HK1159155B - Enzymatic textile bleaching compositions and methods of use thereof - Google Patents

Description

Enzymatic textile bleaching compositions and methods of use thereof

Priority

The present application claims priority from U.S. provisional patent application serial nos. 61/095,807, 61/099,020, and 61/156,593, filed on 10/9/2008, 22/9/2008, and 3/2/2009, respectively, each of which is incorporated by reference in its entirety.

Technical Field

The present compositions and methods relate to enzymatic bleaching of textiles.

Background

In the processing of textile fibres, yarns and fabrics, a pre-treatment or pre-treatment step is often required to prepare these natural materials appropriately for further use, especially in the dyeing, printing and/or finishing stages that are often required for commercial goods. These textile treatment steps remove impurities and color bodies that are naturally present or added to the fibers and/or fabrics during spinning or weaving.

Textile production typically includes a number of treatments and stages, most commonly: desizing (i.e., removal of sizing agents, such as starch, by enzymatic, alkaline, or oxidizing agent soaking); scouring (i.e., removal of grease, oil, wax, pectic substances, particulates, proteins and fats by contact with sodium hydroxide solution at near boiling temperature); and bleaching (i.e., removal and bleaching of color bodies from textiles by conventional use of oxidizing agents such as hydrogen peroxide, hypochlorite, and chlorine dioxide, or by use of reducing agents such as sulfur dioxide or bisulfite). A common bleaching technique involves bleaching with alkaline hydrogen peroxide at temperatures in excess of 95 ℃. This high temperature and strong bleaching system requires high energy input and typically produces high pH effluent, which is undesirable from an environmental sustainability standpoint.

There is a need for an efficient enzymatic textile bleaching process that minimizes environmental footprint and textile mill costs, provides improved fabric strength retention and reduced fiber damage compared to conventional textile bleaching processes. The enzymatic bleaching process preferably operates at lower pH and lower temperature, reduces the use of corrosive chemicals, and is environmentally friendly over conventional processes.

Summary of The Invention

The compositions and methods of the present invention relate to enzymatic bleaching of textiles. Use of the textile bleaching compositions and methods of the present invention results in bleached textiles with reduced textile damage, bulkier softer handle, and/or increased dye uptake as compared to chemical bleaching of textiles.

In one aspect, the present invention provides an enzymatic textile bleaching composition comprising: (i) perhydrolases (perhydrolases); (ii) an ester substrate of said perhydrolase; (iii) a source of hydrogen peroxide; (iv) surfactants and/or emulsifiers; (v) a peroxide stabilizer; (vi) sequestering agents (sequestrant agents); and (vii) a buffer to maintain a pH of about 6 to about 8.

In some embodiments, the perhydrolase comprises SEQ ID NO:1, or a variant or homologue thereof. In particular embodiments, the perhydrolase is SEQ ID NO:1 (i.e., a variant of SEQ ID NO:1 with the substitution S54V). In some embodiments, the perhydrolase enzyme comprises (i.e., exhibits) a perhydrolysis to hydrolysis ratio greater than 1. In some embodiments, the perhydrolase enzyme is present at a concentration of about 1 to about 2.5ppm, e.g., about 1.7 ppm.

In some embodiments, the ester substrate is selected from the group consisting of propylene glycol diacetate, ethylene glycol diacetate, triacetin, ethyl acetate, and tributyrin. In a particular embodiment, the ester substrate is propylene glycol diacetate. In some embodiments, the propylene glycol diacetate is present in the composition in an amount from about 2,000 to about 4,000ppm, for example about 3,000 ppm.

In some embodiments, the hydrogen peroxide source is hydrogen peroxide. In some embodiments, the hydrogen peroxide is present at a concentration of about 1,000 to about 3,000ppm, for example about 2,100 ppm.

In some embodiments, the surfactant and/or emulsifier comprises a nonionic surfactant. In one embodiment, the nonionic surfactant is an alcohol ethoxylate. In one embodiment, the surfactant and/or emulsifier comprises isotridecanol ethoxylate. In one embodiment, the surfactant and/or emulsifier comprises an alcohol ethoxylate and an isotridecanol ethoxylate. In one embodiment, the composition comprises a surfactant and an emulsifier.

In some embodiments, the enzymatic textile bleaching compositions comprise a peroxide stabilizer and/or a sequestering agent. In one embodiment, the peroxide stabilizer is a phosphonic acid. In one embodiment, the sequestering agent is polyacrylic acid.

In some embodiments, the composition further comprises a bioscouring enzyme. In some embodiments, the bioscouring enzyme is selected from pectinases, cutinases, cellulases, hemicellulases, proteases, and lipases. In one embodiment, the bioscouring enzyme is a pectinase.

In another aspect, the present invention provides a method for bleaching a textile, comprising contacting the textile with an enzymatic textile bleaching composition as described herein for a length of time and under conditions that allow measurable whitening of the textile, thereby producing a bleached textile, wherein the bleached textile comprises at least one of reduced textile damage, bulkier softer handle, and increased dye uptake when compared to a chemical textile bleaching process comprising contacting the textile with a chemical textile bleaching composition that does not comprise a perhydrolase enzyme. In some embodiments, the method further comprises hydrolyzing the hydrogen peroxide with catalase enzyme after producing the bleached textile. In one embodiment, the liquid ratio is about 10: 1. In some embodiments, the process is carried out as a batch process or an exhaust process.

In some embodiments, the methods of the present invention provide any of at least about 10, 20, 30, 40, or 50% less weight loss compared to a chemical bleaching composition that does not comprise a perhydrolase enzyme.

In some embodiments, the methods of the present invention provide a textile that has an increased dye uptake as compared to a textile treated with a chemical bleaching composition that does not contain a perhydrolase enzyme to produce a dyed textile having an increased dye depth of at least about any of 5, 10, 15, 20, 25, or 30%.

In some embodiments, the present methods provide for a textile that: it exhibits (i.e., exhibits or has) a reduced tendency to pilling as compared to textiles treated with a chemical bleaching composition that does not contain a perhydrolase enzyme.

In some embodiments, the textile is contacted with the enzymatic textile bleaching compositions of the present invention at a bleaching temperature of from about 60 ℃ to about 70 ℃ for a treatment time of from about 40 to about 60 minutes. In some embodiments, the temperature of the enzymatic textile bleaching composition is increased from a starting temperature of about 20 ℃ to about 40 ℃ at about 3 ℃ per minute until the bleaching temperature is reached. In one embodiment, the bleaching temperature is about 65 ℃ and the treatment time is about 50 minutes.

In some embodiments, the bleached textile is rinsed with an aqueous composition at a rinse temperature of from about 40 ℃ to about 60 ℃ to remove the textile enzymatic bleaching composition. In one embodiment, the rinse temperature is about 50 ℃. In one embodiment, rinsing comprises rinsing the bleached textile twice for about 10 minutes each. In some embodiments, the aqueous composition comprises a catalase enzyme to hydrolyze hydrogen peroxide.

In another aspect, the present invention provides the use of an enzymatic textile bleaching composition for bleaching cellulose-containing textiles, the composition comprising an enzymatic textile bleaching composition as described herein, characterized in that treatment of textiles with the composition provides improved dye uptake, bulkier softer handle, and/or reduced textile damage compared to treatment with chemical bleaching.

Detailed Description

The present compositions and methods relate to enzymatic bleaching of textiles using perhydrolases. The enzymatic process results in a textile that has a bulkier, softer hand, increased dye uptake, and/or reduced textile damage compared to chemical bleaching processes. The process of the present invention is typically carried out at lower temperatures and with lower rinsing requirements than conventional chemical bleaching processes, resulting in energy and water savings. The effluent from the enzymatic bleaching process also has a lower pH (i.e., < 8) than the effluent from a conventional chemical bleaching process (i.e., about 13), thereby reducing the environmental impact of textile bleaching.

The practice of the compositions and methods of this invention will employ, unless otherwise indicated, conventional techniques in the following fields: molecular biology (including recombinant techniques), microbiology, cell biology, and biochemistry, and such conventional techniques are known to those skilled in the art. These techniques are described in the literature, for example, Molecular Cloning: a Laboratory Manual, 2 nd edition (Sambrook et al, 1989); oligonucleotide Synthesis (M.J. Gate edition, 1984; Current Protocols in molecular Biology (F.M. Ausubel et al edition, 1994); PCR: The polymerasechamin Reaction (Mullis et al edition, 1994); and Gene Transfer and Expression: A Laboratory Manual (Kriegler, 1990); are described in FIGS.

Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art. Singleton et al, Dictionary of Microbiology and molecular Biology, 2 nd edition, John Wiley and Sons, New York (1994) and Hale & Markham, The Harper Collins Dictionary of Biology, Harper perennial, NY (1991) provide a general Dictionary for review.

Unless otherwise indicated, a numerical range covers the numbers defining the range, with nucleic acids written from left to right in the 5 'to 3' direction and amino acid sequences written from left to right in the amino to carboxyl direction. The articles "a," "an," and "the" include reference to singular and plural referents unless the context clearly dictates otherwise. Unless otherwise indicated, any methods and materials similar or equivalent to those described herein can be used in the practice or testing of the compositions and methods of this invention. All references cited herein are hereby incorporated by reference.

Definition of

For clarity, the following terms and phrases are defined:

as used herein, the term "bleaching" refers to a process of treating textile materials, such as fibers, yarns, fabrics, garments, or nonwovens, to produce lighter colors. Bleaching involves whitening the textile by removing, modifying or masking the color-causing compounds in the cellulose or other textile material. Thus, "bleaching" refers to treating a textile under suitable pH and temperature conditions for a sufficient period of time to cause whitening (i.e., whitening) of the textile. Bleaching may be performed using chemical bleaching agents and/or enzymatically generated bleaching agents. Examples of suitable bleaching agents include, but are not limited to, ClO₂、H₂O₂Peracid, NO2, and the like.

As used herein, the term "bleaching agent" includes any moiety/chemical capable of bleaching a textile. Bleaching agents may require the presence of a bleach activator. Examples of suitable chemical bleaching agents are sodium peroxide, sodium perborate, potassium permanganate and peracids. H2O2 may be considered a chemical bleaching agent when it has been enzymatically generated in situ. A "chemical bleaching composition" comprises one or more chemical bleaching agents.

As used herein, an enzyme is a protein (polypeptide) with catalytic activity.

As used herein, an "enzymatic bleach system" or "enzymatic bleach composition" includes one or more enzymes and substrates capable of enzymatically producing a bleach. For example, the enzymatic bleaching system may contain a perhydrolase enzyme for producing a peracid bleach, an ester substrate, and a hydrogen peroxide source.

As used herein, "ester substrate" with respect to a perhydrolase-containing enzymatic bleaching system refers to a perhydrolase substrate that contains an ester linkage. Esters comprising aliphatic and/or aromatic carboxylic acids and alcohols may be used as substrates for perhydrolases. In some embodiments, the ester source is an acetate ester. In some embodiments, the ester source is selected from one or more of propylene glycol diacetate, ethylene glycol diacetate, triacetin, ethyl acetate, and tributyrin. In some embodiments, the ester source is selected from esters of one or more of the following acids: formic acid, acetic acid, propionic acid, butyric acid, valeric acid, caproic acid, caprylic acid, pelargonic acid, capric acid, dodecanoic acid, myristic acid, palmitic acid, stearic acid, and oleic acid.

As used herein, the term "perhydrolase enzyme" refers to an enzyme capable of catalyzing perhydrolysis reactions that result in the production of sufficiently high amounts of peracid suitable for use in the textile bleaching process. Generally, perhydrolases exhibit high perhydrolysis to hydrolysis ratios. In some embodiments, the perhydrolase comprises SEQ ID NO:1, or a variant or homologue thereof, or consisting essentially of, or consisting of. In some embodiments, the perhydrolase enzyme comprises acyltransferase activity and catalyzes an aqueous acyltransferase reaction.

As used herein, a "peracid" is an organic acid of the formula RC (═ O) OOH.

As used herein, the term "hydrogen peroxide source" refers to hydrogen peroxide added to a textile treatment bath, which may be derived from an external (i.e., external or outside) source, or generated in situ by the action of a hydrogen peroxide-generating oxidase enzyme on a substrate. "sources of hydrogen peroxide" include hydrogen peroxide and components of systems that can spontaneously or enzymatically produce hydrogen peroxide as a reaction product.

The phrase "perhydrolysis to hydrolysis ratio" refers to the ratio of the amount of peracid enzymatically produced by a perhydrolase enzyme to the amount of acid enzymatically produced under specified conditions and for a specified time. In some embodiments, the assay provided in WO 05/056782 is used to determine the amount of peracid and acid produced by the enzyme.

As used herein, the term "acyl" refers to an organic group having the general formula RCO-, which is obtainable by removing the-OH group from an organic acid. In general, the acyl name ends with the suffix "-oyl", e.g. formyl chloride CH₃CO-Cl is an acid chloride formed from CH3CO-OH formate.

As used herein, the term "acylation" refers to a chemical transformation in which one substituent of a molecule is replaced with an acyl group, or a process in which an acyl group is introduced into a molecule.

As used herein, the term "transferase" refers to an enzyme that catalyzes the transfer of a functional group from one substrate to another. For example, acyltransferases may transfer an acyl group from an ester substrate to a hydrogen peroxide substrate to form a peracid.

As used herein, the term "hydrogen peroxide-producing oxidase" means an enzyme that catalyzes an oxidation/reduction reaction involving molecular oxygen (O2) as an electron acceptor. In this reaction, oxygen is reduced to water (H2O) or hydrogen peroxide (H2O 2). An oxidase suitable for use herein is one that produces hydrogen peroxide (rather than water) on its substrate. Examples of hydrogen peroxide-producing oxidases and substrates thereof suitable for use herein are glucose oxidase and glucose. Other oxidases that can be used to produce hydrogen peroxide include alcohol oxidase, ethylene glycol oxidase, glycerol oxidase, amino acid oxidase, and the like. In some embodiments, the hydrogen peroxide-producing oxidase is a carbohydrate oxidase.

As used herein, the term "textile" refers to fibers, yarns, fabrics, garments, and nonwoven fabrics. The term includes textiles made from natural, synthetic (e.g., manufactured), and various blends of natural and synthetic (blends). Thus, the term "textile" refers to both raw and processed fibers, yarns, woven or knitted fabrics, nonwoven fabrics, and garments. In some embodiments, the textile comprises cellulose.

As used herein, the phrase "textile in need of processing" refers to a textile that requires desizing and/or scouring and/or bleaching, or may require other treatments such as biopolishing.

As used herein, the phrase "textile in need of bleaching" refers to a textile in need of bleaching regardless of other possible treatments. These textiles may have been otherwise treated or may not have been. Similarly, these textiles may or may not require subsequent treatment.

As used herein, the term "fabric" refers to an assembly of fibers and/or yarns produced that has a substantial surface area associated with its thickness and sufficient cohesion to impart useful mechanical strength to the assembly.

As used herein, the phrase "effective amount of perhydrolase" refers to the amount of perhydrolase necessary to achieve/produce the enzymatic activity required for the process or method of the present invention. The effective amount is readily determined by one of ordinary skill in the art and depends on many factors, such as the particular enzyme variant used, the pH used, the temperature used, and the like, as well as the desired result (e.g., level of whiteness).

As used herein, the term "oxidizing chemical" refers to a chemical that has the ability to bleach textiles. The oxidizing chemical is present in an amount, pH and temperature suitable for bleaching. The term includes, but is not limited to, hydrogen peroxide and peracids.

As used herein, "oxidative stability" refers to the ability of a protein to function under oxidative conditions. In particular, the term refers to the ability of a protein to function in the presence of various concentrations of H2O2 and/or peracids. Stability under various oxidation conditions can be measured by standard procedures known to those skilled in the art. A substantial change in oxidative stability may be reflected in an increase or decrease (in most embodiments, preferably an increase) in the half-life of the enzymatic activity of at least about 5% or more, as compared to the enzymatic activity exhibited in the absence of the oxidizing compound.

As used herein, the term "pH stability" with respect to a protein refers to the ability of the protein to function and/or maintain activity at a particular pH. Generally, most enzymes have a limited pH range for their function and remain stable. In addition to enzymes that function at intermediate ranges of pH (i.e., about pH 7), there are enzymes that are capable of operating at very high or very low pH conditions. Stability at various pH can be measured by standard procedures known to those skilled in the art. A substantial change in pH stability may be reflected by an increase or decrease in the half-life of the enzymatic activity by at least about 5% or more (in most embodiments, preferably an increase) compared to the enzymatic activity at the enzyme's optimum pH. However, the inventive processes, methods and/or compositions described herein are not intended to be limited to any pH stability level and pH range.

As used herein, "thermostability" with respect to a protein refers to the ability of the protein to function and/or maintain activity at a particular temperature. Generally, most enzymes have a limited temperature range for their function and remain active. In addition to enzymes that operate at temperatures in the mid range (e.g., room temperature), there are enzymes that are capable of operating at extremely high or low temperatures. Thermal stability can be measured by known procedures. A substantial change in thermostability can be reflected in an increase or decrease in the half-life of the catalytic activity of the mutant by at least about 5% or more when exposed to a different temperature (i.e., higher or lower) than the temperature optimum for enzymatic activity. However, the inventive processes, methods and/or compositions described herein are not intended to be limited to any temperature stability level and temperature range.

As used herein, the term "chemical stability" with respect to a protein refers to the stability that a protein (e.g., an enzyme) exhibits with respect to chemicals that negatively affect its activity. In some embodiments, the chemical includes, but is not limited to, hydrogen peroxide, peracids, anionic surfactants, cationic surfactants, nonionic surfactants, chelating agents, and the like. However, the inventive processes, methods and/or compositions described herein are not intended to be limited to any particular level or range of chemical stability.

As used herein, the terms "purified" and "isolated" refer to the removal of impurities from a sample, and/or to the separation of a substance (e.g., a protein, nucleic acid, cell, etc.) from at least one component with which it is naturally associated. For example, these terms may refer to a substance that is substantially or essentially free of components with which it is normally associated in its native state (e.g., an intact biological system).

As used herein, the term "polynucleotide" refers to a polymeric form of nucleotides of any length and of any three-dimensional structure, which may be single-stranded or multi-stranded (e.g., single-stranded, double-stranded, triple-helical, etc.), which may contain deoxyribonucleotides, ribonucleotides, and/or analogs or modified forms of deoxyribonucleotides or ribonucleotides, including modified nucleotides or bases or analogs thereof. Because of the degeneracy of the genetic code, more than one codon may be used to encode a particular amino acid, and thus the compositions and methods of the invention encompass polynucleotides that encode a particular amino acid sequence. Any type of modified nucleotide or nucleotide analog can be used, including modifications that increase nuclease resistance (e.g., deoxy, 2' -O-Me, phosphorothioate, etc.), so long as the polynucleotide retains the desired functionality under the conditions of use. Labels, such as radioactive or non-radioactive labels or anchors, such as biotin, may also be incorporated for detection or capture purposes. The term polynucleotide also includes Peptide Nucleic Acids (PNA). The polynucleotide may be naturally occurring or non-naturally occurring. The terms "polynucleotide" and "nucleic acid" and "oligonucleotide" are used interchangeably herein. The polynucleotide may comprise RNA, DNA, or both, and/or modified forms and/or analogs thereof. The sequence of nucleotides may be interrupted by non-nucleotide components. One or more phosphodiester linkages may be substituted with alternative linking groups. These alternative linking groups include, but are not limited to, embodiments in which the phosphate ester is replaced by p (O) S ("thioester"), p (S) S ("dithioester"), (O) NR2 ("amidate"), p (O) R, P (O) OR ', CO, OR CH2 ("formacetal"), wherein each R OR R' is independently H OR substituted OR unsubstituted alkyl (1-20C), optionally containing an ether (-O-) linkage, aryl, alkenyl, cycloalkyl, cycloalkenyl, OR araldyl. Not all linkages in a polynucleotide need be identical. The polynucleotide may be linear or circular, or comprise a combination of linear and circular portions.

As used herein, the term "polypeptide" refers to any composition consisting of amino acids and recognized as a protein by those skilled in the art. The conventional single or three letter codes for amino acid residues are used herein. The terms "polypeptide" and "protein" are used interchangeably herein to refer to a polymer of amino acids of any length. The polymer may be linear or branched, it may comprise modified amino acids, and it may be interrupted by non-amino acids. The term also encompasses amino acid polymers that have been modified either naturally or by intervention; for example, disulfide bond formation, glycosylation, lipidation, acetylation, phosphorylation or any other manipulation or modification, for example conjugation to a labeling component. The definition also includes, for example, polypeptides containing one or more analogs of an amino acid (including, for example, unnatural amino acids, etc.), as well as other modifications known in the art.

As used herein, the term "related proteins" refers to functionally and/or structurally similar proteins. In some embodiments, these proteins are derived from different genera and/or species, including differences between classes of organisms (e.g., bacterial proteins and fungal proteins). In further embodiments, the related proteins are provided by the same species. Indeed, the processes, methods, and/or compositions described herein are not intended to be limited to any particular source of the protein of interest. Furthermore, the term "related proteins" encompasses tertiary structural homologues and primary sequence homologues. In further embodiments, the term encompasses proteins that are immunologically cross-reactive.

As used herein, the term "derivative" refers to a protein derived from the addition of one or more amino acids to one or both of the C-terminus and N-terminus of the protein, the substitution of one or more amino acids at one or several different positions in the amino acid sequence, and/or the deletion of one or more amino acids at either or both ends of the protein or at one or more positions in the amino acid sequence, and/or the insertion of one or more amino acids at one or more positions in the amino acid sequence. The preparation of protein derivatives can be accomplished by altering the DNA sequence encoding the native protein, transforming the DNA sequence into a suitable host, and expressing the altered DNA sequence to form a derivative protein.

As used herein, the term "variant protein" refers to related proteins and derived proteins. In some embodiments, a variant protein differs from a parent (or parent) protein, e.g., a wild-type protein, in that different amino acid residues are present at a few amino acid positions. The number of different amino acid residues may be one or more, for example 1, 2, 3, 4,5, 10, 15, 20, 30, 40, 50 or more amino acid residues. The number of different amino acids may be 1 to 10. A variant protein can have a defined level of sequence identity, e.g., at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or even at least 99% amino acid sequence identity to a reference protein (e.g., a parent protein). Alternatively or in addition, the variant protein may differ from the reference protein or parent protein in the number of significant regions (i.e., domains, epitopes, or similar structural or functional portions). For example, in some embodiments, a variant protein has 1, 2, 3, 4,5, or 10 corresponding significant regions that differ from the parent protein. Methods suitable for producing variants of the enzymes described herein are known in the art and include, but are not limited to, site-saturation mutagenesis, scanning mutagenesis, insertion mutagenesis, random mutagenesis, site-directed mutagenesis, and directed mutagenesis, as well as various other recombinant methods and combinatorial approaches.

As used herein, the term "similar sequence" refers to a sequence that provides similar function, tertiary structure, and/or conserved residues in a protein as compared to a reference protein (e.g., a protein of interest having a desired structure or function). For example, in epitope regions comprising alpha-helical or beta-sheet structures, the replacement amino acids in similar sequences preferably retain the same specific structure. The term also refers to nucleotide sequences as well as amino acid sequences. In some embodiments, similar sequences are developed such that substitution of amino acids results in variant enzymes that exhibit similar or improved function. In some embodiments, the tertiary structure and/or conserved residues of these amino acids in the protein of interest are located at or adjacent to the segment or fragment of interest. Thus, when the segment or fragment of interest contains, for example, an alpha-helix or beta-sheet structure, the substituted amino acids preferably retain that particular structure.

As used herein, the term "homologous protein" refers to a protein (e.g., a perhydrolase) that has a similar effect and/or structure as a reference protein (e.g., a protein of interest from another source, e.g., a perhydrolase). Homology does not mean necessarily evolutionarily related. Thus, the term is intended to encompass the same or similar enzymes (i.e., in structural and functional respects) obtained from different species. In some embodiments, it is desirable to identify a homolog that has a similar quaternary, tertiary, and/or primary structure as the protein of interest, because replacing a segment or fragment in the protein of interest with a similar segment from the homolog will reduce the disruption of the alteration. In some embodiments, the homologous protein induces a similar immune response(s) as the protein of interest. In some embodiments, homologous proteins are engineered to produce enzymes having the desired activity(s).

As used herein, the terms "wild-type" and "native" with respect to proteins and nucleic acids refer to those occurring in nature. The terms "wild-type sequence" and "wild-type gene" are used interchangeably herein to refer to a sequence (protein or nucleic acid) that is native or naturally occurring in a host cell. In some embodiments, the wild-type sequence refers to a sequence of interest as a starting point for protein engineering. Genes encoding naturally occurring proteins can be obtained according to general methods known to those skilled in the art. The methods generally involve synthesizing a labeled probe having a putative sequence of a region encoding a protein of interest, preparing a genomic library from an organism expressing the protein, and screening the library for the gene of interest by hybridization to the probe. Positively hybridizing clones are then mapped and sequenced.

The degree of homology between the columns can be determined using any suitable method known in the art (see, e.g., Smith and Waterman (1981) adv.Appl.Math.2: 482; Needleman and Wunsch (1970) J.mol.biol.48: 443; Pearson and Lipman (1988) Proc.Natl.Acad.Sci.USA 85: 2444; programs such as GAP, BESTFIT, FASTA and TFASTA in Wisconsin Genetics software Package (Genetics Computer Group, Madison, Wis.), and Devereux et al (1984) Nucleic Acids Res.12: 95).

For example, PILEUP is a useful program for determining the level of sequence homology. PILEUP creates a multiple sequence alignment from a set of related sequences using progressive, pairwise sequence alignments. It can also draw a tree that is used to build the display cluster relationships of the alignment. PILEUP uses a simplified Feng and Doolittle progressive alignment method (Feng and Doolittle (1987) J.mol.Evol.35: 351-60). This method is similar to that described by Higgins and Sharp (1989) CABIOS 5: 151-53). Useful PILEUP parameters include a default gap weight of 3.00, a default gap length weight of 0.10, and weighted end gaps. Another useful example of an algorithm is the BLAST algorithm described by Altschul et al (1990) J.mol.biol., 215: 403-10; and Karlin et al (1993) Proc.Natl.Acad.Sci.USA 90: 5873-87). A particularly useful BLAST program is the WU-BLAST-2 program (Altschul et al (1996) meth. enzymol.266: 460-80). The parameters "W", "T" and "X" determine the sensitivity and speed of the alignment. The BLAST program uses a word length (W) of 11, BLOSUM62 scoring matrix (Henikoff and Henikoff (1989) Proc. Natl. Acad. Sci. USA 89: 10915), alignment (B) of 50, expectation (E) of 10, M '5, N' -4 and two-strand comparison as defaults.

As used herein, the terms "substantially similar" and "substantially identical" in relation to at least two nucleic acids or polypeptides generally mean that the polynucleotide or polypeptide comprises a sequence having at least about 40% identity, at least about 50% identity, at least about 60% identity, at least about 75% identity, at least about 80% identity, at least about 90% identity, at least about 91% identity, at least about 92% identity, at least about 93% identity, at least about 94% identity, at least about 95% identity, at least about 96% identity, at least about 97% identity, at least about 98% identity, at least about 99% identity, as compared to a reference (i.e., wild-type) sequence. Sequence identity can be determined using standard parameters using known programs such as BLAST, ALIGN and CLUSTAL (see, e.g., Altschul et al (1990) J.mol.biol.215: 403-10; Henikoff et al (1989) Proc.Natl.Acad.Sci.USA 89: 10915; Karin et al (1993) Proc.Natl.Acad.Sci USA 90: 5873; and Higgins et al (1988) Gene 73: 237-44). Software for performing BLAST analysis is publicly available through the national center for biotechnology information. At the same time, the database can be searched using FASTA (Pearson et al (1988) Proc. Natl. Acad. Sci. USA 85: 2444-48). One indication that two polypeptides are substantially identical is that the first polypeptide is immunologically cross-reactive with the second polypeptide. Generally, polypeptides that differ by conservative amino acid substitutions are immunologically cross-reactive. Thus, for example, a first polypeptide is substantially identical to a second polypeptide when the two peptides differ only by conservative substitutions. Another indication that two nucleic acid sequences are substantially identical is that the two molecules hybridize to each other under stringent conditions (e.g., in the range of moderate to high stringency).

As used herein, the term "size" or "sizing" refers to a compound used in the textile industry to improve weaving performance by increasing the abrasion resistance and strength of the yarn. The slurry is typically made of, for example, starch or amyloid.

As used herein, the term "desizing" or "desizing" refers to a process of removing size (typically starch) from a textile, typically before a particular finish, dye or bleach is applied.

As used herein, the term "desizing enzyme" refers to an enzyme used for enzymatic removal of slurry. Exemplary enzymes are amylases, cellulases and mannanases.

As used herein, the terms "perhydrolysis", or "perhydrolysis" refer to a reaction in which a peracid is produced from an ester and a hydrogen peroxide substrate. In one embodiment, the perhydrolysis reaction is catalyzed by a perhydrolase enzyme, such as an acyltransferase or an arylate enzyme. In some embodiments, the peracid is produced by reacting formula R in the presence of hydrogen peroxide (H2O2)₁C(＝O)OR₂(wherein R is₁And R₂Are the same or different organic moieties) is produced by perhydrolysis of an ester substrate. In one embodiment, -OR₂is-OH. In one embodiment, -OR₂is-NH₂And (6) replacing. In some embodiments, the peracid is generated by perhydrolysis of a carboxylic acid or amide substrate.

As used herein, the term "peracid" refers to a molecule derived from a carboxylic acid ester that has reacted with hydrogen peroxide to form a highly reactive product capable of transferring one of its oxygen atoms. It is this ability to transfer oxygen atoms that enables peracids (e.g., peracetic acid) to function as bleaching agents.

As used herein, the term "scouring" refers to the removal of impurities, such as most non-cellulosic compounds (e.g., pectin, proteins, waxes, dust, etc.), that are naturally present in cotton or other textiles. In addition to natural non-cellulosic impurities, scouring may also remove residual materials introduced by the manufacturing process, such as spinning, spooling or slashing lubricants. In some embodiments, bleaching may be employed to remove impurities from the textile.

As used herein, the term "bioscouring enzyme" refers to an enzyme that is capable of removing at least a portion of impurities in cotton or other textiles.

As used herein, the term "dust" refers to unwanted impurities, such as cotton seed pieces, leaves, stems, and other plant parts, that stick to the fibers even after the mechanical ginning process.

As used herein, the term "greige" (pronounced gray) refers to a textile that has not undergone any bleaching, dyeing or finishing treatments after manufacture. For example, any woven or knitted fabric that leaves the loom that has not been finished (desized, scoured, etc.), bleached, or dyed is referred to as a greige textile. The textiles used in the examples below are greige textiles.

As used herein, the term "dyeing" refers to, inter alia, applying color to, for example, a textile by immersion in a dyeing solution.

As used herein, the term "non-cotton cellulosic" fiber, yarn or fabric means a fiber, yarn or fabric consisting essentially of a non-cotton cellulose-based composition. Examples of such compositions include flax (lichen), ramie, jute, flax (flax), rayon, lyocell, cellulose acetate, bamboo, other similar compositions derived from non-cotton cellulose.

As used herein, the term "pectate lyase (pectate lyase)" refers to a pectinase. Pectinases are a group of enzymes that cleave the glycosidic bond of pectic substances, primarily poly (1, 4-alpha-D-galacturonide) and its derivatives (see Sakai et al (1993) Advances in applied microbiology 39: 213-294). Preferably, pectinases catalyze the random cleavage of alpha-1, 4-glycosidic bonds in pectic acids (also known as polygalacturonic acids) by elimination in trans, such as enzymes of the polygalacturonic acid lyase class (PGL; EC 4.2.2.2), also known as poly (1, 4-alpha-D-galacturonide) lyase or pectate lyase.

As used herein, the term "pectin" refers to pectic acids, polygalacturonic acids and pectins, which may be esterified to a higher or lower degree.

As used herein, the term "cutinase" refers to a lipolytic enzyme of plant, bacterial or fungal origin for use in textile processing. Cutinases are capable of hydrolyzing the substrate cutin. Cutinases can degrade fatty acid esters and other oil-based compositions that need to be removed in textile processing (e.g., scouring). In some embodiments, the cutinase has significant plant cutinase activity. In a particular embodiment, the cutinase has hydrolytic activity on the biopolyester polymer cutin on the plant leaves. Suitable cutinases can be isolated from many different plant, fungal and bacterial sources.

As used herein, the term "alpha-amylase" refers to an enzyme that cleaves the alpha (1-4) glycosidic bond of amylose to produce a maltose molecule (a disaccharide of alpha-glucose). Amylases are digestive enzymes found in saliva, which are also produced by many plants. Amylases break down long chain carbohydrates (e.g., starch) into smaller units. An "oxidatively stable" alpha-amylase is an alpha-amylase that is resistant to degradation by oxidative means when compared to a non-oxidatively stable alpha-amylase, particularly when compared to the form of the non-oxidatively stable alpha-amylase from which the oxidatively stable alpha-amylase is derived.

As used herein, the term "protease" refers to a protein capable of catalyzing the cleavage of peptide bonds.

As used herein, "catalase (catalase)" refers to an enzyme that catalyzes the decomposition of hydrogen peroxide into hydrogen and oxygen.

As used herein, the term "wicking" refers to the passage of liquid along or through the textile material or textile components of the sized fabric, or along the interstices formed by the textile components of the sized fabric and the sized polymer. Wicking involves spontaneous transport of liquid into a porous system driven by capillary forces.

As used herein, the phrase "degree of polymerization" refers to the number of repeat units in a single macromolecule in a polymer. The degree of polymerization may be based on mass (weight) or number average.

As used herein, the term "fastness" or "color fastness" refers to the ability of a material to resist discoloration, i.e., retain its original color, especially when wetted, washed, cleaned, or stored under normal conditions when exposed to light, heat, or other influences, without losing color, fading, or changing color.

As used herein, the term "handle" or "hand" refers to the quality of a textile material (e.g., fabric or yarn) that is evaluated by a reaction derived from the sense of touch. This is relevant for judgments such as roughness, smoothness, harshness, flexibility, thickness and other tactile parameters.

As used herein, the term "pilling" refers to the entanglement of the textile fibers during washing, dry cleaning, testing or wearing to form globules or pellets protruding from the fabric surface and having a density such that light cannot pass through them, for example, such that they cast shadows. Pilling that occurs during normal wear can be simulated, for example, on laboratory test machines by controlled friction of an elastic pad having specifically selected mechanical properties. The degree of pilling can be assessed on any scale from 5 (indicating no pilling) to 1 (indicating very severe pilling) relative to the standard.

As used herein, the term "surfactant" refers to a substance that reduces the surface tension of a liquid.

As used herein, the term "emulsifier" refers to a substance that facilitates the suspension of one liquid in another liquid.

As used herein, the term "sequestering agent" refers to a substance capable of reacting with a metal ion by forming a water-soluble complex in which the metal is retained in a non-ionizable form.

As used herein, the terms "batch process", "batch process" or "batch process" refer to batch or batch processing of textiles, wherein all of a batch is processed at once in a process or one process stage.

As used herein, the term "finishing process" refers to a batch process wherein the pretreatment chemicals and/or enzymatic pretreatment compositions and dyes are added simultaneously or sequentially to a single textile treatment bath.

As used herein, the term "liquor ratio" refers to the ratio of the weight of liquor (liquid) used in a textile treatment process to the weight of the textile being treated.

Enzymatic textile bleaching compositions

In one aspect of the compositions and methods of the present invention, enzymatic bleaching compositions and methods of bleaching textiles with these compositions are provided. Textiles include cellulose-containing textiles, such as textiles made from cotton, linen, hemp (hemp), ramie, cellulose, acetate, lyocell, viscose rayon, bamboo and various cellulose blends (blends), and textiles made from polyamide, polyacrylic, wool or blends thereof. In some embodiments, the textile comprises a blend having elasticity. The enzymatic bleaching compositions and methods of the present invention are particularly useful for bleaching textiles containing fibers that are sensitive to high pH and high temperature conditions. The enzymatic bleaching compositions and methods of the present invention are particularly useful in batch, exhaust, or batch processes.

The enzymatic bleaching compositions of the present invention comprise a perhydrolase enzyme, an ester substrate for the perhydrolase enzyme suitable for generating a peracid in the presence of hydrogen peroxide in a catalyzed reaction of the perhydrolase enzyme on the substrate, a hydrogen peroxide source, a surfactant and/or emulsifier, a peroxide stabilizer, a sequestering agent, and a buffering agent to maintain a pH of about 6 to about 8 during a textile bleaching process in which the enzymatic bleaching composition is used. The enzymatic bleaching composition may also optionally comprise bioscouring agents or enzymes and/or desizing agents or enzymes.

The enzymatic bleaching compositions of the present invention, when used in a textile pretreatment process, advantageously produce bleached textiles that exhibit increased dye uptake, reduced textile damage due to the bleaching process, and/or bulkier, softer hand compared to pretreatment with a chemical bleaching composition that does not contain a perhydrolase enzyme. In some embodiments, the enzymatic bleaching compositions, when used in a textile pretreatment process, produce textiles with a reduced tendency to pilling.

Perhydrolase

The enzymatic bleaching compositions of the present invention comprise one or more perhydrolases. In some embodiments, the perhydrolase enzyme is naturally-occurring (i.e., a perhydrolase enzyme encoded by the genome of a cell). In some embodiments, the perhydrolase enzyme comprises, consists essentially of, or consists of an amino acid sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or even at least about 99.5% identical to the amino acid sequence of a naturally-occurring perhydrolase enzyme.

In some embodiments, the perhydrolase enzyme is a naturally-occurring mycobacterium smegmatis perhydrolase. In some embodiments, the perhydrolase comprises SEQ ID NO:1 or a variant or analogue thereof, or consisting essentially of or consisting of. In some embodiments, the perhydrolase comprises a substitution with SEQ ID NO:1, at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or even at least about 99.5% identical, or consists essentially of, an amino acid sequence.

The amino acid sequence of Mycobacterium smegmatis perhydrolase (SEQ ID NO: 1) is:

MAKRILCFGDSLTWGWVPVEDGAPTERFAPDVRWTGVLAQQLGADFEVIEEGLSA

RTTNIDDPTDPRLNGASYLPSCLATHLPLDLVHMLGTNDTKAYFRRTPLDIALGMSV

LVTQVLTSAGGVGTTYPAPKVLVVSPPPLAPMPHPWFQLIFEGGEQKTTELARVYS

ALASFMKVPFFDAGSVISTDGVDGIHFTEANNRDLGVALAEQVRSLL

the corresponding polynucleotide sequence (SEQ ID NO: 2) encoding Mycobacterium smegmatis perhydrolase is:

5′-ATGGCCAAGCGAATTCTGTGTTTCGGTGATTCCCTGACCTGGGGCTGGGTCC

CCGTCGAAGACGGGGCACCCACCGAGCGGTTCGCCCCCGACGTGCGCTGGACC

GGTGTGCTGGCCCAGCAGCTCGGAGCGGACTTCGAGGTGATCGAGGAGGGACT

GAGCGCGCGCACCACCAACATCGACGACCCCACCGATCCGCGGCTCAACGGCG

CGAGCTACCTGCCGTCGTGCCTCGCGACGCACCTGGCCGCTCGACCTGGTGATCA

TCATGCTGGGCACCAACGACACCAAGGCCTACTTCCGGCGCACCCCGCTCGACA

TCGCGCTGGGCATGTCGGTGCTCGTCACGCAGTTGCCTCACCAGCGCGGGCGGCG

TCCGGCACCACGTACCCGGCACCCAAGGTGCTGGTGGTGTCGCCGCCACCGCTGG

CGCCCATGCCGCACCCCTGGTTCCAGTTGATCTTCGAGGGCGGCGAGCAGAAGA

CCACTGAGCTCGCCCGCGTGTACAGCGCCGCTCGCGTCGTTCATGAAGGTGCCGT

TCTTCGACGCGGGTTCGGTGATCAGCACCGACGGCGTCGACGGAATCCACTTGCA

CCGAGGCCAACAATCGCGATCTCGGGGTGGCCCTCGCGGAACAGGTGCGGAGC

CTGCTGTAA-3′

in some embodiments, the perhydrolase enzyme is a compound that hybridizes to SEQ ID NO:1 comprising one or more substitutions at one or more amino acid positions equivalent to position(s) in the mycobacterium smegmatis perhydrolase amino acid sequence. In some embodiments, the perhydrolase comprises any substitution or any combination of substitutions of amino acids selected from the group consisting of:

m1, K3, R4, I5, L6, C7, D10, S11, L12, T13, W14, W16, G15, V17, P18, V19, D21, G22, a 22, P22, T22, E22, R22, F22, a 22, P22, D22, V22, R22, W22, T22, G22, L22, Q22, D22, L22, G22, a 22, F22, E22, V22, I22, E22, G22, L22, S22, a 22, R22, T22, N22, I22, D22, P22, G22, L22, N22, L22, N22, P22, L22, N22, L22, P22, L22, P122, N22, P122, L22, P22, N22, L122, P22, N22, P22, N22, L22, P22, L36.

In some embodiments, the perhydrolase enzyme is a compound that hybridizes to SEQ ID NO:1 comprising one or more of the following substitutions at one or more amino acid positions equivalent to position(s) in the mycobacterium smegmatis perhydrolase amino acid sequence set forth in seq id no: L12C, Q or G; T25S, G or P; L53H, Q, G or S; S54V, L A, P, T, or R; A55G or T; R67T, Q, N, G, E, L or F; K97R; V125S, G, R, A or P; F154Y; F196G.

In some embodiments, the perhydrolase enzyme is a compound that hybridizes to SEQ ID NO:1 comprising a combination of amino acid substitutions at amino acid positions equivalent to the amino acid positions in the mycobacterium smegmatis perhydrolase amino acid sequence set forth in seq id no: L12I S54V; L12M S54T; L12T S54V; L12Q T25S S54V; l53HS 54V; S54P V125R; S54V V125G; S54V F196G; S54V K97R V125G; or a55G R67T K97R V125G.

In some embodiments, the perhydrolase enzyme has a perhydrolysis of at least 1: hydrolysis ratio. In some embodiments, the perhydrolase enzyme has a perhydrolysis greater than 1: hydrolysis ratio.

In some embodiments, the perhydrolase enzyme is provided in the enzymatic bleaching composition at a concentration of about 1 to about 2.5pm, about 1.5 to about 2.0ppm, or about 1.7 ppm.

Ester substrates

The enzymatic bleaching compositions of the present invention also include esters, which act as substrates for perhydrolases to produce peracids in the presence of hydrogen peroxide. In some embodiments, the ester substrate is an ester of an aliphatic and/or aromatic carboxylic acid or alcohol. In some embodiments, the ester substrate is an ester of one or more of the following acids: formic acid, acetic acid, propionic acid, butyric acid, valeric acid, caproic acid, caprylic acid, pelargonic acid, capric acid, dodecanoic acid, myristic acid, palmitic acid, stearic acid, and oleic acid. In some embodiments, triacetin, tributyrin, and other esters are used as acyl donors for peracid formation. In some embodiments, the ester substrate is propylene glycol diacetate, ethylene glycol diacetate, or ethyl acetate. In one embodiment, the ester substrate is propylene glycol diacetate.

In some embodiments, the ester substrate is provided at a concentration of about 2,000 to about 4,000ppm, about 2,500 to about 3,500ppm, about 2,800ppm to about 3,200ppm, or about 3,000 ppm.

Hydrogen peroxide source

The enzymatic bleaching compositions of the present invention further comprise a source of hydrogen peroxide. The hydrogen peroxide can be added directly in portions or generated continuously "in situ" by chemical, electrochemical and/or enzymatic methods.

In some embodiments, the hydrogen peroxide source is hydrogen peroxide. In some embodiments, the source of hydrogen peroxide is a solid compound that generates hydrogen peroxide upon addition of water. Such compounds include adducts of hydrogen peroxide with various inorganic or organic compounds, among which sodium carbonate perhydrate, also known as sodium percarbonate, is the most widely used.

Inorganic perhydrate salts are one preferred embodiment of the source of hydrogen peroxide. Examples of inorganic perhydrate salts include perborate, percarbonate, perphosphate, persulfate and persilicate salts. The inorganic perhydrate salts are normally the alkali metal salts.

Other hydrogen peroxide adducts useful in the compositions of the present invention include adducts of hydrogen peroxide with a zeolite-type compound, or adducts of urea with hydrogen peroxide.

The hydrogen peroxide source compound may be included in crystalline and/or substantially pure solid form without additional protection. However, for certain particulate perhydrate salts, it is preferred that the coating be in a form which provides better storage stability. Suitable coating materials include inorganic salts such as alkali metal silicates, carbonates or borates or mixtures thereof, or organic substances such as waxes, oils or fatty soaps.

In some embodiments, the hydrogen peroxide source is an enzymatic hydrogen peroxide generation system. In one embodiment, the enzymatic hydrogen peroxide generation system comprises an oxidase and a substrate therefor. Suitable oxidases include, but are not limited to: glucose oxidase, sorbitol oxidase, hexose oxidase, choline oxidase, alcohol oxidase, glycerol oxidase, cholesterol oxidase, pyranose oxidase, carboxyl alcohol oxidase, L-amino acid oxidase, glycine oxidase, pyruvate oxidase, glutamate oxidase, sarcosine oxidase, lysine oxidase, lactate oxidase, vanillyl oxidase, glycolate oxidase, galactose oxidase, uricase, oxalate oxidase, and xanthine oxidase.

An example of a coupled system for the enzymatic generation of hydrogen peroxide is provided by the following formula:

the compositions and methods of the present invention are not intended to be limited to any particular enzyme, as any enzyme that produces H2O2 with a suitable substrate may be used. For example, lactate oxidase from Lactobacillus (Lactobacillus) species, which is known to produce H2O2 from lactic acid and oxygen, may be used. One advantage of enzymatically generating an acid (e.g., gluconic acid in the above example) is that this can lower the pH of the alkaline solution to the pH range at which the peracid is most effective in performing bleaching (i.e., at or below pKa). Other enzymes that can produce hydrogen peroxide (e.g., alcohol oxidase, ethylene glycol oxidase, glycerol oxidase, amino acid oxidase, etc.) can also be used in combination with perhydrolase and ester substrates to produce peracids.

Hydrogen peroxide can also be produced electrochemically, for example using a fuel cell supplied with oxygen and hydrogen gas.

In some embodiments, the hydrogen peroxide source is hydrogen peroxide provided at a concentration of about 1,000 to about 3,000ppm, about 1,500 to about 2,500ppm, about 2,000ppm to about 2,200ppm, or about 2,100 ppm.

Surfactants and emulsifiers

The enzymatic textile bleaching compositions of the present invention may further comprise one or more, i.e. at least one, surfactants and/or emulsifiers. Suitable surfactants include, but are not limited to, nonionic (see, e.g., U.S. Pat. No. 4,565,647, which is incorporated herein by reference); an anionic type; a cationic type; and zwitterionic surfactants (see, e.g., U.S. Pat. No. 3,929,678). Anionic surfactants include, but are not limited to, linear alkylbenzene sulfonates, alpha-olefin sulfonates, alkyl sulfates (fatty alcohol sulfates), alcohol ethoxy sulfates, secondary alkyl sulfonates, alpha-sulfo fatty acid methyl esters, alkyl or alkenyl succinic acids, and soaps. Nonionic surfactants include, but are not limited to, alcohol ethoxylates, nonylphenol ethoxylates, alkylpolyglycosides, alkyldimethylamine oxides, ethoxylated fatty acid monoethanolamine, polyhydroxyalkyl fatty acid amides, and N-acyl N-alkyl derivatives of glucosamine ("glucamides").

In some embodiments, the enzymatic bleach composition contains a nonionic surfactant. In one embodiment, the nonionic surfactant is an alcohol ethoxylate.

The surfactant may be present at a concentration of about 5% to about 40%, about 20% to about 30%, or about 5% to about 10% (w/w).

In some embodiments, the enzymatic bleach composition contains an ethoxylated isotridecanol at a concentration of from about 5% to about 30%, from about 10% to about 25%, or from about 15% to about 20% (w/w).

Peroxide stabilizers

The enzymatic bleaching compositions of the present invention may also comprise a peroxide stabilizer. Examples of peroxide stabilizers include, but are not limited to, sodium silicate, sodium carbonate, acrylic acid polymers, magnesium salts, and phosphonic acids. In one embodiment, the peroxide stabilizer is a phosphonic acid.

The peroxide stabilizer may be present in the enzymatic bleach composition at a concentration of from about 1% to about 5%, from about 1% to about 10%, or from about 2% to about 8% (w/w).

Sequestering agents

The enzymatic bleaching compositions of the present invention may also comprise a sequestering agent. Examples of sequestering agents include, but are not limited to, amino carboxylates, amino phosphonates, polyfunctionally-substituted aromatic chelants, polyhydroxy-carboxylic acids, amino polycarboxylic acids, polyphosphonates, and polyacrylic acids, and mixtures thereof. Specific aminocarboxylates useful as sequestering agents include ethylenediaminetetraacetate, N-hydroxyethylethylenediaminetriacetate, nitrilotriacetate, ethylenediaminetetrapropionate, and triethylenetetraminehexaacetate.

Polyfunctional substituted aromatic sequestering agents are also useful in the compositions of the present invention (see, e.g., U.S. Pat. No. 3,812,044 to Connor et al, issued 5/21 1974). Such preferred compounds in acid form are dihydroxydisulfobenzenes such as 1, 2-dihydroxy-3, 5-disulfobenzene diethylene triamine pentaacetate, and ethanoldiglycine, alkali metal, ammonium, and substituted ammonium salts, and mixtures thereof.

Aminophosphonates are also suitable for use as sequestering agents in the compositions of the invention, especially when at least low total phosphorus levels are permitted.

Biodegradable sequestrants suitable for use herein are ethylenediamine disuccinate ("EDDS"), particularly the [ S, S ] isomer described in U.S. patent No. 4,704,233(Hartman and Perkins, issued 11/3 in 1987).

In one embodiment, the sequestering agent is polyacrylic acid.

The sequestering agent may be present in the enzymatic bleach compositions described herein at a concentration of from about 1% to about 15%, from about 5% to about 10%, or from about 3% to about 10% (w/w).

Buffering agent

The enzymatic bleach composition of the present invention may include a buffering agent capable of maintaining the pH of the composition from about 6 to about 8. In one embodiment, the buffer is a phosphate buffer, for example 100mM phosphate buffer (pH 8).

Enzymatic textile bleaching process

Another aspect of the compositions and methods of the present invention provides a method of bleaching a textile using any of the enzymatic bleaching compositions described herein. Typically, the textile to be bleached is contacted with the enzymatic bleaching textile composition described herein under conditions and for a length of time suitable to allow measurable whitening of the textile.

Textiles include cellulose-containing textiles such as those made from cotton, linen, hemp, ramie, cellulose, acetate, lyocell, viscose rayon, bamboo and various cellulose blends, and those made from polyamide, polyacrylic, wool or blends thereof. In some embodiments, the textile comprises a blend having elasticity. The enzymatic bleaching compositions and methods are particularly useful for bleaching textiles containing fibers that are sensitive to high pH and high temperature conditions.

Advantageously, treating textiles according to the methods of the present invention results in bleached textiles having increased dye uptake, reduced textile damage, and/or bulkier softer hand when compared to chemical bleaching processes with chemical bleaching compositions that do not contain perhydrolase enzymes. In some embodiments, textiles with reduced pilling processes are produced as compared to chemical bleaching processes without perhydrolase enzymes.

The enzymatic bleaching of the present invention also advantageously requires less energy due to the lower processing temperatures employed compared to typical chemical bleaching processes. Furthermore, less rinsing is required than in chemical bleaching processes, resulting in lower water usage. The process of the present invention also results in lower pH effluent (< 8) than chemical bleaching (about 13), resulting in reduced adverse environmental impact.

Generally, the present process utilizes a liquor ratio of from about 6: 1 to about 15: 1, for example about 10: 1. In some embodiments, the method is performed as a batch, a complete, or a batch textile bleaching process.

The textile is contacted with the enzymatic bleaching composition at a temperature of from about 40 ℃ to about 70 ℃, for example from about 60 ℃ to about 70 ℃, for a treatment time of from about 40 to about 60 minutes. In one embodiment, the bleaching temperature is about 65 ℃ and the treatment time is about 50 minutes. In some embodiments, the temperature of the enzymatic bleach composition is increased from a starting temperature of about 20 ℃ to about 50 ℃ at about 3 ℃ per minute until the processing temperature for bleaching is reached.

In some embodiments, after incubating the textile in the enzymatic bleaching composition, one or more rinsing steps are performed to remove the bleaching composition. Typically, the textile is rinsed with an aqueous composition (water or an aqueous composition). In some embodiments, the rinse temperature is from about 40 ℃ to about 60 ℃, e.g., about 50 ℃. In some embodiments, the aqueous rinse composition contains a catalase enzyme to hydrolyze the hydrogen peroxide. In one embodiment, the textile is rinsed twice with an aqueous composition containing catalase enzyme, each rinse for about 10 minutes.

In some embodiments, textiles bleached using the present methods comprise softer, bulkier, and more natural hand compared to textiles treated with chemical bleaching compositions that do not contain perhydrolase enzymes. This bulkier, softer hand generally results in improved sewability (needle resistance) and stretch. Moreover, this durable, bulkier, softer hand often results in improved wrinkle recovery, e.g., less risk of wrinkle formation in cloth and garment processing.

In some embodiments, the elastic properties are enhanced using the enzymatic bleaching methods herein as compared to bleaching with a chemical process that does not contain a perhydrolase enzyme.

In some embodiments, the enzymatic bleaching process herein produces natural fibers with less swelling and avoids the channeling effect in a package dyeing machine as compared to a chemical bleaching process without perhydrolase.

Biological scouring enzyme

In some embodiments, the present compositions and methods for enzymatic bleaching of textiles comprise one or more bioscouring enzymes. The bioscouring enzyme can be included in the enzymatic textile bleaching composition, or the textile can be treated with the bioscouring enzyme in a subsequent processing step after the textile is pretreated in the enzymatic textile bleaching composition. Exemplary bioscouring enzymes are described below.

Pectinase

Any pectolytic enzyme capable of degrading a pectin component, e.g., in a plant cell wall, can be used in the compositions and methods of the invention. Suitable pectinases include, but are not limited to, those of fungal or bacterial origin. The pectinase may be of natural origin or produced recombinantly, and/or may be chemically or genetically modified. In some embodiments, the pectinase is a single component enzyme.

Pectinases can be classified according to their preferred substrates, i.e., high methyl esterified pectin or low methyl esterified pectin and polygalacturonic acid (pectic acid), and their reaction mechanism, i.e., beta-elimination or hydrolysis. Pectinases can be mainly endo-acting, cleaving the polymer at random sites within the chain to give a mixture of oligomers, or they can be exo-acting, attacking and producing monomers or dimers starting from one end of the polymer. Several pectinase activities acting on the smooth region of pectin are included in the Enzyme classification provided by Enzyme Nomenclature (1992), for example, pectate lyase (EC 4.2.2.2), pectin lyase (EC 4.2.2.10), polygalacturonase (EC 3.2.1.15), exo-polygalacturonase (EC 3.2.1.67), exo-polygalacturonate-lyase (EC 4.2.2.9), and exo-poly-alpha-galacturonosidase (EC 3.2.1.82). In a preferred embodiment, the method of the invention utilizes a pectate lyase.

Pectate lyase enzymatic activity as used herein refers to the random cleavage of alpha-1, 4-glycosidic bonds in pectic acids (also known as polygalacturonic acids) catalyzed by elimination in trans. Pectate lyases are also known as polygalacturonate lyases and poly (1, 4-D-galacturonide) lyases. For the purposes of the compositions and Methods of the present invention, pectate lyase enzymatic activity is that activity determined by measuring the increase in absorbance at 235nm of a 0.1% w/v solution of sodium polygalacturonate in 0.1M glycine buffer, pH 10 (see Collmer et al (1988) Methods Enzymol 161: 329-35). The enzyme activity is usually expressed as x mol/min, i.e.the amount of enzyme catalyzing the formation of x mol of product/min. An alternative assay measures the viscosity reduction of a 5% w/v solution of sodium polygalacturonate in 0.1M glycine buffer at pH 10, as measured by a vibratile viscometer (APSU units). It is understood that any pectate lyase may be used in the practice of the compositions and methods of the invention.

Non-limiting examples of pectate lyases that may be used in the compositions and methods of the invention include pectate lyases that have been cloned from different bacterial genera such as Erwinia (Erwinia), Pseudomonas (Pseudomonas), Bacillus (Bacillus), Klebsiella (Klebsiella), and Xanthomonas (Xanthomonas). Pectate lyases suitable for use herein may be from Bacillus subtilis (Nasser et al (1993) FEBS letters.335: 319-26) and Bacillus species YA-14(Kim et al (1994) biosci.Biotech.biochem.58: 947-49). Other pectate lyases produced by Bacillus pumilus (Dave and Vaughn (1971) J.Bacteriol.108: 166-74), Bacillus polymyxa (B.polymyxa) (Nagel and Vaughn (1961) Arch.biochem.Biophys.93: 344-52), Bacillus stearothermophilus (B.stearothermophilus) (Karbassi and Vaughn (1980) Can.J.Microbiol.26: 377-84), Bacillus species (Hasegawa and Nagel (1966) J.food Sci.31: 838-45) and Bacillus species RK9(Kelly and Fogarty (1978) Can.J.Microbiol.24: 1164-72) have also been described and may be considered for use in the compositions and methods of the invention. Any of the above-described and divalent cation independent and/or heat stable pectate lyases may be used in the practice of the compositions and methods of the present invention. In some embodiments, the pectate lyase comprises, for example, those described in WO 04/090099 (divesa) or WO 03/095638 (Novozymes).

The effective amount of pectolyase to be used in accordance with the compositions and methods of the present invention depends on a variety of factors, but according to the compositions and methods of the present invention, the concentration of pectolyase in the aqueous medium may be from about 0.0001% to about 1% μ g enzyme protein by weight of the fabric, for example from about 0.0005% to about 0.2% enzyme protein by weight of the fabric, or from about 0.001% to about 0.05% enzyme protein by weight of the fabric.

Enzymes for hydrolyzing polyester substrates

Any enzyme that hydrolyzes a polyester substrate is suitable for use in the compositions and methods of the present invention, for example, a cutinase or lipase, including, for example, an enzyme derived from Humicola insolens strain DSM1800, as described in example 2 of U.S. Pat. No. 4,810,414, or in one embodiment, an enzyme derived from Pseudomonas mendocina (Pseudomonas mendocina), variants and/or equivalents thereof, described in U.S. Pat. No. 5,512,203. Suitable variants are described, for example, in WO 03/76580. These documents are incorporated herein by reference.

Suitable bacterial enzymes may be derived from Pseudomonas (Pseudomonas) or Acinetobacter (Acinetobacter) species, preferably from Pseudomonas sp (P.stutzeri), Pseudomonas alcaligenes (P.alcaligenes), Pseudomonas pseudoalcaligenes (P.pseudoalcaligenes), Pseudomonas aeruginosa (P.aeruginosa) or Acinetobacter calcoaceticus (A.calcomaceticus), most preferably from Pseudomonas sp strains Thai IV 17-1(CBS 461.85), PG-1-3(CBS 137.89), PG-1-4(CBS 138.89), PG-II-11.1(CBS 139.89) or PG-II-11.2(CBS 140.89), Pseudomonas aeruginosa PAO (ATCC 15692), Pseudomonas alcaligenes DSM 42, Pseudomonas alcaligenes INII-5 (CBS.468), Pseudomonas pseudoalcaligenes M-1(CBS 140.85) or Acinetobacter calcoaceticus GrV (CBS 50385). With respect to the use of plant-derived enzymes, enzymes that hydrolyze polyester substrates are known to be present in the pollen of many plants, and these enzymes will be useful in the processes, methods, and compositions of the present invention. Enzymes that hydrolyze polyester substrates may also be derived from fungi, such as Absidia species (Absidia spp.); acremonium species (Acremonium spp.); agaricus species (Agaricus spp.); anaeromyces spp; aspergillus species (Aspergillus spp.) including Aspergillus aculeatus (a. auriculatus), Aspergillus awamori (a. awamori), Aspergillus flavus (a. flavus), Aspergillus foetidus (a. foetidus), a.fumaricus, Aspergillus fumigatus (a. fumigatus), Aspergillus nidulans (a. nidulans), Aspergillus niger (a.niger), Aspergillus oryzae (a.oryzae), Aspergillus terreus (a.terreus) and Aspergillus versicolor (a.versicolor); aureobasidium species (Aeurobasidium spp.); cephalosporium species (cephalosporium spp.); chaetomium spp; coprinus species (Coprinus spp.); dactyllum spp.; fusarium species (Fusarium spp.) including f.conglomeratans, Fusarium polytrichum (f.decemcellulare), Fusarium javanicum (F.j avanicom), Fusarium linonense (f.lini), Fusarium oxysporum (f.oxysporum), and Fusarium solani (f.solani); gliocladium spp; humicola species, including specific huminase (h.insolens) and humicola lanuginosa (h.lanuginosa); mucor (Mucor spp.); neurospora species (Neurospora pp.), including Neurospora crassa (N.crassa) and Neurospora sitophila (N.sitophila); neocallimastix spp.; orpinomyes spp.; penicillium species (Penicillium spp.); protochaeta spp (Phanerochaete spp.); phlebia species (Phlebia spp.); piromyces spp.; pseudomonas sp (Pseudomonas spp.); rhizopus species (Rhizopus spp.); schizophyllum species (Schizophyllum spp.); trametes species (tramessespp.); trichoderma species (Trichoderma spp.) including Trichoderma reesei (t. reesei), Trichoderma longibrachiatum (t. reesei), and Trichoderma viride (t. viride); and species of the genus A. (Zygorhynchus spp.). Similarly, it is contemplated that enzymes that hydrolyze polyester substrates may be present in bacteria, such as bacillus species; cellulomonas species (Cellulomonas spp.); clostridium species (Clostridium spp.); myceliophthora spp (Myceliophthora spp.); pseudomonas species (Pseudomonas spp), including Pseudomonas mendocina and Pseudomonas putida (p.putida); thermomomonas (Thermomonospora spp.); thermomyces species (Thermomyces spp.) including Thermomyces lanuginose (t. lanuginose); streptomyces species (spp.) including Streptomyces olivaceus (s. olivochromogenes); and in fiber degrading ruminal bacteria, such as filamentous bacillus succinogenes (Fibrobacter succinogenes); and among yeasts, including Candida species (Candida spp.), including Candida antarctica (c.antarctica), Candida rugosa (c.rugosa), c.torresii; candida parapsilosis (c. parapaslosis); candida sake (c.sake); candida salivarius (c. zeaylanoides); pichia pastoris (Pichia minuta); rhodotorula glutinis (Rhodotorula glutinis); rhodotorula mucilaginosa (r. mucolaginosa); and asymmetric Sporobolomyces persicaria.

In some embodiments, the enzyme that hydrolyzes the polyester substrate, e.g., cutinase and/or lipase, is added to the enzymatic bleach composition in an amount of enzyme protein from about 0.00001% to about 2% by weight of the fabric, e.g., in an amount of enzyme protein from about 0.0001% to about 1% by weight of the fabric, or in an amount of enzyme protein from 0.005% to 0.5% by weight of the fabric, typically in an amount of enzyme protein from about 0.001% to about 0.5% by weight of the fabric.

Cellulase enzymes

Cellulases can be added to the compositions and methods of the invention, for example, to facilitate bioscouring. Cellulases are classified into a series of enzyme families, encompassing both endo-and exo-activities as well as cellobiose hydrolyzing ability. Cellulases may be derived from microorganisms known to produce cellulolytic enzymes, such as species of humicola, thermomyces, bacillus, trichoderma, fusarium, myceliophthora, phanerochaete, Irpex, Scytalidium, schizophyllum, penicillium, aspergillus, or geotrichum. Known species capable of producing cellulolytic enzymes include humicola insolens, fusarium oxysporum or trichoderma reesei. U.S. Pat. nos. 4,435,307; european patent application No. 0495257; PCT patent application Nos. WO 91/17244; and European patent application No. EP-A2-271004, which are incorporated herein by reference.

Cellulases are also useful in the biopolishing of textiles. Cotton and other natural fibers based on cellulose can be modified by enzymatic biopolishing to produce fabrics with a smoother and more shiny appearance. This treatment can be used to remove "fuzz", i.e., micro-fiber bundles protruding from the yarn surface. Fluff balls are known in the textile industry as "pilling". After biopolishing, fuzz and pilling are reduced. Other benefits of pile removal are softer and smoother hand and superior color brightness.

In some embodiments of the compositions and methods of the present invention, the cellulase may be used at a concentration of from about 0.0001% to about 1% by weight of the enzyme protein by weight of the fabric, for example from about 0.0001% to about 0.05% by weight of the enzyme protein by weight of the fabric, or from about 0.0001% to about 0.01% by weight of the enzyme protein by weight of the fabric.

In some embodiments, one or more cellulases is comprised in a textile enzymatic bleaching composition as described herein and a system for removing hydrogen peroxide, such as a catalase enzyme, is added after the bleached and biopolished textile is produced.

In some embodiments, a method of combined bleaching and biopolishing a textile is provided comprising (i) contacting the textile with an enzymatic bleaching composition as described herein and a biopolishing enzyme (e.g., a cellulase) for a suitable condition and length of time to allow measurable whitening of the textile and the biopolished textile, wherein the bleached and biopolished textile comprises at least one of reduced textile damage, bulkier softer handle and increased dye uptake when compared to a chemical bleaching process comprising contacting the textile with a chemical bleaching composition for the textile that does not contain a perhydrolase enzyme; and (ii) hydrolyzing the hydrogen peroxide after producing the bleached and biopolished textile using a system for removing hydrogen peroxide (e.g., catalase).

Determination of cellulase activity (ECU): cellulolytic activity can be determined in endo-cellulase units (ECU) by measuring the ability of the enzyme to reduce the viscosity of a carboxymethyl cellulose (CMC) solution. The ECU test quantifies the amount of catalytic activity present in a sample by measuring the ability of the sample to reduce the viscosity of a carboxymethyl cellulose (CMC) solution. The test can be performed as follows: in a vibrating viscometer (e.g., MIVI 3000 from Sofraser, France) at 40 ℃; pH 7.5; 0.1M phosphate buffer; the time period was 30 minutes, relative enzyme standards were used to reduce the CHIC substrate viscosity (Hercules 7LED), and the enzyme concentration was approximately 0.15 ECU/mi. This major standard (arch standard) was defined as 8,200 ECU/g. One ECU is the amount of enzyme that reduces the viscosity to half under these conditions.

Other Bioscouring enzymes

The compositions and methods of the present invention are not limited to the use of the enzymes discussed above for bioscouring. Other enzymes may be used alone or in combination with each other or with the enzymes listed above. For example, proteases may be used in the compositions and methods of the invention. Suitable proteases include those of animal, vegetable or microbial origin, preferably of microbial origin. The protease may be a serine protease or a metalloprotease, preferably an alkaline microbial protease or a trypsin-like protease. Examples of proteases include aminopeptidases including prolylaminopeptidase (3.4.11.5), X-pro aminopeptidase (3.4.11.9), bacterial leucyl aminopeptidase (3.4.11.10), thermophilic aminopeptidase (3.4.11.12), lysyl aminopeptidase (3.4.11.15), tryptophanyl aminopeptidase (3.4.11.17), and methionyl aminopeptidase (3.4.11.18); serine endopeptidases including chymotrypsin (3.4.21.1), trypsin (3.4.21.4), cucumisin (3.4.21.25), brachyurin (3.4.21.32), cerevisin (3.4.21.48) and subtilisin (3.4.21.62); cysteine endopeptidases including papain (3.4.22.2), ficin (3.4.22.3), chymopapain (3.4.22.6), proteinase (3.4.22.7), actinidin (3.4.22.14), papain (3.4.22.30) and ananain (3.4.22.31); aspartic endopeptidases including pepsin a (3.4.23.1), aspergillus opepsin I (3.4.23.18), Penicillopepsin (3.4.23.20) and Saccharopepsin (3.4.23.25); and metalloendopeptidases including bacillus lysin (3.4.24.28).

Non-limiting examples of subtilisins include subtilisin BPN', subtilisin amylosuchritus, subtilisin 168, subtilisin mesterizopeptidase, subtilisin Carisberg, subtilisin DY, subtilisin 309, subtilisin 147, thermitase, aqualysin, Bacillus subtilis PB92 protease, protease K, protease TW7, and protease TW 3.

Commercially available proteases include ALCALASE^TM、SAVINASE^TM、PRIMASE.^TM、DURALASE.^TM、ESPERASE^TM、KANNASE^TMAnd DURAZYM^TM(Novo Nordisk A/S)、MAXATASE.^TM、MAXACAL.^TM、MAXAPEM^TM、PROPERASE^TM、Purafect ^TM、PURAFECT OXP ^TM、FN2·^TMAnd FN3^TM(Genencor Division，Danisco US Inc.)。

Protease variants may also be used in the compositions and methods of the invention, for example in the patent or published patent applications EP 130,756(Genentech), EP 214,435(Henkel), WO 87/04461(Amgen), WO 87/05050(Genex), EP 251,446(Genencor), EP 260,105(Genencor), Thomas et al (1985) Nature 318: 375-76, Thomas et al (1987) J.mol.biol.193: 803-13, Russel et al (1987) Nature 328: 496 + 500, WO88/08028(Genex), WO 88/08033(Amgen), WO 89/06279(Novo Nordisk A/S), WO 91/00345(Novo Nordisk A/S), EP 525610(Solvay) and WO94/02618(Gist-Brocades N.V.), all of which are incorporated herein by reference.

The protease activity can be determined as described in "Methods of enzymic Analysis", 3 rd edition, 1984, VerlaggChemie, Weinheim, volume 5.

In other embodiments, it is contemplated that lipases can be used, alone or in combination with other bioscouring enzymes, in the textile bioscouring of the present compositions and methods. Suitable lipases (also known as carboxylic ester hydrolases) include, but are not limited to, those of bacterial or fungal origin, including triacylglycerol lipases (3.1.1.3) and phospholipase a2 (3.1.1.4.). Lipases include, but are not limited to, lipases from the genus Humicola (synonymous with Thermomyces), e.g.from Humicola lanuginosus (T.lanuginosus), as described in patents or published patent applications EP 258,068 and EP305,216, or from Humicola insolens, as described in WO 96/13580; pseudomonas lipases, for example from Pseudomonas alcaligenes or Pseudomonas pseudoalcaligenes (EP 218,272), Pseudomonas cepacia (P.cepacia) (EP 331,376), Pseudomonas plecoglosa (GB 1,372,034), Pseudomonas fluorescens (P.fluorosceens), Pseudomonas species strain SD 705(WO 95/06720 and WO 96/27002), P.wisconsinensis (WO 96/12012); bacillus lipases, e.g., from Bacillus subtilis (Dartois et al (1993) biochem. Biophys. acta 1131: 253-360); bacillus stearothermophilus (JP 64/744992) or Bacillus pumilus (WO 91/16422), all of which are incorporated herein by reference. Further examples are lipase variants, e.g.in WO 92/05249, WO 94/01541, EP407225, EP 260105, WO 95/35381, WO 96/00292, WO 95/30744, WO 94/25578, WO 95/14783, WO 95/22615, WO 97/04079 and WO 97/07202, all of which are incorporated herein by reference. Preferred commercially available lipases include LIPOLASE^TMAnd LIPOLASE ULTRA^TM、LIPOZYME ^TM、PALATASE ^TM、NOVOZYM ^TM435 and LECITASE^TM(both available from Novo Nordisk A/S). The lipase activity can be determined as described in "Methods of enzymic Analysis", 3 rd edition, 1984, Verlag Chemie, Weinhein, volume 4.

It is understood that any enzyme exhibiting bioscouring activity can be used in the practice of the compositions and methods of the present invention. That is, bioscouring enzymes derived from other organisms, or bioscouring enzymes derived from the above-listed enzymes in which one or more amino acids have been added, deleted or substituted, including hybrid polypeptides, may be used, provided that the resulting polypeptide exhibits bioscouring activity. Variants can be generated using conventional mutagenesis methods and identified using, for example, high throughput screening techniques such as agar plate screening. For example, pectate lyase activity may be measured by: the test solution (containing 0.7% w/v sodium polygalacturonate (Sigma P1879)) was applied to 4mm wells punched out of an agar plate (e.g., LB agar). The plates are then incubated for 6 hours at a specific temperature (e.g., 75 ℃). The plates were then immersed (i) in 1M CaCl2 for 0.5 hours or (ii) in 1% mixed alkyltrimethylammonium bromide (MTAB, Sigma M-7635) for 1 hour. Both procedures resulted in the precipitation of polygalacturonate in agar. Pectate lyase activity may be detected by the appearance of a clear zone in the background of precipitated polygalacturonate. The sensitivity of the assay was calibrated using dilutions of the pectate lyase standard preparation.

Desizing enzyme

In some embodiments, the enzymatic textile bleaching methods described herein comprise one or more desizing enzymes. The one or more desizing enzymes may be included in the enzymatic textile bleaching composition or the textile may be treated with the desizing enzymes in a subsequent processing step after pretreatment of the textile in the enzymatic textile bleaching composition.

Any suitable desizing enzyme may be used in the compositions and methods of the present invention. In some embodiments, the desizing enzyme is an amylolytic enzyme. Mannanases and glucoamylases may also be used. In some embodiments, the desizing enzyme is an alpha-or beta-amylase and combinations thereof.

Amylase

Suitable alpha and beta amylases in the compositions and methods of the invention include those of bacterial or fungal origin. Chemically or genetically modified mutants of such amylases are also included in this connection. Preferred alpha amylases include, for example, alpha amylases obtainable from Bacillus species. Useful amylases include, but are not limited to, OPTISIZE 40^TM、OPTISIZE 160^TM、OPTISIZE HT260^TM、OPTISIZE HT 520^TM、OPTISIZE HT Plus^TM、OPTISIZE FLEX^TM(all from Genencor), DURAMYL^TM、TERMAMYL ^TM、FUNGAMYL^TMAnd BAN^TM(all available from Novozymes A/S, Bagsvaerd, Denmark). Other preferred amylolytic enzymes are CGTases (cyclodextrin glucanotransferase, EC 2.4.1.19), such as those obtained from bacillus, thermoanaerobacter or thermoanaerobacter species.

OPTISIZE 40^TMAnd OPTISIZE 160^TMExpressed as RAU per gram of product. One RAU is the amount of enzyme that converts 1g of starch to soluble sugars within 1 hour under standard conditions. OPTISIZE HT 260^TM、OPTISIZE HT 520^TMAnd OPTISIZE HT Plus^TMIs expressed as TTAU/g. One TTAU is the amount of enzyme required to hydrolyze 100mg of starch to soluble sugars per hour under standard conditions. OPTISIZE FLEX^TMThe activity of (A) was determined as TSAU/g. One TSAU is the amount of enzyme required to convert 1mg of starch to soluble sugars within 1 minute under standard conditions.

The dosage of amylase varies depending on the type of process. A smaller dose will require more time than a larger dose of the same enzyme. However, there is no upper limit to the amount of desizing amylase, except as may be controlled by the physical characteristics of the solution. Excessive enzyme does not damage the fabric; which allows for shorter processing times. According to the foregoing and the enzymes used, the following minimum doses are suggested for desizing:

amylase product	Minimum dose (per liter desizing liquor)	Typical range (per liter desizing liquor)
			OPTISIZE 40TM	1,000RAU	2,000-70,000RAU
OPTISIZE 160TM	1,000RAU	2,000-70,000RAU
			OPTISIZE HT 26TM 0	1,000TTAU	3,000-100,000TTAU
OPTISIZE HT 520TM	1,000TTAU	3,000-100,000TTAU
			OPTISIZE HT Plus TM	1,000TTAU	3,000-100,000TTAU
OPTISIZE FLEX TM	5,000TSAU	13,000-65,000TSAU

Desizing enzymes may be derived from the enzymes listed above and wherein one or more amino acids have been added, deleted or substituted, including hybrid polypeptides, provided that the resulting polypeptide exhibits desizing activity. Variants useful in practicing the compositions and methods of the invention can be generated using conventional mutagenesis procedures and identified using, for example, high throughput screening techniques such as agar plate screening.

The desizing enzyme is added to the aqueous solution (i.e., the treatment composition) in an amount effective to desize the textile material. Typically, a desizing enzyme, such as an alpha-amylase, is added to the treatment composition in an amount from about 0.00001% to about 2% enzyme protein by weight of the fabric, preferably in an amount from about 0.0001% to about 1% enzyme protein by weight of the fabric, more preferably in an amount from about 0.001% to about 0.5% enzyme protein by weight of the fabric, even more preferably in an amount from about 0.01% to about 0.2% enzyme protein by weight of the fabric.

Textile product

The compositions and methods of the present invention provide textiles, e.g., bleached textiles, produced according to any of the enzymatic bleaching methods described herein. Bleached textiles produced by incubation with an enzymatic textile bleaching composition as described herein exhibit at least one of reduced textile damage, increased dye uptake, and bulkier softer hand when compared to bleached textiles prepared with a chemical bleaching composition that does not contain a perhydrolase enzyme. The compositions and methods of the present invention also provide dyed textiles produced from bleached textiles that have been produced according to the enzymatic bleaching methods herein.

In some embodiments, the bleached textiles and/or bleached and dyed textiles are cellulose-containing textiles including, but not limited to, cotton, linen, hemp, ramie, cellulose acetate, lyocell, viscose rayon, bamboo, and various cellulose blends. In some embodiments, the bleached textile and/or bleached and dyed textile is a polyamide, polyacrylic or wool textile, or blends thereof.

Reagent kit

The compositions and methods of the invention may be provided in the form of a kit of parts (i.e., a kit). In one embodiment, the kit provides a perhydrolase enzyme, and instructions for using the perhydrolase enzyme in the enzymatic textile bleaching compositions and/or enzymatic textile bleaching methods described herein. The present invention provides suitable packaging. As used herein, "package" refers to a solid matrix or material typically used in systems that is capable of containing the components of a kit as described herein (e.g., perhydrolase) within fixed boundaries.

The instructions may be provided in printed form, or in the form of an electronic medium such as a floppy disk, CD or DVD, or a website address where the instructions are available.

The following examples are intended to illustrate, but not limit, the compositions and methods of the present invention.

Examples

Example 1

Enzymatic bleach pretreatment of 100% cotton Jersey (Single Jersey) material

The comparison between enzymatic and chemical bleaching processes was performed in a batch process using cotton knit textile material in a Mathis AG Lab Jet plant.

Bleaching composition

The compositions shown in table 1 were used in the experiments described below.

TABLE 1

Bleaching composition

Comprises the following components:

0.5% (w/w) [ [ (phosphonomethyl) imino ] bis [2, 1-ethanediylbis (methylene) ] ] tetra-phosphonic acid, sodium salt

5-10% (w/w) alkyl ethoxylates

15-20% (w/w) ethoxylated isotridecanol

< 5% (w/w) polyacrylic acid, sodium salt.

The phosphate buffer contained 10% soda ash.

The pectinase is 10% BIOPREP^TM3000L (from Novozymes).

Perhydrolase at a stock concentration of 1.7g/l of SEQ ID NO: 1S 54V variant.

Pretreatment process

Approximately 120g of fabric was incubated in each pretreatment composition at a liquor ratio of approximately 10: 1. The MathisAG Laboratory Jet machine increased the bath temperature at 3 ℃ per minute from room temperature to a target temperature of 65 ℃. The bath was then held at 65 ℃ for 50 minutes.

Rinsing twice at 50 ℃ each timeThe time is 10 minutes. Each rinse contained 25% CATALASE T100^TM(available from Genencor). The peroxide concentrations before and after rinsing are shown in table 2 for each bleaching composition tested. Peroxide concentration was evaluated using indicator strips from Merck, inc.

TABLE 2

Peroxide concentration before and after rinsing with Catalase

Bleaching composition		1	2	3	4
						Before one	ppm	25	25	20	15
After that	ppm	0	0	0	0

Rewetting

Rewet of fabrics treated with each of the bleaching compositions described above was evaluated using a modified wicking test. Deionized water was placed in a beaker, a strip of fabric was placed in the beaker, just touching the water, and then the time for the water to travel 1cm was measured. A low rewet rate (expressed in cm/sec) indicates a better hydrophobicity. The results are shown in Table 3.

TABLE 3

Rewet value

Bleaching composition	1	2	3	4
					Hydrophobicity (cm/sec)	1.5	2.7	104	80

Whiteness degree

Whiteness was quantified using 4 different test methods. The results are shown in Table 4.

TABLE 4

Whiteness degree

Bleaching composition	1	2	3	4
					Ganz	50	46	25	24
ISO/Tappi	86.0	85.4	80.1	78.9
					CIE	73	71	60	58
Berger	72	69	59	58

Fabric damage evaluation

The degree of polymerization of the fabrics treated with each of the bleaching compositions described above was evaluated. The degree of polymerisation was determined using the Swiss EWNNMethod (Swiss Standard SNV 195598). According to the formula of o.eisenhut, a damage factor (S) is determined, correlating fiber damage to changes in the degree of polymerization before and after pretreatment.

The results are shown in Table 5. For comparison, the degree of polymerization of the greige 100% cotton knit (knitgood) was 2380.

TABLE 5

Fabric damage evaluation

Bleaching composition	1	2	3	4
					Degree of polymerization	2060	2110	2280	2230
Injury factor	S：0.17	S：0.15	S：0.05	S：0.08

Dyeing and color fastness

The fabrics treated with the bleaching composition described above were used at 60 ℃ in a MathisAG Labomat apparatusFN 3G, 3% (w/w) staining for 90 min. The staining depth, color deviation and chromaticity deviation were evaluated.

The results of the colorimetric evaluation are shown in table 6. Colorimetric evaluation is based on the colorimetric method CIE-Lab (Munsell), and color deviation indicates the difference in color (red-green and blue-yellow) and chromaticity deviation indicates the difference in brightness.

TABLE 6

Evaluation of colorimetric analysis

The fastness was evaluated in terms of rubbing fastness, washing fastness, water fastness, and acid and alkali perspiration fastness. The wet/dry crocking fastness (crocking) was evaluated according to test method ISO 105-X12. The fastness to washing was evaluated at 60 ℃ according to test method ISO 105-C06. The water fastness was evaluated according to test method ISO 105-E01. The acid/alkali perspiration fastness was evaluated according to test method ISO 105-E04. Similar results were obtained for the chemical bleaching compositions (1 and 2) and the enzymatic bleaching compositions (3 and 4) for all these parameters.

Hand feeling

Before and after dyeing, it was observed that the fabrics pretreated in the enzymatic bleaching compositions (3 and 4) had a bulkier, softer fabric hand compared to the fabrics pretreated in the chemical bleaching compositions (1 and 2).

Although the foregoing compositions and methods of this invention have been described in some detail by way of illustration and example for purposes of clarity of understanding, it will be apparent to those of ordinary skill in the art that certain changes and modifications may be practiced without departing from the scope and spirit of the invention. Accordingly, the description should not be construed as limiting the scope of the invention.

All publications, patents, and patent applications cited herein are hereby incorporated by reference in their entirety for all purposes to the same extent as if each individual publication, patent, or patent application were specifically and individually indicated to be incorporated by reference.

Claims (42)

1. An enzymatic textile bleaching composition comprising:

(i) a perhydrolase;

(ii) an ester substrate of said perhydrolase;

(iii) a source of hydrogen peroxide;

(iv) surfactants and/or emulsifiers;

(v) phosphonic acid as a peroxide stabilizer;

(vi) polyacrylic acid as a sequestering agent; and

(vii) a buffer to maintain a pH of 6 to 8.

2. The enzymatic textile bleaching composition of claim 1, wherein the perhydrolase enzyme comprises the amino acid sequence depicted in SEQ ID NO. 1, or a variant or homologue thereof.

3. The enzymatic textile bleaching composition of claim 2, wherein the perhydrolase enzyme is the S54V variant of seq id No. 1.

4. The enzymatic textile bleaching composition of any of claims 1-3, wherein the perhydrolase enzyme exhibits a perhydrolysis to hydrolysis ratio of greater than 1.

5. The enzymatic textile bleaching composition of any of preceding claims 1-3, wherein the perhydrolase enzyme is present at a concentration of 1 to 2.5 ppm.

6. The enzymatic textile bleaching composition of any of the preceding claims 1-3, wherein the ester substrate is selected from the group consisting of propylene glycol diacetate, ethylene glycol diacetate, triacetin, ethyl acetate, and tributyrin.

7. The enzymatic textile bleaching composition of claim 6, wherein the ester substrate is propylene glycol diacetate.

8. The enzymatic textile bleaching composition of claim 7, wherein the propylene glycol diacetate is present in the composition in an amount of from 2,000 to 4,000 ppm.

9. The enzymatic textile bleaching composition of claim 1, wherein the hydrogen peroxide source is hydrogen peroxide.

10. The enzymatic textile bleaching composition of claim 9, wherein the hydrogen peroxide is present at a concentration of 1,000 to 3,000 ppm.

11. The enzymatic textile bleaching composition of claim 1, wherein the surfactant and/or emulsifier comprises a nonionic surfactant.

12. The enzymatic textile bleaching composition of claim 11, wherein the nonionic surfactant is an alcohol ethoxylate.

13. The enzymatic textile bleaching composition of claim 1, wherein the surfactant and/or emulsifier comprises an isotridecanol ethoxylate.

14. The enzymatic textile bleaching composition of claim 1, wherein the surfactant and/or emulsifier comprises an alkyl ethoxylate and an isotridecyl alcohol ethoxylate.

15. The enzymatic textile bleaching composition of claim 1, comprising a surfactant and an emulsifier.

16. The enzymatic textile bleaching composition of claim 1, further comprising a bioscouring enzyme.

17. The enzymatic textile bleaching composition of claim 16, wherein the bioscouring enzyme is selected from the group consisting of pectinases, cutinases, cellulases, hemicellulases, proteases, and lipases.

18. The enzymatic textile bleaching composition of claim 17, wherein the bioscouring enzyme is a pectinase.

19. A method for bleaching a textile, comprising contacting the textile with the enzymatic textile bleaching composition of any of claims 1-18 under conditions and for a length of time that allow measurable whitening of the textile, thereby producing a bleached textile, wherein the bleached textile comprises at least one of reduced textile damage, bulkier softer handle, and increased dye uptake when compared to a chemical textile bleaching process comprising contacting the textile with a chemical textile bleaching composition that does not contain a perhydrolase enzyme.

20. The method of claim 19, further comprising hydrolyzing the hydrogen peroxide with catalase enzyme after producing the bleached textile.

21. The method of claim 19, wherein the liquor ratio is 10: 1.

22. The method of claim 19, which is carried out in a batch process or a run-down process.

23. The method of any one of claims 19-22, wherein the method provides at least 10% less weight loss compared to a chemical bleaching composition that does not comprise a perhydrolase enzyme.

24. The process of claim 23, wherein the process provides at least 20% less weight loss.

25. The process of claim 24, wherein the process provides at least 30% less weight loss.

26. The process of claim 25, wherein the process provides at least 40% less weight loss.

27. The process of claim 26, wherein the process provides at least 50% less weight loss.

28. The method of any of claims 19-22, wherein the method provides a textile capable of increasing dye uptake to produce a dyed textile having at least a 5% increase in dye depth as compared to a textile treated with a chemical bleaching composition that does not contain a perhydrolase enzyme.

29. The method of claim 28, wherein the method provides a textile that is capable of increasing dye uptake to produce a dyed textile having at least a 10% increase in dye depth.

30. The method of claim 29, wherein the method provides a textile that is capable of increasing dye uptake to produce a dyed textile having at least a 15% increase in dye depth.

31. The method of claim 30, wherein the method provides a textile that is capable of increasing dye uptake to produce a dyed textile with at least a 20% increase in dye depth.

32. The method of claim 31, wherein the method provides a textile that is capable of increasing dye uptake to produce a dyed textile with at least a 25% increase in dye depth.

33. The method of claim 32, wherein the method provides a textile that is capable of increasing dye uptake to produce a dyed textile having at least a 30% increase in dye depth.

34. The method according to any one of claims 19-22, wherein the method provides a textile exhibiting a reduced tendency to pilling as compared to a textile treated with a chemical bleaching composition that does not comprise a perhydrolase enzyme.

35. The method of any of claims 19-22, wherein the textile is contacted with the textile enzymatic bleaching composition at a bleaching temperature of 60 ℃ to 70 ℃ for a treatment time of 40 to 60 minutes.

36. The method of claim 35, wherein the temperature of the enzymatic textile bleaching composition is increased at a starting temperature of from 20 ℃ to 40 ℃ at 3 ℃ per minute until the bleaching temperature is reached.

37. The method of claim 35, wherein the bleaching temperature is 65 ℃ and the treatment time is 50 minutes.

38. The method of any of claims 19-22, wherein the bleached textile is rinsed with an aqueous composition at a rinsing temperature of 40 ℃ to 60 ℃ to remove the textile enzymatic bleaching composition.

39. The method of claim 38, wherein the rinse temperature is 50 ℃.

40. The method of claim 38, wherein said rinsing comprises rinsing said bleached textile twice for 10 minutes each.

41. The process of any of the preceding claims 38-40, wherein the aqueous composition comprises a catalase enzyme to hydrolyze the hydrogen peroxide.

42. Use of an enzymatic textile bleaching composition for bleaching cellulose-containing textiles, said composition comprising an enzymatic textile bleaching composition according to any of claims 1 to 18, characterized in that the treatment of textiles with said composition provides improved dye uptake, bulkier softer handle and/or reduced textile damage compared to treatment with chemical bleaching.