheating agent of the present invention is ethylenediamine disuccinate (" EDDS "), [S, S] isomer particularly, and this is open in the US4704233 of the Hartman of promulgation on November 3rd, 1987 and Perkin.

Composition of the present invention can also contain as sequestrant or with for example undissolved washing assistant, for example water-soluble methylglycine oxalic acid (MGDA) salt (or acid) of the common washing assistant that uses together such as zeolite, layered silicate.

If use, these sequestrants generally account for about 0.1%-about 15% of washing composition of the present invention and/or Fabrid care composition weight.More preferably, if use, these sequestrants are about 0.1%-about 3.0% of said composition weight.

Suds suppressor

The component of another selection is a suds suppressor, for example polysiloxane and silicon dioxide-poly-mixture of siloxanes.Polysiloxane can be expressed as alkylating polysiloxane material usually, and silicon-dioxide uses with the broken form of fine powder usually, for example various types of silicon-dioxide aerosols, xerogel and water drain silica.These materials can be used as particulate and add, and wherein suds suppressor advantageously water-soluble or water is dispersible, go up substantially and discharge the ground adding in the impermeable carrier by the nonsurfactant washing composition.In addition, suds suppressor solubilized or be dispersed in the liquid vehicle adds by being sprayed on one or more other components.

Preferred polysiloxane Foam Control is open in the US3933672 of Bartollota etc.Other suds suppressor that especially is suitable for is the self-emulsifying polysiloxane suds suppressor of describing among the disclosed German patent application DTOS2646126 on April 28th, 1977.This examples for compounds is DC-544, is commercially obtained by Dow Corning, and it is polysiloxane-glycol copolymer.Especially preferred Foam Control is the suds suppressor system that contains the mixture of polysiloxane oil and 2-alkyl chain triacontanol.Suitable 2-alkyl-alkanol is a 2-butyl octanol, and it obtains with trade(brand)name Isofol12R commercial.

This suds suppressor system is described not examining in the european patent application 92870174.7 of application on November 10th, 1992.

Especially preferred polysiloxane Foam Control is described in the european patent application of not examining 92201649.8, and described composition can contain and sootiness atresia silicon-dioxide, for example Aerosil ^RBonded polysiloxane/silica mixture.

Usually to press composition weight meter 0.001%-2%, the content of preferred 0.01%-1% by weight uses above-mentioned suds suppressor.

Other

Can use other component that is used for washing composition and/or Fabrid care composition, for example soil-suspending agent, stain remover, white dyes, abrasive material, sterilant, tarnish inhibitor, tinting material and/or encapsulate capsule or do not wrap capsular spices.

Especially suitable encapsulate capsule material is a water-soluble capsule, and described in GB1464616, it is made up of the matrix of polysaccharide and polyol.Other suitable water-soluble encapsulate capsule material comprises the dextrin that the starch acid esters by the not gelation of the dicarboxylic acid that replaces described in US3455838 obtains.These acid esters dextrin are preferably by starch, and for example nephrite rice, soft Chinese sorghum, sago, tapioca (flour) and potato prepare.The suitable example of described encapsulate capsule material comprises the N-Lok by National Starch preparation.N-Lok encapsulate capsule material is made up of modified corn starch and glucose.Starch is by adding simple function group substituted radical, for example octenyl succinic acid anhydride modification.

Suitable anti-redeposition and the soil-suspending agent of the present invention comprises derivatived cellulose, for example methylcellulose gum, carboxymethyl cellulose and Natvosol and poly carboxylic acid or its salt all or multipolymer.Such polymkeric substance comprises previously mentioned as the polyacrylic ester of washing assistant and the multipolymer of maleic anhydride-acrylic copolymer and maleic anhydride and ethene, methyl ethylene ester or methacrylic acid, and maleic anhydride accounts at least 20% mole of multipolymer.These materials are usually with 0.5%-10% by weight, and more preferably 0.75%-8% most preferably presses the amount of composition weight meter 1%-6% and uses.

Preferred white dyes is the negatively charged ion feature, the example is 4,4 '-two-(2-alcohol amido-4-aniline-s-triazine-6-base is amino) stilbene-2:2 ' disulfonic acid disodium, 4,4 '-two-(2-morpholino base-4-aniline-s-triazine-6-base is amino) stilbene-2:2 ' disulfonic acid disodium, 4,4 '-two-(2,4-pentanoic-s-triazine-6-base is amino) stilbene-2:2 ' disulfonic acid disodium, 4 '; 4 "-two-(2,4-pentanoic-s-triazine-6-base is amino) stilbene-2:2 ' disulfonate sodium, 4,4 '-two-(2-aniline-4-(N-methyl-N-2-hydroxyethyl amino)-s-triazine-6-base is amino) stilbene-2:2 ' disulfonic acid disodium, 4,4 '-two-(4-phenyl-2,1,3-triazole-2-yl) stilbene-2:2 ' disulfonic acid disodium, 4,4 '-two-(2-aniline-4-(1-methyl-2-hydroxyethyl amino)-s-triazine-6-base is amino) stilbene-2:2 ' disulfonic acid disodium, 2-(stilbene radicals-4 "-(naphtho--1 '; 2 ': 4; 5)-1; 2; 3-triazole-2 "-sodium sulfonate and 4,4 '-two (2-sulfo group styryl) biphenyl.Preferred whitening agent is a disclosed concrete whitening agent among the EP753567.

Other useful polymkeric substance is a polyoxyethylene glycol, and especially molecular weight is 1000-10000, more preferably 2000-8000, most preferably from about 4000 polymkeric substance.They are with 0.20%-5% by weight, and more preferably the amount of 0.25%-2.5% is used.The polycarboxylate of these polymkeric substance and homopolymerization mentioned above or copolymerization is improving whiteness maintenance, fabric dust deposit and is being valuable aspect the scourability of earth, protein and oxidable dirt in the presence of transition metal impurity.

The stain remover that is used for the present composition is the conventional terephthalic acid and the multipolymer or the trimer of ethylene glycol and/or the unitary different arrangement modes of propylene glycol.The example of this polymkeric substance the common US4116885 that transfers the possession of and 4711730 and EP0272033 in describe, according to EP-A-0272033, especially preferred polymkeric substance has following formula: (CH ₃(PEG) ₄₃) _0.75(POH) _0.25[T-PO) _2.8(T-PEG) _0.4] T (POH) _0.25((PEG) ₄₃CH ₃) _0.75Wherein PEG is-(OC ₂H ₄) O-, P0 is (0C ₃H ₆O) and T be (pcOC ₆H ₄CO).

Equally of great use be modified poly ester, the random copolymers of dimethyl terephthalate (DMT), sulfoisophthalic acid dimethyl ester, ethylene glycol and 1-2 propylene glycol for example, end group mainly are made up of sulfosalicylic acid ester and less important monoesters by ethylene glycol and/or propylene glycol.Target is to obtain at two ends by the end capped polymkeric substance of sulfosalicylic acid ester, and in the present invention, most of above-mentioned multipolymer of the present invention " mainly " is by sulfosalicylic acid ester end-blocking.Yet some multipolymer will not be end capped fully, thereby their end group can be made up of the ethylene glycol and/or the third 1-2 glycol, and its " inferior strategic point " forms this material.

The polyester that the present invention selects contains about by weight 46% dimethyl terephthalate (DMT), by weight about 16% the third-1,2 glycol, about 10% ethylene glycol, about 13% sulfosalicylic acid dimethyl ester and about by weight 15% sulfoisophthalic acid by weight by weight, its molecular weight is about 3000.Polyester and their preparation method describe in detail in EPA311342.

The enzyme that promptly exists in the inactivation detergent composition of the known free chlorine that in tap water, exists of people in the prior art.Therefore, use chlorine scavenger to press the about content more than 0.1% of general composition weight meter in prescription, for example perborate, ammonium sulfate, S-WAT or poly-inferior ethyliminum will provide the improvement stability of detergent enzyme in washing process.The composition that contains chlorine scavenger is described in the european patent application 92870018.6 of application on January 31st, 1992.

Oxyalkylated polycarboxylate by the polyacrylate preparation is used for the present invention so that the additional performance of deoiling to be provided.This material is described in WO91/08281 and PCT90/01815 page 4, classifies this paper reference as.Contain the polyacrylate of the oxyethyl group side chain in every 7-8 acrylate unit at these materials chemically.Side chain has formula-(CH ₂CH ₂O) _m(CH ₂) _nCH ₃, wherein m is 2-3, n is 6-12.The side chain ester is connected in polyacrylate " skeleton " so that " comb shape " polymer-type structure to be provided.Molecular weight can change, but is generally about 2000-about 50000.This alkoxylate polycarboxylate can account for about 0.05%-about 10% of present composition weight.

Dispersion agent

Washing composition of the present invention and/or Fabrid care composition can contain dispersion agent: suitable water-soluble organic salt is all or co-polymeric acids or its salt, and wherein poly carboxylic acid contains and is no more than at least two carboxyls that two carbon atoms separate.Such polymkeric substance is open in GB-A-1596756.The example of this salt is the multipolymer of the polyacrylate of MW2000-5000 and they and maleic anhydride, and this multipolymer has the molecular weight of 1000-100000.

Especially molecular weight is 4000 the acrylate and the multipolymer of methacrylate, and for example 480N can add by the content of composition weight meter 0.5-20% in washing composition of the present invention and/or the Fabrid care composition.

Composition of the present invention can contain calcium soap peptizing agent compound, and it has as what give a definition and is no more than 8, preferably is no more than 7, is most preferably not exceeding 6 lime soap dispersing power (LSDP).Calcium soap peptizing agent compound preferably exists with the amount of 0%-20% by weight.

The various measuring results of calcium soap peptizing agent effect provide by calcium soap peptizing agent ability (LSDP), it is by using at article H.C.Borghetty and C.A.Bergman, and the calcium soap distributed test of describing in the 88-90 page or leaf of J.Am.Oil.Chem.Soc. the 27th volume is measured.This calcium soap distributed test method is extensive use of by the practitioner in this area, and is relevant with for example following comment; W.N.Linfield, Surfactant science Series, the 7th volume, page 3; W.N.Linfield, Tenside surf.det. the 27th volume, 159-163 page or leaf (1990) and M.K.Nagarajan, W.F.Masler, Cosmetics and Toiletries, the 104th volume, 71-73 page or leaf (1989).LSDP is at 333ppm lime carbonate (calcium: magnesium=3: 2) disperse the required dispersion agent of the calcium soap precipitates that formed by the 0.025g sodium oleate and the % weight ratio of sodium oleate in the 30ml water of equivalent hardness.

Tensio-active agent with good calcium soap peptizing agent ability comprises some amine oxide, trimethyl-glycine, sultaine, alkyl ethoxy sulfate and ethoxylated alcohol.

Having LSDP is no more than 8 the example that is used for tensio-active agent of the present invention and comprises C ₁₆-C ₁₈Dimethyl oxidation amine, average degree of ethoxylation are the C of 1-5 ₁₂-C ₁₈Alkyl ethoxy sulfate, especially average degree of ethoxylation are 3 C ₁₂-C ₁₅Alkyl ethoxy sulfate surfactant (LSDP=4) and average degree of ethoxylation are the C of 12 (LSDP=6) or 30 ₁₄-C ₁₅Alkyl ethoxylated alcohol is sold with trade(brand)name Lutensol AO12 and Lutensol A030 respectively by BASF GmbH.

Be applicable to polymerization calcium soap peptizing agent of the present invention M.K.Nagarajan W.F.Masler at Cosmetics and Toiletries, the 104th the volume, describe in the article in the 71-73 page or leaf (1989).

The hydrophobic bleach agent is the amino caproyl of 4-[N-capryloyl-6-for example] benzene sulfonate; the amino caproyl of 4-[N-nonanoyl-6-] benzene sulfonate, the amino caproyl of 4-[N-decanoyl-6-] benzene sulfonate and their mixture and nonanoly acyloxy benzene sulfonate also can be used as calcium soap peptizing agent compound with the hydrophilic/hydrophobic SYNTHETIC OPTICAL WHITNER.

Dye transfer suppresses

Washing composition of the present invention and/or Fabrid care composition also contain and are suppressed at the dyestuff that comprises the dissolving that runs in the washing operation of being with yarn dyed fabric and suspension is transferred to another kind of fabric from a kind of fabric compound.

The polymeric dye transfer inhibitor

Washing composition of the present invention and/or Fabrid care composition contain 0.001%-10% by weight usually, preferred 0.01%-2%, more preferably 0.05%-1% polymeric dye transfer inhibitor.In detergent composition, add described polymeric dye transfer inhibitor so as the dye transfer of inhibition zone yarn dyed fabric to other fabric that therewith washs.In washing process, before the dyestuff that DYED FABRICS washes out had an opportunity to contact other article, this polymkeric substance had the ability of complexing or absorbing dye.

Especially suitable polymeric dye transfer inhibitor is multipolymer, polyvinylpyrrolidonepolymers polymers, Ju Yi Xi oxazolin ketone and the polyvinyl imidazol or their mixture of polyamine N-oxide pllymers, N-vinyl pyrrolidone and N-vinyl imidazole.

Add this polymkeric substance and improved the performance of enzyme of the present invention.

A) polyamine N-oxide pllymers

The polyamine N-oxide pllymers that is suitable for contains the unit with following structural formula: Wherein P is polymerisable unit, and the R-N-O group can the coupled or wherein polymerisable unit of R-N-O group component part or both combinations.A is -O-,-S-,-N-; X is 0 or 1;

R is aliphatic series, aliphatic, the aromatic series of ethoxylation, heterocycle or alicyclic group or its any combination, the part that the nitrogen of N-O group can nitrogen coupled or wherein N-O group is these groups.

The N-O group can be represented by following general structural formula:

Wherein R1, R2 and R3 are aliphatic group, aromatic series, heterocycle or carbon ring group or their combination, x or/and y or/and z is 0 or 1, the part that the nitrogen of N-O group can be connected with it or wherein the nitrogen of N-O group constitutes these groups wherein.

The N-O group can be the part of polymerizable unit (P) or can link to each other with polymeric skeleton or both combinations.

Wherein the suitable polyamine N-oxide of the part of N-O group formation polymerizable unit comprises polyamine N-oxide, and wherein R is selected from aliphatic series, aromatic series, carbocyclic ring or heterocyclic group.

The above-mentioned polyamine N-oxide of one class comprises one group of polyamine N-oxide, and wherein the nitrogen of N-O group constitutes the part of R-group.Preferred polyamine N-oxide compound is that wherein R is a heterocyclic group, for example the compound of pyridine, pyrroles, imidazoles, tetramethyleneimine, piperidines, quinoline, acridine and their derivative.

Another kind of above-mentioned polyamine N-oxide comprises one group of polyamine N-oxide, and wherein the nitrogen of N-O group is connected in the R group.

Other suitable polyamine N-oxide is the polyamine oxide compound that is connected with polymerizable unit of N-O group wherein.

These polyamine N-oxide of preferred class are the polyamine N-oxide with general formula (I), and wherein R is aromatic series, heterocycle or carbon ring group, and wherein the nitrogen of N-O functional group is the part of described R group.

The example of this compounds is the polyamine oxide compound, and wherein R is a heterogeneous ring compound, for example pyridine, pyrroles, imidazoles and their derivative.

The polyamine N-oxide of another preferred class is the polyamine oxide compound with general formula (I), and wherein R is aromatic series, heterocycle or alicyclic group, and wherein the nitrogen of N-O functional group is connected in described R group.

The example of this compounds is the polyamine oxide compound, and wherein the R group can be aryl, for example phenyl.

Can use any polymer backbone as long as the amine oxide polymers that forms is water miscible and has the dye transfer inhibition activity.The example of suitable polymeric skeleton is polyethylene, polyalkylene, polyester, polyethers, polymeric amide, pi, polyacrylic ester and their mixture.

The ratio that common amine n-oxide polymkeric substance of the present invention has amine and amine n-oxide is 10: 1-1: 1000000.Yet the amount of the amine oxide group that exists in the polyamine oxide polymer can change by suitable copolymerization or by suitable N-degree of oxidation.The ratio of amine and amine n-oxide is preferably 2: 3-1: 1000000, more preferably 1: 4-1: 1000000, most preferably be 1: 7-1: 1000000.In fact polymkeric substance of the present invention comprises random or segmented copolymer, and one of them monomer type is an amine n-oxide, and another monomer type is amine n-oxide or is not.The unitary PKa of the amine oxide of polyamine N-oxide＜10, preferred PKa＜7, more preferably PKa＜6.

The polyamine oxide compound can obtain by almost any extent of polymerization, and extent of polymerization is not crucial, as long as material has required water-soluble and dye suspension ability.

Molecular-weight average is generally 500-1000000; Preferred 1000-50000, more preferably 2000-30000, most preferably 3000-20000.

B) multipolymer of N-vinyl pyrrolidone and N-vinyl imidazole

Be used for N-vinyl imidazole N-vinyl pyrrolidone polymer of the present invention and have 5000-1000000, the molecular-weight average of preferred 5000-200000.

The preferred polymkeric substance that is used for cleaning composition of the present invention comprises the polymkeric substance that is selected from N-vinyl imidazole N-vinylpyrrolidone copolymer, wherein said polymkeric substance has 5000-50000, more preferably 8000-30000, the most preferably molecular-weight average of 10000-20000.

Average molecular weight range is measured by photoscanning described in " modernism of polymer characterization " by as Barth H.G. and Mays J.W. " chemical analysis " 113 volumes.

Preferred N-vinyl imidazole N-nvp copolymer has 5000-50000; More preferably 8000-30000; The molecular-weight average of 10000-20000 most preferably.

With above-mentioned molecular-weight average is that the N-vinyl imidazole N-vinylpyrrolidone copolymer of feature provides fabulous dye transfer inhibition activity, and does not influence the cleaning composition scourability with its preparation unfriendly.

The mol ratio that N-vinyl imidazole N-vinylpyrrolidone copolymer of the present invention has N-vinyl imidazole and N-vinyl pyrrolidone is 1-0.2, more preferably 0.8-0.3, most preferably 0.6-0.4.

C) Polyvinylpyrolidone (PVP)

Washing composition of the present invention and/or Fabrid care composition can also use molecular-weight average to be about 2500-about 400000, preferred about 5000-about 200000, more preferably from about 5000-is about 50000, most preferably from about the Polyvinylpyrolidone (PVP) of 5000-about 15000 (" PVP ").Suitable Polyvinylpyrolidone (PVP) commercial by ISP Corporation, New York, NY and Montreal, Canadian with ProductName PVP K-15 (viscosity molecular weight is 10000), PVP K-30 (molecular-weight average 40000), PVP K-60 (molecular-weight average 160000) and PVP K-90 (molecular-weight average 360000).Can comprise Sokalan HP 165 and Sokalan HP 12 by other suitable Polyvinylpyrolidone (PVP) that BASF Cooperation obtains commercial; Polyvinylpyrolidone (PVP) known to the skilled in detergent applications (referring to for example EP-A-262897 and EP-A-256696).

D) Ju Yi Xi oxazolidinone:

Washing composition of the present invention and/or Fabrid care composition also can use Ju Yi Xi oxazolidinone as the polymeric dye transfer inhibitor, described Ju Yi Xi oxazolidinone has about 2500-about 400000, preferred about 5000-about 200000, more preferably from about 5000-is about 50000, most preferably from about the molecular-weight average of 5000-about 15000.

E) polyvinyl imidazol:

Washing composition of the present invention and/or Fabrid care composition also can use polyvinyl imidazol as the polymeric dye transfer inhibitor, described polyvinyl imidazol has about 2500-about 400000, preferred about 5000-about 200000, more preferably from about 5000-is about 50000, most preferably from about the molecular-weight average of 5000-about 15000.

F) cross-linked polymer:

Cross-linked polymer is that wherein skeleton is interconnected to a certain degree polymkeric substance; These connections can be chemistry or physical properties, can be the active groups on skeleton or the side chain; Cross-linked polymer is described in the 1035-1039 page or leaf at " polymkeric substance meeting will " 22 volumes.

In one embodiment, prepare cross-linked polymer in some way and make them form three-dimensional rigid structure, it can catch dyestuff in the hole that is formed by three-dimensional structure.In another embodiment, cross-linked polymer is caught dyestuff by swelling.This cross-linked polymer is described in the patent application of not examining 94870213.9.

Washing methods

Composition of the present invention can be used for any washing or cleaning method basically, the method that it comprises immersion process, pretreatment process and can add the rinse step of independent rinsing auxiliary composition.

Method of the present invention comprises fabric, tableware or any other crust to contact with washing soln with mode hereinafter described usually.Method of the present invention is carried out in washing process easily.Washing methods especially carries out between 10 ℃-60 ℃ preferably at 5 ℃-95 ℃.The pH of treatment soln is preferably 7-12.

Preferred manual dishwashing method is included in to be used strong solution or soaks in the diluting soln of the large volume of detergent composition on the tableware surface.

Following embodiment is used to illustrate composition of the present invention, rather than limits or define in addition scope of the present invention.In washing composition and/or Fabrid care composition, the content of enzyme represents that with the pure enzyme of general composition weight meter except as otherwise noted, detergent component is to represent by general composition weight meter.The component symbol of abbreviation has following implication: LAS: straight chain C _11-13Sodium alkyl benzene sulfonate TAS: Tallow, beef sodium alkyl sulfate C _XYAS: C _1x-C _1ySodium alkyl sulfate C _XYSAS: C _1x-C _1ySecondary (2,3) sodium alkyl sulfate C _XYE _Z: with the C of average z moles of ethylene oxide condensation _1x-C _1yBasic straight chain primary alcohol C _XYE _ZS: with the C of average z moles of ethylene oxide condensation _1x-C _1ySodium alkyl sulfate QAS: R ₂.N ⁺(CH ₃) ₂(C ₂H ₄OH), R ₂=C ₁₂-C ₁₄QAS1: R ₂.N ⁺(CH ₃) ₂(C ₂H ₄OH), R ₂=C ₈-C ₁₁APA: C _8-10Amido propyl-dimethyl amine soap: the straight-chain alkyl carboxylic acid's sodium nonionogenic tenside that obtains by Tallow, beef and cocinic acid 80/20 mixture: C ₁₃-C ₁₅Mixed ethoxylated/propoxylated fatty alcohol, average degree of ethoxylation 3.8, flat

Equal degree of propoxylation 4.5Neodol 45-13: C ₁₄-C ₁₅The straight chain primary alcohol ethoxylate is sold STS by Shell Chemical CO: toluenesulfonic acid sodium salt CFAA: C ₁₂-C ₁₄Alkyl N-methyl glucose amide TFAA: C ₁₆-C ₁₈Alkyl N-methyl glucose amide TPKFA: C ₁₂-C ₁₄The full cut lipid acid of topping silicate: amorphous sodium silicate (SiO ₂: Na ₂The metasilicate of O ratio=1.6-3.2): Starso (SiO ₂: Na ₂O ratio=1.0) zeolite A: formula Na ₁₂(AlO ₂SiO ₂) ₁₂.27H ₂The hydrated sodium aluminosilicate of O, main particle size is 0.1-10

Micron (weight of representing based on anhydrous benchmark) NaSKS-6: formula δ-Na ₂Si ₂O ₅The crystalline layered silicate Citrate trianion: citrate trisodium dihydrate, active 86.4%, particle size distribution 425-850 micron citric acid: Citric Acid, usp, Anhydrous Powder borate: Sodium Tetraborate carbonate: anhydrous sodium carbonate, particle size 200-900 micron supercarbonate: anhydrous sodium bicarbonate, particle size distribution is a 400-1200 micron vitriol: anhydrous sodium sulphate sal epsom: anhydrous magnesium sulfate STPP: tripoly phosphate sodium STPP TSPP: tetrasodium pyrophosphate MA/AA: 4: 1 acrylate/maleic acid ester random copolymerss, the about 70000-80000MA/AA1 of molecular-weight average: 6: 4 acrylate/maleic acid ester random copolymerss, the about 10000AA of molecular-weight average: molecular-weight average is 4500 sodium polyacrylate polymer P A30: molecular-weight average is the polyacrylic acid 480N of about 4500-8000: 7: 3 acrylate/methacrylate random copolymerss, the about 3500Polygel/carbopol of molecular-weight average: polymer crosslinked polypropylene PB1: anhydrous sodium perborate monohydrate, empirical formula: NaBO ₂.H ₂O ₂PB4: anhydrous sodium perborate tetrahydrate, empirical formula: NaBO ₂.3H ₂O.H ₂O ₂Percarbonate: anhydrous SPC-D, empirical formula: 2Na ₂CO ₃.3H ₂O ₂NaDCC: dichloroisocyanuric acid sodium TAED: tetra acetyl ethylene diamine NOBS: nonanoly acyloxy benzene sulfonate; sodium-salt form NACA-OBS: (the amino caproyl of 6-nonanoyl) oxygen biphenyl sulfonate DTPA: diethylenetriamine pentaacetic acid HEDP: 1; 1-hydroxyl ethane di 2 ethylhexyl phosphonic acid DETPMP: two inferior second triamines five (methylene phosphonic acid); all sell EDDS: quadrol-N by Meng Shan with trade(brand)name Dequest 2060; N '-disuccinic acid; (S; S) isomer; sodium-salt form MnTACN: 1; 4; 7-trimethylammonium-1; 4; the 7-7-triazacyclononane closes manganese photoactivation SYNTHETIC OPTICAL WHITNER: the capsular sulfonation zinc phthalocyanine phthalocyanine photoactivation SYNTHETIC OPTICAL WHITNER 1 of bag in the dextrin dissolved polymers: the capsular sulphonation aluminum phthalocyanine PAAC of bag in the dextrin dissolved polymers: five amine cobaltous acetate (III) salt paraffin: the paraffin oil NaBz that is sold with trade mark Winog by Wintershall: Sodium Benzoate BzP: benzoyl peroxide mannase: Bacillus agaradherens; the mannosans protease enzyme that NICMB40482 obtains: with trade(brand)name Savinase; Alcalase; Durazym (Novo Industries A/S)

The protease that Maxacal, Maxapem (Gist-Brocades) sell and in patent

The proteolytic enzyme amylase of describing among WO91/06637 and/or WO95/10591 and/or the EP251446: at WO94/18314, the trade(brand)name that the Genencor that describes among the WO96/05295 sells

Purafact Ox Am , the Termamyl  that obtains by Novo Nordisk A/S Denmark,

Fungamyl  and Duramyl  and the amylolytic enzyme lipase of in WO95/26397, describing: by Novo Nordisk A/S with trade(brand)name Lipolase, Lipolase Ultra and by

The lipolytic enzyme cellulase that Gist-Brocades sells with Lipomax: by Novo Industries A/S with trade(brand)name Carezymea, Celluzyme and/or

The cellulolytic enzyme CMC that Endolase sells: Xylo-Mucine PVP: polyethylene polymer, molecular-weight average is 60000PVNO: polyvinylpyridine-N-oxide compound, molecular-weight average is 50000PVPVI: vinyl imidazole and vinylpyrrolidone copolymer, molecular-weight average is 20000 whitening agent 1: 4,4 '-two (2-sulfo group styryl) biphenyl disodium whitening agent 2: 4,4 '-two (4-anilinos-6-morpholino-1,3,5-triazine-2-yl) stilbene-2:2 '-disulfonic acid disodium polysiloxane defoamers: with siloxanes-oxyalkylene copolymers is the polydimethylsiloxane Foam Control of dispersion agent, institute

The ratio of stating Foam Control and described dispersion agent is 10: 1-100: 1 suds suppressor: 12% polysiloxane/silicon oxide, 18% stearyl alcohol, 70% starch, particle form opalizer: water base single vinylbenzene latex mixture, sell SRP1 by BASF with trade(brand)name Lytron621: the end capped polyester SRP2 of negatively charged ion: diethoxyization is gathered (1,2 propylidene phthalic ester) short chain block polymer QEA: two ((C ₂H ₅O) (C ₂H ₄O) _n) (CH ₃)-N ⁺-C ₆H ₁₂-N ⁺-(CH ₃) two ((C ₂H ₅O)-(C ₂H ₄O)) _n,

N=20-30PEI wherein: polymine, molecular-weight average 1800, average degree of ethoxylation 7SCS: cumene sodium sulfonate HMWPFO: polyphosphazene polymer oxyethane PEGx: polyoxyethylene glycol, molecular weight xPEO: polyethylene oxide, molecular-weight average 5000TEPAE: tetraethylene-pentamine ethoxylate BTA: benzotriazole pH: 20 ℃ of 1% measured in solution that are used in the distilled water

Embodiment 1

Be prepared as follows high-density laundry detergent composition of the present invention:

I II III IV

LAS 8.0 2.0 6.0 6.0

TAS 0.5 0.5 1.0 0.1

C46(S)AS 2.5 - - -

C25AS - 7.0 4.5 5.5

C68AS 5.0 - - -

C25E5 - 10.0 4.6 4.6

C25E7 3.4 - - -

C25E3S - 2.0 5.0 4.5

QAS 0.8 - - -

QAS?1 - 0.8 0.5 1.0

Zeolite A 18.0 18.1 20.0 18.1

Citric acid-2.5-2.5

Carbonate 13.0 10.0 10.0 13.0

Na-SKS-6 - 10.0 - 10.0

Silicate 1.4 0.3 0.5 0.3

Citrate trianion 1.0 3.0--

Vitriol 26.1 6.0--

Sal epsom-0.2-0.2

MA/AA 0.3 4.0 1.0 1.0

CMC 0.2 0.2 0.4 0.4

PB4 9.0 - - -

Percarbonate--18.0 18.0

TAED 0.4 - 3.9 4.2

NACA-OBS 2.0 - - -

DETPMP 0.25 0.25 - -

SRP1 - 0.2 - 0.2

EDDS 0.25 - 0.5 0.5

CFAA 1.0 2.0 - -

HEDP 0.3 0.3 0.4 0.4

QEA - 0.2 - 0.5

Mannase 0.001 0.02 0.001 0.002

Proteinase-10 .009 0.04 0.05 0.03

Amylase 0.002 0.006 0.008 0.008

Cellulase-0.0007 0.0007 0.0007

Lipase-0.01 0.01 0.01

Photoactivation SYNTHETIC OPTICAL WHITNER (ppm) 15-20 20

PVNO/PVPVI - 0.1 - -

Whitening agent 1 0.09-0.09 0.09

Spices 0.3 0.4 0.4 0.4

Polysiloxane defoamers 0.5-0.3 0.3

Density, g/l 850 850 850 850

Other/minor component to 100%

Embodiment 2

Following granular laundry detergent and/or Fabrid care composition that preparation the present invention especially uses under European washing machine condition:

I II III IV V VI

LAS 5.5 7.5 5.0 5.0 6.0 7.0TAS 1.25 1.9-0.8 0.4 0.3C24AS/C25AS-2.2 5.0 5.0 5.0 2.2C25E3S-0.8 1.0 1.5 3.0 1.0C45E7 3.25----3.0TFAA--2.0---C25E5-5.5----QAS 0.8-----QAS1-0.7 1.0 0.5 1.0 0.7STPP 19.7-----A-19.5 25.0 19.5 20.0 17.0Na-SKS-6/-10.6-10.6-- ( 79∶21Na-SKS-6--9.0-10.0 10.0 6.0 21.4 9.0 10.0 10.0 18.0-2.0 7.0 5.0-2.0 6.8--0.3 0.5---4.0 4.0-- 39.8--5.0-12.0--0.1 0.2 0.2-MA/AA 0.5 1.6 3.0 4.0 1.0 1.0CMC 0.2 0.4 1.0 1.0 0.4 0.4PB4 5.0 12.7--------18.0 15.0TAED 0.5 3.1--5.0-NACA-OBS 1.0 3.5---2.5DETPMP 0.25 0.2 0.3 0.4-0.2HEDP-0.3-0.3 0.3 0.3QEA--1.0 1.0 1.0- 0.001 0.002 0.002 0.001 0.002 0.001 0.009 0.03 0.03 0.05 0.05 0.02 0.003 0.003 0.006 0.006 0.006 0.004 0.0006 0.0006 0.0005 0.0005 0.0007 0.0007 0.002 0.002 0.006 0.006 0.01 0.003PVNO/PVPVI--0.2 0.2--PVP 0.9 1.3---0.9SRP1--0.2 0.2 0.2- ( ppm ) 15 27--20 201 ( ppm ) 15-----1 0.08 0.2--0.09 0.152-0.04---- 0.3 0.5 0.4 0.3 0.4 0.3 0.5 2.4 0.3 0.5 0.3 2.0,g/l 750 750 750 750 750 750/ 100％

Embodiment 3

Be prepared as follows granular detergent composition of the present invention:

I II III IV V VI blows powder

LAS 23.0 8.0 7.0 9.0 7.0 7.0

TAS - - - - 1.0 -

C45AS 6.0 6.0 5.0 8.0 - -

C45AES - 1.0 1.0 1.0 - -

C45E35 - - - - 2.0 4.0

Zeolite A 10.0 18.0 14.0 12.0 10.0 10.0

MA/AA - 0.5 - - - 2.0

MA/AA1 7.0 - - - - -

AA - 3.0 3.0 2.0 3.0 3.0

Vitriol 5.0 6.3 14.3 11.0 15.0 19.3

Silicate 10.0 1.0 1.0 1.0 1.0 1.0

Carbonate 15.0 20.0 10.0 20.7 8.0 6.0

PEG4000 0.4 1.5 1.5 1.0 1.0 1.0

DTPA - 0.9 0.5 - - 0.5

Whitening agent 2 0.3 0.2 0.3-0.1 0.3 spraying liquid

C45E7 - 2.0 - - 2.0 2.0

C25E9 3.0 - - - - -

C23E9 - - 1.5 2.0 - 2.0

Spices 0.3 0.3 0.3 2.0 0.3 0.3 agglomerates

C45AS - 5.0 5.0 2.0 - 5.0

LAS - 2.0 2.0 - - 2.0

Zeolite A-6.5 6.5 7.0-7.5

Carbonate-4.0 4.0 5.0-4.0

The dried additive of PEG4000-0.5 0.5--0.5 minor component (water etc.)-2.0 2.0 2.0-2.0

QAS 1.0 1.0 1.0 1.0 1.0 1.0

Citric acid----2.0-

PB4 - - - - 12.0 1.0

PB1 4.0 1.0 3.0 2.0 - -

Percarbonate----2.0 9.0

Carbonate-5.3 1.8-4.0 4.0

NOBS 4.0 - 6.0 - - 0.6

Methylcellulose gum 0.2-----

Na-SKS-6 8.0 - - - - -

STS - - 2.0 - 1.0 -

Cumene sulfonic acid-1.0---2.0

Mannase 0.001 0.002 0.001 0.02 0.001 0.02

Proteinase-10 .02 0.02 0.02 0.01 0.02 0.02

Lipase 0.004-0.004-0.004 0.008

Amylase 0.003-0.002-0.003-

Cellulase 0.0005 0.0005 0.0005 0.0007 0.0005 0.0005

PVPVI - - - - 0.5 0.1

PVP - - - - 0.5 -

PVNO - - 0.5 0.3 - -

QEA - - - - 1.0 -

SRP1 0.2 0.5 0.3 - 0.2 -

Silicone suds suppressor 0.2 0.4 0.2 0.4 0.1-

Sal epsom--0.2-0.2-trace and a small amount of component to 100% embodiment 4 are prepared as follows detergent composition of the present invention:

I II III blows powder

Zeolite A 14.0 15.0 15.0

Vitriol-5.0-

LAS 3.0 3.0 3.0

QAS 1.0 1.5 1.5

DETPMP 0.4 0.2 0.4

EDDS - 0.4 0.2

CMC 0.4 0.4 0.4

MA/AA 4.0 2.0 2.0 agglomerates

LAS 5.0 5.0 5.0

TAS 2.0 2.0 1.0

Silicate 3.0 3.0 4.0

Zeolite A 8.0 8.0 8.0

Carbonate 8.0 8.0 4.0 spray liquid

Spices 0.3 0.3 0.3

C45E7 2.0 2.0 2.0

C25E3 2.0--dried additive

Citrate trianion 5.0-2.0

Supercarbonate-3.0-

Carbonate 8.0 15.0 10.0

TAED 6.0 2.0 5.0

PB1 14.0 7.0 10.0

PEO - - 0.2

Wilkinite--10.0

Mannase 0.01 0.02 0.0015

Proteinase-10 .03 0.03 0.03

Lipase 0.008 0.008 0.008

Cellulase 0.001 0.001 0.001

Amylase 0.01 0.01 0.01

Polysiloxane defoamers 5.0 5.0 5.0

Vitriol-3.0-density (g/l) 850 850 850 minor components and a small amount of component to 100% embodiment 5 preparations following detergent composition of the present invention:

I II III IVLAS 18.0 14.0 23.0 20.0QAS 0.7 1.0 1.0 0.7TFAA-1.0--C23E56.5--1.0-C45E7-1.0--C45E3S 1.0 2.5 1.0-STPP 32.0 18.0 30.0 22.0 9.0 5.0 9.0 8.0 11.0 7.5 10.0 5.0-7.5--PB1 3.0 1.0--PB4-1.0--NOBS 2.0 1.0--DETPMP-1.0--DTPA 0.5-0.2 0.3SRP1 0.3 0.2-0.1MA/AA 1.0 1.5 2.0 0.5CMC 0.8 0.4 0.4 0.2PEI--0.4- 20.0 10.0 20.0 30.0 0.2-0.4 0.9 0.001 0.02 0.001 0.002 0.03 0.03 0.02 0.02 0.008 0.007-0.004 0.004-0.002- 0.0003--0.0001 30ppm 20ppm-10ppm 0.3 0.3 0.1 0.21/2 0.05 0.02 0.08 0.1 100％6：

I II III IV VC17ES 28.5 27.4 19.2 34.1 34.1 2.6 5.0 2.0 3.0 3.0C12--6.0-- 0.9--2.0 2.0QAS 0.5 0.5 0.8 0.2 0.5 2.0 4.0-2.0-Neodol C11E9--5.0-----6.5 6.5 ( 40％ ) --0.03--TAED---0.06 0.06---1.5 1.5 4.0 5.5 5.5 9.1 9.1----2.3---0.5 1.1 0.06 0.1----1.0--- 3.3-0.7----0.4----0.06-- 0.08-------2.2 2.2---1.1 1.1 200ppm 0.16 0.006-- 0.001 0.005 0.001 0.02 0.02 0.017 0.005 .0035 0.003 0.002 0.18 0.09 0.09 0.2 0.2 100％7：

I II III IV V 0.001 0.001 0.001 0.02 0.02 0.01 0.002 0.005-- 0.05 0.01 0.02-----6.0 6.8---2.5-DTPA---0.2----0.05-EDTA* 0.05 0.05 0.05--/ 2.9 2.9 2.9 1.0-LAS 0.5 0.5 0.5--C12AS 0.5 0.5 0.5--C10AS----1.7C12 ( E ) S 0.5 0..5 0.5--C12,13E6.5 7.0 7.0 7.0--Neodol 23-6.5---12.0-Dobanol 23-3----1.5Dobanol 91-10----1.6C25AE1.8S---6.0-QAS1 0.5 0.4 0.5 0.5 1.0---6.0- 1.0 1.0 1.0 0.5 0.2---1.5----1.0-2-----0.5** 1.0 1.0 1.0--SCS 1.3 1.3 1.3--pH 7-12 7-12 7-12 4- 100％*Na4**8：

Mannase 0.005

Amylase 0.01

Proteinase-10 .01

Sodium octyl sulfate 2.0

Sodium lauryl sulphate 4.0

QAS 1.0

Sodium hydroxide 0.8

Silicate 0.04

Diethylene glycol monobutyl ether * 4.0

Spices 0.35

Water/minor component reaches 100%* Diethylene Glycol single-butyl ether

Sequence table The applicant: Title; The Procter ﹠ Gamble Company Street: One Procter ﹠ Gamble Plaza City: Cincinnati, OHIO Country: USA Postcode: 45202 Denomination of invention: the composition of detergent that contains mannase and cationic surfactant Sequence numbering: 6 The computer reading method: Medium type: disk Computer: compatible IBM PC Operating system: PC-DOS/MS-DOS Software: Patentln Release#1.0 Version 1.25 (EPO) SEQ ID NO:1 Sequence signature: Length: 1407 base-pairs Type: nucleic acid Chain: single Topology: straight chain Molecule type: genomic DNA Primary source Feature: Title/keyword: CDS Position: 1-1482 Sequence description: SEQ ID NO:1 ATGAAAAAAAAGTTATCACAGATTTATCATTTAATTATTTGCACACTTATAATA AGTGTGGGAATAATGGGGATTACAACGTCCCCATCAGCAGCAAGTACAGGC TTTTATGTTGATGGCAATACGTTATATGACGCAAATGGGCAGCCATTTGTCAT GAGAGGTATTAACCATGGACATGCTTGGTATAAAGACACCGCTTCAACAGCT ATTCCTGCCATTGCAGAGCAAGGCGCCAACACGATTCGTATTGTTTTATCAG ATGGCGGTCAATGGGAAAAAGACGACATTGACACCATTCGTGAAGTCATTG AGCTTGCGGAGCAAAATAAAATGGTGGCTGTCGTTGAAGTTCATGATGCCA CGGGTCGCGATTCGCGCAGTGATTTAAATCGAGCCGTTGATTATTGGATAG AAATGAAAGATGCGCTTATCGGTAAAGAAGATACGGTTATTATTAACATTGCA AACGAGTGGTATGGGAGTTGGGATGGCTCAGCTTGGGCCGATGGCTATATT GATGTCATTCCGAAGCTTCGCGATGCCGGCTTAACACACACCTTAATGGTTG ATGCAGCAGGATGGGGGCAATATCCGCAATCTATTCATGATTACGGACAAG ATGTGTTTAATGCAGATCCGTTAAAAAATACGATGTTCTCCATCCATATGTAT GAGTATGCTGGTGGTGATGCTAACACTGTTAGATCAAATATTGATAGAGTCA TAGATCAAGACCTTGCTCTCGTAATAGGTGAATTCGGTCATAGACATACTGA TGGTGATGTTGATGAAGATACAATCCTTAGTTATTCTGAAGAAACTGGCACA GGGTGGCTCGCTTGGTCTTGGAAAGGCAACAGTACCGAATGGGACTATTTA GACCTTTCAGAAGACTGGGCTGGTCAACATTTAACTGATTGGGGGAATAGAA TTGTCCACGGGGCCGATGGCTTACAGGAAACCTCCAAACCATCCACCGTAT TTACAGATGATAACGGTGGTCACCCTGAACCGCCAACTGCTACTACCTTGTA TGACTTTGAAGGAAGCACACAAGGGTGGCATGGAAGCAACGTGACCGGTG GCCCTTGGTCCGTAACAGAATGGGGTGCTTCAGGTAACTACTCTTTAAAAGC CGATGTAAATTTAACCTCAAATTCTTCACATGAACTGTATAGTGAACAAAGTC GTAATCTACACGGATACTCTCAGCTCAACGCAACCGTTCGCCATGCCAATTG GGGAAATCCCGGTAATGGCATGAATGCAAGACTTTACGTGAAAACGGGCTC TGATTATACATGGCATAGCGGTCCTTTTACACGTATCAATAGCTCCAACTCA GGAACAACGTTATCTTTTGATTTAAACAACATCGAAAATAGTCATCATGTTAG GGAAATAGGCGTGCAATTTTCAGCGGCAGATAATAGCAGTGGTCAAACTGC TCTATACGTTGATAACGTTACTTTAAGATAG

SEQ ID NO：2

Sequence signature:

Length: 493 amino acid

Type: amino acid

Topology: straight chain

Molecule type: protein

Sequence description: SEQ ID NO:2 MKKKLSQIYHLIICTLIISVGIMGITTSPSAASTGFYVDGNTLYDANGQPFVMRGI N HGHAWYKDTASTAIPAIAEQGANTIRIVLSDGGQWEKDDIDTIREVIELAEQNKM VAVVEVHDATGRDSRSDLNRAVDYWIEMKDALIGKEDTVIINIANEWYGSWDGS AWADGYIDVIPKLRDAGLTHTLMVDAAGWGQYPQSIHDYGQDVFNADPLKNTM FSIHMYEYAGGDANTVRSNIDRVIDQDLALVIGEFGHRHTDGDVDEDTILSYSEE TGTGWLAWSWKGNSTEWDYLDLSEDWAGQHLTDWGNRIVHGADGLQETSKP STVFTDDNGGHPEPPTATTLYDFEGSTQGWHGSNVTGGPVSVTEWGASGNY SLKADVNLTSNSSHELYSEQSRNLHGYSQLNATVRHANWGNPGNGMNARLYV KTGSDYTWHSGPFTRINSSNSGTTLSFDLNNIENSHHVREIGVQFSAADNSSGQ TALYVDNVTLR

SEQ ID NO：3

Sequence signature:

Length: 1407 base-pairs

Type: nucleic acid

Chain: single

Topology: straight chain

Molecule type: genomic DNA

：SEQ ID NO：3 ATGAAAAAAAAGTTATCACAGATTTATCATTTAATTATTTGCACACTTATAATA AGTGTGGGAATAATGGGGATTACAACGTCCCCATCAGCAGCAAGTACAGGC TTTTATGTTGATGGCAATACGTTATATGACGCAAATGGGCAGCCATTTGTCAT GAGAGGTATTAACCATGGACATGCTTGGTATAAAGACACCGCTTCAACAGCT ATTCCTGCCATTGCAGAGCAAGGCGCCAACACGATTCGTATTGTTTTATCAG ATGGCGGTCAATGGGAAAAAGACGACATTGACACCATTCGTGAAGTCATTG AGCTTGCGGAGCAAAATAAAATGGTGGCTGTCGTTGAAGTTCATGATGCCA CGGGTCGCGATTCGCGCAGTGATTAAATCGAGCCGTTGATTATTGGATAG AAATGAAAGATGCGCTTATCGGTAAAGAAGATACGGTTATTATAACATTGCA AACGAGTGGTATGGGAGTTGGGATGGCTCAGCTTGGGCCGATGGCTATATT GATGTCATTCCGAAGCTTCGCGATGCCGGCTTAACACACACCTTAATGGTTG ATGCAGCAGGATGGGGGCAATATCCGCAATCTATTCATGATTACGGACAAG ATGTGTTTAATGCAGATCCGTTAAAAAATACGATGTTCTCCATCCATATGTAT GAGTATGCTGGTGGTGATGCTAACACTGTTAGATCAAATATTGATAGAGTCA TAGATCAAGACCTTGCTCTCGTAATAGGTGAATTCGGTCATAGACATACTGA TGGTGATGTTGATGAAGATACAATCCTTAGTTATTCTGAAGAAACTGGCACA GGGTGGCTCGCTTGGTCTTGGAA AGGCAACAGTACCGAATGGGACTATTTA GACCTTTCAGAAGACTGGGCTGGTCAACATTTAACTGATTGGGGGAATAGAA TTGTCCACGGGGCCGATGGCTTACAGGAAACCTCCAAACCATCCACCGTAT TTACAGATGATAACGGTGGTCACCCTGAACCGCCAACTGCTACTACCTTGTA TGACTTTGAAGGAAGCACACAAGGGTGGCATGGAAGCAACGTGACCGGTG GCCCTTGGTCCGTAACAGAATGGGGTGCTTCAGGTAACTACTCTTTAAAAGC CGATGTAAATTTAACCTCAAATTCTTCACATGAACTGTATAGTGAACAAAGTC GTAATCTACACGGATACTCTCAGCTCAACGCAACCGTTCGCCATGCCAATTG GGGAAATCCCGGTAATGGCATGAATGCAAGACTTTACGTGAAAACGGGCTC TGATTATACATGGCATAGCGGTCCTTTTACACGTATCAATAGCTCCAACTCA GGAACAACGTTATCTTTTGATTTAAACAACATCGAAAATATCATCATGTTAGG GAAATAG

SEQ ID NO：4

Sequence signature:

Length: 468 amino acid

Type: amino acid

Topology: straight chain

Molecule type: protein

：SEQ ID NO：4 MKKKLSQIYHLIICTLIISVGIMGITTSPSAASTGFYVDGNTLYDANGQPFVMRGIN HGHAWYKDTASTAIPAIAEQGANTIRIVLSDGGQWEKDDIDTIREVIELAEQNKM VAVVQEVHDATGRDSRSDLNRAVDYWIEMKDALIGKEDTVIINIANEWYGSWDGS AWADGYIDVIPKLRDAGLTHTLMVDAAGWGQYPQSIHDYGQDVFNADPLKNTM FSIHMYEYAGGDANTVRSNIDRVIDQDLALVIGEFGHRHTDGDVDEDTILSYSEE TGTGWLAWSWKGNSTEWDYLDLSEDWAGQHLTDWGNRIVHGADGLQETSKP STVFTDDNGGHPEPPTATTLYDFEGSTQGWHGSNVTGGPWSVTEWGASGNY SLKADVNLTSNSSHELYSEQSRNLHGYSQLNATVRHANWGNPGNGMNARLYV KTGSDYTWHSGPFTRINSSNSGTTLSFDLNNIENIIMLGK SEQ ID NO：5 ：：1029 ：：：：DNA ：SEQ ID NO：5 5′ AAT TGG CGC ATA CTG TGT CGC CTG TGA ATC CTA ATG CCC AGC AGA CAA CAA AAA CAG TGA TGA ACT GGC TTG CGC ACC TGC CGA ACC GAA CGG AAA ACA GAG TCC TTT CCG GAG CGT TCG GAG GTT ACA GCC ATG ACA CAT TTT CTA TGG CTG AGG CTG ATA GAA TCC GAA GCG CCA CCG GGC AAT CGC CTG CTA TTT ATG GCT GCG ATT ATG CCA GAG GAT GGC TTG AAA CAG CAA ATA TTG AAG ATT CAA TAG ATG TAA GCT GCA ACG GCG ATT TAA TGT CGT ATT GGA AAA ATG GCG GAA TTC CGC AAA TCA GTT TGC ACC TGG CGA ACC CTG CTT TTC AGT CAG GGC ATT TTA AAA CAC CGA TTA CAA ATG ATC AGT ATA AAA ACA TAT TAG ATT CAG CAA CAG CGG AAG GGA AGC GGC TAA ATG CCA TGC TCA GCA AAA TTG CTG ACG GAC TTC AAG AGT TGG AGA ACC AAG GTG TGC CTG TTC TGT TCA GGC CGC TGC ATG AAA TGA ACG GCG AAT GGT TTT GGT GGG GAC TCA CAT CAT ATA ACC AAA AGG ATA ATG AAA GAA TCT CTC TAT ATA AAC AGC TCT ACA AGA AAA TCT ATC ATT ATA TGA CCG ACA CAA GAG GAC TTG ATC ATT TGA TTT GGG TTT ACT CTC CCG ACG CCA ACC GAG ATT TTA AAA CTG ATT TTT ACC CGG GCG CGT CTT ACG TGG ATA TTG TCG GAT TAG ATG CGT ATT TTC AAG ATG CCT ACT CGA TCA ATG GAT ACG ATC AGC TAA CAG CGC TTA ATA AAC CAT TTG CTT TTA CAG AAG TCG GCC CGC AAA CAG CAA ACG GCA GCT TCG ATT ACA GCC TGT TCA TCA ATG CAA TAA AAC AAA AAT ATC CTA AAA CCA TTT ACT TTC TGG CAT GGA ATG ATG AAT GGA GCG CAG CAG TAA ACA AGG GTG CTT CAG CTT TAT ATC ATG ACA GCT GGA CAC TCA ACA AGG GAG AAA TAT GGA ATG GTG ATT CTT TAA CGC CAA TCG TTG AGT GAA TCC GGG ATC 3′ SEQ ID NO：6 ：：363 ：：：：SEQ ID NO：6 ydhT 1 LFKKHTISLLIIFLLASAVLAKPIEAHTVSPVNPNAQQTTKTVMNWLAHL 50 ydhT 51 PNRTENRVLSGAFGGYSHDTFSMAEADRIRSATGQSPAIYGCDYARGWLE 100 ydhT 101 TANIEDSIDVSCNGDLMSYWKNGGIPQISLHLANPAFQSGHFKTPITNDQ 150 ydhT 151 YKNILDSATAEGKRLNAMLSKIADGLQELENQGVPVLFRPLHEMNGEWFW 200 ydhT 201 WGLTSYNQKDNERISLYKQLYKKIYHYMTDTRGLDHLIWVYSPDANRDFK 250 ydhT 251 TDFYPGASYVDIVGLDAYFQDAYSINGYDQLTALNKPFAFTEVGPQTANG 300 ydhT 301 SFDYSLFINAIKQKYPKTIYFLAWNDEWSAAVNKGASALYHDSWTLNKGE 350 ydhT 351 IWNGDSLTPIVE*.363

Claims

1. washing composition and/or Fabrid care composition, it contains mannase and cats product.

2. the washing composition of claim 1 and/or Fabrid care composition, wherein said mannase to be accounting for 0.0001%-2% by the pure enzyme of general composition weight meter, preferred 0.0005%-0.5%, and more preferably the content of 0.001%-0.02% exists.

3. the washing composition of claim 1-2 and/or Fabrid care composition, wherein cats product is to press general composition weight meter 0.2-25%, and the content of preferred 1%-8% exists.

4. the washing composition of claim 1-3 and/or Fabrid care composition, wherein cats product is selected from ammonium surfactant and/or its ethoxylated derivative.

5. the washing composition of claim 4 and/or Fabrid care composition, wherein ammonium surfactant is alkyl trimethyl ammonium halide, alkyl dimethyl-hydroxyalkyl ammonium halide, the two hydroxyalkyl ammonium halides of alkyl methyl and/or their mixture.

6. the washing composition of claim 5 and/or Fabrid care composition, wherein ammonium surfactant is selected from C12-14 alkyl hydroxy ethyl alkyl dimethyl ammonium chloride, C8-C11 alkyl hydroxy ethyl alkyl dimethyl ammonium chloride and/or their mixture.

7. the washing composition of above-mentioned arbitrary claim and/or Fabrid care composition, it also contains anion surfactant, preferred alkyl vitriol and/or sulfonate.

8. the washing composition of above-mentioned arbitrary claim and/or Fabrid care composition, it also contains nonionogenic tenside, preferably have the C8-C20 chain length, preferred C12-C16 and degree of ethoxylation are 2-9, the alkyl ethoxylated nonionogenic tenside of preferred 3-7; Alkyl chain length is C8-C20, the alkyl methyl glucamide tensio-active agent of preferred C12-C18; And/or their mixture.

9. the washing composition of claim 1-4 and/or Fabrid care composition, wherein said cats product contains two long alkyl chains.

10. with the method for washing composition and/or Fabrid care composition laundering of textile fabrics, tableware and/or the crust of above-mentioned arbitrary claim.