Family 44 xyloglucanase variants

Abstract

The present disclosure relates to xyloglucanase variants. The invention also relates to polynucleotides encoding the variants; nucleic acid constructs, vectors, and host cells comprising the polynucleotides; and methods of using the variants.

Description

[0001] Incorporation by Reference of Sequence Listing

[0002] The instant application contains a Sequence Listing which has been submitted in computer readable form via EFS-Web. The computer readable form, filed in structural form, is incorporated herein by reference. TECHNICAL FIELD

[0003] The present invention relates to xylogucanase variants belonging to glycosyl hydrolase family 44, polynucleotides encoding the variants and methods of producing the variants. BACKGROUND

[0004] Xylogucan is the major structural polysaccharide in the primary (growing) cell walls of plants. Structurally, xylogucan consists of a cellulose-like beta-1,4-linked glucose backbone that is often substituted with a variety of side chains. Xylogucan is believed to function in the primary plant cell wall by cross-linking cellulose microfibrils to form a cellulose-xylogucan network.

[0005] Xylogucanases are capable of catalyzing the solubilization of xylogucan into xylogucan oligosaccharides. Some xylogucanases exhibit only xylogucanase activity, while other xylogucanases exhibit both xylogucanase activity and cellulase activity. Xylogucanases can be classified as EC 3.2.1.4 or EC.3.2.1.151. Enzymes with xylogucanase activity are described, for example, in Vincken et al. (1997) Carbohydrate Research 298(4):299-310, where three different endoglucanases, Endo I, Endo V and Endo VI, from Trichoderma viride (similar to T. reesei) are characterized. Endo I, Endo V and Endo VI belong to glycosyl hydrolase families 5, 7 and 12, respectively, see Henrissat, B. (1991) Biochem. J. 280:309-316 and Henrissat, B. and Bairoch, A. (1993) Biochem. J. 293:781-788. WO 94 / 14953 discloses a family 12 xylogucanase (EG II) cloned from the fungus Aspergillus aculeatus. WO 99 / 02663 discloses a family 12 and a family 5 xylogucanase cloned from Bacillus licheniformis and Bacillus agaradhaerens, respectively. WO 01 / 062903 discloses a family 44 xylogucanase.

[0006] In particular, WO 99 / 02663 and WO 01 / 062903 indicate that xyloglucanases can be used in detergents. WO 2009 / 147210 provides xyloglucanase variants.

[0007] It is an object of the present invention to provide xyloglucanase variants belonging to glycosyl hydrolase family 44. SUMMARY

[0008] The present invention relates to isolated xyloglucanase variants comprising an alteration at one or more positions corresponding to positions selected from the group consisting of 111, 123, 159, 256, 294, 8, 18, 20, 41, 42, 76, 82, 83, 87, 94, 103, 104, 105, 121, 125, 126, 127, 136, 137, 146, 147, 148, 152, 153, 155, 165, 168, 169, 177, 184, 189, 203, 206, 210, 211, 214, 217, 219, 220, 226, 237, 238, 240, 243, 244, 248, 251, 252, 267, 271, 276, 289, 295, 298, 300, 302, 322, 329, 339, 347, 347, 353, 383, 384, 392, 394, 395, 402, 414, 427, 431, 445, 447, 459, 473, 474, 476, 482, 488, 489, 491, 492, 503, and 505 of the polypeptide of SEQ ID NO: 1, wherein the variant has xyloglucanase activity. Preferably, the variant has at least 60%, e.g., 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%, e.g., at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to the polypeptide of SEQ ID NO: 1, SEQ ID NO: 2, or SEQ ID NO: 3 but less than 100% sequence identity, and wherein the variant has xyloglucanase activity.

[0009] The present invention relates to isolated xyloglucanase variants comprising a substitution at a position corresponding to position 129T of the polypeptide of SEQ ID NO: 2, wherein the variant has xyloglucanase activity. Preferably, the variant has at least 99% sequence identity to the polypeptide of SEQ ID NO: 2 but less than 100% sequence identity. In particular, the present invention relates to isolated xyloglucanase variants comprising or consisting of the polypeptide of SEQ ID NO: 3.

[0010] The present application also relates to isolated polynucleotides encoding these variants; nucleic acid constructs, vectors, and host cells comprising the polynucleotides; and methods of producing the variants.

[0011] The present application also relates to cleaning methods and compositions comprising the variants of the present application.

[0012] Sequence

[0013] SEQ ID NO: 1 mature polypeptide obtained from Paenibacillus polymyxa.

[0014] SEQ ID NO: 2 variant polypeptide.

[0015] SEQ ID NO: 3 variant polypeptide.

[0016] SEQ ID NO: 4 protease protein sequence from Bacillus lentus.

[0017] Definitions

[0018] According to this detailed description, the following definitions apply. Note that the singular forms "a," "an," and "the" include plural referents unless the context clearly dictates otherwise.

[0019] Reference herein to "about" an amount or parameter includes aspects that are directed to that amount or parameter per se. For example, reference to "about X" includes the aspect "X."

[0020] Unless otherwise defined or indicated by context, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this application belongs.

[0021] Allelic variant: The term "allelic variant" means any of two or more alternative forms of a gene occupying the same chromosomal locus. Allelic variation is naturally occurring and can result in polymorphism in the population. A gene mutation can be silent (no change in the encoded polypeptide) or can encode polypeptides having altered amino acid sequences. An allelic variant of a polypeptide is a polypeptide encoded by an allelic variant of a gene.

[0022] Amino acid:The term "amino acid" as used herein refers to the standard twenty genetically encoded amino acids and their corresponding stereoisomers in the "d" form (as opposed to the natural "l" form), omega-amino acids, other naturally occurring amino acids, nonstandard amino acids (e.g., alpha, alpha-disubstituted amino acids, N-alkyl amino acids, etc.), and chemically derivatized amino acids. Chemical derivatives of one or more amino acids can be achieved by reaction with functional side groups. Such derivatized molecules include, for example, those in which the free amino group has been derivatized to form an amine salt hydrochloride, a p-toluenesulfonyl group, a carboxyphenoxy group, a t-butoxy carbonyl group, a chloroacetyl group, or a formyl group. The free carboxyl group can be derivatized to form a salt, a methyl and ethyl ester, or other types of esters and hydrazides. The free hydroxyl group can be derivatized to form O-acyl or O-alkyl derivatives. Also included as chemical derivatives are those naturally occurring amino acid derivatives containing the twenty standard amino acids. For example: 4-hydroxyproline can be substituted for proline; 5-hydroxylysine can be substituted for lysine; 3-methylhistidine can be substituted for histidine; homoserine can be substituted for serine and ornithine can be substituted for lysine. Derivatives also include peptides containing one or more additions or deletions, so long as the necessary activity is maintained. Other included modifications are amidation, amino-terminal acylation (e.g., acetylation or mercaptoacetic acid amidation), carboxyl-terminal amidation (e.g., with ammonia or methylamine), and similar terminal modifications.

[0023] When an amino acid is specifically named, such as "alanine" or "Ala" or "A", unless otherwise specifically indicated, the term refers to both the l-alanine and the d-alanine. Other nonstandard amino acids can also be suitable components of the polypeptides of the present application, so long as the desired functional properties are retained by the polypeptide. For the peptides shown, each encoded amino acid residue is represented by a single letter code, where appropriate, corresponding to the common name of the conventional amino acid. In one embodiment, the polypeptides of the present application comprise or consist of l-amino acids

[0024] Cellulolytic enzyme or cellulaseThe term "cellulolytic enzyme" or "cellulase" means one or more (e.g., several) enzymes that hydrolyze cellulose material. Such enzymes include one or more endo-glucanases (e.g., EC 3.2.1.4), one or more cellobiohydrolases, one or more beta-glucosidases, or a combination thereof. Two basic methods for measuring cellulolytic enzyme activity include: (1) measuring total cellulolytic enzyme activity, and (2) measuring individual cellulolytic enzyme activities (endo-glucanases, cellobiohydrolases, and beta-glucosidases), as reviewed in Zhang et al., 2006, Biotechnology Advances 24:452-481. Total cellulolytic enzyme activity can be measured using insoluble substrates, including Whatman® No. 1 filter paper, Avicel® microcrystalline cellulose, bacterial cellulose, algal cellulose, cotton, pretreated lignocellulose, and the like. The most common total cellulolytic activity assay is the filter paper assay, which uses Whatman® No. 1 filter paper as a substrate. This assay was established by the International Union of Pure and Applied Chemistry (IUPAC) (Ghose, 1987, Pure Appl. Chem. 59:257-68).

[0025] Cellulosic material The term "cellulosic material" means any material containing cellulose. The predominant polysaccharide in the primary cell wall of biomass is cellulose, the second most abundant is hemicellulose, and the third most abundant is pectin. The secondary cell wall, produced after the cell has stopped growing, also contains polysaccharides and it is reinforced by polymeric lignin covalently cross-linked with hemicellulose. Cellulose is a homopolymer of anhydroglucopyranose and thus is a linear beta-(1-4)-D-glucan, while hemicelluloses include a variety of compounds with a range of substituents in complex branched structures, such as xylans, xyloglucans, arabinoxylans, and mannans. While cellulose is often polymorphous, it is found primarily as an insoluble crystalline matrix of parallel glucan chains in plant tissue. Hemicelluloses are often hydrogen-bonded to cellulose as well as other hemicelluloses, which helps to stabilize the cell wall matrix.

[0026] cDNA The term "cDNA" means a DNA molecule that can be prepared by reverse transcription from a mature, spliced, mRNA molecule obtained from a eukaryotic or prokaryotic cell. cDNA lacks intron sequences that are present in the corresponding genomic DNA. The initial, primary RNA transcript is a precursor to mRNA that is processed through a series of steps, including splicing, before the mature spliced mRNA is presented.

[0027] Coding sequenceThe term "coding sequence" means a polynucleotide, the direct expression product of which specifies the amino acid sequence of a variant. The boundaries of a coding sequence are typically determined by a start codon (e.g., ATG, GTG, or TTG) at the 5' end and a stop codon (e.g., TAA, TAG, or TGA) at the 3' end. A coding sequence can be genomic DNA, cDNA, synthetic DNA, or combinations thereof.

[0028] Control sequence The term "control sequence" means a nucleic acid sequence necessary for the expression of a polynucleotide encoding a variant of the present application. Each control sequence can be native (i.e., from the same gene) or foreign (i.e., from a different gene) to the polynucleotide encoding the variant or native or foreign to each other. Such control sequences include, but are not limited to, a leader, polyadenylation sequence, prepro-sequence, promoter, signal peptide sequence, and transcription terminator. At a minimum, these control sequences include a promoter, and transcriptional and translational stop signals. The control sequences can be provided with linkers for the purpose of introducing specific restriction sites facilitating ligation of the control sequences with the coding region of the polynucleotide encoding a variant.

[0029] Dishwashing composition: The term "dishwashing composition" as used herein refers to all forms of compositions for cleaning hard surfaces. The present application is not limited to any particular type of dishwashing composition or any particular detergent. Thus, in one embodiment, the dishwashing composition is a liquid dishwashing composition, a powder dishwashing composition, wherein the composition can optionally be in the form of a unit dose.

[0030] Detergent component The term "detergent component" is defined herein to mean the type of chemical that can be used in a detergent composition. Examples of detergent components are surfactants, hydrotropes, builders, co-builders, chelators or chelating agents, bleach systems or bleach components, polymers, fabric hueing agents, fabric conditioning agents, foam boosters, foam inhibitors, dispersants, dye transfer inhibitors, optical brighteners, perfumes, optical brighteners, bactericides, fungicides, soil suspending agents, soil release polymers, anti-redeposition agents, enzyme inhibitors or stabilizers, enzyme activators, antioxidants, and solubilizers. A detergent composition can comprise one or more detergent components of any type.

[0031] Detergent compositionThe term "detergent composition" refers to a composition for removing undesired compounds from articles to be cleaned, such as textiles, dishes, and hard surfaces. The detergent composition can be used, for example, to clean textiles, dishes, and hard surfaces, for both household and industrial cleaning and / or for fabric care. These terms encompass any material / compound selected for the particular type of cleaning composition and product form (e.g., liquid, gel, powder, granulate, paste, or spray composition) desired, and include, but are not limited to, detergent compositions (e.g., liquid and / or solid laundry detergents and fine fabric detergents; hard surface cleaning formulations, such as for glass, wood, plastic, ceramic, and metal countertops and windows; carpet cleaners; oven cleaners; fabric fresheners; fabric softeners; and textile and laundry pre-spotters, along with dishwashing detergents). In addition to containing the enzymes of the present invention, the detergent formulation can also contain one or more additional enzymes (e.g., amylases, proteases, peroxidases, cellulases, beta-glucanases, xyloglucanases, hemicellulases, xanthanases, xanthan lyases, lipases, acyl transferases, phospholipases, esterases, laccases, catalases, aryl esterases, amylases, alpha-amylases, glucoamylases, cutinases, pectinases, pectin lyases, keratinases, reductases, oxidases, phenol oxidases, lipoxygenases, ligninases, carrageenases, pullulanases, tannases, arabinosidases, hyaluronidases, chondroitinases, xyloglucanases, xylanases, pectate acetyl esterases, polygalacturonase, rhamnogalacturonase, endo-beta-mannanases, exo-beta-mannanases (GH5 and / or GH26), licheninases, phosphodiesterases, pectin methyl esterases, cellobiohydrolases, transglutaminases, nucleases, and combinations thereof, or any mixture thereof), and / or components such as surfactants, builders, chelating agents or chelating

[0032] Dishwashing The term "dishwashing" refers to all forms of washing dishes, such as hand dishwashing (HDW) or automatic dishwashing (ADW). Washing dishes includes, but is not limited to, cleaning all forms of tableware, such as plates, cups, glasses, bowls, all forms of cutlery (such as spoons, knives, forks), and serving utensils along with ceramic, plastic, metal, china, glass, and acrylic.

[0033] Enzymatic wash benefitThe term "enzyme wash benefit" is defined herein as the advantageous effect of adding an enzyme to a detergent as compared to the same detergent without the enzyme. Important wash benefits that can be provided by enzymes are stain removal with no or very little visible soil after washing and / or cleaning, prevention or reduction of redeposition of soil released during washing (also referred to as anti-redeposition effect), complete or partial restoration of whiteness of textiles (also referred to as whitening effect) that were originally white but acquired a greyish or yellowish appearance after repeated use and washing. Textile care benefits that are not directly related to catalyzing stain removal or prevention of soil redeposition are also important for enzyme wash benefits. Examples of such textile care benefits are prevention or reduction of dye transfer from one fabric to another fabric or to another part of the same fabric (also referred to as dye transfer inhibition or anti-backstaining effect), removal of protruding or broken fibers from fabric surfaces to reduce pilling tendency or to remove already existing pills or fuzz (also referred to as anti-pilling effect), improvement of fabric softness, color clarification of fabrics and removal of particulate soil trapped in the fibers of a fabric or garment. Enzymatic bleaching is a further enzyme wash benefit, wherein catalytic activity is used to catalyze the formation of a bleaching component, such as hydrogen peroxide or other peroxides.

[0034] Expression The term "expression" includes any step involved in the production of a variant, including but not limited to transcription, post-transcriptional modification, translation, post-translational modification, and secretion.

[0035] Expression vector The term "expression vector" means a linear or circular DNA molecule that comprises a polynucleotide encoding a variant and is operably linked to control sequences providing for its expression.

[0036] Fragment The term "fragment" means a polypeptide having wood po lygalacturonase activity that has one or more (e.g., several) amino acids deleted from the amino and / or carboxyl terminus of a mature polypeptide; in one aspect, a fragment contains at least 445 amino acid residues, at least 471 amino acid residues, or at least 497 amino acid residues.

[0037] Fusion polypeptideThe term "fusion polypeptide" is a polypeptide in which one polypeptide is fused at the N-terminus or C-terminus of a variant of the application. Fusion polypeptides are produced by fusing a polynucleotide encoding another polypeptide to a polynucleotide of the application. Techniques for producing fusion polypeptides are known in the art and include joining the coding sequences encoding the polypeptides such that they are in reading frame, and the expression of the fusion polypeptide is under the control of one or more of the same promoters and terminators. The fusion polypeptide can also be constructed using intein technology, in which the fusion polypeptide is produced post-translationally (Cooper et al., 1993, EMBO J. 12:2575-2583; Dawson et al., 1994, Science 266:776-779). The fusion polypeptide can further comprise a cleavage site between the two polypeptides. At the time of secretion of the fusion protein, the site is cleaved, thereby releasing the two polypeptides. Examples of cleavage sites include, but are not limited to, those disclosed in Martin et al., 2003, J. Ind. Microbiol. Biotechnol. 3:568-576; Svetina et al., 2000, J. Biotechnol. 76:245-251; Rasmussen-Wilson et al., 1997, Appl. Environ. Microbiol. 63:3488-3493; Ward et al., 1995, Biotechnology 13:498-503; and Contreras et al., 1991, Biotechnology 9:378-381; Eaton et al., 1986, Biochemistry 25:505-512; Collins-Racie et al., 1995, Biotechnology 13:982-987; Carter et al., 1989, Proteins: Structure, Function, and Genetics 6:240-248; and Stevens, 2003, Drug Discovery World 4:35-48.

[0038] Hard surface cleaningThe term "hard surface cleaning" is defined herein as cleaning of hard surfaces, which can include floors, tables, walls, roofs, etc., along with surfaces of hard objects, such as cars (car washing) and dishes (dish washing). Dish washing includes, but is not limited to, cleaning of plates, cups, glasses, bowls, cutlery (such as spoons, knives, forks), serving utensils, ceramics, plastics, metals, china, glass, and acrylics.

[0039] Host cell The term "host cell" means any cell type that is susceptible to transformation, transfection, transduction, etc. with a nucleic acid construct or expression vector comprising a polynucleotide of the present application. The term "host cell" encompasses any progeny of the parent cell which is not identical to the parent cell (due to mutations that occur during replication).

[0040] Hybrid polypeptide: The term "hybrid polypeptide" means a polypeptide comprising domains from two or more polypeptides of different origin (e.g., a binding module from one polypeptide and a catalytic domain from another polypeptide). The domains can be fused at the N- or C-terminus. Of particular interest herein are polypeptides comprising a binding module from one polypeptide (which can be naturally occurring or further modified), an engineered linker region (e.g., a proline-rich linker region, which is a synthetic construct), and a catalytic domain from another polypeptide (which can be naturally occurring or further modified).

[0041] Hybridization:The term "hybridization" means the pairing of substantially complementary strands of nucleic acids using standard Southern blot procedures. Hybridization can be performed under medium, medium-high, high, or very high stringency conditions. Medium stringency conditions mean prehybridization and hybridization at 42°C in 5X SSPE, 0.3% SDS, 200 microgram / ml sheared and denatured salmon sperm DNA, and 35% formamide, followed by washing twice in 2X SSC with 0.2% SDS at 50°C, and then twice in 0.2X SSC at 50°C. Medium-high stringency conditions mean prehybridization and hybridization at 42°C in 5X SSPE, 0.3% SDS, 200 microgram / ml sheared and denatured salmon sperm DNA, and 35% formamide, followed by washing twice in 2X SSC with 0.2% SDS at 60°C, and then twice in 0.2X SSC at 60°C. High stringency conditions mean prehybridization and hybridization at 42°C in 5X SSPE, 0.3% SDS, 200 microgram / ml sheared and denatured salmon sperm DNA, and 50% formamide, followed by washing twice in 2X SSC with 0.2% SDS at 65°C, and then twice in 0.2X SSC at 65°C. Very high stringency conditions mean prehybridization and hybridization at 42°C in 5X SSPE, 0.3% SDS, 200 microgram / ml sheared and denatured salmon sperm DNA, and 50% formamide, followed by washing twice in 2X SSC with 0.2% SDS at 70°C, and then twice in 0.2X SSC at 70°C.

[0042] Improved properties The term "improved property" means a characteristic associated with a variant that is improved compared to a reference enzyme / parent enzyme. Such improved properties include, but are not limited to, improved wash performance, improved enzyme wash benefit, improved stability, and / or improved whiteness.

[0043] Improved stability The term "improved stability" means that a variant enzyme exhibits retention of enzyme activity after incubation in the presence of one or more naturally occurring or synthetic chemicals that decrease the enzyme activity of the parent enzyme for a period of time. Improved stability means that the variant enzyme has better stability in the presence of a protease relative to the stability of the reference enzyme / parent enzyme and includes, for example, proteolysis stability, in-detergent storage stability, in-detergent storage stability in the presence of a chelating agent or chelating agent, improved stability during production of a detergent composition, and in-wash stability. In particular, improved detergent stability is an improved stability of xyloglucanase activity when the xyloglucanase variant of the present invention is mixed into a liquid detergent formulation or unit dose detergent formulation and then stored at a temperature of 15°C to 50°C.

[0044] In the present invention, liquid detergents are particularly useful as liquid laundry detergents and / or unit dose laundry detergents.

[0045] The improvement in stability can be quantified, for example, by the detergent stability assay as described in Example 3.

[0046] Improved wash performance The term "improved wash performance" is defined herein as an increased wash performance of an enzyme in a detergent composition, for example, by increased color clarification and / or anti-pilling effect, relative to the wash performance of a reference enzyme / parent enzyme, when evaluated on a new sample and / or after storing the sample under the same conditions. The term "improved wash performance" includes wash performance in laundry washing and also in, for example, hard surface cleaning such as automated dishwashing (ADW).

[0047] Isolated The term "isolated" means a polypeptide, nucleic acid, cell, or other specified material or component that is separated from at least one other material or component with which it is naturally associated in nature, including but not limited to, for example, other proteins, nucleic acids, cells, etc. Isolated polypeptides include, but are not limited to, culture broth containing secreted polypeptides.

[0048] Washing: The term "washing" relates to both domestic washing and industrial washing and means the process of treating a textile with a solution containing a cleaning or detergent composition of the present application. The washing process can be performed, for example, using a domestic or industrial washing machine or can be performed manually.

[0049] Mature polypeptide The term "mature polypeptide" means a polypeptide in its mature form following processing (e.g., removal of a signal peptide) at the N-terminus.

[0050] Mature polypeptide coding sequence The term "mature polypeptide-encoding sequence" means a polynucleotide that encodes a mature polypeptide having xyloglucanase activity.

[0051] Mutant The term "mutant" means a polynucleotide encoding a variant.

[0052] Modification: In the context of the polypeptides of the present application, the term "modification" means altering one or more amino acids within a reference amino acid sequence (i.e., SEQ ID NO: 1, 2, or 3) by substitution with a different amino acid, by insertion of an amino acid, or by deletion (preferably by at least one deletion). The terms "modification", "alteration", and "mutation" can be used interchangeably and constitute the same meaning and purpose.

[0053] Nucleic acid constructThe term "nucleic acid construct" means a nucleic acid molecule, either single- or double-stranded, which is isolated from a naturally occurring gene or which has been modified to contain segments of nucleic acids in a manner that would not otherwise exist in nature, or which is synthetic, which comprises one or more control sequences.

[0054] Operably linked: The term "operably linked" means a configuration in which the control sequences are placed at appropriate positions in relation to the coding sequence of the polynucleotide such that the control sequences direct the expression of the coding sequence.

[0055] Parent or parent xyloglucanase The term "parent" or "parent xyloglucanase" means any polypeptide having xyloglucanase activity, which is altered (e.g., one or more substitutions, one or more insertions, one or more deletions, and / or one or more truncations) to produce a xyloglucanase variant of the present invention. The parent can be a naturally occurring (wild-type) polypeptide or a variant or fragment thereof. The parent can be a naturally occurring (wild-type) polypeptide, such as the enzyme of SEQ ID NO: 1 or a polypeptide having at least 60%, more preferably at least 65%, more preferably at least 70%, more preferably at least 75%, more preferably at least 80%, more preferably at least 85%, even more preferably 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 at least 99% sequence identity thereto. The parent polypeptide can also be a variant of a naturally occurring polypeptide, which has been modified or altered in its amino acid sequence, such as the polypeptide of SEQ ID NO: 2 or SEQ ID NO: 3 herein. The parent can also be an allelic variant, which is a polypeptide encoded by either of two or more alternative forms of the gene occupying the same chromosomal locus.

[0056] PurifiedThe term "purified" means a nucleic acid or polypeptide that is substantially free of other components as determined by analytical techniques well known in the art (e.g., the purified polypeptide or nucleic acid can form a discrete band upon polyacrylamide gel electrophoresis or HPLC or density gradient centrifugation). A purified nucleic acid or polypeptide is at least about 50% pure, typically at least about 60%, about 65%, about 70%, about 75%, about 80%, about 85%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, about 99%, about 99.5%, about 99.6%, about 99.7%, about 99.8%, or more pure (e.g., weight percent on a molar basis). In a related sense, a composition is enriched for a molecule when there is a substantial increase in the concentration of the molecule after application of a purification or enrichment technique. The term "enriched" refers to a compound, polypeptide, cell, nucleic acid, amino acid, or other specified material or component being present in a composition in a relative or absolute concentration greater than that of the starting composition.

[0057] Recombinant The term "recombinant," when used in reference to a cell, nucleic acid, protein, or vector, means that it has been modified from its natural state. Thus, for example, a recombinant cell expresses genes that are not found within the native (non-recombinant) form of the cell, or expresses native genes at different levels, or under different conditions than found in nature. A recombinant nucleic acid differs from a native sequence in that one or more nucleotides and / or is operably linked to a heterologous sequence (e.g., a heterologous promoter in an expression vector). A recombinant protein differs from a native sequence in that it can have one or more amino acids and / or is fused to a heterologous sequence. A vector comprising a nucleic acid encoding a polypeptide is a recombinant vector. The term "recombinant" is synonymous with "genetically modified" and "transgenic."

[0058] Sequence identity The degree of relatedness between two amino acid sequences or between two nucleotide sequences is described by the parameter "sequence identity."

[0059] For the purposes of the present application, the sequence identity between two amino acid sequences is determined using the Needleman-Wunsch algorithm as implemented in the Needle program of the EMBOSS package (EMBOSS: The European Molecular Biology Open Software Suite, Rice et al., 2000, Trends Genet. 16: 276-277) version 5.0.0 or later (preferably EMBOSS: The European Molecular Biology open software suite: EMBOSS version 5.0.0 ed., Rice et al., 2000, European Molecular Biology Open Software Suite (EMBOSS), pp. 1-357, European Molecular Biology Open Software Foundation, ISBN 095743498X). The parameters used are gap open penalty of 10, gap extension penalty of 0.5, and the EBLOSUM62 (EMBOSS version of BLOSUM62) substitution matrix. The output of the Needle program is used as the percent identity and is calculated as the number of exact matches divided by the length of the reference sequence and multiplied by 100. The reference sequence is aligned to itself as the query sequence.

[0060] (exact matches x 100) / (length of query - total number of gaps in the alignment)

[0061] For the purposes of the present application, the sequence identity between two deoxyribonucleotide sequences is determined using the Needleman-Wunsch algorithm as implemented in the Needle program of the EMBOSS package (EMBOSS: The European Molecular Biology Open Software Suite, Rice et al., 2000, supra) version 5.0.0 or later (preferably EMBOSS: The European Molecular Biology open software suite: EMBOSS version 5.0.0 ed., Rice et al., 2000, European Molecular Biology Open Software Suite (EMBOSS), pp. 1-357, European Molecular Biology Open Software Foundation, ISBN 095743498X). The parameters used are gap open penalty of 10, gap extension penalty of 0.5, and the EDNAFULL (EMBOSS version of NCBI NUC4.4) substitution matrix. The output of the Needle program is used as the percent identity and is calculated as the number of exact matches divided by the length of the reference sequence and multiplied by 100.

[0062] (exact matches x 100) / (length of query - total number of gaps in the alignment)

[0063] Subsequence The term "subsequence" means a polynucleotide having one or more nucleotides deleted from the 5' and / or 3' end of a mature polypeptide coding sequence; wherein the subsequence encodes a fragment having xyloglucanase activity.

[0064] TextileThe term "textile" means any textile material, including yarns, yarn intermediates, fibers, nonwoven materials, natural materials, synthetic materials, and any other textile material, fabrics made from these materials, and products made from the fabrics (e.g., garments and other articles). The textile or fabric can be in the form of knitgoods, woven, denim, nonwoven, felt, yarn, and terry. The textile can be cellulose-based, such as natural cellulose articles, including cotton, linen / flax, jute, ramie, sisal, or coir, or man-made cellulose articles (e.g., derived from wood pulp), including viscose / rayon, cellulose acetate fibers (tricell), lyocell, or blends thereof. The textile or fabric can also be non-cellulose-based, such as natural polyamides, including wool, camel hair, cashmere, mohair, rabbit hair, and silk, or synthetic polymers such as nylon, aramid, polyester, acrylate, polypropylene, and spandex / elastane, or blends thereof and blends of cellulose-based and non-cellulose-based fibers. An example of a blend is a blend of cotton and / or rayon / viscose with one or more companion materials such as wool, synthetic fibers (e.g., polyamide fibers, acrylic fibers, polyester fibers, polyvinyl chloride fibers, polyurethane fibers, polyurea fibers, aramid fibers), and / or cellulose-containing fibers (e.g., rayon / viscose, ramie, linen / flax, jute, cellulose acetate fibers, lyocell). The fabric can be a conventional washable laundry, such as a soiled household laundry. When the term fabric or garment is used, it is intended to also include the broader term textile.

[0065] Variant The term "variant" means a polypeptide having xyloglucanase activity that comprises an alteration at one or more (e.g., several) positions and retains the activity of the parent. By substitution is meant replacing the amino acid occupying a certain position with a different amino acid. Variants of the present invention have at least 20% of the xyloglucanase activity of the polypeptide of SEQ ID NO: 1, such as at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, 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%, at least 99%, or at least 100%.

[0066] Wash liquor The term "wash liquor" refers to an aqueous solution containing a detergent composition in diluted form, such as but not limited to a detergent solution containing a laundry detergent composition in diluted form, such as a wash liquor during a laundry process.

[0067] WhitenessThe term "whiteness" is defined herein as a broad term that has different meanings in different fields and for different customers. Loss of whiteness can for example be attributed to graying, yellowing, or removal of optical brighteners / hueing agents. Graying and yellowing can be attributed to soil redeposition, body soils, coloration from e.g. iron and copper ions or dye transfer. Whiteness can include one or several issues from the following list: colorant or dye action; incomplete stain removal (e.g. body soils, sebum, etc.); redeposition (graying, yellowing or other discoloration of the object) (removed soil reassociating with other parts of the textile (soiled or unsoiled)); chemical changes of the textile during application; and clarification or brightening of color.

[0068] Wild type The term "wild type" when referring to an amino acid sequence or a nucleic acid sequence means that the amino acid sequence or nucleic acid sequence is a natural or naturally occurring sequence. As used herein, the term "naturally occurring" refers to any substance (e.g., protein, amino acid, or nucleic acid sequence) found in nature. In contrast, the term "non-naturally occurring" refers to any substance (e.g., recombinant nucleic acid and protein sequences produced in the laboratory, or modifications of wild type sequences) that is not found in nature.

[0069] Xyloglucanase activity The term "xyloglucanase activity" is defined herein as the catalysis of the hydrolysis of xyloglucan by an enzyme. The reaction involves endohydrolysis of 1,4-beta-D- glucosidic linkages in xyloglucan. For the purposes of the present invention, xyloglucanase activity is determined using AZCL-xyloglucan (from Megazyme) as the reaction substrate. The assay can be performed in several ways, for example, as described in Example 2 of the present application, or as described in WO 01 / 62903. One xyloglucanase activity unit (XyloU) is defined with reference to the assay method described in WO 01 / 62903, page 60, lines 3-17.

[0070] Variant nomenclature

[0071] For the purposes of the present invention, the amino acid sequence of the xyloglucanase disclosed in SEQ ID NO: 1 is used to determine the corresponding amino acid residue in another xyloglucanase. The amino acid sequence of another xyloglucanase is aligned with the amino acid sequence of the xyloglucanase disclosed in SEQ ID NO: 1 and based on this alignment, the amino acid position number corresponding to any amino acid residue in the amino acid sequence of the xyloglucanase disclosed in SEQ ID NO: 1 can be determined using the Needleman and Wunsch algorithm (Needleman and Wunsch, 1970, J. Mol. Biol. 48: 443-453) as implemented in the needle program of the EMBOSS software package (EMBOSS: The European Molecular Biology Open Software Suite, Rice et al., 2000, Trends Genet. 16: 276-277), preferably version 5.0.0 or later. The parameters used are a gap open penalty of 10, a gap extension penalty of 0.5, and the EBLOSUM62 (EMBOSS version of BLOSUM62) substitution matrix.

[0072] In describing the variants of the present invention, the following nomenclature as described below has been adapted for ease of reference. The accepted IUPAC single letter or three letter amino acid abbreviations have been used.

[0073] Substitution For amino acid substitutions, the following nomenclature is used: original amino acid, position, substituted amino acid. Accordingly, a substitution of threonine at position 226 with alanine is denoted "Thr226Ala" or "T226A". Multiple mutations are separated by a plus sign ("+"), e.g. "Gly205Arg+Ser411Phe" or "G205R+S411F" represents a substitution of glycine (G) and serine (S) at positions 205 and 411, respectively, with arginine (R) and phenylalanine (F).

[0074] Deletion For amino acid deletions, the following nomenclature is used: original amino acid, position, * . Accordingly, a deletion of glycine at position 195 is denoted "Gly195*" or "G195*". Multiple deletions are separated by a plus sign ("+"), e.g. "Gly195*+Ser411*" or "G195*+S411*".

[0075] Insertion.For amino acid insertions, the following nomenclature is used: original amino acid, position, original amino acid, inserted amino acid. Accordingly, an insertion of lysine after the glycine at position 195 is represented as "Gly195GlyLys" or "G195GK". Multiple amino acid insertions are represented as [original amino acid, position, original amino acid, inserted amino acid #1, inserted amino acid #2; etc.]. For example, an insertion of lysine and alanine after the glycine at position 195 is represented as "Gly195GlyLysAla" or "G195GKA".

[0076] In such cases, the inserted amino acid residue(s) are numbered by adding a lowercase letter to the position number of the amino acid residue preceding the inserted amino acid residue(s). In the above example, the sequence would thus be:

[0077] Parent: ]]> ​ Variant: ]]> ​ 195 195 195a 195b G G-K-A

[0078] Multiple alterations . Variants comprising multiple alterations are separated by a plus sign ("+"), e.g., "Arg170Tyr+Gly195Glu" or "R170Y+G195E" represents substitution of arginine and glycine at positions 170 and 195, respectively, with tyrosine and glutamic acid.

[0079] Different alterations . In cases where different alterations can be introduced at a position, the different alterations are separated by a comma, e.g., "Arg170Tyr,Glu" represents substitution of arginine at position 170 with tyrosine or glutamic acid. Thus, "Tyr167Gly,Ala+Arg170Gly,Ala" designates the following variants:

[0080] "Tyr167Gly+Arg170Gly", "Tyr167Gly+Arg170Ala", "Tyr167Ala+Arg170Gly", and "Tyr167Ala+Arg170Ala". DETAILED DESCRIPTION

[0081] Xyloglucanase variants

[0082] The present invention relates to isolated xyloglucanase variants, which comprise an alteration at one or more positions corresponding to positions selected from the group consisting of 111, 123, 159, 256, 294, 8, 18, 20, 41, 42, 76, 82, 83, 87, 94, 103, 104, 105, 121, 125, 126, 127, 136, 137, 146, 147, 148, 152, 153, 155, 165, 168, 169, 177, 184, 189, 203, 206, 210, 211, 214, 217, 219, 220, 226, 237, 238, 240, 243, 244, 248, 251, 252, 267, 271, 276, 289, 295, 298, 300, 302, 322, 329, 339, 347, 347, 353, 383, 384, 392, 394, 395, 402, 414, 427, 431, 445, 447, 459, 473, 474, 476, 482, 488, 489, 491, 492, 503, and 505 of the polypeptide of SEQ ID NO: 1, wherein the variant has xyloglucanase activity. Preferably, the variant has at least 60%, e.g., 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%, e.g., at least 96%, at least 97%, at least 98%, or at least 99%, but less than 100%, sequence identity to the polypeptide of SEQ ID NO: 1, SEQ ID NO: 2, or SEQ ID NO: 3, and wherein the variant has xyloglucanase activity.

[0083] The present invention relates to isolated xyloglucanase variants, which comprise a substitution at a position corresponding to position 129T of the polypeptide of SEQ ID NO: 2, wherein the variant has xyloglucanase activity. Preferably, the variant has at least 99% sequence identity to the polypeptide of SEQ ID NO: 2, but less than 100% sequence identity. In particular, the present invention relates to isolated xyloglucanase variants, which comprise or consist of the polypeptide of SEQ ID NO: 3.

[0084] In embodiments, the variant has at least 60%, e.g., 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 at least 99%, but less than 100%, sequence identity to the amino acid sequence of the parent xyloglucanase.

[0085] In another embodiment, the variant has at least 60%, e.g., 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%, such as at least 96%, at least 97%, at least 98%, or at least 99%, but less than 100% sequence identity with the polypeptide of SEQ ID NO: 1.

[0086] In another embodiment, the variant has at least 60%, e.g., 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%, such as at least 96%, at least 97%, at least 98%, or at least 99%, but less than 100% sequence identity with the polypeptide of SEQ ID NO: 2.

[0087] In another embodiment, the variant has at least 60%, e.g., 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%, such as at least 96%, at least 97%, at least 98%, or at least 99%, but less than 100% sequence identity with the polypeptide of SEQ ID NO: 3.

[0088] In one aspect, the number of alterations is 1-50, e.g., 1-45, 1-40, 1-35, 1-30, 1-25, 1-20, 1-15, 1-10, or 1-5, such as 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, or 50 alterations.

[0089] In one aspect, the number of substitutions is 1-50, e.g., 1-45, 1-40, 1-35, 1-30, 1-25, 1-20, 1-15, 1-10, or 1-5, such as 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, or 50 substitutions.

[0090] In one aspect, the number of deletions is 1-50, such as 1-45, 1-40, 1-35, 1-30, 1-25, 1-20, 1-15, 1-10, or 1-5, such as 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, or 50 deletions.

[0091] In one aspect, the substituted amino acid residue is different from the naturally occurring amino acid residue at the position. In one embodiment, the substitution is selected from the group consisting of A, C, D, E, F, G, H, I, K, L, M, N, P, Q, R, S, T, V, W, and Y, provided that the substituted amino acid residue is different from the naturally occurring amino acid residue at the position.

[0092] In one embodiment, the xyloglucanase variant of the application is an isolated variant.

[0093] In another aspect, a variant comprises alterations at two or more positions corresponding to positions selected from the group consisting of 111, 123, 159, 256, 294, 8, 18, 20, 41, 42, 76, 82, 83, 87, 94, 103, 104, 105, 121, 125, 126, 127, 136, 137, 146, 147, 148, 152, 153, 155, 165, 168, 169, 177, 184, 189, 203, 206, 210, 211, 214, 217, 219, 220, 226, 237, 238, 240, 243, 244, 248, 251, 252, 267, 271, 276, 289, 295, 298, 300, 302, 322, 329, 339, 347, 347, 353, 383, 384, 392, 394, 395, 402, 414, 427, 431, 445, 447, 459, 473, 474, 476, 482, 488, 489, 491, 492, 503, and 505 of the polypeptide of SEQ ID NO: 1, wherein the variant has xylogucanase activity and wherein the variant has at least 60%, e.g., 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%, e.g., at least 96%, at least 97%, at least 98%, or at least 99%, sequence identity to the polypeptide of SEQ ID NO: 1, SEQ ID NO: 2, or SEQ ID NO: 3, but less than 100% sequence identity.

[0094] In another aspect, a variant comprises alterations at three or more positions corresponding to positions selected from the group consisting of: 111, 123, 159, 256, 294, 8, 18, 20, 41, 42, 76, 82, 83, 87, 94, 103, 104, 105, 121, 125, 126, 127, 136, 137, 146, 147, 148, 152, 153, 155, 165, 168, 169, 177, 184, 189, 203, 206, 210, 211, 214, 217, 219, 220, 226, 237, 238, 240, 243, 244, 248, 251, 252, 267, 271, 276, 289, 295, 298, 300, 302, 322, 329, 339, 347, 347, 353, 383, 384, 392, 394, 395, 402, 414, 427, 431, 445, 447, 459, 473, 474, 476, 482, 488, 489, 491, 492, 503, and 505 of the polypeptide of SEQ ID NO: 1, wherein the variant has xylogucanase activity and wherein the variant has at least 60%, e.g., 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%, e.g., at least 96%, at least 97%, at least 98%, or at least 99%, sequence identity to the polypeptide of SEQ ID NO: 1, SEQ ID NO: 2, or SEQ ID NO: 3, but less than 100% sequence identity.

[0095] In another aspect, a variant comprises alterations at four or more positions corresponding to positions selected from the group consisting of: 111, 123, 159, 256, 294, 8, 18, 20, 41, 42, 76, 82, 83, 87, 94, 103, 104, 105, 121, 125, 126, 127, 136, 137, 146, 147, 148, 152, 153, 155, 165, 168, 169, 177, 184, 189, 203, 206, 210, 211, 214, 217, 219, 220, 226, 237, 238, 240, 243, 244, 248, 251, 252, 267, 271, 276, 289, 295, 298, 300, 302, 322, 329, 339, 347, 347, 353, 383, 384, 392, 394, 395, 402, 414, 427, 431, 445, 447, 459, 473, 474, 476, 482, 488, 489, 491, 492, 503, and 505 of the polypeptide of SEQ ID NO: 1, wherein the variant has xylogucanase activity and wherein the variant has at least 60%, e.g., 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%, e.g., at least 96%, at least 97%, at least 98%, or at least 99%, but less than 100%, sequence identity to the polypeptide of SEQ ID NO: 1, SEQ ID NO: 2, or SEQ ID NO: 3.

[0096] In another aspect, a variant comprises alterations at five or more positions corresponding to positions selected from the group consisting of: 111, 123, 159, 256, 294, 8, 18, 20, 41, 42, 76, 82, 83, 87, 94, 103, 104, 105, 121, 125, 126, 127, 136, 137, 146, 147, 148, 152, 153, 155, 165, 168, 169, 177, 184, 189, 203, 206, 210, 211, 214, 217, 219, 220, 226, 237, 238, 240, 243, 244, 248, 251, 252, 267, 271, 276, 289, 295, 298, 300, 302, 322, 329, 339, 347, 347, 353, 383, 384, 392, 394, 395, 402, 414, 427, 431, 445, 447, 459, 473, 474, 476, 482, 488, 489, 491, 492, 503, and 505 of the polypeptide of SEQ ID NO: 1, wherein the variant has xylogucanase activity and wherein the variant has at least 60%, e.g., 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%, e.g., at least 96%, at least 97%, at least 98%, or at least 99%, but less than 100%, sequence identity to the polypeptide of SEQ ID NO: 1, SEQ ID NO: 2, or SEQ ID NO: 3.

[0097] In another aspect, a variant comprises alterations at six or more positions corresponding to positions selected from the group consisting of: 111, 123, 159, 256, 294, 8, 18, 20, 41, 42, 76, 82, 83, 87, 94, 103, 104, 105, 121, 125, 126, 127, 136, 137, 146, 147, 148, 152, 153, 155, 165, 168, 169, 177, 184, 189, 203, 206, 210, 211, 214, 217, 219, 220, 226, 237, 238, 240, 243, 244, 248, 251, 252, 267, 271, 276, 289, 295, 298, 300, 302, 322, 329, 339, 347, 347, 353, 383, 384, 392, 394, 395, 402, 414, 427, 431, 445, 447, 459, 473, 474, 476, 482, 488, 489, 491, 492, 503, and 505 of the polypeptide of SEQ ID NO: 1, wherein the variant has xylogucanase activity and wherein the variant has at least 60%, e.g., 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%, e.g., at least 96%, at least 97%, at least 98%, or at least 99%, sequence identity to the polypeptide of SEQ ID NO: 1, SEQ ID NO: 2, or SEQ ID NO: 3, but less than 100% sequence identity.

[0098] In another aspect, a variant comprises alterations at seven or more positions corresponding to positions selected from the group consisting of: 111, 123, 159, 256, 294, 8, 18, 20, 41, 42, 76, 82, 83, 87, 94, 103, 104, 105, 121, 125, 126, 127, 136, 137, 146, 147, 148, 152, 153, 155, 165, 168, 169, 177, 184, 189, 203, 206, 210, 211, 214, 217, 219, 220, 226, 237, 238, 240, 243, 244, 248, 251, 252, 267, 271, 276, 289, 295, 298, 300, 302, 322, 329, 339, 347, 347, 353, 383, 384, 392, 394, 395, 402, 414, 427, 431, 445, 447, 459, 473, 474, 476, 482, 488, 489, 491, 492, 503, and 505 of the polypeptide of SEQ ID NO: 1, wherein the variant has xylogucanase activity and wherein the variant has at least 60%, e.g., 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%, e.g., at least 96%, at least 97%, at least 98%, or at least 99%, sequence identity to the polypeptide of SEQ ID NO: 1, SEQ ID NO: 2, or SEQ ID NO: 3, but less than 100% sequence identity.

[0099] In another aspect, a variant comprises alterations at eight or more positions corresponding to positions selected from the group consisting of 111, 123, 159, 256, 294, 8, 18, 20, 41, 42, 76, 82, 83, 87, 94, 103, 104, 105, 121, 125, 126, 127, 136, 137, 146, 147, 148, 152, 153, 155, 165, 168, 169, 177, 184, 189, 203, 206, 210, 211, 214, 217, 219, 220, 226, 237, 238, 240, 243, 244, 248, 251, 252, 267, 271, 276, 289, 295, 298, 300, 302, 322, 329, 339, 347, 347, 353, 383, 384, 392, 394, 395, 402, 414, 427, 431, 445, 447, 459, 473, 474, 476, 482, 488, 489, 491, 492, 503, and 505 of the polypeptide of SEQ ID NO: 1, wherein the variant has xylogucanase activity and wherein the variant has at least 60%, e.g., 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%, e.g., at least 96%, at least 97%, at least 98%, or at least 99%, sequence identity to the polypeptide of SEQ ID NO: 1, SEQ ID NO: 2, or SEQ ID NO: 3, but less than 100% sequence identity.

[0100] In another aspect, a variant comprises alterations at nine or more positions corresponding to positions selected from the group consisting of: 111, 123, 159, 256, 294, 8, 18, 20, 41, 42, 76, 82, 83, 87, 94, 103, 104, 105, 121, 125, 126, 127, 136, 137, 146, 147, 148, 152, 153, 155, 165, 168, 169, 177, 184, 189, 203, 206, 210, 211, 214, 217, 219, 220, 226, 237, 238, 240, 243, 244, 248, 251, 252, 267, 271, 276, 289, 295, 298, 300, 302, 322, 329, 339, 347, 347, 353, 383, 384, 392, 394, 395, 402, 414, 427, 431, 445, 447, 459, 473, 474, 476, 482, 488, 489, 491, 492, 503, and 505 of the polypeptide of SEQ ID NO: 1, wherein the variant has xylogucanase activity and wherein the variant has at least 60%, e.g., 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%, e.g., at least 96%, at least 97%, at least 98%, or at least 99%, but less than 100%, sequence identity to the polypeptide of SEQ ID NO: 1, SEQ ID NO: 2, or SEQ ID NO: 3.

[0101] In another aspect, a variant comprises alterations at ten or more positions corresponding to positions selected from the group consisting of 111, 123, 159, 256, 294, 8, 18, 20, 41, 42, 76, 82, 83, 87, 94, 103, 104, 105, 121, 125, 126, 127, 136, 137, 146, 147, 148, 152, 153, 155, 165, 168, 169, 177, 184, 189, 203, 206, 210, 211, 214, 217, 219, 220, 226, 237, 238, 240, 243, 244, 248, 251, 252, 267, 271, 276, 289, 295, 298, 300, 302, 322, 329, 339, 347, 347, 353, 383, 384, 392, 394, 395, 402, 414, 427, 431, 445, 447, 459, 473, 474, 476, 482, 488, 489, 491, 492, 503, and 505 of the polypeptide of SEQ ID NO: 1, wherein the variant has xylogucanase activity and wherein the variant has at least 60%, e.g., 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%, e.g., at least 96%, at least 97%, at least 98%, or at least 99%, but less than 100%, sequence identity to the polypeptide of SEQ ID NO: 1, SEQ ID NO: 2, or SEQ ID NO: 3.

[0102] In another aspect, a variant comprises alterations at eleven or more positions corresponding to positions selected from the group consisting of 111, 123, 159, 256, 294, 8, 18, 20, 41, 42, 76, 82, 83, 87, 94, 103, 104, 105, 121, 125, 126, 127, 136, 137, 146, 147, 148, 152, 153, 155, 165, 168, 169, 177, 184, 189, 203, 206, 210, 211, 214, 217, 219, 220, 226, 237, 238, 240, 243, 244, 248, 251, 252, 267, 271, 276, 289, 295, 298, 300, 302, 322, 329, 339, 347, 347, 353, 383, 384, 392, 394, 395, 402, 414, 427, 431, 445, 447, 459, 473, 474, 476, 482, 488, 489, 491, 492, 503, and 505 of the polypeptide of SEQ ID NO: 1, wherein the variant has xylogucanase activity and wherein the variant has at least 60%, e.g., 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%, e.g., at least 96%, at least 97%, at least 98%, or at least 99%, sequence identity to the polypeptide of SEQ ID NO: 1, SEQ ID NO: 2, or SEQ ID NO: 3, but less than 100% sequence identity.

[0103] In another aspect, a variant comprises alterations at twelve or more positions of the polypeptide of SEQ ID NO: 1 corresponding to positions selected from the group consisting of 111, 123, 159, 256, 294, 8, 18, 20, 41, 42, 76, 82, 83, 87, 94, 103, 104, 105, 121, 125, 126, 127, 136, 137, 146, 147, 148, 152, 153, 155, 165, 168, 169, 177, 184, 189, 203, 206, 210, 211, 214, 217, 219, 220, 226, 237, 238, 240, 243, 244, 248, 251, 252, 267, 271, 276, 289, 295, 298, 300, 302, 322, 329, 339, 347, 347, 353, 383, 384, 392, 394, 395, 402, 414, 427, 431, 445, 447, 459, 473, 474, 476, 482, 488, 489, 491, 492, 503, and 505, wherein the variant has xylogucanase activity and wherein the variant has at least 60%, e.g., 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%, e.g., at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to the polypeptide of SEQ ID NO: 1, SEQ ID NO: 2, or SEQ ID NO: 3 but less than 100% sequence identity.

[0104] In another aspect, a variant comprises alterations at thirteen or more positions corresponding to positions selected from the group consisting of: 111, 123, 159, 256, 294, 8, 18, 20, 41, 42, 76, 82, 83, 87, 94, 103, 104, 105, 121, 125, 126, 127, 136, 137, 146, 147, 148, 152, 153, 155, 165, 168, 169, 177, 184, 189, 203, 206, 210, 211, 214, 217, 219, 220, 226, 237, 238, 240, 243, 244, 248, 251, 252, 267, 271, 276, 289, 295, 298, 300, 302, 322, 329, 339, 347, 347, 353, 383, 384, 392, 394, 395, 402, 414, 427, 431, 445, 447, 459, 473, 474, 476, 482, 488, 489, 491, 492, 503, and 505 of the polypeptide of SEQ ID NO: 1, wherein the variant has xylogucanase activity and wherein the variant has at least 60%, e.g., 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%, e.g., at least 96%, at least 97%, at least 98%, or at least 99%, but less than 100%, sequence identity to the polypeptide of SEQ ID NO: 1, SEQ ID NO: 2, or SEQ ID NO: 3.

[0105] In another aspect, a variant comprises alterations at fourteen or more positions of the polypeptide of SEQ ID NO: 1 corresponding to positions selected from the group consisting of 111, 123, 159, 256, 294, 8, 18, 20, 41, 42, 76, 82, 83, 87, 94, 103, 104, 105, 121, 125, 126, 127, 136, 137, 146, 147, 148, 152, 153, 155, 165, 168, 169, 177, 184, 189, 203, 206, 210, 211, 214, 217, 219, 220, 226, 237, 238, 240, 243, 244, 248, 251, 252, 267, 271, 276, 289, 295, 298, 300, 302, 322, 329, 339, 347, 347, 353, 383, 384, 392, 394, 395, 402, 414, 427, 431, 445, 447, 459, 473, 474, 476, 482, 488, 489, 491, 492, 503, and 505, wherein the variant has xylogucanase activity and wherein the variant has at least 60%, e.g., 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%, e.g., at least 96%, at least 97%, at least 98%, or at least 99%, sequence identity to the polypeptide of SEQ ID NO: 1, SEQ ID NO: 2, or SEQ ID NO: 3, but less than 100% sequence identity.

[0106] In another aspect, a variant comprises alterations at fifteen or more positions of the polypeptide of SEQ ID NO: 1 corresponding to positions selected from the group consisting of 111, 123, 159, 256, 294, 8, 18, 20, 41, 42, 76, 82, 83, 87, 94, 103, 104, 105, 121, 125, 126, 127, 136, 137, 146, 147, 148, 152, 153, 155, 165, 168, 169, 177, 184, 189, 203, 206, 210, 211, 214, 217, 219, 220, 226, 237, 238, 240, 243, 244, 248, 251, 252, 267, 271, 276, 289, 295, 298, 300, 302, 322, 329, 339, 347, 347, 353, 383, 384, 392, 394, 395, 402, 414, 427, 431, 445, 447, 459, 473, 474, 476, 482, 488, 489, 491, 492, 503, and 505, wherein the variant has xylogucanase activity and wherein the variant has at least 60%, e.g., 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%, e.g., at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to the polypeptide of SEQ ID NO: 1, SEQ ID NO: 2, or SEQ ID NO: 3 but less than 100% sequence identity.

[0107] In another aspect, a variant comprises alterations at sixteen or more positions of the polypeptide of SEQ ID NO: 1 corresponding to positions selected from the group consisting of 111, 123, 159, 256, 294, 8, 18, 20, 41, 42, 76, 82, 83, 87, 94, 103, 104, 105, 121, 125, 126, 127, 136, 137, 146, 147, 148, 152, 153, 155, 165, 168, 169, 177, 184, 189, 203, 206, 210, 211, 214, 217, 219, 220, 226, 237, 238, 240, 243, 244, 248, 251, 252, 267, 271, 276, 289, 295, 298, 300, 302, 322, 329, 339, 347, 347, 353, 383, 384, 392, 394, 395, 402, 414, 427, 431, 445, 447, 459, 473, 474, 476, 482, 488, 489, 491, 492, 503, and 505, wherein the variant has xylogucanase activity and wherein the variant has at least 60%, e.g., 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%, e.g., at least 96%, at least 97%, at least 98%, or at least 99%, sequence identity to the polypeptide of SEQ ID NO: 1, SEQ ID NO: 2, or SEQ ID NO: 3, but less than 100% sequence identity.

[0108] In another aspect, a variant comprises alterations at seventeen or more positions of the polypeptide of SEQ ID NO: 1 corresponding to positions selected from the group consisting of 111, 123, 159, 256, 294, 8, 18, 20, 41, 42, 76, 82, 83, 87, 94, 103, 104, 105, 121, 125, 126, 127, 136, 137, 146, 147, 148, 152, 153, 155, 165, 168, 169, 177, 184, 189, 203, 206, 210, 211, 214, 217, 219, 220, 226, 237, 238, 240, 243, 244, 248, 251, 252, 267, 271, 276, 289, 295, 298, 300, 302, 322, 329, 339, 347, 347, 353, 383, 384, 392, 394, 395, 402, 414, 427, 431, 445, 447, 459, 473, 474, 476, 482, 488, 489, 491, 492, 503, and 505, wherein the variant has xylogucanase activity and wherein the variant has at least 60%, e.g., 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%, e.g., at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to the polypeptide of SEQ ID NO: 1, SEQ ID NO: 2, or SEQ ID NO: 3 but less than 100% sequence identity.

[0109] In another aspect, a variant comprises alterations at eighteen or more positions corresponding to positions selected from the group consisting of: 111, 123, 159, 256, 294, 8, 18, 20, 41, 42, 76, 82, 83, 87, 94, 103, 104, 105, 121, 125, 126, 127, 136, 137, 146, 147, 148, 152, 153, 155, 165, 168, 169, 177, 184, 189, 203, 206, 210, 211, 214, 217, 219, 220, 226, 237, 238, 240, 243, 244, 248, 251, 252, 267, 271, 276, 289, 295, 298, 300, 302, 322, 329, 339, 347, 347, 353, 383, 384, 392, 394, 395, 402, 414, 427, 431, 445, 447, 459, 473, 474, 476, 482, 488, 489, 491, 492, 503, and 505 of the polypeptide of SEQ ID NO: 1, wherein the variant has xylogucanase activity and wherein the variant has at least 60%, e.g., 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%, e.g., at least 96%, at least 97%, at least 98%, or at least 99%, but less than 100%, sequence identity to the polypeptide of SEQ ID NO: 1, SEQ ID NO: 2, or SEQ ID NO: 3.

[0110] In another aspect, a variant comprises alterations at ten or more positions corresponding to positions selected from the group consisting of 111, 123, 159, 256, 294, 8, 18, 20, 41, 42, 76, 82, 83, 87, 94, 103, 104, 105, 121, 125, 126, 127, 136, 137, 146, 147, 148, 152, 153, 155, 165, 168, 169, 177, 184, 189, 203, 206, 210, 211, 214, 217, 219, 220, 226, 237, 238, 240, 243, 244, 248, 251, 252, 267, 271, 276, 289, 295, 298, 300, 302, 322, 329, 339, 347, 347, 353, 383, 384, 392, 394, 395, 402, 414, 427, 431, 445, 447, 459, 473, 474, 476, 482, 488, 489, 491, 492, 503, and 505 of the polypeptide of SEQ ID NO: 1, wherein the variant has xylogucanase activity and wherein the variant has at least 60%, e.g., 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%, e.g., at least 96%, at least 97%, at least 98%, or at least 99%, but less than 100%, sequence identity to the polypeptide of SEQ ID NO: 1, SEQ ID NO: 2, or SEQ ID NO: 3.

[0111] In another aspect, a variant comprises alterations at twenty or more positions of the polypeptide of SEQ ID NO: 1 corresponding to positions selected from the group consisting of 111, 123, 159, 256, 294, 8, 18, 20, 41, 42, 76, 82, 83, 87, 94, 103, 104, 105, 121, 125, 126, 127, 136, 137, 146, 147, 148, 152, 153, 155, 165, 168, 169, 177, 184, 189, 203, 206, 210, 211, 214, 217, 219, 220, 226, 237, 238, 240, 243, 244, 248, 251, 252, 267, 271, 276, 289, 295, 298, 300, 302, 322, 329, 339, 347, 347, 353, 383, 384, 392, 394, 395, 402, 414, 427, 431, 445, 447, 459, 473, 474, 476, 482, 488, 489, 491, 492, 503, and 505, wherein the variant has xylogucanase activity and wherein the variant has at least 60%, e.g., 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%, e.g., at least 96%, at least 97%, at least 98%, or at least 99%, sequence identity to the polypeptide of SEQ ID NO: 1, SEQ ID NO: 2, or SEQ ID NO: 3, but less than 100% sequence identity.

[0112] In another aspect, a variant comprises an alteration at each position corresponding to a position selected from the group consisting of positions 111, 123, 159, 256, 294, 8, 18, 20, 41, 42, 76, 82, 83, 87, 94, 103, 104, 105, 121, 125, 126, 127, 136, 137, 146, 147, 148, 152, 153, 155, 165, 168, 169, 177, 184, 189, 203, 206, 210, 211, 214, 217, 219, 220, 226, 237, 238, 240, 243, 244, 248, 251, 252, 267, 271, 276, 289, 295, 298, 300, 302, 322, 329, 339, 347, 347, 353, 383, 384, 392, 394, 395, 402, 414, 427, 431, 445, 447, 459, 473, 474, 476, 482, 488, 489, 491, 492, 503, and 505 of the polypeptide of SEQ ID NO: 1, wherein the variant has xylogucanase activity and wherein the variant has at least 60%, e.g., 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%, e.g., at least 96%, at least 97%, at least 98%, or at least 99%, sequence identity to the polypeptide of SEQ ID NO: 1, SEQ ID NO: 2, or SEQ ID NO: 3, but less than 100% sequence identity.

[0113] In another aspect, a variant comprises an alteration at each position corresponding to a position selected from the group consisting of 111, 123, 129, 159, 256, 294, 8, 18, 20, 41, 42, 76, 82, 83, 87, 94, 103, 104, 105, 121, 125, 126, 127, 136, 137, 146, 147, 148, 152, 153, 155, 165, 168, 169, 177, 184, 189, 203, 206, 210, 211, 214, 217, 219, 220, 226, 237, 238, 240, 243, 244, 248, 251, 252, 267, 271, 276, 289, 295, 298, 300, 302, 322, 329, 339, 347, 347, 353, 383, 384, 392, 394, 395, 402, 414, 427, 431, 445, 447, 459, 473, 474, 476, 482, 488, 489, 491, 492, 503, and 505 of the polypeptide of SEQ ID NO: 1, wherein the variant has xylogucanase activity and wherein the variant has at least 60%, e.g., 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%, e.g., at least 96%, at least 97%, at least 98%, or at least 99%, sequence identity to the polypeptide of SEQ ID NO: 1, SEQ ID NO: 2, or SEQ ID NO: 3, but less than 100% sequence identity.

[0114] In another aspect, a variant comprises an alteration at each position corresponding to a position selected from the group consisting of 111, 123, 129, 159, 256, 294, 8, 18, 20, 41, 42, 76, 82, 83, 87, 94, 103, 104, 105, 121, 125, 126, 127, 136, 137, 146, 147, 148, 152, 153, 155, 165, 168, 169, 177, 184, 189, 203, 206, 210, 211, 214, 217, 219, 220, 226, 237, 238, 240, 243, 244, 248, 251, 252, 267, 271, 276, 289, 295, 298, 300, 302, 322, 329, 339, 347, 347, 353, 383, 384, 392, 394, 395, 402, 414, 427, 431, 445, 447, 459, 473, 474, 476, 482, 488, 489, 491, 492, 503, and 505 of the polypeptide of SEQ ID NO: 2, wherein the variant has xylogucanase activity and wherein the variant has at least 60%, e.g., 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%, e.g., at least 96%, at least 97%, at least 98%, or at least 99%, sequence identity to the polypeptide of SEQ ID NO: 1, SEQ ID NO: 2, or SEQ ID NO: 3, but less than 100% sequence identity.

[0115] In one aspect, a variant comprises one or more of the following alterations at positions corresponding to positions P111, S123, V159, S256, I294, K8, K18, R20, A41, A42, S76, Q82, A83, K87, S94, G103, T104, Y105, A118, N121, Q125, E126, S127, N136, Q137, F146, Q147, L148, L152, N153, N155, F165, N168, K169, A177, L184, A189, V203, K206, D210, R211, S214, K217, V219, K220, A226, G237, A238, K240, Q243, T244, W248, V251, K252, R267, Q271, R276, A289, R295, N298, V300, N302, K322, Q329, P339, K347, R347, K353, R353, N383, D384, K392, K394, D395, P395, S402, K414, T427, V431, K445, L447, A459, I473, S474, K476, K482, K488, E489, A491, P492, Y503, and V505 of the polypeptide of SEQ ID NO: 1, wherein the variant has xylogucanase activity and wherein the variant has at least 60%, e.g., 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%, e.g., at least 96%, at least 97%, at least 98%, or at least 99%, but less than 100%, sequence identity to the polypeptide of SEQ ID NO: 1, SEQ ID NO: 2, or SEQ ID NO: 3.

[0116] In one aspect, the variant comprises one or more of the following alterations at positions corresponding to positions P111, S123, V159, S256, I294, K8, K18, R20, A41, A42, S76, Q82, A83, K87, S94, G103, T104, Y105, A118, N121, Q125, E126, S127, K129, N136, Q137, F146, Q147, L148, L152, N153, N155, F165, N168, K169, A177, L184, A189, V203, K206, D210, R211, S214, K217, V219, K220, A226, G237, A238, K240, Q243, T244, W248, V251, K252, R267, Q271, R276, A289, R295, N298, V300, N302, K322, Q329, P339, K347, R347, K353, R353, N383, D384, K392, K394, D395, P395, S402, K414, T427, V431, K445, L447, A459, I473, S474, K476, K482, K488, E489, A491, P492, Y503, and V505 of the polypeptide of SEQ ID NO: 1, wherein the variant has xylogucanase activity and wherein the variant has at least 60%, e.g., 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%, e.g., at least 96%, at least 97%, at least 98%, or at least 99%, but less than 100%, sequence identity to the polypeptide of SEQ ID NO: 1, SEQ ID NO: 2, or SEQ ID NO: 3.

[0117] In one aspect, the variant comprises one or more of the following alterations at positions corresponding to positions P111, S123, V159, S256, I294, K8, K18, R20, A41, A42, S76, Q82, A83, K87, S94, G103, T104, Y105, A118, N121, Q125, E126, S127, A129, N136, Q137, F146, Q147, L148, L152, N153, N155, F165, N168, K169, A177, L184, A189, V203, K206, D210, R211, S214, K217, V219, K220, A226, G237, A238, K240, Q243, T244, W248, V251, K252, R267, Q271, R276, A289, R295, N298, V300, N302, K322, Q329, P339, K347, R347, K353, R353, N383, D384, K392, K394, D395, P395, S402, K414, T427, V431, K445, L447, A459, I473, S474, K476, K482, K488, E489, A491, P492, Y503, and V505 of the polypeptide of SEQ ID NO: 2, wherein the variant has xylogucanase activity and wherein the variant has at least 60%, e.g., 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%, e.g., at least 96%, at least 97%, at least 98%, or at least 99%, but less than 100%, sequence identity to the polypeptide of SEQ ID NO: 1, SEQ ID NO: 2, or SEQ ID NO: 3.

[0118] In one aspect, the variant comprises one or more of the following alterations at positions corresponding to the following positions of the polypeptide of SEQ ID NO: 1 : P111Q, S123P, V159M, S256E, S256Q, I294E, I294Q, K8E, K8R, K18E, R20K, A41L, A41E, A41R, A42V, S76E, Q82E, A83E, K87E, S94R, G103V, T104G, T104R, Y105E, A118K, N121E, Q125F, Q125K, Q125L, Q125P, Q125S, E126P, S127H, S127L, S127W, S127D, N136D, Q137E, Q137K, F146D, Q147G, Q147K, L148P, L152*, L152D, L152E, L152P, N153E, N155D, N155E, F165H, N168R, K169E, K169R, A177G, L184M, A189G, V203T, K206E, K206R, D210H, D210R, R211K, S214Q, K217R, K217T, V219A, V219T, K220R, A226D, A226K, G237M, A238S, A238T, K240F, K240L, Q243E, T244E, T244R, W248V, V251E, K252E, R267C, R267H, R267K, Q271D, Q271E, R276K, A289T, R295K, N298D, V300L, N302H, K322E, Q329E, P339S, K347E, K347R, K353R, N383E, N383Q, D384G, K392E, K394R, D395P, S402Q, K414E, T427V, V431E, K445E, L447M, A459P, I473T, S474E, K476R, K482R, K488T, E489K, E489R, A491E, A491V, P492D, Y503L, Y503V, and V505L, wherein the variant has xylogucanase activity and wherein the variant has at least 60%, e.g., 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%, e.g., at least 96%, at least 97%, at least 98%, or at least 99%, sequence identity to the polypeptide of SEQ ID NO: 1, SEQ ID NO: 2, or SEQ ID NO: 3, but less than 100% sequence identity.

[0119] In one aspect, the variant comprises one or more of the following alterations at positions corresponding to the following positions of the polypeptide of SEQ ID NO: 1 : P111Q, S123P, V159M, S256E, S256Q, I294E, I294Q, K8E, K8R, K18E, R20K, A41L, A41E, A41R, A42V, S76E, Q82E, A83E, K87E, S94R, G103V, T104G, T104R, Y105E, A118K, N121E, Q125F, Q125K, Q125L, Q125P, Q125S, E126P, S127H, S127L, S127W, S127D, K129A, K129T, N136D, Q137E, Q137K, F146D, Q147G, Q147K, L148P, L152*, L152D, L152E, L152P, N153E, N155D, N155E, F165H, N168R, K169E, K169R, A177G, L184M, A189G, V203T, K206E, K206R, D210H, D210R, R211K, S214Q, K217R, K217T, V219A, V219T, K220R, A226D, A226K, G237M, A238S, A238T, K240F, K240L, Q243E, T244E, T244R, W248V, V251E, K252E, R267C, R267H, R267K, Q271D, Q271E, R276K, A289T, R295K, N298D, V300L, N302H, K322E, Q329E, P339S, K347E, K347R, K353R, N383E, N383Q, D384G, K392E, K394R, D395P, S402Q, K414E, T427V, V431E, K445E, L447M, A459P, I473T, S474E, K476R, K482R, K488T, E489K, E489R, A491E, A491V, P492D, Y503L, Y503V, and V505L, wherein the variant has xylogucanase activity and wherein the variant has at least 60%, e.g., 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%, e.g., at least 96%, at least 97%, at least 98%, or at least 99%, sequence identity to the polypeptide of SEQ ID NO: 1, SEQ ID NO: 2, or SEQ ID NO: 3, but less than 100% sequence identity.

[0120] In one aspect, the variant comprises one or more of the following alterations at positions corresponding to the following positions of the polypeptide of SEQ ID NO: 2: P111Q, S123P, V159M, S256E, S256Q, I294E, I294Q, K8E, K8R, K18E, R20K, A41L, A41E, A41R, A42V, S76E, Q82E, A83E, K87E, S94R, G103V, T104G, T104R, Y105E, A118K, N121E, Q125F, Q125K, Q125L, Q125P, Q125S, E126P, S127H, S127L, S127W, S127D, A129T, N136D, Q137E, Q137K, F146D, Q147G, Q147K, L148P, L152*, L152D, L152E, L152P, N153E, N155D, N155E, F165H, N168R, K169E, K169R, A177G, L184M, A189G, V203T, K206E, K206R, D210H, D210R, R211K, S214Q, K217R, K217T, V219A, V219T, K220R, A226D, A226K, G237M, A238S, A238T, K240F, K240L, Q243E, T244E, T244R, W248V, V251E, K252E, R267C, R267H, R267K, Q271D, Q271E, R276K, A289T, R295K, N298D, V300L, N302H, K322E, Q329E, P339S, K347E, K347R, K353R, N383E, N383Q, D384G, K392E, K394R, D395P, S402Q, K414E, T427V, V431E, K445E, L447M, A459P, I473T, S474E, K476R, K482R, K488T, E489K, E489R, A491E, A491V, P492D, Y503L, Y503V, and V505L, wherein the variant has xylogucanase activity and wherein the variant has at least 60%, e.g., 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%, e.g., at least 96%, at least 97%, at least 98%, or at least 99%, sequence identity to the polypeptide of SEQ ID NO: 1, SEQ ID NO: 2, or SEQ ID NO: 3, but less than 100% sequence identity.

[0121] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 8. In another aspect, the amino acid at a position corresponding to position 8 is substituted with Ala, Arg, Asn, Asp, Cys, Gin, Glu, Gly, His, lie, Leu, Met, Phe, Pro, Ser, Thr, Trp, Tyr, or Val, preferably Pro. In another aspect, the variant comprises or consists of the substitution K8R or K8E of the polypeptide of SEQ ID NO: 1.

[0122] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 18. In another aspect, the amino acid at a position corresponding to position 18 is substituted with Ala, Arg, Asn, Asp, Cys, Gin, Glu, Gly, His, lie, Leu, Met, Phe, Pro, Ser, Thr, Trp, Tyr, or Val, preferably Pro. In another aspect, the variant comprises or consists of the substitution K18E of the polypeptide of SEQ ID NO: 1.

[0123] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 20. In another aspect, the amino acid at a position corresponding to position 20 is substituted with Ala, Arg, Asn, Asp, Cys, Gin, Glu, Gly, His, lie, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp, Tyr, or Val, preferably Pro. In another aspect, the variant comprises or consists of the substitution R20K of the polypeptide of SEQ ID NO: 1.

[0124] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 41. In another aspect, the amino acid at a position corresponding to position 41 is substituted with Arg, Asn, Asp, Cys, Gin, Glu, Gly, His, lie, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp, Tyr, or Val, preferably Pro. In another aspect, the variant comprises or consists of the substitution A41E or A41L or A41R of the polypeptide of SEQ ID NO: 1.

[0125] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 42. In another aspect, the amino acid at a position corresponding to position 42 is substituted with Arg, Asn, Asp, Cys, Gin, Glu, Gly, His, lie, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp, Tyr, or Val, preferably Pro. In another aspect, the variant comprises or consists of the substitution A42V of the polypeptide of SEQ ID NO: 1.

[0126] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 76. In another aspect, the amino acid at a position corresponding to position 76 is substituted with Ala, Arg, Asn, Asp, Cys, Gin, Glu, Gly, His, lie, Leu, Lys, Met, Phe, Pro, Thr, Trp, Tyr, or Val, preferably Pro. In another aspect, the variant comprises or consists of the substitution S76E of the polypeptide of SEQ ID NO: 1.

[0127] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 82. In another aspect, the amino acid at a position corresponding to position 82 is substituted with Ala, Arg, Asn, Asp, Cys, Glu, Gly, His, lie, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp, Tyr, or Val, preferably Pro. In another aspect, the variant comprises or consists of the substitution Q82E of the polypeptide of SEQ ID NO: 1.

[0128] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 83. In another aspect, the amino acid at a position corresponding to position 83 is substituted with Arg, Asn, Asp, Cys, Gin, Glu, Gly, His, lie, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp, Tyr, or Val, preferably Pro. In another aspect, the variant comprises or consists of the substitution A83E of the polypeptide of SEQ ID NO: 1.

[0129] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 87. In another aspect, the amino acid at a position corresponding to position 87 is substituted with Ala, Arg, Asn, Asp, Cys, Gin, Glu, Gly, His, lie, Leu, Met, Phe, Pro, Ser, Thr, Trp, Tyr, or Val, preferably Pro. In another aspect, the variant comprises or consists of the substitution K87E of the polypeptide of SEQ ID NO: 1.

[0130] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 94. In another aspect, the amino acid at a position corresponding to position 94 is substituted with Ala, Arg, Asn, Asp, Cys, Gin, Glu, Gly, His, lie, Leu, Lys, Met, Phe, Pro, Thr, Trp, Tyr, or Val, preferably Pro. In another aspect, the variant comprises or consists of the substitution S94R of the polypeptide of SEQ ID NO: 1.

[0131] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 103. In another aspect, the amino acid at a position corresponding to position 103 is substituted with Ala, Arg, Asn, Asp, Cys, Gin, Glu, Gly, His, lie, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp, Tyr, or Val, preferably Val. In another aspect, the variant comprises or consists of the substitution G103V of the polypeptide of SEQ ID NO: 1.

[0132] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 104. In another aspect, the amino acid at a position corresponding to position 104 is substituted with Ala, Arg, Asn, Asp, Cys, Gin, Glu, Gly, His, lie, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp, Tyr, or Val, preferably Gly, Arg. In another aspect, the variant comprises or consists of the substitution T104G or T104R of the polypeptide of SEQ ID NO: 1.

[0133] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 105. In another aspect, the amino acid at a position corresponding to position 105 is substituted with Ala, Arg, Asn, Asp, Cys, Gin, Glu, Gly, His, lie, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp, or Val, preferably Pro. In another aspect, the variant comprises or consists of the substitution Y105E of the polypeptide of SEQ ID NO: 1.

[0134] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 111. In another aspect, the amino acid at a position corresponding to position 111 is substituted with Ala, Arg, Asn, Asp, Cys, Gin, Glu, Gly, His, lie, Leu, Lys, Met, Phe, Ser, Thr, Trp, Tyr, or Val, preferably Pro. In another aspect, the variant comprises or consists of the substitution P111Q of the polypeptide of SEQ ID NO: 1.

[0135] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 118. In another aspect, the amino acid at a position corresponding to position 118 is substituted with Arg, Asn, Asp, Cys, Gin, Glu, Gly, His, lie, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp, Tyr, or Val, preferably Pro. In another aspect, the variant comprises or consists of the substitution A118K of the polypeptide of SEQ ID NO: 1.

[0136] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 121. In another aspect, the amino acid at a position corresponding to position 121 is substituted with Ala, Arg, Asp, Cys, Gin, Glu, Gly, His, lie, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp, Tyr, or Val, preferably Pro. In another aspect, the variant comprises or consists of the substitution N121E of the polypeptide of SEQ ID NO: 1.

[0137] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 123. In another aspect, the amino acid at a position corresponding to position 123 is substituted with Ala, Arg, Asn, Asp, Cys, Gin, Glu, Gly, His, lie, Leu, Lys, Met, Phe, Pro, Thr, Trp, Tyr, or Val, preferably Pro. In another aspect, the variant comprises or consists of the substitution S123P of the polypeptide of SEQ ID NO: 1.

[0138] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 125. In another aspect, the amino acid at a position corresponding to position 125 is substituted with Ala, Arg, Asn, Asp, Cys, Gin, Glu, Gly, His, lie, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp, Tyr, or Val, preferably with Phe, Leu, Ser, most preferably Ser. In another aspect, the variant comprises or consists of the substitution Q125F, Q125K, Q125L, Q125P, Q125S, most preferably Q125S, of the polypeptide of SEQ ID NO: 1.

[0139] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 126. In another aspect, the amino acid at a position corresponding to position 126 is substituted with Ala, Arg, Asn, Asp, Cys, Gin, Glu, Gly, His, lie, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp, Tyr, or Val, preferably with Pro. In another aspect, the variant comprises or consists of the substitution E126P of the polypeptide of SEQ ID NO: 1.

[0140] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 127. In another aspect, the amino acid at a position corresponding to position 127 is substituted with Ala, Arg, Asn, Asp, Cys, Gin, Glu, Gly, His, lie, Leu, Lys, Met, Phe, Pro, Thr, Trp, Tyr, or Val, preferably with Pro. In another aspect, the variant comprises or consists of the substitution S127H or S127L or S127W or S127D of the polypeptide of SEQ ID NO: 1.

[0141] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 136. In another aspect, the amino acid at a position corresponding to position 136 is substituted with Ala, Arg, Asp, Cys, Gin, Glu, Gly, His, lie, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp, Tyr, or Val, preferably with Pro. In another aspect, the variant comprises or consists of the substitution N136D of the polypeptide of SEQ ID NO: 1.

[0142] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 137. In another aspect, the amino acid at a position corresponding to position 137 is substituted with Ala, Arg, Asn, Asp, Cys, Glu, Gly, His, lie, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp, Tyr, or Val, preferably Pro. In another aspect, the variant comprises or consists of the substitution Q137E or Q137K of the polypeptide of SEQ ID NO: 1.

[0143] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 146. In another aspect, the amino acid at a position corresponding to position 146 is substituted with Ala, Arg, Asn, Asp, Cys, Gin, Glu, Gly, His, lie, Leu, Lys, Met, Pro, Ser, Thr, Trp, Tyr, or Val, preferably Pro. In another aspect, the variant comprises or consists of the substitution F146D of the polypeptide of SEQ ID NO: 1.

[0144] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 147. In another aspect, the amino acid at a position corresponding to position 147 is substituted with Ala, Arg, Asn, Asp, Cys, Glu, Gly, His, lie, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp, Tyr, or Val, preferably Pro. In another aspect, the variant comprises or consists of the substitution Q147G or Q147K of the polypeptide of SEQ ID NO: 1.

[0145] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 148. In another aspect, the amino acid at a position corresponding to position 148 is substituted with Ala, Arg, Asn, Asp, Cys, Gin, Glu, Gly, His, lie, Lys, Met, Phe, Pro, Ser, Thr, Trp, Tyr, or Val, preferably Pro. In another aspect, the variant comprises or consists of the substitution L148P of the polypeptide of SEQ ID NO: 1.

[0146] In another aspect, the variant comprises or consists of a substitution or deletion at a position corresponding to position 152. In another aspect, the amino acid at a position corresponding to position 152 is substituted with Ala, Arg, Asn, Asp, Cys, Gin, Glu, Gly, His, lie, Lys, Met, Phe, Pro, Ser, Thr, Trp, Tyr, or Val, preferably Pro. In another aspect, the variant comprises or consists of the substitution L152D or L152E or L152P or L152* of the polypeptide of SEQ ID NO: 1.

[0147] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 153. In another aspect, the amino acid at a position corresponding to position 153 is substituted with Ala, Arg, Asp, Cys, Gin, Glu, Gly, His, lie, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp, Tyr, or Val, preferably Pro. In another aspect, the variant comprises or consists of the substitution N153E of the polypeptide of SEQ ID NO: 1.

[0148] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 155. In another aspect, the amino acid at a position corresponding to position 155 is substituted with Ala, Arg, Asp, Cys, Gin, Glu, Gly, His, lie, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp, Tyr, or Val, preferably Pro. In another aspect, the variant comprises or consists of the substitution N155D or N155E of the polypeptide of SEQ ID NO: 1.

[0149] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 159. In another aspect, the amino acid at a position corresponding to position 159 is substituted with Ala, Arg, Asn, Asp, Cys, Gin, Glu, Gly, His, lie, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp, Tyr, or Val, preferably Pro. In another aspect, the variant comprises or consists of the substitution V159M of the polypeptide of SEQ ID NO: 1.

[0150] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 165. In another aspect, the amino acid at a position corresponding to position 165 is substituted with Ala, Arg, Asn, Asp, Cys, Gin, Glu, Gly, His, lie, Leu, Lys, Met, Pro, Ser, Thr, Trp, Tyr, or Val, preferably Pro. In another aspect, the variant comprises or consists of the substitution F165H of the polypeptide of SEQ ID NO: 1.

[0151] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 168. In another aspect, the amino acid at a position corresponding to position 168 is substituted with Ala, Arg, Asp, Cys, Gin, Glu, Gly, His, lie, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp, Tyr, or Val, preferably Pro. In another aspect, the variant comprises or consists of the substitution N168R of the polypeptide of SEQ ID NO: 1.

[0152] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 169. In another aspect, the amino acid at a position corresponding to position 169 is substituted with Ala, Asn, Asp, Cys, Gin, Glu, Gly, His, lie, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp, Tyr, or Val, preferably Pro. In another aspect, the variant comprises or consists of the substitution K169E or K169R of the polypeptide of SEQ ID NO: 1.

[0153] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 177. In another aspect, the amino acid at a position corresponding to position 177 is substituted with Arg, Asn, Asp, Cys, Gin, Glu, Gly, His, lie, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp, Tyr, or Val, preferably Pro. In another aspect, the variant comprises or consists of the substitution A177G of the polypeptide of SEQ ID NO: 1.

[0154] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 184. In another aspect, the amino acid at a position corresponding to position 184 is substituted with Ala, Arg, Asn, Asp, Cys, Gin, Glu, Gly, His, lie, Lys, Met, Phe, Pro, Ser, Thr, Trp, Tyr, or Val, preferably Pro. In another aspect, the variant comprises or consists of the substitution L184M of the polypeptide of SEQ ID NO: 1.

[0155] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 189. In another aspect, the amino acid at a position corresponding to position 189 is substituted with Arg, Asn, Asp, Cys, Gin, Glu, Gly, His, lie, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp, Tyr, or Val, preferably Pro. In another aspect, the variant comprises or consists of the substitution A189G of the polypeptide of SEQ ID NO: 1.

[0156] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 203. In another aspect, the amino acid at a position corresponding to position 203 is substituted with Ala, Arg, Asn, Asp, Cys, Gin, Glu, Gly, His, lie, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp, Tyr, or Val, preferably Pro. In another aspect, the variant comprises or consists of the substitution V203T of the polypeptide of SEQ ID NO: 1.

[0157] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 206. In another aspect, the amino acid at a position corresponding to position 206 is substituted with Ala, Asn, Asp, Cys, Gin, Glu, Gly, His, lie, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp, Tyr, or Val, preferably Pro. In another aspect, the variant comprises or consists of the substitution K206E or K206R of the polypeptide of SEQ ID NO: 1.

[0158] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 210. In another aspect, the amino acid at a position corresponding to position 210 is substituted with Ala, Arg, Asn, Asp, Cys, Gin, Glu, Gly, His, lie, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp, Tyr, or Val, preferably Pro. In another aspect, the variant comprises or consists of the substitution D210H or D210R of the polypeptide of SEQ ID NO: 1.

[0159] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 211. In another aspect, the amino acid at a position corresponding to position 211 is substituted with Ala, Asn, Asp, Cys, Gin, Glu, Gly, His, lie, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp, Tyr, or Val, preferably Pro. In another aspect, the variant comprises or consists of the substitution R211K of the polypeptide of SEQ ID NO: 1.

[0160] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 214. In another aspect, the amino acid at a position corresponding to position 214 is substituted with Ala, Arg, Asn, Asp, Cys, Gin, Glu, Gly, His, lie, Leu, Lys, Met, Phe, Pro, Thr, Trp, Tyr, or Val, preferably Pro. In another aspect, the variant comprises or consists of the substitution S214Q of the polypeptide of SEQ ID NO: 1.

[0161] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 217. In another aspect, the amino acid at a position corresponding to position 217 is substituted with Ala, Asn, Asp, Cys, Gin, Glu, Gly, His, lie, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp, Tyr, or Val, preferably Pro. In another aspect, the variant comprises or consists of the substitution K217R or K217T of the polypeptide of SEQ ID NO: 1.

[0162] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 219. In another aspect, the amino acid at a position corresponding to position 219 is substituted with Ala, Arg, Asn, Asp, Cys, Gin, Glu, Gly, His, lie, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp, Tyr, or Val, preferably Pro. In another aspect, the variant comprises or consists of the substitution V219A or V219T of the polypeptide of SEQ ID NO: 1.

[0163] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 220. In another aspect, the amino acid at a position corresponding to position 220 is substituted with Ala, Asn, Asp, Cys, Gin, Glu, Gly, His, lie, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp, Tyr, or Val, preferably Pro. In another aspect, the variant comprises or consists of the substitution K220R of the polypeptide of SEQ ID NO: 1.

[0164] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 226. In another aspect, the amino acid at a position corresponding to position 226 is substituted with Ala, Arg, Asn, Asp, Cys, Gin, Glu, Gly, His, lie, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp, Tyr, or Val, preferably Pro. In another aspect, the variant comprises or consists of the substitution A226D or A226K of the polypeptide of SEQ ID NO: 1.

[0165] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 237. In another aspect, the amino acid at a position corresponding to position 237 is substituted with Ala, Arg, Asn, Asp, Cys, Gin, Glu, His, lie, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp, Tyr, or Val, preferably Pro. In another aspect, the variant comprises or consists of the substitution G237M of the polypeptide of SEQ ID NO: 1.

[0166] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 238. In another aspect, the amino acid at a position corresponding to position 238 is substituted with Ala, Arg, Asn, Asp, Cys, Gin, Glu, Gly, His, lie, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp, Tyr, or Val, preferably Pro. In another aspect, the variant comprises or consists of the substitution A238S or A238T of the polypeptide of SEQ ID NO: 1.

[0167] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 240. In another aspect, the amino acid at a position corresponding to position 240 is substituted with Ala, Asn, Asp, Cys, Gin, Glu, Gly, His, lie, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp, Tyr, or Val, preferably Pro. In another aspect, the variant comprises or consists of the substitution K240L or K240F of the polypeptide of SEQ ID NO: 1.

[0168] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 243. In another aspect, the amino acid at a position corresponding to position 243 is substituted with Ala, Arg, Asn, Asp, Cys, Glu, Gly, His, lie, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp, Tyr, or Val, preferably Pro. In another aspect, the variant comprises or consists of the substitution Q243E of the polypeptide of SEQ ID NO: 1.

[0169] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 244. In another aspect, the amino acid at a position corresponding to position 244 is substituted with Ala, Arg, Asn, Asp, Cys, Gin, Glu, Gly, His, lie, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp, Tyr, or Val, preferably Pro. In another aspect, the variant comprises or consists of the substitution T244E or T244R of the polypeptide of SEQ ID NO: 1.

[0170] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 248. In another aspect, the amino acid at a position corresponding to position 248 is substituted with Ala, Arg, Asn, Asp, Cys, Gin, Glu, Gly, His, lie, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp, Tyr, or Val, preferably Pro. In another aspect, the variant comprises or consists of the substitution W248V of the polypeptide of SEQ ID NO: 1.

[0171] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 251. In another aspect, the amino acid at a position corresponding to position 251 is substituted with Ala, Arg, Asn, Asp, Cys, Gin, Glu, Gly, His, lie, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp, Tyr, or Val, preferably Pro. In another aspect, the variant comprises or consists of the substitution V251E of the polypeptide of SEQ ID NO: 1.

[0172] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 252. In another aspect, the amino acid at a position corresponding to position 252 is substituted with Ala, Asn, Asp, Cys, Gin, Glu, Gly, His, lie, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp, Tyr, or Val, preferably Pro. In another aspect, the variant comprises or consists of the substitution K252E of the polypeptide of SEQ ID NO: 1.

[0173] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 256. In another aspect, the amino acid at a position corresponding to position 256 is substituted with Ala, Arg, Asn, Asp, Cys, Gin, Glu, Gly, His, lie, Leu, Lys, Met, Phe, Pro, Thr, Trp, Tyr, or Val, preferably Pro. In another aspect, the variant comprises or consists of the substitution S256E or S256Q of the polypeptide of SEQ ID NO: 1.

[0174] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 267. In another aspect, the amino acid at a position corresponding to position 267 is substituted with Ala, Asn, Asp, Cys, Gin, Glu, Gly, His, lie, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp, Tyr, or Val, preferably Pro. In another aspect, the variant comprises or consists of the substitution R267C or R267H or R267K of the polypeptide of SEQ ID NO: 1.

[0175] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 271. In another aspect, the amino acid at a position corresponding to position 271 is substituted with Ala, Arg, Asn, Asp, Cys, Glu, Gly, His, lie, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp, Tyr, or Val, preferably Pro. In another aspect, the variant comprises or consists of the substitution Q271D or Q271E of the polypeptide of SEQ ID NO: 1.

[0176] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 276. In another aspect, the amino acid at a position corresponding to position 276 is substituted with Ala, Asn, Asp, Cys, Gin, Glu, Gly, His, lie, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp, Tyr, or Val, preferably Pro. In another aspect, the variant comprises or consists of the substitution R276K of the polypeptide of SEQ ID NO: 1.

[0177] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 289. In another aspect, the amino acid at a position corresponding to position 289 is substituted with Arg, Asn, Asp, Cys, Gin, Glu, Gly, His, lie, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp, Tyr, or Val, preferably Pro. In another aspect, the variant comprises or consists of the substitution A289T of the polypeptide of SEQ ID NO: 1.

[0178] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 294. In another aspect, the amino acid at a position corresponding to position 294 is substituted with Ala, Arg, Asn, Asp, Cys, Gin, Glu, Gly, His, lie, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp, Tyr, or Val, preferably Pro. In another aspect, the variant comprises or consists of the substitution I294E or I294Q of the polypeptide of SEQ ID NO: 1.

[0179] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 295. In another aspect, the amino acid at a position corresponding to position 295 is substituted with Ala, Asn, Asp, Cys, Gin, Glu, Gly, His, lie, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp, Tyr, or Val, preferably Pro. In another aspect, the variant comprises or consists of the substitution R295K of the polypeptide of SEQ ID NO: 1.

[0180] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 298. In another aspect, the amino acid at a position corresponding to position 298 is substituted with Ala, Arg, Asp, Cys, Gin, Glu, Gly, His, lie, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp, Tyr, or Val, preferably Pro. In another aspect, the variant comprises or consists of the substitution N298D of the polypeptide of SEQ ID NO: 1.

[0181] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 300. In another aspect, the amino acid at a position corresponding to position 300 is substituted with Ala, Arg, Asn, Asp, Cys, Gin, Glu, Gly, His, lie, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp, Tyr, or Val, preferably Pro. In another aspect, the variant comprises or consists of the substitution V300L of the polypeptide of SEQ ID NO: 1.

[0182] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 302. In another aspect, the amino acid at a position corresponding to position 302 is substituted with Ala, Arg, Asn, Asp, Cys, Gin, Glu, Gly, His, lie, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp, Tyr, or Val, preferably Pro. In another aspect, the variant comprises or consists of the substitution N302H of the polypeptide of SEQ ID NO: 1.

[0183] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 322. In another aspect, the amino acid at a position corresponding to position 322 is substituted with Ala, Asn, Asp, Cys, Gin, Glu, Gly, His, lie, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp, Tyr, or Val, preferably Pro. In another aspect, the variant comprises or consists of the substitution K322E of the polypeptide of SEQ ID NO: 1.

[0184] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 329. In another aspect, the amino acid at a position corresponding to position 329 is substituted with Ala, Arg, Asn, Asp, Cys, Glu, Gly, His, lie, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp, Tyr, or Val, preferably Pro. In another aspect, the variant comprises or consists of the substitution Q329E of the polypeptide of SEQ ID NO: 1.

[0185] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 339. In another aspect, the amino acid at a position corresponding to position 339 is substituted with Ala, Arg, Asn, Asp, Cys, Gin, Glu, Gly, His, lie, Leu, Lys, Met, Phe, Ser, Thr, Trp, Tyr, or Val, preferably Pro. In another aspect, the variant comprises or consists of the substitution P339S of the polypeptide of SEQ ID NO: 1.

[0186] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 347. In another aspect, the amino acid at a position corresponding to position 347 is substituted with Ala, Asn, Asp, Cys, Gin, Glu, Gly, His, lie, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp, Tyr, or Val, preferably Pro. In another aspect, the variant comprises or consists of the substitution K347E or K347R of the polypeptide of SEQ ID NO: 1.

[0187] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 353. In another aspect, the amino acid at a position corresponding to position 353 is substituted with Ala, Arg, Asn, Asp, Cys, Gin, Glu, Gly, His, lie, Leu, Met, Phe, Pro, Ser, Thr, Trp, Tyr, or Val, preferably Pro. In another aspect, the variant comprises or consists of the substitution K353R of the polypeptide of SEQ ID NO: 1.

[0188] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 383. In another aspect, the amino acid at a position corresponding to position 383 is substituted with Ala, Arg, Asn, Asp, Cys, Gin, Glu, Gly, His, lie, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp, Tyr, or Val, preferably Pro. In another aspect, the variant comprises or consists of the substitution N383E or N383Q of the polypeptide of SEQ ID NO: 1.

[0189] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 384. In another aspect, the amino acid at a position corresponding to position 384 is substituted with Ala, Arg, Asn, Cys, Gin, Glu, Gly, His, lie, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp, Tyr, or Val, preferably Pro. In another aspect, the variant comprises or consists of the substitution D384G of the polypeptide of SEQ ID NO: 1.

[0190] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 392. In another aspect, the amino acid at a position corresponding to position 392 is substituted with Ala, Arg, Asn, Asp, Cys, Gin, Glu, Gly, His, lie, Leu, Met, Phe, Pro, Ser, Thr, Trp, Tyr, or Val, preferably Pro. In another aspect, the variant comprises or consists of the substitution K392E of the polypeptide of SEQ ID NO: 1.

[0191] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 394. In another aspect, the amino acid at a position corresponding to position 394 is substituted with Ala, Arg, Asn, Asp, Cys, Gin, Glu, Gly, His, lie, Leu, Met, Phe, Pro, Ser, Thr, Trp, Tyr, or Val, preferably Pro. In another aspect, the variant comprises or consists of the substitution K394R of the polypeptide of SEQ ID NO: 1.

[0192] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 395. In another aspect, the amino acid at a position corresponding to position 395 is substituted with Ala, Arg, Asn, Cys, Gin, Glu, Gly, His, lie, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp, Tyr, or Val, preferably Pro. In another aspect, the variant comprises or consists of the substitution D395P of the polypeptide of SEQ ID NO: 1.

[0193] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 402. In another aspect, the amino acid at a position corresponding to position 402 is substituted with Ala, Arg, Asn, Asp, Cys, Gin, Glu, Gly, His, lie, Leu, Lys, Met, Phe, Pro, Thr, Trp, Tyr, or Val, preferably Pro. In another aspect, the variant comprises or consists of the substitution S402Q of the polypeptide of SEQ ID NO: 1.

[0194] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 414. In another aspect, the amino acid at a position corresponding to position 414 is substituted with Ala, Arg, Asn, Asp, Cys, Gin, Glu, Gly, His, lie, Leu, Met, Phe, Pro, Ser, Thr, Trp, Tyr, or Val, preferably Pro. In another aspect, the variant comprises or consists of the substitution K414E of the polypeptide of SEQ ID NO: 1.

[0195] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 427. In another aspect, the amino acid at a position corresponding to position 427 is substituted with Ala, Arg, Asn, Asp, Cys, Gin, Glu, Gly, His, lie, Leu, Lys, Met, Phe, Pro, Ser, Trp, Tyr, or Val, preferably Pro. In another aspect, the variant comprises or consists of the substitution T427V of the polypeptide of SEQ ID NO: 1.

[0196] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 431. In another aspect, the amino acid at a position corresponding to position 431 is substituted with Ala, Arg, Asn, Asp, Cys, Gin, Glu, Gly, His, lie, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp, or Tyr, preferably Pro. In another aspect, the variant comprises or consists of the substitution V431E of the polypeptide of SEQ ID NO: 1.

[0197] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 445. In another aspect, the amino acid at a position corresponding to position 445 is substituted with Ala, Arg, Asn, Asp, Cys, Gin, Glu, Gly, His, lie, Leu, Met, Phe, Pro, Ser, Thr, Trp, Tyr, or Val, preferably Pro. In another aspect, the variant comprises or consists of the substitution K445E of the polypeptide of SEQ ID NO: 1.

[0198] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 447. In another aspect, the amino acid at a position corresponding to position 447 is substituted with Ala, Arg, Asn, Asp, Cys, Gin, Glu, Gly, His, lie, Lys, Met, Phe, Pro, Ser, Thr, Trp, Tyr, or Val, preferably Pro. In another aspect, the variant comprises or consists of the substitution L447M of the polypeptide of SEQ ID NO: 1.

[0199] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 459. In another aspect, the amino acid at a position corresponding to position 459 is substituted with Arg, Asn, Asp, Cys, Gin, Glu, Gly, His, lie, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp, Tyr, or Val, preferably Pro. In another aspect, the variant comprises or consists of the substitution A459P of the polypeptide of SEQ ID NO: 1.

[0200] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 473. In another aspect, the amino acid at a position corresponding to position 473 is substituted with Ala, Arg, Asn, Asp, Cys, Gin, Glu, Gly, His, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp, Tyr, or Val, preferably Pro. In another aspect, the variant comprises or consists of the substitution I473T of the polypeptide of SEQ ID NO: 1.

[0201] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 474. In another aspect, the amino acid at a position corresponding to position 474 is substituted with Ala, Arg, Asn, Asp, Cys, Gin, Glu, Gly, His, lie, Leu, Lys, Met, Phe, Pro, Thr, Trp, Tyr, or Val, preferably Pro. In another aspect, the variant comprises or consists of the substitution S474E of the polypeptide of SEQ ID NO: 1.

[0202] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 476. In another aspect, the amino acid at a position corresponding to position 476 is substituted with Ala, Arg, Asn, Asp, Cys, Gin, Glu, Gly, His, lie, Leu, Met, Phe, Pro, Ser, Thr, Trp, Tyr, or Val, preferably Pro. In another aspect, the variant comprises or consists of the substitution K476R of the polypeptide of SEQ ID NO: 1.

[0203] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 482. In another aspect, the amino acid at a position corresponding to position 482 is substituted with Ala, Arg, Asn, Asp, Cys, Gin, Glu, Gly, His, lie, Leu, Met, Phe, Pro, Ser, Thr, Trp, Tyr, or Val, preferably Pro. In another aspect, the variant comprises or consists of the substitution K482R of the polypeptide of SEQ ID NO: 1.

[0204] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 488. In another aspect, the amino acid at a position corresponding to position 488 is substituted with Ala, Arg, Asn, Asp, Cys, Gin, Glu, Gly, His, lie, Leu, Met, Phe, Pro, Ser, Thr, Trp, Tyr, or Val, preferably Pro. In another aspect, the variant comprises or consists of the substitution K488T of the polypeptide of SEQ ID NO: 1.

[0205] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 489. In another aspect, the amino acid at a position corresponding to position 489 is substituted with Ala, Arg, Asn, Asp, Cys, Gin, Gly, His, lie, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp, Tyr, or Val, preferably Pro. In another aspect, the variant comprises or consists of the substitution E489K or E489R of the polypeptide of SEQ ID NO: 1.

[0206] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 491. In another aspect, the amino acid at a position corresponding to position 491 is substituted with Arg, Asn, Asp, Cys, Gin, Glu, Gly, His, lie, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp, Tyr, or Val, preferably Pro. In another aspect, the variant comprises or consists of the substitution A491E or A491V of the polypeptide of SEQ ID NO: 1.

[0207] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 492. In another aspect, the amino acid at a position corresponding to position 492 is substituted with Ala, Arg, Asn, Asp, Cys, Gin, Glu, Gly, His, lie, Leu, Lys, Met, Phe, Ser, Thr, Trp, Tyr, or Val, preferably Pro. In another aspect, the variant comprises or consists of the substitution P492D of the polypeptide of SEQ ID NO: 1.

[0208] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 503. In another aspect, the amino acid at a position corresponding to position 503 is substituted with Ala, Arg, Asn, Asp, Cys, Gin, Glu, Gly, His, lie, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp, or Val, preferably Pro. In another aspect, the variant comprises or consists of the substitution Y503L or Y503V of the polypeptide of SEQ ID NO: 1.

[0209] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 505. In another aspect, the amino acid at a position corresponding to position 505 is substituted with Ala, Arg, Asn, Asp, Cys, Gin, Glu, Gly, His, lie, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp, or Tyr, preferably Pro. In another aspect, the variant comprises or consists of the substitution V505L of the polypeptide of SEQ ID NO: 1.

[0210] In one aspect, the variant comprises an alteration at a position corresponding to a position selected from the group consisting of: E126P, S127H, T104G, Q125K, Q125P, Q125S, D395P, G103V, T104R, Q125L, A41E, A41R, Q125F, S127L, A226K, A41L, A226D, K394R, S127D, R211K, S123P, K488T, S256Q, K476R, K217R, Q271D, S214Q, L447M, K482R, K169R, L152D, R267K, L152E, D210R, L152*, R295K, N155D, Q137K, N155E, Q147K, R276K, V203T, S94R, K18E, K252E, V219T, Q243E, K414E, K445E, V159M, K392E, Q82E, S76E, A83E, Q271E, S256E, I294E, Q329E, V431E of the polypeptide of SEQ ID NO: 1, wherein the variant has xylogucanase activity and wherein the variant has at least 60%, e.g., 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%, e.g., at least 96%, at least 97%, at least 98%, or at least 99%, sequence identity, but less than 100%, sequence identity, to the polypeptide of SEQ ID NO: 1, SEQ ID NO: 2, or SEQ ID NO: 3.

[0211] In one aspect, the variant comprises an alteration at a position corresponding to a position selected from the group consisting of: E126P, S127H, T104G, Q125K, Q125P, Q125S, K129T, K129A, D395P, G103V, T104R, Q125L, A41E, A41R, Q125F, S127L, A226K, A226D, K394R, S127D, R211K, S123P, K488T, S256Q, K476R, K217R, Q271D, S214Q, L447M, K482R, K169R, L152D, R267K, L152E, D210R, L152*, R295K, N155D, Q137K, N155E, Q147K, R276K, V203T, S94R, K18E, K252E, V219T, Q243E, K414E, K445E, V159M, K392E, Q82E, S76E, A83E, Q271E, S256E, I294E, Q329E, V431E, of the polypeptide of SEQ ID NO: 1, wherein the variant has xylogucanase activity and wherein the variant has at least 60%, e.g., 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%, e.g., at least 96%, at least 97%, at least 98%, or at least 99%, sequence identity, but less than 100%, sequence identity, to the polypeptide of SEQ ID NO: 1, SEQ ID NO: 2, or SEQ ID NO: 3.

[0212] In one aspect, the variant comprises an alteration at a position corresponding to a position selected from the group consisting of: E126P, S127H, T104G, Q125K, Q125P, Q125S, A129T, D395P, G103V, T104R, Q125L, A41E, A41R, Q125F, S127L, A226K, A41L, A226D, K394R, S127D, R211K, S123P, K488T, S256Q, K476R, K217R, Q271D, S214Q, L447M, K482R, K169R, L152D, R267K, L152E, D210R, L152*, R295K, N155D, Q137K, N155E, Q147K, R276K, V203T, S94R, K18E, K252E, V219T, Q243E, K414E, K445E, V159M, K392E, Q82E, S76E, A83E, Q271E, S256E, I294E, Q329E, V431E of the polypeptide of SEQ ID NO: 2, wherein the variant has xylogucanase activity and wherein the variant has at least 60%, e.g., 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%, e.g., at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to the polypeptide of SEQ ID NO: 1, SEQ ID NO: 2, or SEQ ID NO: 3 but less than 100% sequence identity.

[0213] In one aspect, the variant comprises an alteration at a position corresponding to a position selected from the group consisting of: A118K+S123P, R267C+T427V, R20K+S123P, S123P+K206R, S123P+K347R, S123P+D395P, S123P+S127D, E489R+P492D, Y503L+V505L, Y503V+V505L, L184M+V219A of the polypeptide of SEQ ID NO: 1, wherein the variant has xylogucanase activity and wherein the variant has at least 60%, e.g., 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%, e.g., at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to the polypeptide of SEQ ID NO: 1, SEQ ID NO: 2, or SEQ ID NO: 3 but less than 100% sequence identity.

[0214] In an aspect, the variant comprises alterations at positions corresponding to positions selected from the group consisting of: A118K+S123P+K129T, R267C+T427V+K129T, R20K+S123P+K129T, S123P+K206R+K129T, S123P+K347R+K129T, S123P+D395P+K129T, S123P+S127D+K129T, E489R+P492D+K129T, Y503L+V505L+K129T, Y503V+V505L+K129T, L184M+V219A+K129T of the polypeptide of SEQ ID NO: 1, wherein the variant has xylogucanase activity and wherein the variant has at least 60%, e.g., 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%, e.g., at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to the polypeptide of SEQ ID NO: 1, SEQ ID NO: 2, or SEQ ID NO: 3 but less than 100% sequence identity.

[0215] In an aspect, the variant comprises alterations at positions corresponding to positions selected from the group consisting of: A118K+S123P+A129T, R267C+T427V+A129T, R20K+S123P+A129T, S123P+K206R+A129T, S123P+K347R+A129T, S123P+D395P+A129T, S123P+S127D+A129T, E489R+P492D+A129T, Y503L+V505L+A129T, Y503V+V505L+A129T, L184M+V219A+A129T of the polypeptide of SEQ ID NO: 2, wherein the variant has xylogucanase activity and wherein the variant has at least 60%, e.g., 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%, e.g., at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to the polypeptide of SEQ ID NO: 1, SEQ ID NO: 2, or SEQ ID NO: 3 but less than 100% sequence identity.

[0216] In an aspect, the variant comprises alterations at positions corresponding to positions selected from the group consisting of: R20K+S123P+R211K, R20K+S123P+K220R, P111Q+S123P+V159M, K8E+P111Q+V159M, S94R+P111Q+V159M, P111Q+Q137K+V159M, P111Q+Q147K+V159M of the polypeptide of SEQ ID NO: 1, wherein the variant has xylogucanase activity and wherein the variant has at least 60%, e.g., 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%, e.g., at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to the polypeptide of SEQ ID NO: 1, SEQ ID NO: 2, or SEQ ID NO: 3 but less than 100% sequence identity.

[0217] In an aspect, the variant comprises alterations at positions corresponding to positions selected from the group consisting of: R20K+S123P+R211K+K129T, R20K+S123P+K220R+K129T, P111Q+S123P+V159M+K129T, K8E+P111Q+V159M+K129T, S94R+P111Q+V159M+K129T, P111Q+Q137K+V159M+K129T, P111Q+Q147K+V159M+K129T of the polypeptide of SEQ ID NO: 1, wherein the variant has xylogucanase activity and wherein the variant has at least 60%, e.g., 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%, e.g., at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to the polypeptide of SEQ ID NO: 1, SEQ ID NO: 2, or SEQ ID NO: 3 but less than 100% sequence identity.

[0218] In an aspect, the variant comprises alterations at positions corresponding to positions selected from the group consisting of: R20K+S123P+R211K+A129T, R20K+S123P+K220R+A129T, P111Q+S123P+V159M+A129T, K8E+P111Q+V159M+A129T, S94R+P111Q+V159M+A129T, P111Q+Q137K+V159M+A129T, P111Q+Q147K+V159M+A129T of the polypeptide of SEQ ID NO: 2, wherein the variant has xylogucanase activity and wherein the variant has at least 60%, e.g., 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%, e.g., at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to the polypeptide of SEQ ID NO: 1, SEQ ID NO: 2, or SEQ ID NO: 3 but less than 100% sequence identity.

[0219] In an aspect, the variant comprises alterations at positions corresponding to positions selected from the group consisting of: S123P+K347R+K353R+D395P, R20K+Y105E+S123P+R267K, R20K+Y105E+S123P+N136D, R20K+S123P+Q137K+Q147K, S94R+P111Q+S123P+V159M, K87E+P111Q+S123P+V159M, P111Q+S123P+V159M+S402Q, P111Q+Q147K+V159M+K220R, P111Q+V159M+K206E+I294Q, K8E+P111Q+N155D+V159M, P111Q+Q137K+Q147K+V159M, R20K+S123P+K220R+I294E, R20K+S123P+N155D+K220R, R20K+S123P+V203T+V219 of the polypeptide of SEQ ID NO: 1, wherein the variant has xylogucanase activity and wherein the variant has at least 60%, e.g., 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%, e.g., at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to the polypeptide of SEQ ID NO: 1, SEQ ID NO: 2, or SEQ ID NO: 3 but less than 100% sequence identity.

[0220] In an aspect, the variant comprises alterations at positions corresponding to positions selected from the group consisting of: S123P+K347R+K353R+D395P+K129T of the polypeptide of SEQ ID NO: 1, R20K+Y105E+S123P+R267K+K129T, R20K+Y105E+S123P+N136D+K129T, R20K+S123P+Q137K+Q147K+K129T, S94R+P111Q+S123P+V159M+K129T, K87E+P111Q+S123P+V159M+K129T, P111Q+S123P+V159M+S402Q+K129T, P111Q+Q147K+V159M+K220R+K129T, P111Q+V159M+K206E+I294Q+K129T, K8E+P111Q+N155D+V159M+K129T, P111Q+Q137K+Q147K+V159M+K129T, R20K+S123P+K220R+I294E+K129T, R20K+S123P+N155D+K220R+K129T, R20K+S123P+V203T+V219T+K129T, wherein the variant has xylogucanase activity and wherein the variant has at least 60%, e.g., 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%, e.g., at least 96%, at least 97%, at least 98%, or at least 99%, but less than 100%, sequence identity to the polypeptide of SEQ ID NO: 1, SEQ ID NO: 2, or SEQ ID NO: 3.

[0221] In one aspect, the variant comprises alterations at positions corresponding to positions selected from the group consisting of: S123P+K347R+K353R+D395P+A129T, R20K+Y105E+S123P+R267K+A129T, R20K+Y105E+S123P+N136D+A129T, R20K+S123P+Q137K+Q147K+A129T, S94R+P111Q+S123P+V159M+A129T, K87E+P111Q+S123P+V159M+A129T, P111Q+S123P+V159M+S402Q+A129T, P111Q+Q147K+V159M+K220R+A129T, P111Q+V159M+K206E+I294Q+A129T, K8E+P111Q+N155D+V159M+A129T, P111Q+Q137K+Q147K+V159M+A129T, R20K+S123P+K220R+I294E+A129T, R20K+S123P+N155D+K220R+A129T, R20K+S123P+V203T+V219T+A129T of the polypeptide of SEQ ID NO: 2, wherein the variant has xylogucanase activity and wherein the variant has at least 60%, e.g., 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%, e.g., at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to the polypeptide of SEQ ID NO: 1, SEQ ID NO: 2, or SEQ ID NO: 3, but less than 100% sequence identity.

[0222] In an aspect, the variant comprises alterations at positions corresponding to positions selected from the group consisting of: S123P+R211K+K217R+S256Q+K488T, R20K+Y105E+S123P+Q147K+R267K, K87E+P111Q+S123P+V159M+S402Q, K87E+P111Q+V159M+I294Q+I473T, K8E+K18E+P111Q+V159M+K206E, R20K+P111Q+S123P+S127D+V159M, P111Q+V159M+K206E+I294Q+K347E, P111Q+Q137K+Q147K+V159M+K252E, P111Q+Q137K+Q147K+N155D+K252E, P111Q+Q137K+L152D+V203T+K217R, P111Q+S123P+Q137K+V159M+K488T, P111Q+S123P+Q137K+V159M+S256Q, K87E+P111Q+Q147K+L152D+V159M, R20K+A83E+S123P+K220R+S256E, R20K+A83E+S123P+K220R+K252E, R20K+Q82E+S123P+Q147K+K220R, R20K+A83E+S123P+K220R+S256Q, R20K+Q82E+S123P+N155D+K220R, R20K+S123P+V203T+K220R+K252E, R20K+A41L+Q82E+S123P+K220R, R20K+A42V+S76E+S123P+K220R, R20K+A42V+A83E+S123P+K220R, A83E+P111Q+S123P+V159M+K252E, of the polypeptide of SEQ ID NO: 1, wherein the variant has xylogucanase activity and wherein the variant has at least 60%, e.g., 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%, e.g., at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to the polypeptide of SEQ ID NO: 1, SEQ ID NO: 2, or SEQ ID NO: 3, but less than 100% sequence identity.

[0223] In one aspect, the variant comprises alterations at positions corresponding to positions selected from the group consisting of: S123P+R211K+K217R+S256Q+K488T+K129T of the polypeptide of SEQ ID NO: 1, R20K+Y105E+S123P+Q147K+R267K+K129T, K87E+P111Q+S123P+V159M+S402Q+K129T, K87E+P111Q+V159M+I294Q+I473T+K129T, K8E+K18E+P111Q+V159M+K206E+K129T, R20K+P111Q+S123P+S127D+V159M+K129T, P111Q+V159M+K206E+I294Q+K347E+K129T, P111Q+Q137K+Q147K+V159M+K252E+K129T, P111Q+Q137K+Q147K+N155D+K252E+K129T, P111Q+Q137K+L152D+V203T+K217R+K129T, P111Q+S123P+Q137K+V159M+K488T+K129T, P111Q+S123P+Q137K+V159M+S256Q+K129T, K87E+P111Q+Q147K+L152D+V159M+K129T, R20K+A83E+S123P+K220R+S256E+K129T, R20K+A83E+S123P+K220R+K252E+K129T, R20K+Q82E+S123P+Q147K+K220R+K129T, R20K+A83E+S123P+K220R+S256Q+K129T, R20K+Q82E+S123P+N155D+K220R+K129T, R20K+S123P+V203T+K220R+K252E+K129T, R20K+A41L+Q82E+S123P+K220R+K129T, R20K+A42V+S76E+S123P+K220R+K129T, R20K+A42V+A83E+S123P+K220R+K129T, A83E+P111Q+S123P+V159M+K252E+K129T, where the variant has xyloglucanase activity and where the variant has at least 60%, e.g., 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%, at least 99%, or 100% identity to the polypeptide of SEQ ID NO: 1, SEQ ID NO: 2, or SEQ ID NO: 3.For example, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity but less than 100% sequence identity.

[0224] In one aspect, the variant comprises alterations at positions corresponding to positions selected from the group consisting of: S123P+R211K+K217R+S256Q+K488T+A129T of the polypeptide of SEQ ID NO: 2, R20K+Y105E+S123P+Q147K+R267K+A129T, K87E+P111Q+S123P+V159M+S402Q+A129T, K87E+P111Q+V159M+I294Q+I473T+A129T, K8E+K18E+P111Q+V159M+K206E+A129T, R20K+P111Q+S123P+S127D+V159M+A129T, P111Q+V159M+K206E+I294Q+K347E+A129T, P111Q+Q137K+Q147K+V159M+K252E+A129T, P111Q+Q137K+Q147K+N155D+K252E+A129T, P111Q+Q137K+L152D+V203T+K217R+A129T, P111Q+S123P+Q137K+V159M+K488T+A129T, P111Q+S123P+Q137K+V159M+S256Q+A129T, K87E+P111Q+Q147K+L152D+V159M+A129T, R20K+A83E+S123P+K220R+S256E+A129T, R20K+A83E+S123P+K220R+K252E+A129T, R20K+Q82E+S123P+Q147K+K220R+A129T, R20K+A83E+S123P+K220R+S256Q+A129T, R20K+Q82E+S123P+N155D+K220R+A129T, R20K+S123P+V203T+K220R+K252E+A129T, R20K+A41L+Q82E+S123P+K220R+A129T, R20K+A42V+S76E+S123P+K220R+A129T, R20K+A42V+A83E+S123P+K220R+A129T, A83E+P111Q+S123P+V159M+K252E+A129T, wherein the variant has xyloglucanase activity and wherein the variant has at least 60%, e.g., 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%, at least 99%, or 100% identity to the polypeptide of SEQ ID NO: 1, SEQ ID NO: 2, or SEQ ID NO: 3.at least 97%, at least 98%, or at least 99% sequence identity, but less than 100% sequence identity.

[0225] In one aspect, the variant comprises alterations at positions corresponding to positions selected from the group consisting of: Y105E+A118K+S123P+K206R+K220R+R267K, A41L+P111Q+S123P+Q147K+V159M+V203T, A41L+P111Q+S123P+Q147K+V159M+K217R, P111Q+S123P+Q147K+V159M+I294Q+S402Q, P111Q+Q137K+Q147K+V159M+V203T+K217R, K87E+P111Q+L152D+V159M+V203T+I294Q, R20K+S76E+A83E+S123P+K220R+K252E, R20K+A42V+S123P+K220R+K252E+I294E, R20K+A83E+S123P+V203T+K220R+K252E, R20K+A42V+S76E+S123P+V203T+K220R, A83E+P111Q+S123P+V159M+S256E+I294E, Q82E+P111Q+S123P+V159M+S256E+I294E, Q82E+P111Q+S123P+Q147K+V159M+I294E, K87E+P111Q+S123P+V159M+K217T+I294E, A83E+P111Q+S123P+V159M+K240F+K252E, K87E+P111Q+S123P+V159M+S256Q+I294E, K87E+P111Q+S123P+V159M+K347E+N383E, Q82E+P111Q+S123P+N155D+S256E+I294E, of the polypeptide of SEQ ID NO: 1, wherein the variant has xylogucanase activity and wherein the variant has at least 60%, e.g., 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%, e.g., at least 96%, at least 97%, at least 98%, or at least 99% sequence identity, but less than 100% sequence identity, to the polypeptide of SEQ ID NO: 1, SEQ ID NO: 2, or SEQ ID NO: 3.

[0226] In an aspect, the variant comprises alterations at positions corresponding to positions selected from the group consisting of: Y105E+A118K+S123P+K206R+K220R+R267K+K129T, A41L+P111Q+S123P+Q147K+V159M+V203T+K129T, A41L+P111Q+S123P+Q147K+V159M+K217R+K129T, P111Q+S123P+Q147K+V159M+I294Q+S402Q+K129T, P111Q+Q137K+Q147K+V159M+V203T+K217R+K129T, K87E+P111Q+L152D+V159M+V203T+I294Q+K129T, R20K+S76E+A83E+S123P+K220R+K252E+K129T, R20K+A42V+S123P+K220R+K252E+I294E+K129T, R20K+A83E+S123P+V203T+K220R+K252E+K129T, R20K+A42V+S76E+S123P+V203T+K220R+K129T, A83E+P111Q+S123P+V159M+S256E+I294E+K129T, Q82E+P111Q+S123P+V159M+S256E+I294E+K129T, Q82E+P111Q+S123P+Q147K+V159M+I294E+K129T, K87E+P111Q+S123P+V159M+K217T+I294E+K129T, A83E+P111Q+S123P+V159M+K240F+K252E+K129T, K87E+P111Q+S123P+V159M+S256Q+I294E+K129T, K87E+P111Q+S123P+V159M+K347E+N383E+K129T, Q82E+P111Q+S123P+N155D+S256E+I294E+K129T of the polypeptide of SEQ ID NO: 1, wherein the variant has xylogucanase activity and wherein the variant has at least 60%, e.g., 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%, e.g., at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to the polypeptide of SEQ ID NO: 1, SEQ ID NO: 2, or SEQ ID NO: 3, but less than 100% sequence identity.

[0227] In an aspect, the variant comprises alterations at positions corresponding to positions selected from the group consisting of: Y105E+A118K+S123P+K206R+K220R+R267K+A129T of the polypeptide of SEQ ID NO: 2, A41L+P111Q+S123P+Q147K+V159M+V203T+A129T, A41L+P111Q+S123P+Q147K+V159M+K217R+A129T, P111Q+S123P+Q147K+V159M+I294Q+S402Q+A129T, P111Q+Q137K+Q147K+V159M+V203T+K217R+A129T, K87E+P111Q+L152D+V159M+V203T+I294Q+A129T, R20K+S76E+A83E+S123P+K220R+K252E+A129T, R20K+A42V+S123P+K220R+K252E+I294E+A129T, R20K+A83E+S123P+V203T+K220R+K252E+A129T, R20K+A42V+S76E+S123P+V203T+K220R+A129T, A83E+P111Q+S123P+V159M+S256E+I294E+A129T, Q82E+P111Q+S123P+V159M+S256E+I294E+A129T, Q82E+P111Q+S123P+Q147K+V159M+I294E+A129T, K87E+P111Q+S123P+V159M+K217T+I294E+A129T, A83E+P111Q+S123P+V159M+K240F+K252E+A129T, K87E+P111Q+S123P+V159M+S256Q+I294E+A129T, K87E+P111Q+S123P+V159M+K347E+N383E+A129T, Q82E+P111Q+S123P+N155D+S256E+I294E+A129T, wherein the variant has xyloglucanase activity and wherein the variant has at least 60%, e.g., 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%, e.g., at least 96%, at least 97%, at least 98%, or at least 99% sequence identity, but less than 100% sequence identity, to the polypeptide of SEQ ID NO: 1, SEQ ID NO: 2, or SEQ ID NO: 3.

[0228] In an aspect, the variant comprises alterations at positions corresponding to positions selected from the group consisting of: P111Q+S123P+Q137K+Q147K+V159M+S256Q+S402Q, A41L+P111Q+Q137K+V159M+N168R+Q271D+K488T, S76E+Q82E+K87E+P111Q+S123P+V159M+V203T, Q82E+P111Q+S123P+Q147K+V159M+S256E+I294E, K8E+Q82E+P111Q+S123P+V159M+S256E+I294E, Q82E+P111Q+S123P+V159M+K240F+S256E+I294E, Q82E+P111Q+S123P+V159M+G237M+V251E+I294E, of the polypeptide of SEQ ID NO: 1, wherein the variant has xylogucanase activity and wherein the variant has at least 60%, e.g., 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%, e.g., at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to the polypeptide of SEQ ID NO: 1, SEQ ID NO: 2, or SEQ ID NO: 3 but less than 100% sequence identity.

[0229] In an aspect, the variant comprises alterations at positions corresponding to positions selected from the group consisting of: P111Q+S123P+Q137K+Q147K+V159M+S256Q+S402Q+K129T of the polypeptide of SEQ ID NO: 1, A41L+P111Q+Q137K+V159M+N168R+Q271D+K488T+K129T, S76E+Q82E+K87E+P111Q+S123P+V159M+V203T+K129T, Q82E+P111Q+S123P+Q147K+V159M+S256E+I294E+K129T, K8E+Q82E+P111Q+S123P+V159M+S256E+I294E+K129T, Q82E+P111Q+S123P+V159M+K240F+S256E+I294E+K129T, Q82E+P111Q+S123P+V159M+G237M+V251E+I294E+K129T, wherein the variant has xylogucanase activity and wherein the variant has at least 60%, e.g., 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%, e.g., at least 96%, at least 97%, at least 98%, or at least 99%, but less than 100%, sequence identity to the polypeptide of SEQ ID NO: 1, SEQ ID NO: 2, or SEQ ID NO: 3.

[0230] In an aspect, the variant comprises alterations at positions corresponding to positions selected from the group consisting of: P111Q+S123P+Q137K+Q147K+V159M+S256Q+S402Q+A129T of the polypeptide of SEQ ID NO: 2, A41L+P111Q+Q137K+V159M+N168R+Q271D+K488T+A129T, S76E+Q82E+K87E+P111Q+S123P+V159M+V203T+A129T, Q82E+P111Q+S123P+Q147K+V159M+S256E+I294E+A129T, K8E+Q82E+P111Q+S123P+V159M+S256E+I294E+A129T, Q82E+P111Q+S123P+V159M+K240F+S256E+I294E+A129T, Q82E+P111Q+S123P+V159M+G237M+V251E+I294E+A129T, wherein the variant has xylogucanase activity and wherein the variant has at least 60%, e.g., 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%, e.g., at least 96%, at least 97%, at least 98%, or at least 99%, sequence identity to the polypeptide of SEQ ID NO: 1, SEQ ID NO: 2, or SEQ ID NO: 3, but less than 100% sequence identity.

[0231] In an aspect, the variant comprises alterations at positions corresponding to positions selected from the group consisting of: A83E+P111Q+S123P+Q137K+Q147K+V159M+S256Q+S402Q, P111Q+S123P+Q137K+Q147K+N155D+S256Q+A289T+N302H, P111Q+S123P+Q137K+Q147K+V159M+K252E+S256Q+S402Q, P111Q+S123P+Q137K+Q147K+N155D+S256Q+I294E+S402Q, S76E+P111Q+S123P+Q137K+Q147K+V159M+S256Q+S402Q, K8R+P111Q+S123P+Q137K+Q147K+V159M+S256Q+S402Q, S123P+S127D+N136D+Q137K+Q147K+L152E+N153E+N155E, R20K+S123P+K169R+K217T+K240F+S256Q+R267H+I294E, Q82E+P111Q+S123P+V159M+V203T+G237M+S256E+I294E, Q82E+P111Q+S123P+N155D+K169R+G237M+S256E+I294E, Q82E+P111Q+S123P+Q137K+V159M+G237M+S256E+I294E, Q82E+P111Q+S123P+Q137K+V159M+K240F+S256E+I294E, Q82E+P111Q+S123P+Q147K+N155D+K240F+S256E+I294E, A83E+P111Q+S123P+Q147K+V159M+S256E+I294E+Q329E, Q82E+P111Q+S123P+V159M+K169R+S256E+I294E+V431E, A41L+Q82E+P111Q+S123P+V159M+S256E+I294E+N383E of the polypeptide of SEQ ID NO: 1, wherein the variant has xylogucanase activity and wherein the variant has at least 60%, e.g., 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%, e.g., at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to the polypeptide of SEQ ID NO: 1, SEQ ID NO: 2, or SEQ ID NO: 3, but less than 100% sequence identity.

[0232] In one aspect, the variant comprises alterations at positions corresponding to positions selected from the group consisting of: A83E+P111Q+S123P+Q137K+Q147K+V159M+S256Q+S402Q+K129T, P111Q+S123P+Q137K+Q147K+N155D+S256Q+A289T+N302H+K129T, P111Q+S123P+Q137K+Q147K+V159M+K252E+S256Q+S402Q+K129T, P111Q+S123P+Q137K+Q147K+N155D+S256Q+I294E+S402Q+K129T, S76E+P111Q+S123P+Q137K+Q147K+V159M+S256Q+S402Q+K129T, K8R+P111Q+S123P+Q137K+Q147K+V159M+S256Q+S402Q+K129T, S123P+S127D+N136D+Q137K+Q147K+L152E+N153E+N155E+K129T, R20K+S123P+K169R+K217T+K240F+S256Q+R267H+I294E+K129T, Q82E+P111Q+S123P+V159M+V203T+G237M+S256E+I294E+K129T, Q82E+P111Q+S123P+N155D+K169R+G237M+S256E+I294E+K129T, Q82E+P111Q+S123P+Q137K+V159M+G237M+S256E+I294E, Q82E+P111Q+S123P+Q137K+V159M+K240F+S256E+I294E+K129T, Q82E+P111Q+S123P+Q147K+N155D+K240F+S256E+I294E+K129T, A83E+P111Q+S123P+Q147K+V159M+S256E+I294E+Q329E+K129T, Q82E+P111Q+S123P+V159M+K169R+S256E+I294E+V431E+K129T, A41L+Q82E+P111Q+S123P+V159M+S256E+I294E+N383E+K129T, of the polypeptide of SEQ ID NO: 1, wherein the variant has xylogucanase activity and wherein the variant has at least 60%,For example, 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%, for example, at least 96%, at least 97%, at least 98%, or at least 99%, but less than 100% sequence identity.

[0233] In one aspect, the variant comprises alterations at positions corresponding to positions selected from the group consisting of: A83E+P111Q+S123P+Q137K+Q147K+V159M+S256Q+S402Q+A129T, P111Q+S123P+Q137K+Q147K+N155D+S256Q+A289T+N302H+A129T, P111Q+S123P+Q137K+Q147K+V159M+K252E+S256Q+S402Q+A129T, P111Q+S123P+Q137K+Q147K+N155D+S256Q+I294E+S402Q+A129T, S76E+P111Q+S123P+Q137K+Q147K+V159M+S256Q+S402Q+A129T, K8R+P111Q+S123P+Q137K+Q147K+V159M+S256Q+S402Q+A129T, S123P+S127D+N136D+Q137K+Q147K+L152E+N153E+N155E+A129T, R20K+S123P+K169R+K217T+K240F+S256Q+R267H+I294E+A129T, Q82E+P111Q+S123P+V159M+V203T+G237M+S256E+I294E+A129T, Q82E+P111Q+S123P+N155D+K169R+G237M+S256E+I294E+A129T, Q82E+P111Q+S123P+Q137K+V159M+G237M+S256E+I294E+A129T, Q82E+P111Q+S123P+Q137K+V159M+K240F+S256E+I294E+A129T, Q82E+P111Q+S123P+Q147K+N155D+K240F+S256E+I294E+A129T, A83E+P111Q+S123P+Q147K+V159M+S256E+I294E+Q329E+A129T, Q82E+P111Q+S123P+V159M+K169R+S256E+I294E+V431E+A129T, A41L+Q82E+P111Q+S123P+V159M+S256E+I294E+N383E+A129T, of the polypeptide of SEQ ID NO: 2, wherein the variant has xylogucanase activity and wherein the variant has at least 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to the polypeptide of SEQ ID NO: 1, SEQ ID NO: 2, or SEQ ID NO: 3.For example, 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%, for example, at least 96%, at least 97%, at least 98%, or at least 99%, but less than 100% sequence identity.

[0234] In an aspect, the variant comprises alterations at positions corresponding to positions selected from the group consisting of: S76E+P111Q+S123P+Q137E+Q147K+V159M+K252E+S256Q+S402Q, P111Q+N121E+Q137K+Q147K+V159M+K169R+S256Q+I294E+S402Q, P111Q+S123P+Q137K+Q147K+V159M+K169R+S256Q+I294E+S402Q, P111Q+S123P+Q137K+Q147K+V159M+L184M+V219T+S256Q+S402Q, A83E+P111Q+S123P+Q137K+Q147K+V159M+L184M+S256Q+S402Q, Q82E+P111Q+S123P+Q137K+Q147K+V159M+K252E+S256Q+S402Q, S123P+S127D+N136D+Q137K+Q147K+L152E+N153E+N155E+A491E, Q82E+P111Q+S123P+V159M+A177G+K240F+S256E+I294E+D384G, K8R+Q82E+P111Q+S123P+V159M+K169R+K240F+S256E+I294E, R20K+S123P+Q137K+K169R+K217T+K240F+S256Q+R267H+I294E, R20K+S123P+N155D+K169R+K217T+K240F+S256Q+R267H+I294E, R20K+S123P+Q147K+K169R+V219T+K240F+S256Q+R267H+I294E, Q82E+P111Q+S123P+V159M+V203T+G237M+K252E+S256E+I294E, S76E+Q82E+P111Q+S123P+V159M+V203T+G237M+S256E+I294E, Q82E+P111Q+S123P+Q147K+V159M+V203T+G237M+S256E+I294E, Q82E+P111Q+S123P+V159M+V203T+G237M+T244R+S256E+I294E of the polypeptide of SEQ ID NO: 1, wherein the variant has xylogucanase activity and wherein the variant has at least 60%,For example, 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%, for example, at least 96%, at least 97%, at least 98%, or at least 99%, but less than 100% sequence identity.

[0235] In an aspect, the variant comprises alterations at positions corresponding to positions selected from the group consisting of: S76E+P111Q+S123P+Q137E+Q147K+V159M+K252E+S256Q+S402Q+K129T, P111Q+N121E+Q137K+Q147K+V159M+K169R+S256Q+I294E+S402Q+K129T, P111Q+S123P+Q137K+Q147K+V159M+K169R+S256Q+I294E+S402Q+K129T, P111Q+S123P+Q137K+Q147K+V159M+L184M+V219T+S256Q+S402Q+K129T, A83E+P111Q+S123P+Q137K+Q147K+V159M+L184M+S256Q+S402Q+K129T, Q82E+P111Q+S123P+Q137K+Q147K+V159M+K252E+S256Q+S402Q+K129T, S123P+S127D+N136D+Q137K+Q147K+L152E+N153E+N155E+A491E+K129T, Q82E+P111Q+S123P+V159M+A177G+K240F+S256E+I294E+D384G+K129T, K8R+Q82E+P111Q+S123P+V159M+K169R+K240F+S256E+I294E+K129T, R20K+S123P+Q137K+K169R+K217T+K240F+S256Q+R267H+I294E+K129T, R20K+S123P+N155D+K169R+K217T+K240F+S256Q+R267H+I294E, R20K+S123P+Q147K+K169R+V219T+K240F+S256Q+R267H+I294E+K129T, Q82E+P111Q+S123P+V159M+V203T+G237M+K252E+S256E+I294E+K129T, S76E+Q82E+P111Q+S123P+V159M+V203T+G237M+S256E+I294E+K129T, Q82E+P111Q+S123P+Q147K+V159M+V203T+G237M+S256E+I294E+K129T, Q82E+P111Q+S123P+V159M+V203T+G237M+T244R+S256E+I294E+K129T, of the polypeptide of SEQ ID NO: 1.wherein the variant has xyloglucanase activity and wherein the variant has at least 60%, e.g., 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%, e.g., at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to the polypeptide of SEQ ID NO: 1, SEQ ID NO: 2, or SEQ ID NO: 3 but less than 100% sequence identity.

[0236] In one aspect, the variant comprises alterations at positions corresponding to positions selected from the group consisting of: S76E+P111Q+S123P+Q137E+Q147K+V159M+K252E+S256Q+S402Q+A129T, P111Q+N121E+Q137K+Q147K+V159M+K169R+S256Q+I294E+S402Q+A129T, P111Q+S123P+Q137K+Q147K+V159M+K169R+S256Q+I294E+S402Q+A129T, P111Q+S123P+Q137K+Q147K+V159M+L184M+V219T+S256Q+S402Q+A129T, A83E+P111Q+S123P+Q137K+Q147K+V159M+L184M+S256Q+S402Q+A129T, Q82E+P111Q+S123P+Q137K+Q147K+V159M+K252E+S256Q+S402Q+A129T, S123P+S127D+N136D+Q137K+Q147K+L152E+N153E+N155E+A491E+A129T, Q82E+P111Q+S123P+V159M+A177G+K240F+S256E+I294E+D384G+A129T, K8R+Q82E+P111Q+S123P+V159M+K169R+K240F+S256E+I294E+A129T, R20K+S123P+Q137K+K169R+K217T+K240F+S256Q+R267H+I294E+A129T, R20K+S123P+N155D+K169R+K217T+K240F+S256Q+R267H+I294E, R20K+S123P+Q147K+K169R+V219T+K240F+S256Q+R267H+I294E+A129T, Q82E+P111Q+S123P+V159M+V203T+G237M+K252E+S256E+I294E+A129T, S76E+Q82E+P111Q+S123P+V159M+V203T+G237M+S256E+I294E+A129T, Q82E+P111Q+S123P+Q147K+V159M+V203T+G237M+S256E+I294E+A129T, Q82E+P111Q+S123P+V159M+V203T+G237M+T244R+S256E+I294E+A129T, of the polypeptide of SEQ ID NO: 2.wherein the variant has xyloglucanase activity and wherein the variant has at least 60%, e.g., 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%, e.g., at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to the polypeptide of SEQ ID NO: 1, SEQ ID NO: 2, or SEQ ID NO: 3 but less than 100% sequence identity.

[0237] In an aspect, the variant comprises alterations at positions corresponding to positions selected from the group consisting of: A83E+P111Q+S123P+Q137K+Q147K+V159M+L184M+S256Q+I294E+S402Q, A83E+P111Q+S123P+Q137K+Q147K+N155D+L184M+S256Q+I294E+S402Q, R20K+S123P+Q147K+N155D+K169R+K217T+K240F+S256Q+R267H+I294E, R20K+S123P+N155D+K169R+K217T+K240F+S256Q+R267H+I294E+S474E, R20K+A41L+S123P+L152P+K169R+K217T+K240F+S256Q+R267H+I294E, K8R+R20K+S123P+K169R+D210H+K217T+K240F+S256Q+R267H+I294E, P111Q+S123P+Q137K+Q147K+V159M+G237M+T244R+S256Q+I294E+S402Q, P111Q+S123P+Q137K+Q147K+V159M+K169R+V203T+S256Q+I294E+S402Q, Q82E+P111Q+S123P+Q137K+V159M+V203T+D210H+G237M+S256E+I294E, Q82E+P111Q+S123P+V159M+K169R+V203T+G237M+T244R+S256E+I294E of the polypeptide of SEQ ID NO: 1, wherein the variant has xylogucanase activity and wherein the variant has at least 60%, e.g., 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%, e.g., at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to the polypeptide of SEQ ID NO: 1, SEQ ID NO: 2, or SEQ ID NO: 3 but less than 100% sequence identity.

[0238] In an aspect, the variant comprises alterations at positions corresponding to positions selected from the group consisting of: A83E+P111Q+S123P+Q137K+Q147K+V159M+L184M+S256Q+I294E+S402Q+K129T, A83E+P111Q+S123P+Q137K+Q147K+N155D+L184M+S256Q+I294E+S402Q+K129T, R20K+S123P+Q147K+N155D+K169R+K217T+K240F+S256Q+R267H+I294E+K129T, R20K+S123P+N155D+K169R+K217T+K240F+S256Q+R267H+I294E+S474E+K129T, R20K+A41L+S123P+L152P+K169R+K217T+K240F+S256Q+R267H+I294E+K129T, K8R+R20K+S123P+K169R+D210H+K217T+K240F+S256Q+R267H+I294E+K129T, P111Q+S123P+Q137K+Q147K+V159M+G237M+T244R+S256Q+I294E+S402Q+K129T, P111Q+S123P+Q137K+Q147K+V159M+K169R+V203T+S256Q+I294E+S402Q+K129T, Q82E+P111Q+S123P+Q137K+V159M+V203T+D210H+G237M+S256E+I294E+K129T, Q82E+P111Q+S123P+V159M+K169R+V203T+G237M+T244R+S256E+I294E+K129T, of the polypeptide of SEQ ID NO: 1, wherein the variant has xylogucanase activity and wherein the variant has at least 60%, e.g., 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%, e.g., at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to the polypeptide of SEQ ID NO: 1, SEQ ID NO: 2, or SEQ ID NO: 3, but less than 100% sequence identity.

[0239] In an aspect, the variant comprises alterations at positions corresponding to positions selected from the group consisting of: A83E+P111Q+S123P+Q137K+Q147K+V159M+L184M+S256Q+I294E+S402Q+A129T of the polypeptide of SEQ ID NO: 2, A83E+P111Q+S123P+Q137K+Q147K+N155D+L184M+S256Q+I294E+S402Q+A129T, R20K+S123P+Q147K+N155D+K169R+K217T+K240F+S256Q+R267H+I294E+A129T, R20K+S123P+N155D+K169R+K217T+K240F+S256Q+R267H+I294E+S474E+A129T, R20K+A41L+S123P+L152P+K169R+K217T+K240F+S256Q+R267H+I294E+A129T, K8R+R20K+S123P+K169R+D210H+K217T+K240F+S256Q+R267H+I294E+A129T, P111Q+S123P+Q137K+Q147K+V159M+G237M+T244R+S256Q+I294E+S402Q+A129T, P111Q+S123P+Q137K+Q147K+V159M+K169R+V203T+S256Q+I294E+S402Q+A129T, Q82E+P111Q+S123P+Q137K+V159M+V203T+D210H+G237M+S256E+I294E+A129T, Q82E+P111Q+S123P+V159M+K169R+V203T+G237M+T244R+S256E+I294E+A129T, wherein the variant has xylogucanase activity and wherein the variant has at least 60%, e.g., 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%, e.g., at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to the polypeptide of SEQ ID NO: 1, SEQ ID NO: 2, or SEQ ID NO: 3, but less than 100% sequence identity.

[0240] In one aspect, the variant comprises alterations at positions corresponding to positions selected from the group consisting of: Q82E+P111Q+S123P+Q137K+Q147K+V159M+L184M+A238S+S256Q+I294E+S402Q, A83E+P111Q+S123P+Q137K+Q147K+N155D+L184M+G237M+S256Q+I294E+S402Q, A83E+P111Q+S123P+Q137K+Q147K+V159M+L184M+G237M+S256E+I294E+S402Q, R20K+S123P+N155D+K169R+K217T+K240F+S256Q+R267H+I294E+D384G+E489R, R20K+S123P+K169R+K217T+K240F+S256Q+R267H+I294E+Q329E+V431E+E489R, P111Q+S123P+Q137K+Q147K+V159M+K169R+K252E+S256Q+I294E+K322E+S402Q, P111Q+S123P+Q137K+Q147K+V159M+K169R+K240F+S256Q+I294E+K322E+S402Q, S76E+P111Q+S123P+Q137K+Q147K+V159M+K169R+G237M+S256Q+I294E+S402Q, P111Q+S123P+Q137K+Q147K+V159M+K169R+G237M+T244R+S256Q+I294E+S402Q, P111Q+S123P+Q137K+Q147K+V159M+K169R+V219T+K240F+S256Q+I294E+S402Q, P111Q+S123P+Q137K+Q147K+V159M+V251E+S256E+Q271E+I294E+Q329E+S402Q, Q82E+P111Q+S123P+L152P+V159M+K169R+V203T+G237M+T244R+S256E+I294E of the polypeptide of SEQ ID NO: 1, wherein the variant has xylogucanase activity and wherein the variant has at least 60%, e.g., 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%, at least 99%, or 100% sequence identity to the polypeptide of SEQ ID NO: 1, SEQ ID NO: 2, or SEQ ID NO: 3.For example, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity but less than 100% sequence identity.

[0241] In an aspect, the variant comprises alterations at positions corresponding to positions selected from the group consisting of: Q82E+P111Q+S123P+Q137K+Q147K+V159M+L184M+A238S+S256Q+I294E+S402Q+K129T, A83E+P111Q+S123P+Q137K+Q147K+N155D+L184M+G237M+S256Q+I294E+S402Q+K129T, A83E+P111Q+S123P+Q137K+Q147K+V159M+L184M+G237M+S256E+I294E+S402Q+K129T, R20K+S123P+N155D+K169R+K217T+K240F+S256Q+R267H+I294E+D384G+E489R+K129T, R20K+S123P+K169R+K217T+K240F+S256Q+R267H+I294E+Q329E+V431E+E489R+K129T, P111Q+S123P+Q137K+Q147K+V159M+K169R+K252E+S256Q+I294E+K322E+S402Q+K129T, P111Q+S123P+Q137K+Q147K+V159M+K169R+K240F+S256Q+I294E+K322E+S402Q+K129T, S76E+P111Q+S123P+Q137K+Q147K+V159M+K169R+G237M+S256Q+I294E+S402Q+K129T, P111Q+S123P+Q137K+Q147K+V159M+K169R+G237M+T244R+S256Q+I294E+S402Q+K129T, P111Q+S123P+Q137K+Q147K+V159M+K169R+V219T+K240F+S256Q+I294E+S402Q+K129T, P111Q+S123P+Q137K+Q147K+V159M+V251E+S256E+Q271E+I294E+Q329E+S402Q+K129T, Q82E+P111Q+S123P+L152P+V159M+K169R+V203T+G237M+T244R+S256E+I294E+K129T of the polypeptide of SEQ ID NO: 1, wherein the variant has xylogucanase activity and wherein the variant has at least 60%,For example, 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%, for example, at least 96%, at least 97%, at least 98%, or at least 99%, but less than 100% sequence identity.

[0242] In an aspect, the variant comprises alterations at positions corresponding to positions selected from the group consisting of: Q82E+P111Q+S123P+Q137K+Q147K+V159M+L184M+A238S+S256Q+I294E+S402Q+A129T of the polypeptide of SEQ ID NO: 2, A83E+P111Q+S123P+Q137K+Q147K+N155D+L184M+G237M+S256Q+I294E+S402Q+A129T, A83E+P111Q+S123P+Q137K+Q147K+V159M+L184M+G237M+S256E+I294E+S402Q+A129T, R20K+S123P+N155D+K169R+K217T+K240F+S256Q+R267H+I294E+D384G+E489R+A129T, R20K+S123P+K169R+K217T+K240F+S256Q+R267H+I294E+Q329E+V431E+E489R+A129T, P111Q+S123P+Q137K+Q147K+V159M+K169R+K252E+S256Q+I294E+K322E+S402Q+A129T, P111Q+S123P+Q137K+Q147K+V159M+K169R+K240F+S256Q+I294E+K322E+S402Q+A129T, S76E+P111Q+S123P+Q137K+Q147K+V159M+K169R+G237M+S256Q+I294E+S402Q+A129T, P111Q+S123P+Q137K+Q147K+V159M+K169R+G237M+T244R+S256Q+I294E+S402Q+A129T, P111Q+S123P+Q137K+Q147K+V159M+K169R+V219T+K240F+S256Q+I294E+S402Q+A129T, P111Q+S123P+Q137K+Q147K+V159M+V251E+S256E+Q271E+I294E+Q329E+S402Q+A129T, Q82E+P111Q+S123P+L152P+V159M+K169R+V203T+G237M+T244R+S256E+I294E+A129T, wherein the variant has xylogucanase activity and wherein the variant has at least 60%,For example, 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%, for example, at least 96%, at least 97%, at least 98%, or at least 99%, but less than 100% sequence identity.

[0243] In an aspect, the variant comprises alterations at positions corresponding to positions selected from the group consisting of: A83E+P111Q+S123P+Q137K+F146D+Q147G+L148P+V159M+L184M+S256Q+I294E+S402Q, A83E+P111Q+S123P+Q137K+Q147K+V159M+L184M+S256Q+I294E+N298D+V300L+S402Q, A83E+P111Q+S123P+Q137K+Q147K+V159M+K169R+L184M+K240L+S256Q+I294E+S402Q, A83E+P111Q+S123P+Q137K+Q147K+V159M+L184M+G237M+S256Q+I294E+S402Q+K488T, A83E+P111Q+S123P+Q137K+Q147K+V159M+L184M+V203T+K240F+S256Q+I294E+S402Q, A83E+P111Q+S123P+Q137K+Q147K+V159M+L184M+K240F+S256Q+I294E+S402Q+E489R, A83E+P111Q+S123P+Q137K+Q147K+V159M+F165H+L184M+S256Q+I294E+S402Q+V431E, A83E+P111Q+S123P+Q137K+Q147K+V159M+L184M+V251E+Q271E+I294E+Q329E+S402Q, P111Q+S123P+Q137K+Q147K+V159M+K169R+D210H+K240F+S256E+I294E+S402Q+K488T, S76E+P111Q+S123P+Q137K+Q147K+V159M+K169R+V203T+G237M+S256Q+I294E+S402Q, P111Q+S123P+Q137K+Q147K+V159M+K169R+G237M+S256Q+I294E+P339S+S402Q+K488T, S76E+Q82E+P111Q+S123P+V159M+V203T+G237M+S256E+I294E+S474E+E489R+P492D, P111Q+S123P+Q137K+Q147K+V159M+A238T+V251E+S256Q+Q271E+I294E+Q329E+S402Q,A83E+P111Q+S123P+Q137K+Q147K+V159M+L184M+S256E+Q271D+I294E+Q329E+S402Q, Q82E+P111Q+S123P+Q137K+V159M+K169R+A189G+V203T+G237M+T244R+S256E+I294E, Q82E+P111Q+S123P+Q137K+V159M+K169R+V203T+G237M+T244R+V251E+S256E+I294E, Q82E+P111Q+S123P+V159M+K169R+V203T+G237M+T244R+S256E+I294E+N383Q+V431E, S76E+Q82E+S94R+P111Q+S123+V159M+K169R+V203T+G237M+T244R+S256E+I294E, wherein the variant has xylogucanase activity and wherein the variant has at least 60%, e.g., 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%, e.g., at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to the polypeptide of SEQ ID NO: 1, SEQ ID NO: 2, or SEQ ID NO: 3 but less than 100% sequence identity.

[0244] In an aspect, the variant comprises alterations at positions corresponding to positions selected from the group consisting of: A83E+P111Q+S123P+Q137K+F146D+Q147G+L148P+V159M+L184M+S256Q+I294E+S402Q+K129T, A83E+P111Q+S123P+Q137K+Q147K+V159M+L184M+S256Q+I294E+N298D+V300L+S402Q+K129T, A83E+P111Q+S123P+Q137K+Q147K+V159M+K169R+L184M+K240L+S256Q+I294E+S402Q+K129T, A83E+P111Q+S123P+Q137K+Q147K+V159M+L184M+G237M+S256Q+I294E+S402Q+K488T+K129T, A83E+P111Q+S123P+Q137K+Q147K+V159M+L184M+V203T+K240F+S256Q+I294E+S402Q+K129T, A83E+P111Q+S123P+Q137K+Q147K+V159M+L184M+K240F+S256Q+I294E+S402Q+E489R+K129T, A83E+P111Q+S123P+Q137K+Q147K+V159M+F165H+L184M+S256Q+I294E+S402Q+V431E+K129T, A83E+P111Q+S123P+Q137K+Q147K+V159M+L184M+V251E+Q271E+I294E+Q329E+S402Q, P111Q+S123P+Q137K+Q147K+V159M+K169R+D210H+K240F+S256E+I294E+S402Q+K488T+K129T, S76E+P111Q+S123P+Q137K+Q147K+V159M+K169R+V203T+G237M+S256Q+I294E+S402Q+K129T, P111Q+S123P+Q137K+Q147K+V159M+K169R+G237M+S256Q+I294E+P339S+S402Q+K488T+K129T, S76E+Q82E+P111Q+S123P+V159M+V203T+G237M+S256E+I294E+S474E+E489R+P492D+K129T, of the polypeptide of SEQ ID NO: 1.P111Q+S123P+Q137K+Q147K+V159M+A238T+V251E+S256Q+Q271E+I294E+Q329E+S402Q+K129T, A83E+P111Q+S123P+Q137K+Q147K+V159M+L184M+S256E+Q271D+I294E+Q329E+S402Q+K129T, Q82E+P111Q+S123P+Q137K+V159M+K169R+A189G+V203T+G237M+T244R+S256E+I294E+K129T, Q82E+P111Q+S123P+Q137K+V159M+K169R+V203T+G237M+T244R+V251E+S256E+I294E+K129T, Q82E+P111Q+S123P+V159M+K169R+V203T+G237M+T244R+S256E+I294E+N383Q+V431E+K129T, S76E+Q82E+S94R+P111Q+S123+V159M+K169R+V203T+G237M+T244R+S256E+I294E+K129T, wherein the variant has xylogucanase activity and wherein the variant has at least 60%, e.g., 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%, e.g., at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to the polypeptide of SEQ ID NO: 1, SEQ ID NO: 2, or SEQ ID NO: 3 but less than 100% sequence identity.

[0245] In an aspect, the variant comprises alterations at positions corresponding to positions selected from the group consisting of: A83E+P111Q+S123P+Q137K+F146D+Q147G+L148P+V159M+L184M+S256Q+I294E+S402Q+A129T of the polypeptide of SEQ ID NO: 2, A83E+P111Q+S123P+Q137K+Q147K+V159M+L184M+S256Q+I294E+N298D+V300L+S402Q+A129T, A83E+P111Q+S123P+Q137K+Q147K+V159M+K169R+L184M+K240L+S256Q+I294E+S402Q+A129T, A83E+P111Q+S123P+Q137K+Q147K+V159M+L184M+G237M+S256Q+I294E+S402Q+K488T+A129T, A83E+P111Q+S123P+Q137K+Q147K+V159M+L184M+V203T+K240F+S256Q+I294E+S402Q+A129T, A83E+P111Q+S123P+Q137K+Q147K+V159M+L184M+K240F+S256Q+I294E+S402Q+E489R+A129T, A83E+P111Q+S123P+Q137K+Q147K+V159M+F165H+L184M+S256Q+I294E+S402Q+V431E+A129T, A83E+P111Q+S123P+Q137K+Q147K+V159M+L184M+V251E+Q271E+I294E+Q329E+S402Q, P111Q+S123P+Q137K+Q147K+V159M+K169R+D210H+K240F+S256E+I294E+S402Q+K488T+A129T, S76E+P111Q+S123P+Q137K+Q147K+V159M+K169R+V203T+G237M+S256Q+I294E+S402Q+A129T, P111Q+S123P+Q137K+Q147K+V159M+K169R+G237M+S256Q+I294E+P339S+S402Q+K488T+A129T, S76E+Q82E+P111Q+S123P+V159M+V203T+G237M+S256E+I294E+S474E+E489R+P492D+A129T, of the polypeptide of SEQ ID NO: 2.P111Q+S123P+Q137K+Q147K+V159M+A238T+V251E+S256Q+Q271E+I294E+Q329E+ S402Q+A129T, A83E+P111Q+S123P+Q137K+Q147K+V159M+L184M+S256E+Q271D+I294E+ Q329E+S402Q+A129T, Q82E+P111Q+S123P+Q137K+V159M+K169R+A189G+V203T+G237M+ T244R+S256E+I294E+A129T, Q82E+P111Q+S123P+Q137K+V159M+K169R+V203T+G237M+ T244R+V251E+S256E+I294E+A129T, Q82E+P111Q+S123P+V159M+K169R+V203T+G237M+ T244R+S256E+I294E+N383Q+V431E+A129T, S76E+Q82E+S94R+P111Q+S123+V159M+K169R+ V203T+G237M+T244R+S256E+I294E+A129T, wherein the variant has xylogucanase activity and wherein the variant has at least 60%, e.g., 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%, e.g., at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to the polypeptide of SEQ ID NO: 1, SEQ ID NO: 2, or SEQ ID NO: 3 but less than 100% sequence identity.

[0246] In an aspect, the variant comprises alterations at positions corresponding to positions selected from the group consisting of: A83E+P111Q+S123P+Q137K+Q147K+V159M+L184M+V251E+S256Q+Q271E+I294E+Q329E+S402Q, A83E+P111Q+S123P+Q137K+Q147K+V159M+L184M+K252E+S256Q+Q271E+I294E+Q329E+S402Q, Q82E+P111Q+S123P+V159M+K169R+V203T+G237M+T244R+S256E+Q271E+I294E+Q329E+N383E, Q82E+P111Q+S123P+Q137K+N155D+V159M+K169R+V203T+G237M+T244R+S256E+I294E+K445E, K8E+A41L+S76E+Q82E+P111Q+S123P+V159M+K169R+V203T+G237M+T244R+S256E+I294E of the polypeptide of SEQ ID NO: 1, wherein the variant has xylogucanase activity and wherein the variant has at least 60%, e.g., 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%, e.g., at least 96%, at least 97%, at least 98%, or at least 99%, sequence identity, but less than 100%, sequence identity, to the polypeptide of SEQ ID NO: 1, SEQ ID NO: 2, or SEQ ID NO: 3.

[0247] In an aspect, the variant comprises alterations at positions corresponding to positions selected from the group consisting of: A83E+P111Q+S123P+Q137K+Q147K+V159M+L184M+V251E+S256Q+Q271E+I294E+Q329E+S402Q+K129T, A83E+P111Q+S123P+Q137K+Q147K+V159M+L184M+K252E+S256Q+Q271E+I294E+Q329E+S402Q+K129T, Q82E+P111Q+S123P+V159M+K169R+V203T+G237M+T244R+S256E+Q271E+I294E+Q329E+N383E+K129T, Q82E+P111Q+S123P+Q137K+N155D+V159M+K169R+V203T+G237M+T244R+S256E+I294E+K445E+K129T, K8E+A41L+S76E+Q82E+P111Q+S123P+V159M+K169R+V203T+G237M+T244R+S256E+I294E+K129T of the polypeptide of SEQ ID NO: 1, wherein the variant has xylogucanase activity and wherein the variant has at least 60%, e.g., 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%, e.g., at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to the polypeptide of SEQ ID NO: 1, SEQ ID NO: 2, or SEQ ID NO: 3, but less than 100% sequence identity.

[0248] In an aspect, the variant comprises alterations at positions corresponding to positions selected from the group consisting of: A83E+P111Q+S123P+Q137K+Q147K+V159M+L184M+V251E+S256Q+Q271E+I294E+Q329E+S402Q+A129T of the polypeptide of SEQ ID NO: 2, A83E+P111Q+S123P+Q137K+Q147K+V159M+L184M+K252E+S256Q+Q271E+I294E+Q329E+S402Q+A129T of the polypeptide of SEQ ID NO: 2, Q82E+P111Q+S123P+V159M+K169R+V203T+G237M+T244R+S256E+Q271E+I294E+Q329E+N383E+A129T of the polypeptide of SEQ ID NO: 2, Q82E+P111Q+S123P+Q137K+N155D+V159M+K169R+V203T+G237M+T244R+S256E+I294E+K445E+A129T of the polypeptide of SEQ ID NO: 2, K8E+A41L+S76E+Q82E+P111Q+S123P+V159M+K169R+V203T+G237M+T244R+S256E+I294E+A129T of the polypeptide of SEQ ID NO: 2, wherein the variant has xylogucanase activity and wherein the variant has at least 60%, e.g., 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%, e.g., at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to the polypeptide of SEQ ID NO: 1, SEQ ID NO: 2, or SEQ ID NO: 3 but less than 100% sequence identity.

[0249] In an aspect, the variant comprises alterations at positions corresponding to positions selected from the group consisting of: S76E+A83E+P111Q+S123P+Q137K+Q147K+V159M+L184M+V251E+S256Q+Q271E+I294E+Q329E+S402Q of the polypeptide of SEQ ID NO: 1, A83E+P111Q+S123P+Q137K+Q147K+V159M+K169R+L184M+K252E+S256Q+Q271E+I294E+Q329E+S402Q, A83E+P111Q+S123P+Q137K+Q147K+V159M+L184M+V203T+A238S+S256E+Q271E+I294E+Q329E+S402Q, A83E+P111Q+S123P+Q137K+Q147K+V159M+L184M+V251E+S256Q+Q271E+I294E+Q329E+S402Q+V431E, A83E+P111Q+S123P+Q137K+Q147K+V159M+L184M+V251E+S256Q+Q271E+I294E+Q329E+K347E+S402Q, A83E+P111Q+S123P+Q137K+L152P+V159M+L184M+V251E+S256Q+Q271E+I294E+Q329E+S402Q+V431E, K8E+A83E+P111Q+S123P+Q137K+Q147K+V159M+L184M+V251E+S256Q+Q271E+I294E+Q329E+S402Q, A83E+P111Q+S123P+Q137K+Q147K+N155D+L184M+G237M+V251E+S256Q+Q271E+I294E+Q329E+S402Q, A41L+A83E+P111Q+S123P+Q137K+Q147K+V159M+L184M+V251E+S256Q+Q271E+I294E+Q329E+S402Q, A83E+P111Q+S123P+Q137K+Q147K+V159M+L184M+V251E+S256Q+Q271E+I294E+Q329E+N383Q+S402Q, R20K+S76E+Q82E+P111Q+S123P+V159M+K169R+V203T+G237M+T244R+S256E+I294E+Q329E+P492D,S76E+Q82E+P111Q+S123P+V159M+K169R+V203T+G237M+T244R+S256E+I294E+ Q329E+E489R+P492D, wherein the variant has xylogucanase activity and wherein the variant has at least 60%, e.g., 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%, e.g., at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to the polypeptide of SEQ ID NO: 1, SEQ ID NO: 2, or SEQ ID NO: 3 but less than 100% sequence identity.

[0250] In an aspect, the variant comprises alterations at positions corresponding to positions selected from the group consisting of: S76E+A83E+P111Q+S123P+Q137K+Q147K+V159M+L184M+V251E+S256Q+Q271E+I294E+Q329E+S402Q+K129T of the polypeptide of SEQ ID NO: 1, A83E+P111Q+S123P+Q137K+Q147K+V159M+K169R+L184M+K252E+S256Q+Q271E+I294E+Q329E+S402Q+K129T, A83E+P111Q+S123P+Q137K+Q147K+V159M+L184M+V203T+A238S+S256E+Q271E+I294E+Q329E+S402Q+K129T, A83E+P111Q+S123P+Q137K+Q147K+V159M+L184M+V251E+S256Q+Q271E+I294E+Q329E+S402Q+V431E+K129T, A83E+P111Q+S123P+Q137K+Q147K+V159M+L184M+V251E+S256Q+Q271E+I294E+Q329E+K347E+S402Q+K129T, A83E+P111Q+S123P+Q137K+L152P+V159M+L184M+V251E+S256Q+Q271E+I294E+Q329E+S402Q+V431E+K129T, K8E+A83E+P111Q+S123P+Q137K+Q147K+V159M+L184M+V251E+S256Q+Q271E+I294E+Q329E+S402Q+K129T, A83E+P111Q+S123P+Q137K+Q147K+N155D+L184M+G237M+V251E+S256Q+Q271E+I294E+Q329E+S402Q+K129T, A41L+A83E+P111Q+S123P+Q137K+Q147K+V159M+L184M+V251E+S256Q+Q271E+I294E+Q329E+S402Q+K129T, A83E+P111Q+S123P+Q137K+Q147K+V159M+L184M+V251E+S256Q+Q271E+I294E+Q329E+N383Q+S402Q+K129T,R20K+S76E+Q82E+P111Q+S123P+V159M+K169R+V203T+G237M+T244R+S256E+I294E+Q329E+P492D+K129T, S76E+Q82E+P111Q+S123P+V159M+K169R+V203T+G237M+T244R+S256E+I294E+Q329E+E489R+P492D+K129T, wherein the variant has xylogucanase activity and wherein the variant has at least 60%, e.g., 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%, e.g., at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to the polypeptide of SEQ ID NO: 1, SEQ ID NO: 2, or SEQ ID NO: 3 but less than 100% sequence identity.

[0251] In an aspect, the variant comprises alterations at positions corresponding to positions selected from the group consisting of: S76E+A83E+P111Q+S123P+Q137K+Q147K+V159M+L184M+V251E+S256Q+Q271E+I294E+Q329E+S402Q+A129T of the polypeptide of SEQ ID NO: 2, A83E+P111Q+S123P+Q137K+Q147K+V159M+K169R+L184M+K252E+S256Q+Q271E+I294E+Q329E+S402Q+A129T, A83E+P111Q+S123P+Q137K+Q147K+V159M+L184M+V203T+A238S+S256E+Q271E+I294E+Q329E+S402Q+A129T, A83E+P111Q+S123P+Q137K+Q147K+V159M+L184M+V251E+S256Q+Q271E+I294E+Q329E+S402Q+V431E+A129T, A83E+P111Q+S123P+Q137K+Q147K+V159M+L184M+V251E+S256Q+Q271E+I294E+Q329E+K347E+S402Q+A129T, A83E+P111Q+S123P+Q137K+L152P+V159M+L184M+V251E+S256Q+Q271E+I294E+Q329E+S402Q+V431E+A129T, K8E+A83E+P111Q+S123P+Q137K+Q147K+V159M+L184M+V251E+S256Q+Q271E+I294E+Q329E+S402Q+A129T, A83E+P111Q+S123P+Q137K+Q147K+N155D+L184M+G237M+V251E+S256Q+Q271E+I294E+Q329E+S402Q+A129T, A41L+A83E+P111Q+S123P+Q137K+Q147K+V159M+L184M+V251E+S256Q+Q271E+I294E+Q329E+S402Q+A129T, A83E+P111Q+S123P+Q137K+Q147K+V159M+L184M+V251E+S256Q+Q271E+I294E+Q329E+N383Q+S402Q+A129T,R20K+S76E+Q82E+P111Q+S123P+V159M+K169R+V203T+G237M+T244R+S256E+I294E+Q329E+P492D+A129T, S76E+Q82E+P111Q+S123P+V159M+K169R+V203T+G237M+T244R+S256E+I294E+Q329E+E489R+P492D+A129T, wherein the variant has xylogucanase activity and wherein the variant has at least 60%, e.g., 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%, e.g., at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to the polypeptide of SEQ ID NO: 1, SEQ ID NO: 2, or SEQ ID NO: 3 but less than 100% sequence identity.

[0252] In an aspect, the variant comprises alterations at positions corresponding to positions selected from the group consisting of: A83E+P111Q+S123P+Q137K+Q147K+V159M+L184M+V219T+G237M+V251E+S256Q+Q271E+I294E+Q329E+S402Q of the polypeptide of SEQ ID NO: 1, S76E+A83E+P111Q+S123P+Q137K+Q147K+V159M+L184M+V203T+A238S+S256E+Q271E+I294E+Q329E+S402Q, A83E+P111Q+S123P+Q137K+Q147K+V159M+L184M+D210H+T244R+K252E+S256Q+Q271E+I294E+Q329E+S402Q, A83E+P111Q+S123P+Q137K+Q147K+V159M+L184M+T244E+V251E+S256Q+Q271E+I294E+Q329E+S402Q+V431E, K8E+A83E+S94R+P111Q+S123P+Q137K+Q147K+V159M+L184M+V251E+S256Q+Q271E+I294E+Q329E+S402Q, A83E+P111Q+S123P+Q137K+Q147K+V159M+L184M+V251E+S256Q+Q271E+I294E+Q329E+S402Q+V431E+K445E, A83E+P111Q+S123P+Q137K+Q147K+V159M+L184M+D210H+K240F+V251E+S256Q+Q271E+I294E+Q329E+S402Q, A41L+A83E+P111Q+S123P+Q137K+Q147K+V159M+L184M+K240F+V251E+S256Q+Q271E+I294E+Q329E+S402Q, Q82E+P111Q+S123P+Q137K+Q147K+V159M+L184M+V203T+D210H+V251E+S256Q+Q271E+I294E+Q329E+S402Q, A41L+A83E+P111Q+S123P+Q137K+Q147K+V159M+L184M+V251E+S256Q+Q271E+I294E+Q329E+N383Q+S402Q,A83E+P111Q+S123P+Q137K+Q147K+V159M+L184M+T244R+W248V+V251E+S256Q+Q271E+I294E+Q329E+S402Q, A83E+P111Q+S123P+Q137K+Q147K+V159M+L184M+V203T+T244R+V251E+S256Q+Q271E+I294E+Q329E+S402Q, A83E+P111Q+S123P+Q137K+Q147K+V159M+L184M+V251E+S256Q+Q271E+R295K+N298D+Q329E+S402Q+L447M, A83E+P111Q+S123P+Q137K+Q147K+V159M+L184M+S256E+Q271E+I294E+Q329E+P339S+N383E+S402Q+V431E, wherein the variant has xylogucanase activity and wherein the variant has at least 60%, e.g., 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%, e.g., at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to the polypeptide of SEQ ID NO: 1, SEQ ID NO: 2, or SEQ ID NO: 3 but less than 100% sequence identity.

[0253] In an aspect, the variant comprises alterations at positions corresponding to positions selected from the group consisting of: A83E+P111Q+S123P+Q137K+Q147K+V159M+L184M+V219T+G237M+V251E+S256Q+Q271E+I294E+Q329E+S402Q+K129T of the polypeptide of SEQ ID NO: 1, S76E+A83E+P111Q+S123P+Q137K+Q147K+V159M+L184M+V203T+A238S+S256E+Q271E+I294E+Q329E+S402Q+K129T, A83E+P111Q+S123P+Q137K+Q147K+V159M+L184M+D210H+T244R+K252E+S256Q+Q271E+I294E+Q329E+S402Q+K129T, A83E+P111Q+S123P+Q137K+Q147K+V159M+L184M+T244E+V251E+S256Q+Q271E+I294E+Q329E+S402Q+V431E+K129T, K8E+A83E+S94R+P111Q+S123P+Q137K+Q147K+V159M+L184M+V251E+S256Q+Q271E+I294E+Q329E+S402Q+K129T, A83E+P111Q+S123P+Q137K+Q147K+V159M+L184M+V251E+S256Q+Q271E+I294E+Q329E+S402Q+V431E+K445E+K129T, A83E+P111Q+S123P+Q137K+Q147K+V159M+L184M+D210H+K240F+V251E+S256Q+Q271E+I294E+Q329E+S402Q+K129T, A41L+A83E+P111Q+S123P+Q137K+Q147K+V159M+L184M+K240F+V251E+S256Q+Q271E+I294E+Q329E+S402Q+K129T, Q82E+P111Q+S123P+Q137K+Q147K+V159M+L184M+V203T+D210H+V251E+S256Q+Q271E+I294E+Q329E+S402Q+K129T, A41L+A83E+P111Q+S123P+Q137K+Q147K+V159M+L184M+V251E+S256Q+Q271E+I294E+Q329E+N383Q+S402Q+K129T,A83E+P111Q+S123P+Q137K+Q147K+V159M+L184M+T244R+W248V+V251E+S256Q+Q271E+I294E+Q329E+S402Q+K129T, A83E+P111Q+S123P+Q137K+Q147K+V159M+L184M+V203T+T244R+V251E+S256Q+Q271E+I294E+Q329E+S402Q+K129T, A83E+P111Q+S123P+Q137K+Q147K+V159M+L184M+V251E+S256Q+Q271E+R295K+N298D+Q329E+S402Q+L447M+K129T, A83E+P111Q+S123P+Q137K+Q147K+V159M+L184M+S256E+Q271E+I294E+Q329E+P339S+N383E+S402Q+V431E+K129T, wherein the variant has xylogucanase activity and wherein the variant has at least 60%, e.g., 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%, e.g., at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to the polypeptide of SEQ ID NO: 1, SEQ ID NO: 2, or SEQ ID NO: 3 but less than 100% sequence identity.

[0254] In an aspect, the variant comprises alterations at positions corresponding to positions selected from the group consisting of: A83E+P111Q+S123P+Q137K+Q147K+V159M+L184M+V219T+G237M+V251E+S256Q+Q271E+I294E+Q329E+S402Q+A129T of the polypeptide of SEQ ID NO: 2, S76E+A83E+P111Q+S123P+Q137K+Q147K+V159M+L184M+V203T+A238S+S256E+Q271E+I294E+Q329E+S402Q+A129T, A83E+P111Q+S123P+Q137K+Q147K+V159M+L184M+D210H+T244R+K252E+S256Q+Q271E+I294E+Q329E+S402Q+A129T, A83E+P111Q+S123P+Q137K+Q147K+V159M+L184M+T244E+V251E+S256Q+Q271E+I294E+Q329E+S402Q+V431E+A129T, K8E+A83E+S94R+P111Q+S123P+Q137K+Q147K+V159M+L184M+V251E+S256Q+Q271E+I294E+Q329E+S402Q+A129T, A83E+P111Q+S123P+Q137K+Q147K+V159M+L184M+V251E+S256Q+Q271E+I294E+Q329E+S402Q+V431E+K445E+A129T, A83E+P111Q+S123P+Q137K+Q147K+V159M+L184M+D210H+K240F+V251E+S256Q+Q271E+I294E+Q329E+S402Q+A129T, A41L+A83E+P111Q+S123P+Q137K+Q147K+V159M+L184M+K240F+V251E+S256Q+Q271E+I294E+Q329E+S402Q+A129T, Q82E+P111Q+S123P+Q137K+Q147K+V159M+L184M+V203T+D210H+V251E+S256Q+Q271E+I294E+Q329E+S402Q+A129T, A41L+A83E+P111Q+S123P+Q137K+Q147K+V159M+L184M+V251E+S256Q+Q271E+I294E+Q329E+N383Q+S402Q+A129T,A83E+P111Q+S123P+Q137K+Q147K+V159M+L184M+T244R+W248V+V251E+S256Q+Q271E+I294E+Q329E+S402Q+A129T, A83E+P111Q+S123P+Q137K+Q147K+V159M+L184M+V203T+T244R+V251E+S256Q+Q271E+I294E+Q329E+S402Q+A129T, A83E+P111Q+S123P+Q137K+Q147K+V159M+L184M+V251E+S256Q+Q271E+R295K+N298D+Q329E+S402Q+L447M+A129T, A83E+P111Q+S123P+Q137K+Q147K+V159M+L184M+S256E+Q271E+I294E+Q329E+P339S+N383E+S402Q+V431E+A129T, wherein the variant has xylogucanase activity and wherein the variant has at least 60%, e.g., 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%, e.g., at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to the polypeptide of SEQ ID NO: 1, SEQ ID NO: 2, or SEQ ID NO: 3 but less than 100% sequence identity.

[0255] In one aspect, the variant comprises alterations at positions corresponding to positions selected from the group consisting of: A83E+P111Q+S123P+Q137K+Q147K+V159M+L184M+G237M+V251E+S256Q+Q271E+I294E+Q329E+S402Q+E489R+V505L, A83E+P111Q+S123P+Q137K+Q147K+V159M+L184M+K252E+S256Q+Q271E+I294E+Q329E+N383E+S402Q+V431E+A491V, A83E+P111Q+S123P+Q137K+Q147K+V159M+L184M+V203T+V251E+S256Q+Q271E+I294E+Q329E+N383Q+S402Q+V431E, A83E+P111Q+S123P+Q137K+Q147K+V159M+L184M+V219T+G237M+S256Q+Q271E+I294E+Q329E+S402Q+V431E+S474E, A83E+P111Q+S123P+Q137K+Q147K+V159M+L184M+V251E+S256Q+Q271E+I294E+Q329E+N383E+S402Q+V431E+P492D, A83E+P111Q+S123P+Q137K+Q147K+V159M+L184M+A189G+V251E+S256Q+Q271E+I294E+Q329E+N383Q+S402Q+V431E, A83E+P111Q+S123P+Q137K+Q147K+V159M+L184M+V251E+S256Q+Q271D+I294E+Q329E+N383E+S402Q+V431E+A459P, K8R+A83E+P111Q+S123P+Q137K+Q147K+V159M+L184M+V251E+S256Q+Q271E+I294E+Q329E+S402Q+V431E+E489K, A83E+P111Q+S123P+Q137K+Q147K+V159M+L184M+V251E+S256Q+Q271E+I294E+Q329E+N383E+S402Q+V431E+A491V, of the polypeptide of SEQ ID NO: 1, wherein the variant has xylogucanase activity and wherein the variant has at least 60%,For example, 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%, for example, at least 96%, at least 97%, at least 98%, or at least 99%, but less than 100% sequence identity.

[0256] In one aspect, the variant comprises alterations at positions corresponding to positions selected from the group consisting of: A83E+P111Q+S123P+Q137K+Q147K+V159M+L184M+G237M+V251E+S256Q+Q271E+I294E+Q329E+S402Q+E489R+V505L+K129T, A83E+P111Q+S123P+Q137K+Q147K+V159M+L184M+K252E+S256Q+Q271E+I294E+Q329E+N383E+S402Q+V431E+A491V+K129T, A83E+P111Q+S123P+Q137K+Q147K+V159M+L184M+V203T+V251E+S256Q+Q271E+I294E+Q329E+N383Q+S402Q+V431E+K129T, A83E+P111Q+S123P+Q137K+Q147K+V159M+L184M+V219T+G237M+S256Q+Q271E+I294E+Q329E+S402Q+V431E+S474E+K129T, A83E+P111Q+S123P+Q137K+Q147K+V159M+L184M+V251E+S256Q+Q271E+I294E+Q329E+N383E+S402Q+V431E+P492D+K129T, A83E+P111Q+S123P+Q137K+Q147K+V159M+L184M+A189G+V251E+S256Q+Q271E+I294E+Q329E+N383Q+S402Q+V431E+K129T, A83E+P111Q+S123P+Q137K+Q147K+V159M+L184M+V251E+S256Q+Q271D+I294E+Q329E+N383E+S402Q+V431E+A459P+K129T, K8R+A83E+P111Q+S123P+Q137K+Q147K+V159M+L184M+V251E+S256Q+Q271E+I294E+Q329E+S402Q+V431E+E489K+K129T, A83E+P111Q+S123P+Q137K+Q147K+V159M+L184M+V251E+S256Q+Q271E+I294E+Q329E+N383E+S402Q+V431E+A491V+K129T, of the polypeptide of SEQ ID NO: 1.wherein the variant has xyloglucanase activity and wherein the variant has at least 60%, e.g., 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%, e.g., at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to the polypeptide of SEQ ID NO: 1, SEQ ID NO: 2, or SEQ ID NO: 3 but less than 100% sequence identity.

[0257] In an aspect, the variant comprises alterations at positions corresponding to positions selected from the group consisting of: A83E+P111Q+S123P+Q137K+Q147K+V159M+L184M+G237M+V251E+S256Q+Q271E+I294E+Q329E+S402Q+E489R+V505L+A129T, A83E+P111Q+S123P+Q137K+Q147K+V159M+L184M+K252E+S256Q+Q271E+I294E+Q329E+N383E+S402Q+V431E+A491V+A129T, A83E+P111Q+S123P+Q137K+Q147K+V159M+L184M+V203T+V251E+S256Q+Q271E+I294E+Q329E+N383Q+S402Q+V431E+A129T, A83E+P111Q+S123P+Q137K+Q147K+V159M+L184M+V219T+G237M+S256Q+Q271E+I294E+Q329E+S402Q+V431E+S474E+A129T, A83E+P111Q+S123P+Q137K+Q147K+V159M+L184M+V251E+S256Q+Q271E+I294E+Q329E+N383E+S402Q+V431E+P492D+A129T, A83E+P111Q+S123P+Q137K+Q147K+V159M+L184M+A189G+V251E+S256Q+Q271E+I294E+Q329E+N383Q+S402Q+V431E+A129T, A83E+P111Q+S123P+Q137K+Q147K+V159M+L184M+V251E+S256Q+Q271D+I294E+Q329E+N383E+S402Q+V431E+A459P+A129T, K8R+A83E+P111Q+S123P+Q137K+Q147K+V159M+L184M+V251E+S256Q+Q271E+I294E+Q329E+S402Q+V431E+E489K+A129T, A83E+P111Q+S123P+Q137K+Q147K+V159M+L184M+V251E+S256Q+Q271E+I294E+Q329E+N383E+S402Q+V431E+A491V+A129T, of the polypeptide of SEQ ID NO: 2.wherein the variant has xyloglucanase activity and wherein said variant has at least 60%, e.g., 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%, e.g., at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to the polypeptide of SEQ ID NO: 1, SEQ ID NO: 2, or SEQ ID NO: 3 but less than 100% sequence identity.

[0258] In one aspect, the variant comprises alterations at positions corresponding to positions selected from the group consisting of: A83E+P111Q+S123P+Q137K+Q147K+V159M+L184M+D210H+G237M+V251E+S256Q+Q271E+I294E+Q329E+S402Q+E489R+P492D of the polypeptide of SEQ ID NO: 1, S76E+Q82E+P111Q+S123P+Q137K+Q147K+V159M+L184M+V203T+V251E+S256Q+Q271E+I294E+Q329E+N383Q+S402Q+V431E, K8E+A41L+A83E+P111Q+S123P+Q137K+Q147K+V159M+L184M+V251E+S256Q+Q271E+I294E+Q329E+K394R+S402Q+V431E, A83E+P111Q+S123P+Q137K+Q147K+V159M+L184M+V251E+S256Q+Q271E+I294E+Q329E+K347E+N383E+S402Q+V431E+A491V, K8E+A83E+P111Q+S123P+Q137K+Q147K+V159M+L184M+V251E+S256Q+Q271E+I294E+Q329E+N383E+S402Q+V431E+A491V, wherein the variant has xyloglucanase activity and wherein said variant has at least 60%, e.g., 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%, e.g., at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to the polypeptide of SEQ ID NO: 1, SEQ ID NO: 2, or SEQ ID NO: 3 but less than 100% sequence identity.

[0259] In an aspect, the variant comprises alterations at positions corresponding to positions selected from the group consisting of: A83E+P111Q+S123P+Q137K+Q147K+V159M+L184M+D210H+G237M+V251E+S256Q+Q271E+I294E+Q329E+S402Q+E489R+P492D+K129T of the polypeptide of SEQ ID NO: 1, S76E+Q82E+P111Q+S123P+Q137K+Q147K+V159M+L184M+V203T+V251E+S256Q+Q271E+I294E+Q329E+N383Q+S402Q+V431E+K129T, K8E+A41L+A83E+P111Q+S123P+Q137K+Q147K+V159M+L184M+V251E+S256Q+Q271E+I294E+Q329E+K394R+S402Q+V431E+K129T, A83E+P111Q+S123P+Q137K+Q147K+V159M+L184M+V251E+S256Q+Q271E+I294E+Q329E+K347E+N383E+S402Q+V431E+A491V+K129T, K8E+A83E+P111Q+S123P+Q137K+Q147K+V159M+L184M+V251E+S256Q+Q271E+I294E+Q329E+N383E+S402Q+V431E+A491V+K129T, wherein the variant has xylogucanase activity and wherein the variant has at least 60%, e.g., 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%, e.g., at least 96%, at least 97%, at least 98%, or at least 99%, but less than 100%, sequence identity to the polypeptide of SEQ ID NO: 1, SEQ ID NO: 2, or SEQ ID NO: 3.

[0260] In one aspect, the variant comprises alterations at positions corresponding to positions selected from the group consisting of: A83E+P111Q+S123P+Q137K+Q147K+V159M+L184M+D210H+G237M+V251E+S256Q+Q271E+I294E+Q329E+S402Q+E489R+P492D+A129T of the polypeptide of SEQ ID NO: 2, S76E+Q82E+P111Q+S123P+Q137K+Q147K+V159M+L184M+V203T+V251E+S256Q+Q271E+I294E+Q329E+N383Q+S402Q+V431E+A129T, K8E+A41L+A83E+P111Q+S123P+Q137K+Q147K+V159M+L184M+V251E+S256Q+Q271E+I294E+Q329E+K394R+S402Q+V431E+A129T, A83E+P111Q+S123P+Q137K+Q147K+V159M+L184M+V251E+S256Q+Q271E+I294E+Q329E+K347E+N383E+S402Q+V431E+A491V+A129T, K8E+A83E+P111Q+S123P+Q137K+Q147K+V159M+L184M+V251E+S256Q+Q271E+I294E+Q329E+N383E+S402Q+V431E+A491V+A129T, wherein the variant has xylogucanase activity and wherein the variant has at least 60%, e.g., 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%, e.g., at least 96%, at least 97%, at least 98%, or at least 99%, but less than 100%, sequence identity to the polypeptide of SEQ ID NO: 1, SEQ ID NO: 2, or SEQ ID NO: 3.

[0261] In an aspect, the variant comprises alterations at positions corresponding to positions selected from the group consisting of: A83E+P111Q+S123P+Q137K+Q147K+V159M+L184M+V203T+K240F+V251E+S256Q+Q271E+I294E+Q329E+N383E+S402Q+V431E+A491V of the polypeptide of SEQ ID NO: 1, S76E+A83E+P111Q+S123P+Q137K+Q147K+N155D+L184M+G237M+V251E+S256Q+Q271E+I294E+Q329E+N383E+S402Q+V431E+A491V, wherein the variant has xylogucanase activity and wherein the variant has at least 60%, e.g., 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%, e.g., at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to the polypeptide of SEQ ID NO: 1, SEQ ID NO: 2, or SEQ ID NO: 3 but less than 100% sequence identity.

[0262] In an aspect, the variant comprises alterations at positions corresponding to positions selected from the group consisting of: A83E+P111Q+S123P+Q137K+Q147K+V159M+L184M+V203T+K240F+V251E+S256Q+Q271E+I294E+Q329E+N383E+S402Q+V431E+A491V+K129T of the polypeptide of SEQ ID NO: 1, S76E+A83E+P111Q+S123P+Q137K+Q147K+N155D+L184M+G237M+V251E+S256Q+Q271E+I294E+Q329E+N383E+S402Q+V431E+A491V+K129T, wherein the variant has xylogucanase activity and wherein the variant has at least 60%, e.g., 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%, e.g., at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to the polypeptide of SEQ ID NO: 1, SEQ ID NO: 2, or SEQ ID NO: 3 but less than 100% sequence identity.

[0263] In an aspect, the variant comprises alterations at positions corresponding to positions selected from the group consisting of: A83E+P111Q+S123P+Q137K+Q147K+V159M+L184M+V203T+K240F+V251E+S256Q+Q271E+I294E+Q329E+N383E+S402Q+V431E+A491V of the polypeptide of SEQ ID NO: 2, S76E+A83E+P111Q+S123P+Q137K+Q147K+N155D+L184M+G237M+V251E+S256Q+Q271E+I294E+Q329E+N383E+S402Q+V431E+A491V of the polypeptide of SEQ ID NO: 2, wherein the variant has xylogucanase activity and wherein the variant has at least 60%, e.g., 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%, e.g., at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to the polypeptide of SEQ ID NO: 1, SEQ ID NO: 2, or SEQ ID NO: 3 but less than 100% sequence identity.

[0264] In an aspect, the variant comprises alterations or combinations of alterations selected from the group consisting of:

[0265] K394R of the polypeptide of SEQ ID NO: 1,

[0266] S127D,

[0267] R211K,

[0268] S123P,

[0269] K488T,

[0270] S256Q,

[0271] K476R,

[0272] K217R,

[0273] Q271D,

[0274] S214Q,

[0275] L447M,

[0276] K482R,

[0277] K169R,

[0278] L152D,

[0279] R267K,

[0280] L152E,

[0281] D210R,

[0282] L152*,

[0283] R295K,

[0284] S127W,

[0285] E126P,

[0286] S127H,

[0287] T104G,

[0288] Q125K,

[0289] Q125P,

[0290] Q125S,

[0291] D395P,

[0292] G103V,

[0293] T104R,

[0294] Q125L,

[0295] A41E,

[0296] A41R,

[0297] Q125F,

[0298] S127L,

[0299] A226K,

[0300] A41L,

[0301] A226D,

[0302] A118K + S123P,

[0303] N155D,

[0304] Q137K,

[0305] N155E,

[0306] Q147K,

[0307] R276K,

[0308] V203T,

[0309] S94R,

[0310] K18E,

[0311] K252E,

[0312] V219T,

[0313] R267C+T427V,

[0314] Q243E,

[0315] K414E,

[0316] K445E,

[0317] R20K+S123P,

[0318] S123P+K206R,

[0319] R20K+S123P+R211K,

[0320] S123P+K347R,

[0321] S123P+K347R+K353R+D395P,

[0322] S123P+D395P,

[0323] S123P+S127D,

[0324] V159M,

[0325] K392E,

[0326] E489R+P492D,

[0327] Y503L+V505L,

[0328] Y503V+V505L,

[0329] L184M+V219A,

[0330] S123P+R211K+K217R+S256Q+K488T,

[0331] Q82E,

[0332] S76E,

[0333] A83E,

[0334] Q271E,

[0335] S256E,

[0336] I294E,

[0337] Q329E,

[0338] V431E,

[0339] R20K + Y105E + S123P + Q147K + R267K,

[0340] R20K + Y105E + S123P + R267K,

[0341] R20K + S123P + K220R,

[0342] R20K + Y105E + S123P + N136D,

[0343] R20K + S123P + Q137K + Q147K,

[0344] Y105E + A118K + S123P + K206R + K220R + R267K,

[0345] P111Q + S123P + V159M,

[0346] S94R + P111Q + S123P + V159M,

[0347] K87E + P111Q + S123P + V159M + S402Q,

[0348] K87E + P111Q + S123P + V159M,

[0349] P111Q + S123P + V159M + S402Q,

[0350] K87E + P111Q + V159M + I294Q + I473T,

[0351] K8E + P111Q + V159M,

[0352] K8E + K18E + P111Q + V159M + K206E,

[0353] R20K + P111Q + S123P + S127D + V159M,

[0354] P111Q+Q137K+Q147K+V159M+K252E, P111Q+Q137K+Q147K+N155D+K252E, P111Q+Q137K+L152D+V203T+K217R, P111Q+Q137K+V159M, K8E+P111Q+N155D+V159M, A41L+P111Q+S123P+Q147K+V159M+V203T, A41L+P111Q+S123P+Q147K+V159M+K217R, P111Q+S123P+Q137K+Q147K+V159M+S256Q+S402Q, P111Q+S123P+Q147K+V159M+I294Q+S402Q, A41L+P111Q+Q137K+V159M+N168R+Q271D+K488T, P111Q+Q137K+Q147K+V159M+V203T+K217R, P111Q+Q147K+V159M, P111Q+Q137K+Q147K+V159M, P111Q+S123P+Q137K+V159M+K488T, P111Q+S123P+Q137K+V159M+S256Q, K87E+P111Q+L152D+V159M+V203T+I294Q, K87E+P111Q+Q147K+L152D+V159M, R20K+A83E+S123P+K220R+S256E, R20K+S76E+A83E+S123P+K220R+K252E, R20K+A83E+S123P+K220R+K252E, R20K+A42V+S123P+K220R+K252E+I294E, R20K+Q82E+S123P+Q147K+K220R, R20K+A83E+S123P+K220R+S256Q, R20K+Q82E+S123P+N155D+K220R, R20K+S123P+V203T+K220R+K252E, R20K+S123P+K220R+I294E, R20K+A83E+S123P+V203T+K220R+K252E, R20K+A41L+Q82E+S123P+K220R,

[0355] R20K+S123P+N155D+K220R,

[0356] R20K + A42V + S76E + S123P + K220R,

[0357] R20K + S123P + V203T + V219T,

[0358] R20K + A42V + A83E + S123P + K220R,

[0359] R20K + A42V + S76E + S123P + V203T + K220R,

[0360] A83E + P111Q + S123P + V159M + S256E + I294E,

[0361] Q82E + P111Q + S123P + V159M + S256E + I294E,

[0362] Q82E + P111Q + S123P + Q147K + V159M + I294E,

[0363] S76E + Q82E + K87E + P111Q + S123P + V159M + V203T,

[0364] A83E + P111Q + S123P + Q137K + Q147K + V159M + S256Q + S402Q,

[0365] P111Q + S123P + Q137K + Q147K + N155D + S256Q + A289T + N302H,

[0366] S76E + P111Q + S123P + Q137E + Q147K + V159M + K252E + S256Q + S402Q, P111Q + S123P + Q137K + Q147K + V159M + K252E + S256Q + S402Q,

[0367] P111Q + S123P + Q137K + Q147K + N155D + S256Q + I294E + S402Q,

[0368] Q82E + P111Q + S123P + Q137K + Q147K + V159M + L184M + A238S + S256Q + I294E + S402Q,

[0369] A83E + P111Q + S123P + Q137K + Q147K + V159M + L184M + S256Q + I294E + S402Q,

[0370] P111Q + N121 E+ Q137K+ Q147K+ V159M+ K169R+ S256Q+ I294E+ S402Q, P111Q + S123P+ Q137K+ Q147K+ V159M+ L184M+ V219T+ S256Q+ S402Q, P111Q + S123P+ Q137K+ Q147K+ V159M+ L184M+ K252E+ S256Q+ S402Q,

[0371] A83E+ P111Q+ S123P+ Q137K+ Q147K+ V159M+ L184M+ S256Q+ S402Q, Q82E+ P111Q+ S123P+ Q137K+ Q147K+ V159M+ K252E+ S256Q+ S402Q, S76E+ P111Q+ S123P+ Q137K+ Q147K+ V159M+ S256Q+ S402Q,

[0372] K8R+ P111Q+ S123P+ Q137K+ Q147K+ V159M+ S256Q+ S402Q,

[0373] S123P+ S127D+ N136D+ Q137K+ Q147K+ L152E+ N153E+ N155E,

[0374] S123P+ S127D+ N136D+ Q137K+ Q147K+ L152E+ N153E+ N155E+ A491E, R20K+ S123P+ K169R+ K217T+ K240F+ S256Q+ R267H+ I294E,

[0375] K87E+ P111Q+ S123P+ V159M+ K217T+ I294E,

[0376] A83E+ P111Q+ S123P+ V159M+ K240F+ K252E,

[0377] A83E+ P111Q+ S123P+ V159M+ K252E,

[0378] K87E+ P111Q+ S123P+ V159M+ S256Q+ I294E,

[0379] K87E+ P111Q+ S123P+ V159M+ K347E+ N383E,

[0380] Q82E+ P111Q+ S123P+ V159M+ V203T+ G237M+ S256E+ I294E,

[0381] Q82E+P111Q+S123P+N155D+S256E+I294E,

[0382] Q82E+P111Q+S123P+Q137K+V159M+G237M+S256E+I294E,

[0383] Q82E+P111Q+S123P+Q137K+V159M+K240F+S256E+I294E,

[0384] Q82E+P111Q+S123P+Q147K+N155D+K240F+S256E+I294E,

[0385] K8E+Q82E+P111Q+S123P+V159M+S256E+I294E,

[0386] A83E+P111Q+S123P+Q147K+V159M+S256E+I294E+Q329E,

[0387] Q82E+P111Q+S123P+V159M+K240F+S256E I294E,

[0388] Q82E+P111Q+S123P+V159M+A177G+K240F+S256E+I294E+D384G,

[0389] K8R+Q82E+P111Q+S123P+V159M+K169R+K240F+S256E+I294E,

[0390] Q82E+P111Q+S123P+V159M+K169R+S256E+I294E+V431E,

[0391] Q82E+P111Q+S123P+V159M+G237M+V251E+I294E,

[0392] A41L+Q82E+P111Q+S123P+V159M+S256E+I294E+N383E,

[0393] Q82E+P111Q+S123P+V159M+K240F+S256E+I294E,

[0394] Q82E+P111Q+S123P+V159M+K240F+S256E+I294E,

[0395] A83E+P111Q+S123P+Q137K+F146D+Q147G+L148P+V159M+L184M+S256Q+I294E+S402Q、

[0396] A83E+P111Q+S123P+Q137K+Q147K+V159M+L184M+S256Q+I294E+N298D+V300L+S402Q、

[0397] A83E+P111Q+S123P+Q137K+Q147K+V159M+K169R+L184M+K240L+S256Q+I294E+S402Q、

[0398] A83E+P111Q+S123P+Q137K+Q147K+V159M+L184M+G237M+S256Q+I294E+S402Q+K488T、

[0399] A83E+P111Q+S123P+Q137K+Q147K+N155D+L184M+G237M+S256Q+I294E+S402Q、

[0400] A83E+P111Q+S123P+Q137K+Q147K+V159M+L184M+V203T+K240F+S256Q+I294E+S402Q、

[0401] A83E+P111Q+S123P+Q137K+Q147K+V159M+L184M+G237M+S256E+I294E+S402Q、

[0402] A83E+P111Q+S123P+Q137K+Q147K+V159M+L184M+K240F+S256Q+I294E+S402Q+E489R、

[0403] A83E+P111Q+S123P+Q137K+Q147K+V159M+F165H+L184M+S256Q+I294E+S402Q+V431E、

[0404] A83E+P111Q+S123P+Q137K+Q147K+V159M+L184M+V251E+Q271E+I294E+Q329E+S402Q、

[0405] A83E+P111Q+S123P+Q137K+Q147K+V159M+L184M+V251E+S256Q+Q271E+I294E+Q329E+S402Q,

[0406] A83E+P111Q+S123P+Q137K+Q147K+N155D+L184M+S256Q+I294E+S402Q,

[0407] R20K+S123P+Q147K+N155D+K169R+K217T+K240F+S256Q+R267H+I294E,

[0408] R20K+S123P+Q137K+K169R+K217T+K240F+S256Q+R267H+I294E,

[0409] R20K+S123P+N155D+K169R+K217T+K240F+S256Q+R267H+I294E+D384G+E489R,

[0410] R20K+S123P+N155D+K169R+K217T+K240F+S256Q+R267H+I294E+S474E,

[0411] R20K+S123P+N155D+K169R+K217T+K240F+S256Q+R267H+I294E,

[0412] R20K+S123P+Q147K+K169R+V219T+K240F+S256Q+R267H+I294E,

[0413] R20K+A41L+S123P+L152P+K169R+K217T+K240F+S256Q+R267H+I294E,

[0414] K8R+R20K+S123P+K169R+D210H+K217T+K240F+S256Q+R267H+I294E,

[0415] R20K+S123P+K169R+K217T+K240F+S256Q+R267H+I294E+Q329E+V431E+E489R,

[0416] P111Q + S123P + Q137K + Q147K + V159M + K169R + D210H + K240F + S256E + I294E + S402Q + K488T,

[0417] P111Q + S123P + Q137K + Q147K + V159M + K169R + K240F + S256Q + I294E + K322E + S402Q,

[0418] P111Q + S123P + Q137K + Q147K + V159M + K169R + K240F + S256Q + I294E + K322E + S402Q,

[0419] S76E + P111Q + S123P + Q137K + Q147K + V159M + K169R + G237M + S256Q + I294E + S402Q,

[0420] S76E + P111Q + S123P + Q137K + Q147K + V159M + K169R + V203T + G237M + S256Q + I294E + S402Q,

[0421] P111Q + S123P + Q137K + Q147K + V159M + K169R + G237M + S256Q + I294E + P339S + S402Q + K488T,

[0422] P111Q + S123P + Q137K + Q147K + V159M + K169R + G237M + T244R + S256Q + I294E + S402Q,

[0423] P111Q + S123P + Q137K + Q147K + V159M + G237M + T244R + S256Q + I294E + S402Q,

[0424] P111Q + S123P + Q137K + Q147K + V159M + K169R + V219T + K240F + S256Q + I294E + S402Q,

[0425] P111Q + S123P + Q137K + Q147K + V159M + K169R + V203T + S256Q + I294E + S402Q,

[0426] Q82E + P111Q + S123P + Q137K + V159M + V203T + D210H + G237M + S256E + I294E,

[0427] Q82E+P111Q+S123P+V159M+V203T+G237M+K252E+S256E+I294E,

[0428] S76E+Q82E+P111Q+S123P+V159M+V203T+G237M+S256E+I294E,

[0429] Q82E+P111Q+S123P+Q147K+V159M+V203T+G237M+S256E+I294E,

[0430] S76E+Q82E+P111Q+S123P+V159M+V203T+G237M+S256E+I294E+S474E+E489R+P492D,

[0431] Q82E+P111Q+S123P+V159M+K169R+V203T+G237M+T244R+S256E+I294E,

[0432] Q82E+P111Q+S123P+V159M+V203T+G237M+T244R+S256E+I294E,

[0433] S76E+A83E+P111Q+S123P+Q137K+Q147K+V159M+L184M+V251E+S256Q+Q271E+I294E+Q329E+S402Q,

[0434] A83E+P111Q+S123P+Q137K+Q147K+V159M+L184M+V219T+G237M+V251E+S256Q+Q271E+I294E+Q329E+S402Q,

[0435] A83E+P111Q+S123P+Q137K+Q147K+V159M+K169R+L184M+K252E+S256Q+Q271E+I294E+Q329E+S402Q,

[0436] S76E+A83E+P111Q+S123P+Q137K+Q147K+V159M+L184M+V203T+A238S+S256E+Q271E+I294E+Q329E+S402Q,

[0437] A83E+P111Q+S123P+Q137K+Q147K+V159M+L184M+V203T+A238S+S256E+Q271E+I294E+Q329E+S402Q、

[0438] A83E+P111Q+S123P+Q137K+Q147K+V159M+L184M+G237M+V251E+S256Q+Q271E+I294E+Q329E+S402Q+E489R+V505L、

[0439] A83E+P111Q+S123P+Q137K+Q147K+V159M+L184M+D210H+G237M+V251E+S256Q+Q271E+I294E+Q329E+S402Q+E489R+P492D、

[0440] A83E+P111Q+S123P+Q137K+Q147K+V159M+L184M+D210H+T244R+K252E+S256Q+Q271E+I294E+Q329E+S402Q、

[0441] A83E+P111Q+S123P+Q137K+Q147K+V159M+L184M+V251E+S256Q+Q271E+I294E+Q329E+S402Q+V431E、

[0442] A83E+P111Q+S123P+Q137K+Q147K+V159M+L184M+T244E+V251E+S256Q+Q271E+I294E+Q329E+S402Q+V431E、

[0443] A83E+P111Q+S123P+Q137K+Q147K+V159M+L184M+V251E+S256Q+Q271E+I294E+Q329E+K347E+S402Q、

[0444] K8E+A83E+S94R+P111Q+S123P+Q137K+Q147K+V159M+L184M+V251E+S256Q+Q271E+I294E+Q329E+S402Q、

[0445] A83E+P111Q+S123P+Q137K+L152P+V159M+L184M+V251E+S256Q+Q271E+I294E+Q329E+S402Q+V431E、

[0446] A83E+P111Q+S123P+Q137K+Q147K+V159M+L184M+V251E+S256Q+Q271E+I294E+Q329E+S402Q+V431E+K445E、

[0447] A83E+P111Q+S123P+Q137K+Q147K+V159M+L184M+K252E+S256Q+Q271E+I294E+Q329E+S402Q、

[0448] K8E+A83E+P111Q+S123P+Q137K+Q147K+V159M+L184M+V251E+S256Q+Q271E+I294E+Q329E+S402Q、

[0449] A83E+P111Q+S123P+Q137K+Q147K+V159M+L184M+D210H+K240F+V251E+S256Q+Q271E+I294E+Q329E+S402Q、

[0450] A83E+P111Q+S123P+Q137K+Q147K+N155D+L184M+G237M+V251E+S256Q+Q271E+I294E+Q329E+S402Q、

[0451] A41L+A83E+P111Q+S123P+Q137K+Q147K+V159M+L184M+K240F+V251E+S256Q+Q271E+I294E+Q329E+S402Q、

[0452] Q82E+P111Q+S123P+Q137K+Q147K+V159M+L184M+V203T+D210H+V251E+S256Q+Q271E+I294E+Q329E+S402Q、

[0453] A41L+A83E+P111Q+S123P+Q137K+Q147K+V159M+L184M+V251E+S256Q+Q271E+I294E+Q329E+S402Q、

[0454] A41L+A83E+P111Q+S123P+Q137K+Q147K+V159M+L184M+V251E+S256Q+Q271E+I294E+Q329E+N383Q+S402Q、

[0455] P111Q + S123P + Q137K + Q147K + V159M + V251E + S256E + Q271E + I294E + Q329E + S402Q,

[0456] P111Q + S123P + Q137K + Q147K + V159M + A238T + V251E + S256Q + Q271E + I294E + Q329E + S402Q,

[0457] A83E + P111Q + S123P + Q137K + Q147K + V159M + L184M + V251E + S256Q + Q271E + I294E + Q329E + N383Q + S402Q,

[0458] A83E + P111Q + S123P + Q137K + Q147K + V159M + L184M + T244R + W248V + V251E + S256Q + Q271E + I294E + Q329E + S402Q,

[0459] A83E + P111Q + S123P + Q137K + Q147K + V159M + L184M + V203T + T244R + V251E + S256Q + Q271E + I294E + Q329E + S402Q,

[0460] A83E + P111Q + S123P + Q137K + Q147K + V159M + L184M + V251E + S256Q + Q271E + R295K + N298D + Q329E + S402Q + L447M,

[0461] A83E + P111Q + S123P + Q137K + Q147K + V159M + L184M + S256E + Q271D + I294E + Q329E + S402Q,

[0462] Q82E + P111Q + S123P + V159M + K169R + V203T + G237M + T244R + S256E + Q271E + I294E + Q329E + N383E,

[0463] Q82E + P111Q + S123P + Q137K + V159M + K169R + A189G + V203T + G237M + T244R + S256E + I294E,

[0464] Q82E+P111Q+S123P+Q137K+V159M+K169R+V203T+G237M+T244R+V251 E+S256E+I294E,

[0465] Q82E+P111Q+S123P+L152P+V159M+K169R+V203T+G237M+T244R+S256E+I294E,

[0466] Q82E+P111Q+S123P+V159M+K169R+V203T+G237M+T244R+S256E+I294E+N383Q+V431E,

[0467] R20K+S76E+Q82E+P111Q+S123P+V159M+K169R+V203T+G237M+T244R+S256E+I294E+Q329E+P492D,

[0468] S76E+Q82E+P111Q+S123P+V159M+K169R+V203T+G237M+T244R+S256E+I294E+Q329E+E489R+P492D,

[0469] Q82E+P111Q+S123P+Q137K+N155D+V159M+K169R+V203T+G237M+T244R+S256E+I294E+K445E,

[0470] S76E+Q82E+S94R+P111Q+S123P+V159M+K169R+V203T+G237M+T244R+S256E+I294E,

[0471] K8E+A41L+S76E+Q82E+P111Q+S123P+V159M+K169R+V203T+G237M+T244R+S256E+I294E,

[0472] A83E+P111Q+S123P+Q137K+Q147K+V159M+L184M+S256E+Q271E+I294E+Q329E+P339S+N383E+S402Q+V431E,

[0473] S76E+Q82E+P111Q+S123P+Q137K+Q147K+V159M+L184M+V203T+V251E+S256Q+Q271E+I294E+Q329E+N383Q+S402Q+V431E,

[0474] A83E+P111Q+S123P+Q137K+Q147K+V159M+L184M+K252E+S256Q+Q271E+I294E+Q329E+N383E+S402Q+V431E+A491V,

[0475] A83E+P111Q+S123P+Q137K+Q147K+V159M+L184M+V203T+V251E+S256Q+Q271E+I294E+Q329E+N383Q+S402Q+V431E,

[0476] A83E+P111Q+S123P+Q137K+Q147K+V159M+L184M+V203T+K240F+V251E+S256Q+Q271E+I294E+Q329E+N383E+S402Q+V431E+A491V,

[0477] A83E+P111Q+S123P+Q137K+Q147K+V159M+L184M+V219T+G237M+S256Q+Q271E+I294E+Q329E+S402Q+V431E+S474E,

[0478] A83E+P111Q+S123P+Q137K+Q147K+V159M+L184M+V251E+S256Q+Q271E+I294E+Q329E+N383E+S402Q+V431E+P492D,

[0479] S76E+A83E+P111Q+S123P+Q137K+Q147K+N155D+L184M+G237M+V251E+S256Q+Q271E+I294E+Q329E+N383E+S402Q+V431E+A491V,

[0480] A83E+P111Q+S123P+Q137K+Q147K+V159M+L184M+A189G+V251E+S256Q+Q271E+I294E+Q329E+N383Q+S402Q+V431E,

[0481] K8E+A41 L+A83E+P111Q+S123P+Q137K+Q147K+V159M+L184M+V251E+S256Q+Q271E+I294E+Q329E+K394R+S402Q+V431E,

[0482] A83E+P111Q+S123P+Q137K+Q147K+V159M+L184M+V251E+S256Q+Q271D+I294E+Q329E+N383E+S402Q+V431E+A459P,

[0483] A83E+P111Q+S123P+Q137K+Q147K+V159M+L184M+V251E+S256Q+Q271E+I294E+Q329E+K347E+N383E+S402Q+V431E+A491V,

[0484] K8E+A83E+P111Q+S123P+Q137K+Q147K+V159M+L184M+V251E+S256Q+Q271E+I294E+Q329E+N383E+S402Q+V431E+A491V,

[0485] K8R+A83E+P111Q+S123P+Q137K+Q147K+V159M+L184M+V251E+S256Q+Q271E+I294E+Q329E+S402Q+V431E+E489K,

[0486] A83E+P111Q+S123P+Q137K+Q147K+V159M+L184M+V251E+S256Q+Q271E+I294E+Q329E+N383E+S402Q+V431E+A491V, wherein the variant has xylogucanase activity and wherein the variant has at least 60%, e.g., 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%, e.g., at least 96%, at least 97%, at least 98%, or at least 99% sequence identity, but less than 100% sequence identity, to the polypeptide of SEQ ID NO: 1, SEQ ID NO: 2, or SEQ ID NO: 3.

[0487] The variant can further comprise a substitution at one or more other positions, e.g., those described in WO 2009 / 147210.

[0488] Amino acid changes can be of a minor nature, that is conservative amino acid substitutions or insertions that do not significantly affect the folding and / or activity of the protein; small deletions, typically of 1-30 amino acids; small amino- or carboxyl-terminal extensions, such as an amino-terminal methionine residue; a small linker peptide of up to 20-25 residues; or a small extension that facilitates purification by changing net charge or another function, such as a poly-histidine tract, an antigenic epitope or a binding domain.

[0489] Examples of conservative substitutions are within the group of basic amino acids (arginine, lysine and histidine), acidic amino acids (glutamic acid and aspartic acid), polar amino acids (glutamine and asparagine), hydrophobic amino acids (leucine, isoleucine and valine), aromatic amino acids (phenylalanine, tryptophan and tyrosine), and small amino acids (glycine, alanine, serine, threonine and methionine). Amino acid substitutions that do not generally alter specific activity are known in the art and are described, for example, by H. Neurath and R.L. Hill, 1979, In, The Proteins, Academic Press, New York. Common substitutions are Ala / Ser, Val / lle, Asp / Glu, Thr / Ser, Ala / Gly, Ala / Thr, Ser / Asn, Ala / Val, Ser / Gly, Tyr / Phe, Ala / Pro, Lys / Arg, Asp / Asn, Leu / lle, Leu / Val, Ala / Glu, and Asp / Gly.

[0490] Alternatively, these amino acid changes are of a such nature that they lead to a change in the physico-chemical properties of the polypeptide. For example, the amino acid changes can improve the thermal stability of the polypeptide, change the substrate specificity, change the pH optimum, etc.

[0491] Essential amino acids in a polypeptide can be identified according to procedures known in the art, such as site-directed mutagenesis or alanine scanning mutagenesis (Cunningham and Wells, 1989, Science 244: 1081-1085). In the latter technique, single alanine mutations are introduced at every residue in the molecule, and the resultant mutant molecules are tested for xylanase activity to identify amino acid residues that are critical to the activity of the molecule. See also, Hilton et al., 1996, J. Biol. Chem. 271 : 4699-4708. The active site of an enzyme or other biological interaction can also be determined by physical analysis of structure, as determined by such techniques as nuclear magnetic resonance, crystallography, electron diffraction, or photoaffinity labeling, in conjunction with mutation of putative contact site amino acids. See, for example, de Vos et al., 1992, Science 255: 306-312; Smith et al., 1992, J. Mol. Biol. 224: 899-904; Wlodaver et al., 1992, FEBS Lett. 309: 59-64. The identity of amino acids that are essential for the activity of the related polypeptide can also be inferred from an alignment with a related polypeptide and are described, for example, in WO 2009 / 147210.

[0492] The variants can consist of 445 to 524 amino acids, such as 471 to 524 amino acids, 497 to 524 amino acids.

[0493] In embodiments, the variant has improved wash performance compared to the parent enzyme.

[0494] In embodiments, the variant has improved enzyme wash benefit compared to the parent enzyme.

[0495] In embodiments, the variant has improved stability compared to the parent enzyme.

[0496] In embodiments, the variant has improved thermostability compared to the parent enzyme.

[0497] In embodiments, the variant has improved storage stability in detergent compared to the parent enzyme.

[0498] In embodiments, the variant has improved whiteness compared to the parent enzyme.

[0499] Parent xylanase

[0500] The parent xylanase can be a polypeptide having at least 60% sequence identity to the polypeptide of SEQ ID NO: 1.

[0501] In one aspect, a parent has at least 60%, e.g., 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%, at least 99%, or 100% sequence identity to the polypeptide of SEQ ID NO: 1, and the parent has xylogucanase activity. In one aspect, the amino acid sequence of the parent differs by up to 10 amino acids, e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10, from the polypeptide of SEQ ID NO: 1. In another aspect, the parent comprises or consists of the amino acid sequence of SEQ ID NO: 1. In another aspect, the parent is a fragment of the polypeptide of SEQ ID NO: 1 containing at least 445 amino acid residues, e.g., at least 471 amino acid residues, and at least 497 amino acid residues.

[0502] In one aspect, a parent has at least 60%, e.g., 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%, at least 99%, or 100% sequence identity to the polypeptide of SEQ ID NO: 2, and the parent has xylogucanase activity. In one aspect, the amino acid sequence of the parent differs by up to 10 amino acids, e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10, from the polypeptide of SEQ ID NO: 2. In another aspect, the parent comprises or consists of the amino acid sequence of SEQ ID NO: 2. In another aspect, the parent is a fragment of the polypeptide of SEQ ID NO: 2 containing at least 445 amino acid residues, e.g., at least 471 amino acid residues, and at least 497 amino acid residues.

[0503] In one aspect, the parent polypeptide has at least 60%, e.g., 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%, at least 99%, or 100% sequence identity to the polypeptide of SEQ ID NO: 3, the parent having xylogucanase activity. In one aspect, the amino acid sequence of the parent differs by up to 10 amino acids, e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10, from the polypeptide of SEQ ID NO: 3. In another aspect, the parent comprises or consists of the amino acid sequence of SEQ ID NO: 3. In another aspect, the parent is a fragment of the polypeptide of SEQ ID NO: 3 containing at least 445 amino acid residues, e.g., at least 471 amino acid residues, and at least 497 amino acid residues.

[0504] The parent polypeptide can be a hybrid polypeptide in which a region of one polypeptide is fused at the N- or C-terminus of another polypeptide.

[0505] The parent can be a fusion polypeptide or a cleavable fusion polypeptide in which another polypeptide is fused at the N- or C-terminus of a polypeptide of the application. Fusion polypeptides are produced by fusing a polynucleotide encoding another polypeptide to a polynucleotide of the application. Techniques for producing fusion polypeptides are known in the art and include joining coding sequences that encode polypeptides such that they are in reading frame, and the expression of the fusion polypeptide is under the control of one or more of the same promoters and terminators. Fusion polypeptides can also be constructed using intein technology, in which the fusion polypeptide is produced post-translationally (Cooper et al., 1993, EMBO J. 12:2575-2583; Dawson et al., 1994, Science 266:776-779).

[0506] The fusion polypeptide can further comprise a cleavage site between the two polypeptides. The site is cleaved when the fusion protein is secreted, thereby releasing the two polypeptides. Examples of cleavage sites include, but are not limited to, those disclosed in Martin et al., 2003, J. Ind. Microbiol. Biotechnol. 3:568-576; Svetina et al., 2000, J. Biotechnol. 76:245-251; Rasmussen-Wilson et al., 1997, Appl. Environ. Microbiol. 63:3488-3493; Ward et al., 1995, Biotechnology 13:498-503; and Contreras et al., 1991, Biotechnology 9:378-381; Eaton et al., 1986, Biochemistry 25:505-512; Collins-Racie et al., 1995, Biotechnology 13:982-987; Carter et al., 1989, Proteins: Structure, Function, and Genetics 6:240-248; and Stevens, 2003, Drug Discovery World 4:35-48.

[0507] The parent can be obtained from a microorganism of any genus. For the purposes of the present invention, the term "obtained from" as used herein in connection with a given source shall mean that the parent polypeptide is produced by the source or is isolated from a member of the source. In one aspect, the parent is excreted extracellularly.

[0508] In another aspect, the parent xyloglucanase can be a bacterial xyloglucanase. For example, the xyloglucanase can be a gram-positive bacterial polypeptide, such as a Bacillus, preferably from the Bacilli / Lactobacilll subphylum, preferably from a species of Paenibacillus, especially Paenibacillus polymyxa, such as Paenibacillus polymyxa ATCC 832, preferably the xyloglucanase is a family 44 xyloglucanase, such as described in WO 01 / 62903.

[0509] It is understood that for the foregoing species, the present application encompasses both the complete and incomplete stages as well as other taxonomic equivalents, e.g., anamorphs, regardless of their known species names. Those of ordinary skill in the art will readily identify appropriate equivalents.

[0510] Strains of these species are readily available to the public in a number of culture collections, such as the American Type Culture Collection (ATCC), the Deutsche Sammlung von Mikroorganismen und Zellkulturen GmbH (DSMZ), the Centraalbureau Voor Schimmelcultures (CBS), and the Agricultural Research Service Patent Culture Collection, Northern Regional Research Center (NRRL).

[0511] The above probes can be used to identify and obtain the parent from other sources, including microorganisms isolated from nature (e.g., soil, compost, water, etc.) or DNA samples obtained directly from natural material (e.g., soil, compost, water, etc.). Techniques for isolating microorganisms and DNA directly from natural habitats are known in the art. A genomic DNA or cDNA library of another microorganism or a mixture of DNA from other sources can then be similarly screened to detect and isolate the polynucleotide encoding the parent. Once the polynucleotide encoding the parent has been detected using one or more probes, the polynucleotide can be isolated or cloned by using techniques known to those of ordinary skill in the art (see, e.g., Sambrook et al., 1989, supra).

[0512] Preparation of variants

[0513] The present invention also relates to methods for obtaining variants having xyloglucanase activity, the methods comprising: (a) introducing alterations into a parent xyloglucanase at one or more positions corresponding to positions selected from the group consisting of: 111, 123, 159, 256, 294, 8, 18, 20, 41, 42, 76, 82, 83, 87, 94, 103, 104, 105, 121, 125, 126, 127, 136, 137, 146, 147, 148, 152, 153, 155, 165, 168, 169, 177, 184, 189, 203, 206, 210, 211, 214, 217, 219, 220, 226, 237, 238, 240, 243, 244, 248, 251, 252, 267, 271, 276, 289, 295, 298, 300, 302, 322, 329, 339, 347, 347, 353, 383, 384, 392, 394, 395, 402, 414, 427, 431, 445, 447, 459, 473, 474, 476, 482, 488, 489, 491, 492, 503, and 505 of the polypeptide of SEQ ID NO: 1, wherein the variant has xyloglucanase activity; and (b) recovering the variant.

[0514] Variants can be prepared using any mutagenesis procedure known in the art, such as site-directed mutagenesis, synthetic gene construction, semi-synthetic gene construction, random mutagenesis, shuffling, and the like.

[0515] Site-directed mutagenesis is a technique that introduces one or more (e.g., several) mutations at one or more defined sites in a polynucleotide encoding the parent.

[0516] Site-directed mutagenesis can be accomplished in vitro by procedures involving PCR using oligonucleotide primers containing the desired mutations. Site-directed mutagenesis can also be performed in vitro by cassette mutagenesis, which involves cleavage by restriction enzymes at sites in a plasmid containing a polynucleotide encoding the parent and subsequent ligation of an oligonucleotide containing the mutation into the polynucleotide. Typically, the restriction enzymes that digest the plasmid and the oligonucleotide are the same, allowing the cohesive ends of the plasmid and the insert to ligate to each other. See, e.g., Scherer and Davis, 1979, Proc. Natl. Acad. Sci. USA 76:4949-4955; and Barton et al., 1990, Nucleic Acids Res. 18:7349-4966.

[0517] Site-directed mutagenesis can also be accomplished in vivo by methods known in the art. See, e.g., U.S. Patent Application Publication No. 2004 / 0171154; Storici et al., 2001, Nature Biotechnol. 19:773-776; Kren et al., 1998, Nat. Med. 4:285-290; and Calissano and Macino, 1996, Fungal Genet. Newslett. 43:15-16.

[0518] Any site-directed mutagenesis procedure can be used in the present application. There are many commercially available kits available for making variants.

[0519] Synthetic gene construction requires in vitro synthesis of a designed polynucleotide molecule to encode a polypeptide of interest. Gene synthesis can be performed using a variety of techniques, such as the multiplex microchip-based technology described by Tian et al. (2004, Nature 432:1050-1054), and similar technologies in which oligonucleotides are synthesized and assembled on a photo-programmable microfluidic chip.

[0520] Single or multiple amino acid substitutions, deletions, and / or insertions can be made and tested using known methods of mutagenesis, recombination, and / or shuffling, followed by appropriate screening procedures. Such procedures are known in the art, see, e.g., Reidhaar-Olson and Sauer, 1988, Science 241 :53-57; Bowie and Sauer, 1989, Proc. Natl. Acad. Sci. USA 86:2152-2156; WO 95 / 17413; or WO 95 / 22625. Other methods that can be used include error-prone PCR, phage display (e.g., Lowman et al., 1991, Biochemistry 30:10832-10837; U.S. Patent No. 5,223,409; WO 92 / 06204), and region-directed mutagenesis (Derbyshire et al., 1986, Gene 46:145; Ner et al., 1988, DNA 7:127).

[0521] Mutagenesis / shuffling methods can be combined with high-throughput, automated screening methods to detect the activities of cloned, mutagenized polypeptides expressed by host cells (Ness et al., 1999, Nature Biotechnology 17:893-896). Mutagenized DNA molecules that encode active polypeptides can be recovered from the host cells and rapidly sequenced using standard methods in the art. These methods allow for the rapid determination of the importance of each amino acid residue in a polypeptide.

[0522] Semi-synthetic gene construction is achieved by combining aspects of synthetic gene construction, and / or site-directed mutagenesis, and / or random mutagenesis, and / or shuffling. Semi-synthetic construction is typically a process that utilizes synthetic polynucleotide fragments in combination with PCR techniques. Thus, defined regions of a gene can be synthesized de novo, while other regions can be amplified using site-specific mutagenic primers, while still other regions can be subjected to error-prone or non-error prone PCR amplification. The polynucleotide subsequences can then be shuffled.

[0523] Polynucleotide

[0524] The present application also relates to polynucleotides encoding the variants of the present application.

[0525] Nucleic acid construct

[0526] The present application also relates to nucleic acid constructs comprising a polynucleotide encoding the variants of the present application operably linked to one or more control sequences that direct the expression of the coding sequence in a suitable host cell under conditions compatible with the control sequences.

[0527] The polynucleotide can be manipulated in a variety of ways to provide for the expression of the variants. Depending on the expression vector utilized, the polynucleotide can be manipulated either before or after insertion into the vector. Techniques for manipulating polynucleotides using recombinant DNA methods are well known in the art.

[0528] The control sequence can be a promoter, a polynucleotide which is recognized by a host cell for expression of a polynucleotide. The promoter comprises a transcription control sequence that mediates the transcription of a variant. The promoter can be any polynucleotide that shows transcriptional activity in the host cell including mutant, truncated, and hybrid promoters, and can be obtained from genes encoding proteins or from non-coding segments of DNA. The control sequence can be an operator, a site at which a repressor can bind to prevent transcription of a polynucleotide.

[0529] Examples of suitable promoters for directing the transcription of the nucleic acid constructs of the application in a bacterial host cell are the promoters obtained from the Bacillus amyloliquefaciens alpha-amylase gene (amyQ), Bacillus licheniformis alpha-amylase gene (amyL), Bacillus licheniformis penicillinase gene (penP), Bacillus stearothermophilus maltogenic amylase gene (amyM), Bacillus subtilis levan sucrose gene (sacB), Bacillus subtilis xylA and xylB genes, a Bacillus thuringiensis crylllA gene (Agaisse and Lereclus, 1994, Molecular Microbiology 13: 97-107), E. coli lac, lldA and trp, the E. coli trc promoter (Egon et al., 1988, Gene 69: 301-315), the Streptomyces coelicolor agarase gene (dagA), and the prokaryotic beta-lactamase gene (Villa-Kamaroff et al., 1978, Proc. Natl. Acad. Sci. USA 75: 3727-3731), as well as the tac promoter (DeBoer et al., 1983, Proc. Natl. Acad. Sci. USA 80: 21-25). Further promoters are described in Gilbert et al., 1980, Scientific American 242: 74-94; and in Sambrook et al., 1989, supra. Examples of tandem promoters are disclosed in WO 99 / 43835.

[0530] Examples of suitable promoters for directing the transcription of the nucleic acid constructs of the present application in a filamentous fungal host cell are promoters obtained from the genes for Aspergillus nidulans acetamidase, Aspergillus niger neutral alpha-amylase, Aspergillus niger acid stable alpha-amylase, Aspergillus niger or Aspergillus awamori glucoamylase (glaA), Aspergillus oryzae TAKA amylase, Aspergillus oryzae alkaline protease, Aspergillus oryzae triose phosphate isomerase, Fusarium oxysporum trypsin-like protease (WO 96 / 00787), Fusarium venenatum amyloglucosidase (WO 00 / 56900), Fusarium venenatum Daria (WO 00 / 56900), Fusarium venenatum Quinn (WO 00 / 56900), Rhizomucor miehei lipase, Rhizomucor miehei aspartic proteinase, Trichoderma reesei beta-glucosidase, Trichoderma reesei cellobiohydrolase I, Trichoderma reesei cellobiohydrolase II, Trichoderma reesei endoxylanase I, Trichoderma reesei endoxylanase II, Trichoderma reesei endoxylanase III, Trichoderma reesei endoxylanase IV, Trichoderma reesei endoxylanase V, Trichoderma reesei xylanase I, Trichoderma reesei xylanase II, Trichoderma reesei beta-xylosidase, as well as the NA2-tpi promoter (a modified version of the Aspergillus neutral alpha-amylase gene, wherein the untranslated leader sequence of the Aspergillus triose phosphate isomerase gene has replaced the untranslated leader sequence), and mutant, truncated, and hybrid promoters thereof. Non-limiting examples of genes are described in U.S. Patent No. 6, view transformations 677, 6,677, 375; and WO 00 / 07088.

[0531] In yeast host cells, useful promoters include promoters from the gene for Saccharomyces cerevisiae enolase (ENO-1), Saccharomyces cerevisiae galactokinase (GAL1), Saccharomyces cerevisiae alcohol dehydrogenase / glyceraldehyde-3-phosphate dehydrogenase (ADH1, ADH2 / GAP), Saccharomyces cerevisiae triose phosphate isomerase (TPI), Saccharomyces cerevisiae metallothionein (CUP1), and Saccharomyces cerevisiae 3-phosphoglycerate kinase. Other useful promoters of yeast host cells are described by Romanos et al., 1992, Yeast 8:423-488.

[0532] The control sequence can also be a transcription terminator, a transcription terminator region that is recognized by a host cell for termination of transcription. The terminator sequence is operably linked to the 3'-terminus of the polynucleotide encoding the variant. Any terminator that is functional in the host cell can be used.

[0533] Preferred terminators for bacterial host cells are obtained from the genes for Bacillus clausii alkaline protease (aprH), Bacillus licheniformis alpha-amylase (amyL), and Escherichia coli ribosomal RNA (rrnB).

[0534] Preferred terminators for fungal host cells are obtained from the genes for Aspergillus nidulans anthranilate synthase, Aspergillus niger glucoamylase, Aspergillus niger alpha- glucosidase, Aspergillus oryzae TAKA amylase, and Aspergillus awamori trypsin-like protease.

[0535] Preferred terminators for yeast host cells are obtained from the genes for Saccharomyces cerevisiae enolase, Saccharomyces cerevisiae cytochrome C (CYC1), and Saccharomyces

[0536] The control sequence can also be an mRNA stabilizer region downstream of the promoter and upstream of the coding sequence of a gene which increases expression of the gene.

[0537] Examples of appropriate mRNA stabilizer regions are obtained from the 3' untranslated region of Bacillus thuringiensis crylllIA gene (WO94 / 25610) and Bacillus subtilis SP82 gene (Hue et al., 1995, Journal of Bacteriology 177: 3465-3471).

[0538] The control sequence can also be a leader sequence, a nontranslated region of an mRNA that is important for translation by the host cell. The leader sequence is operably linked to the 5'-terminus of the polynucleotide encoding the variant. Any leader sequence that is functional in the host cell can be used.

[0539] Preferred leader sequences for filamentous fungal host cells are obtained from the genes for Aspergillus niger glucoamylase, Aspergillus niger alpha-glucosidase, Aspergillus oryzae TAKA amylase, and Aspergillus awamori trypsin-like protease.

[0540] Suitable leader sequences for yeast host cells are obtained from the genes for Saccharomyces cerevisiae enolase (ENO-l), Saccharomyces cerevisiae 3-phosphoglycerate kinase, Saccharomyces cerevisiae alpha-factor, and Saccharomyces cerevisiae alcohol dehydrogenase / glyceraldehyde-3-phosphate dehydrogenase (ADH2 / GAP).

[0541] The control sequence can also be a polyadenylation sequence, a sequence operably linked to the 3'-terminus of the variant coding sequence and which, when transcribed, is recognized by the host cell as a signal to add polyadenosine residues to transcribed mRNA. Any polyadenylation sequence that is functional in the host cell can be used.

[0542] Preferred polyadenylation sequences for filamentous fungal host cells are obtained from the genes for Aspergillus nidulans anthranilate synthase, Aspergillus niger glucoamylase, Aspergillus niger alpha-glucosidase, Aspergillus oryzae TAKA amylase, and Fusarium oxysporum trypsin-like protease.

[0543] Useful polyadenylation sequences for yeast host cells are described by Guo and Sherman, 1995, Mol. Cellular Biol. 15:5983-5990.

[0544] The control sequence can also be a signal peptide coding region that codes for an amino acid signal peptide linked to the N-terminus of a variant and directs secretion of the variant from a host cell into which it is expressed. The 5'-end of the coding sequence of the polynucleotide can inherently contain a signal peptide coding region naturally linked in translation reading frame to the segment of the coding sequence encoding the variant. Alternatively, the 5'-end of the coding sequence can contain a signal peptide coding region that is foreign to the coding sequence. A foreign signal peptide coding region can be required, for example, when the coding sequence is originally derived from a gene that does not naturally produce a secreted product. Alternatively, a foreign signal peptide coding region can simply be used to enhance secretion of the variant. However, any signal peptide coding region that directs the expressed variant into the secretory pathway of a host cell can be used.

[0545] Useful signal peptide coding regions for bacterial host cells are the signal peptide coding regions obtained from the genes for Bacillus NCIB 11837 maltogenic amylase, Bacillus licheniformis subtilisin, Bacillus licheniformis beta-lactamase, Bacillus stearothermophilus alpha-amylase, Bacillus stearothermophilus neutral proteases (nprT, nprS, nprM), and Bacillus subtilis prsA. Further signal peptides are described by Simonen and Palva, 1993, Microbiological Reviews 57: 109-137.

[0546] Useful signal peptide coding sequences for filamentous fungal host cells are the signal peptide coding sequences obtained from the genes for Aspergillus niger neutral amylase, Aspergillus niger glucoamylase, Aspergillus oryzae TAKA amylase, Humicola insolens cellulase, Humicola insolens endoglucanase V, Humicola lanuginosa lipase, and Rhizomucor miehei aspartic proteinase.

[0547] Useful signal peptides for yeast host cells are obtained from the genes for Saccharomyces cerevisiae alpha-factor and Saccharomyces cerevisiae invertase. Other useful signal peptide coding sequences are described by Romanos et al. (1992, supra).

[0548] The control sequence can also be a presequence coding sequence as set forth in WO 88 / 04692. The inclusion of a presequence coding sequence will result in the formation of a precursor protein which contains the variant. The presequence coding sequence of choice will direct the expression of the precursor protein into the periplasm of the cell. The actual signal peptide of choice will depend on the host cell selected and possibly its origin.

[0549] Where both a signal peptide sequence and a presequence sequence are present, the presequence sequence is positioned immediately N-terminal to the variant and the signal peptide sequence is positioned immediately N-terminal to the presequence sequence.

[0550] It can also be desirable to add a regulatory sequence that regulates expression of the variant relative to the growth of the host cell. Examples of regulatory systems are those that respond to a chemical or physical stimulus including the presence of a regulatory compound. Regulatory systems in prokaryotic systems include the lac, tac, and trp operator systems. In yeast, the ADH2 system or GAL1 system can be used. In filamentous fungi, the Aspergillus niger glucoamylase promoter, Aspergillus oryzae TAKA amylase promoter, and Aspergillus niger beta-glucosidase promoter can be used. Other examples of regulatory sequences are those that allow for

[0551] Expression Vectors

[0552] The present application also relates to recombinant expression vectors comprising a polynucleotide encoding a variant of the present application, a promoter, and transcriptional and translational stop signals. The various nucleotide and control sequences can be joined together to produce the recombinant expression vectors, which can include one or more convenient restriction sites to allow insertion or substitution of the polynucleotide encoding the variant at such sites. Alternatively, the polynucleotide can be expressed by inserting the polynucleotide or a nucleic acid construct comprising the polynucleotide into an appropriate vector for expression. When generating the expression vector, the coding sequence is positioned in the vector such that it is operably linked to the appropriate control sequences for expression.

[0553] The recombinant expression vector can be any vector (e.g., a plasmid or virus) that can conveniently be subjected to recombinant DNA procedures and can bring about the expression of the polynucleotide. The choice of vector will typically depend on the compatibility of the vector with the host cell into which the vector is to be introduced. The vector can be linear or closed circular. The vector can be an autonomously replicating vector, i.e., a vector that exists as an extrachromosomal entity, the replication of which is independent of chromosomal replication, e.g., a plasmid, a non-chromosomal element, a mini-chromosome or an artificial chromosome. The vector can contain any means for assuring self-replication. Alternatively, the vector can be one that, when introduced into a host cell, is integrated into the genome and replicated together with the chromosome into which it has been integrated. Furthermore, a purpose-built vector or plasmid, or two or more vectors or plasmids, which together contain the total DNA to be introduced into the host cell genome, can be used, or a transposon can be used.

[0554] The vector can be a self-replicating vector, i.e., a vector that exists as an extrachromosomal entity, the replication of which is independent of chromosomal replication, e.g., a plasmid, an extra-chromosomal element, a mini-chromosome or an artificial chromosome. The vector can contain any means for assuring self-replication. Alternatively, the vector can be one that, when introduced into a host cell, is integrated into the genome and replicated together with the chromosome into which it has been integrated. Furthermore, a purpose-built vector or plasmid, or two or more vectors or plasmids, which together contain the total DNA to be introduced into the host cell genome, can be used, or a transposon can be used.

[0555] The vector preferably contains one or more selectable markers, which permit easy selection of transformed, transfected, transduced, or the like cells. A selectable marker is a gene the product of which provides a biocide resistance or viral resistance, resistance to heavy metals, prototrophy to auxotrophs, and the like.

[0556] Examples of bacterial selectable markers are Bacillus licheniformis or Bacillus subtilis dal genes, or markers that confer antibiotic resistance (such as ampicillin, chloramphenicol, kanamycin, neomycin, spectinomycin, or tetracycline resistance). Suitable markers for yeast host cells include, but are not limited to, ADE2, HIS3, LEU2, LYS2, MET3, TRP1, and URA3. Selectable markers for use in a filamentous fungal host cell include, but are not limited to, amdS (acetamidase), argB (ornithine carbamoyltransferase), bar (phosphinothricin acetyltransferase), hph (hygromycin phosphotransferase), niaD (nitrate reductase), pyrG (orotidine-5'-phosphate decarboxylase), sC (sulfate adenyltransferase), and trpC (anthranilate synthase), as well as equivalents thereof. Preferred for use in an Aspergillus cell are Aspergillus nidulans or Aspergillus oryzae amdS and pyrG genes and a Streptomyces hygroscopicus bar gene.

[0557] The vector preferably contains one or more elements that permit integration of the vector into the host cell's genome or autonomous replication of the vector in the cell independent of the genome.

[0558] For integration into the host cell genome, the vector can rely on a polynucleotide sequence encoding the variant or any other element of the vector for integration of the vector into the genome by homologous or non-homologous recombination. Alternatively, the vector can contain another polynucleotide that encodes a selectable marker, the product of which complements the host cell and confers selectable traits, such as tetracycline resistance to the host cell. Another example of a selectable marker is the dihydrofolate reductase gene that confers resistance to methotrexate. These methods of gene transfer are well known in the art. To increase the likelihood of integration at a precise location, the integrational elements should have homology with the

[0559] For autonomous replication, the vector can further comprise an origin of replication enabling the vector to replicate autonomously in the host cell in question. The origin of replication can be any plasmid or plasmid replicon, mediating autonomous replication in a cell. The term "origin of replication" or "plasmid replicon" means a polynucleotide that enables a plasmid or vector to replicate in vivo.

[0560] Examples of origins of replication for bacterial cells are the origins of replication of the plasmids pBR322, pUC19, pACYC177, and pACYC184, which are replicable in E. coli, and the origins of replication of the plasmids pUBHO, pE194, pTA1060, and pAMb1, which are replicable in Bacillus species.

[0561] Examples of origins of replication for use in yeast host cells are the 2 micron origin of replication, ARS1, ARS4, a combination of ARS1 and CEN3, and a combination of ARS4 and CEN6.

[0562] Examples of origins of replication useful in filamentous fungal cells are AMA1 and ANSI (Gems et al., 1991, Gene 98: 61-67; Cullen et al., 1987, Nucleic Acids Res. 15: 9163-9175; WO 00 / 24883). Isolation of the AMA1 gene and construction of plasmids or vectors comprising the gene can be accomplished according to the methods disclosed in WO 00 / 24883.

[0563] More than one copy of a polynucleotide of the present application can be inserted into a host cell to increase production of a variant. An increase in the copy number of the polynucleotide can be obtained by integrating at least one additional copy of the sequence into the host cell genome or by including an amplifiable selectable marker gene with the polynucleotide where cells containing amplified copies of the selectable marker gene, and thereby additional copies of the polynucleotide, can be selected for by cultivating the cells in the presence of the appropriate selectable agent.

[0564] The procedures used to ligate the elements described above to construct recombinant expression vectors of the present application are well known to those skilled in the art (cf., e.g., Sambrook et al., 1989, supra).

[0565] Host Cells

[0566] The present application also relates to recombinant host cells comprising a polynucleotide encoding a variant of the present application operably linked to one or more control sequences that direct the production of the variant of the present application. The construct or vector comprising the polynucleotide is introduced into a host cell so that the construct or vector is maintained as a chromosomal integrant or as a self-replicating extra-chromosomal vector as described earlier. The term "host cell" encompasses any progeny of the parent cell which is not identical to the parent cell (due to possible mutations that will occur during replication).

[0567] The host cell can be any cell useful in the recombinant production of a variant, such as prokaryotic or eukaryotic.

[0568] The prokaryotic host cell can be any Gram-positive or Gram-negative bacteria. Gram-positive bacteria include, but are not limited to, Bacillus, Clostridium, Enterococcus, Geobacillus, Lactobacillus, Lactococcus, Oceanobacillus, Staphylococcus, Streptococcus, and Streptomyces. Gram-negative bacteria include, but are not limited to, Campylobacter, Escherichia coli, Flavobacterium, Fusobacterium, Helicobacter, Ilyobacter, Neisseria, Pseudomonas, Salmonella, and Ureaplasma.

[0569] The bacterial host cell can be any Bacillus cell, including, but not limited to, Bacillus alkalophilus, Bacillus amyloliquefaciens, Bacillus brevis, Bacillus circulans, Bacillus clausii, Bacillus coagulans, Bacillus firmus, Bacillus lautus, Bacillus lentus, Bacillus licheniformis, Bacillus megaterium, Bacillus pumilus, Bacillus stearothermophilus, Bacillus subtilis, and Bacillus thuringiensis cells.

[0570] The bacterial host cell can also be any Streptococcus cell, including, but not limited to, Streptococcus equisimilis, Streptococcus pyogenes, Streptococcus uberis, and Streptococcus equi subsp. Zooepidemicus cells.

[0571] The bacterial host cell can also be any Streptomyces cell including, but not limited to, Streptomyces achromogenes, Streptomyces avermitilis, Streptomyces coelicolor, Streptomyces griseus, and Streptomyces lividans cells.

[0572] Introduction of DNA into a Bacillus cell can be effected by protoplast transformation (see, e.g., Chang and Cohen, 1979, Mol. Gen. Genet. 168: 111-115), competent cell transformation (see, e.g., Young and Spizizen, 1961, J. Bacteriol. 81: 823-829; or Dubnau and Davidoff-Abelson, 1971, J. Mol. Biol. 56: 209-221), electroporation (see, e.g., Shigekawa and Dower, 1988, Biotechniques 6: 742-751), or conjugation (see, e.g., Koehler and Thorne, 1987, J. Bacteriol. 169: 5271-5278). Introduction of DNA into an E. coli cell can be effected by calcium chloride treatment (see, e.g., Hanahan, 1983, J. Mol. Biol. 166: 557-580) or electroporation (see, e.g., Dower et al., 1988, Nucleic Acids Res. 16: 6127-6145). Introduction of DNA into a Streptomyces cell can be effected by protoplast transformation, electroporation (see, e.g., Gong et al., 2004, Folia Microbiol. (Praha) 49: 399-405), conjugation (see, e.g., Mazodier et al., 1989, J. Bacteriol. 171: 3583-3585), or transduction (see, e.g., Burke et al., 2001, Proc. Natl. Acad. Sci. USA 98: 6289-6294). Introduction of DNA into a Pseudomonas cell can be effected by electroporation (see, e.g., Choi et al., 2006, J. Microbiol. Methods 64: 391-397) or conjugation (see, e.g., Pinedo and Smets, 2005, Appl. Environ. Microbiol. 71: 51-57).Introduction of DNA into Streptococcus cells can be effected by natural competence (see, e.g., Perry and Kuramitsu, 1981, Infect. Immun. 32: 1295-1297), protoplast transformation (see, e.g., Catt and Jollick, 1991, Microbios 68: 189-207), electroporation (see, e.g., Buckley et al., 1999, Appl. Environ. Microbiol. 65: 3800-3804), or conjugation (see, e.g., Clewell, 1981, Microbiol. Rev. 45: 409-436). However, any method known in the art for introducing DNA into a host cell can be used.

[0573] The host cell can also be a eukaryotic organism, such as a mammal, an insect, a plant, or a fungal cell.

[0574] The host cell can be a fungal cell. As used herein, “fungal” includes the Ascomycota, Basidiomycota, Chytridiomycota, and Zygomycota, as well as the Oomycota and all mitosporic fungi (as defined by Hawksworth et al., in Ainsworth and Bisby’s Dictionary of The Fungi, 8th ed., 1995, CAB International, University Press, Cambridge, UK).

[0575] The fungal host cell can be a yeast cell. "Yeast" as used herein includes ascosporogenous yeast (Endomycetales), basidiosporogenous yeast and yeast belonging to the Fungi Imperfecti (Blastomycetes). Since the classification of yeast can change in the future, for the purposes of this application, yeast shall be defined as described in Biology and Activities of Yeast, Skinner, Passmore and Davenport, eds., Soc. App. Bacteriol. Symposium Series No. 9, 1980.

[0576] The yeast host cell can be a Candida, Hansenula, Kluyveromyces, Pichia, Saccharomyces, Schizosaccharomyces, or Yarrowia cell, such as a Kluyveromyces lactis, Saccharomyces carlsbergensis, Saccharomyces cerevisiae, Saccharomyces diastaticus, Saccharomyces douglasii, Saccharomyces kluyveri, Saccharomyces norbensis, Saccharomyces oviformis, or Yarrowia lipolytica cell.

[0577] The fungal host cell can be a filamentous fungus cell. "Filamentous fungi" include all filamentous forms of the subdivision Eumycota and subdivision Deuteromycota (as defined by Hawksworth et al., 1995, In: Ainsworth and Bisby's Dictionary of The Fungi, 8th edition, CAB International, University Press, Cambridge, p.21). The filamentous fungi are generally characterized by a mycelial wall composed of chitin, cellulose, glucan, chitosan, mannan, and other complex polysaccharides. Vegetative growth is by hyphal elongation and carbon catabolism is obligately aerobic. In contrast, vegetative growth of yeasts such as Saccharomyces cerevisiae is by cell budding and carbon catabolism can be fermentative.

[0578] The host cells of filamentous fungi can be *Acremonium*, *Aspergillus*, *Aureobasidium*, *Bjerkandera*, *Ceriporiopsis*, *Chrysosporium*, *Coprinus*, *Coriolus*, *Cryptococcus*, *Filibasidium*, *Fusarium*, *Humicola*, *Magnaporthe*, *Mucor*, and *Myceliop*. hthora), Neocallimastix, Neurospora, Paecilomyces, Penicillium, Phanerochaete, Phlebia, Piromyces, Pleurotus, Schizophyllum, Bassella, Thermoascus, Thievia, Tolypocladium, Trametes, or Trichoderma cells.

[0579] For example, the filamentous fungal host cell can be an Aspergillus awamori, Aspergillus foetidus, Aspergillus fumigatus, Aspergillus japonicus, Aspergillus nidulans, Aspergillus niger, Aspergillus oryzae, Bjerkandera adusta, Ceriporiopsis aneirina, Ceriporiopsis caregiea, Ceriporiopsis gilvescens, Ceriporiopsis pannocinta, Ceriporiopsis subvermispora, Chrysosporium inops, Chrysosporium keratinophilum, Chrysosporium lucknowense, Chrysosporium merdarium, Chrysosporium pannicola, Chrysosporium queenslandicum, Chrysosporium tropicum, Chrysosporium zonatum, Coprinus cinereus, Coriolus hirsutus, Fusarium bactridioides, Fusarium cerealis, Fusarium crookwellense, Fusarium culmorum, Fusarium graminearum, Fusarium graminum, Fusarium heterosporum, Fusarium negundi, Fusarium oxysporum, Fusarium reticulatum, Fusarium roseum, Fusarium sambucinum, Fusarium sarcochroum, Fusarium sporotrichioides, Fusarium sulphureum, Fusarium torulosum, Fusarium trichothecioides, Fusanum verticilliodes, Humicola insolens, Humicola lanuginosa, Mucor miehei, Myceliopthora thermophila, Neurospora crassa, Penicillium purpurogenum, Phanerochaete chrysosporium, Promyces clavus, Rhizomucor miehei, Schizophyllum commune, Trichoderma harzianum, Trichoderma longibrachiatum, Trichoderma reesei, or Trichoderma viride cell.Fusarium sambucinum), Fusarium sarcochroum, Fusarium sporotrichioides, Fusarium sulphureum, Fusarium torulosum, Fusarium torulosum, Fusarium venenatum, Humicola insolens, Humicola lanuginosa, Mucor miehei, Myceliopthora thermophila, Neurospora crassa, Penicillium purpurogenum, Phanerochaete chrysosporium, Phlebia radiata, Pleurotus eryngii, Thielavia terrestris, Trametes villosa, Trametes versicolor, Trichoderma harzianum, Trichoderma koningii, Trichoderma longibrachiatum, Trichoderma reesei, or Trichoderma viride cell.

[0580] Fungal cells can be transformed by a process involving protoplast formation, transformation of the protoplasts, and regeneration of the cell wall, in a manner known per se. Suitable procedures for transformation of Aspergillus and Trichoderma host cells are described in EP 238023 and Yelton et al., 1984, Proc. Natl. Acad. Sci. USA 81 : 1470-1474, and Christensen et al., 1988, Bio / Technology 6: 1419-1422. Suitable methods for transforming Fusarium species are described by Malardier et al., 1989, Gene 78: 147-156, and WO 96 / 00787. Yeast can be transformed using the procedure described by Becker and Guarente, in Abelson, J. N. and Simon, M. I., eds., Guide to Yeast Genetics and Molecular Biology, Methods in Enzymology, Vol. 194, pp. 182-187, Academic Press, Inc., New York; Ito et al., 1983, J. Bacteriol. 153: 163; and Hinnen et al., 1978, Proc. Natl. Acad. Sci. USA 75: 1920.

[0581] Production methods

[0582] The present application also relates to methods of producing the variants, comprising: (a) cultivating the recombinant host cell of the application under conditions suitable for expression of the variant; and optionally (b) recovering the variant.

[0583] The recombinant host cells are cultivated in a nutrient medium using methods known in the art. For example, cells can be cultivated by shake flask cultivation, or in a bioreactor at suitable conditions for expression and / or isolation of the variant. Using known procedures, the cultivation happens in a suitable nutrient medium containing carbohydrates, nitrogen, and inorganic salts and vitamins, as described in the art. Suitable media are available from commercial suppliers or can be prepared according to published compositions (e.g., in catalogues of the American Type Culture Collection). If the variant is secreted into the nutrient medium, the variant can be recovered directly from the medium. If the variant is not secreted, it can be recovered from cell lysates.

[0584] The variant can be detected using methods known in the art that are specific for the variant. These detection methods include, but are not limited to, the use of specific antibodies, the formation of an enzyme product, or the disappearance of an enzyme substrate. For example, an enzyme assay can be used to determine the activity of the variant.

[0585] The variant can be recovered using methods known in the art. For example, the variant can be recovered from the nutrient medium by conventional procedures including, but not limited to, centrifugation, filtration, extraction, spray-drying, evaporation, or precipitation.

[0586] The variant can be purified by a variety of procedures known in the art including, but not limited to, chromatography (e.g., ion exchange, affinity, hydrophobic, chromatofocusing, and size exclusion chromatography), electrophoretic procedures (e.g., preparative isoelectric focusing), differential solubility (e.g., ammonium sulfate precipitation), SDS-PAGE, or extraction (see, e.g., Protein Purification, Janson and Ryden editors, VCH Publishers, New York, 1989).

[0587] In alternative aspects, the variant is not recovered, but rather the host cell of the application expressing the variant is used as a source of the variant.

[0588] Fermentation broth preparations or cell compositions

[0589] The present application also relates to fermentation broth preparations or cell compositions comprising the variant of the present application. Fermentation broth products further comprise additional components used in the fermentation process, such as, for example, cells (including host cells containing a gene encoding the variant of the present application, which are used to produce the variant of interest), cell debris, biomass, fermentation media, and / or fermentation products. In some embodiments, the composition is a cell-killed whole broth containing one or more organic acids, killed cells and / or cell debris, and media.

[0590] As used herein, the term "fermentation broth" refers to a preparation produced by cell fermentation that has undergone or undergone minimal recovery and / or purification. For example, a fermentation broth is produced when a microbial culture is incubated under carbon-limiting conditions such that protein synthesis (e.g., expression of an enzyme by a host cell) and secretion of the protein into the cell culture medium occurs to saturation. The fermentation broth can contain unfractionated or fractionated contents of the fermentation material derived at the end of fermentation. Typically, the fermentation broth is unfractionated and includes spent media and cell debris present after removal of microbial cells (e.g., filamentous fungal cells) by, for example, centrifugation. In some embodiments, the fermentation broth contains spent cell media, extracellular enzymes, and viable and / or non-viable microbial cells.

[0591] In embodiments, the fermentation broth formulation and cell composition comprise a first organic acid component comprising at least one 1-5 carbon organic acid and / or salt thereof and a second organic acid component comprising at least one 6 or more carbon organic acid and / or salt thereof. In particular embodiments, the first organic acid component is acetic acid, formic acid, propionic acid, a salt thereof, or a mixture of two or more of the foregoing; and the second organic acid component is benzoic acid, cyclohexanecarboxylic acid, 4-methylpentanoic acid, phenylacetic acid, a salt thereof, or a mixture of two or more of the foregoing.

[0592] In one aspect, the composition contains one or more organic acids, and optionally further contains killed cells and / or cell debris. In one embodiment, the killed cells and / or cell debris are removed from the cell-killed whole broth to provide a composition free of these components.

[0593] The fermentation broth formulations or cell compositions can further comprise a preservative and / or antimicrobial (e.g., bacteriostatic) agent, including but not limited to sorbitol, sodium chloride, potassium sorbate, and other agents known in the art.

[0594] The cell-killed whole broth or composition can contain unfractionated contents of the fermentation material derived at the end of fermentation. Typically, the cell-killed whole broth or composition contains spent media and cell debris present after microbial cells (e.g., filamentous fungal cells) are incubated to saturation under carbon-limited conditions that allow for protein synthesis. In some embodiments, the cell-killed whole broth or composition contains spent cell culture media, extracellular enzymes, and killed filamentous fungal cells. In some embodiments, the microbial cells present in the cell-killed whole broth or composition can be permeabilized and / or lysed using methods known in the art.

[0595] The whole broth or cell composition as described herein is typically a liquid, but can contain insoluble components, such as killed cells, cell debris, media components, and / or one or more insoluble enzymes. In some embodiments, the insoluble components can be removed to provide a clarified liquid composition.

[0596] The whole broth formulations and cell compositions of the present application can be produced by the methods described in WO 90 / 15861 or WO 2010 / 096673.

[0597] Compositions

[0598] In one embodiment, the present application relates to cleaning compositions, such as detergent compositions, comprising a xyloglucanase variant of the present application in combination with one or more additional cleaning composition components and, preferably, a detergent adjunct ingredient as described herein. The selection of additional components is within the skill of the artisan and includes conventional ingredients, including the exemplary non-limiting components set forth below. The detergent compositions can be used to clean an article to be cleaned, such as a textile, dishware, and hard surface, to pre-treat a stain on the article, to prevent, reduce, or remove redeposition of soil during a wash cycle, to maintain or improve whiteness of the article.

[0599] Preferably, the compositions are enriched for such variants. The term "enriched" indicates that the xyloglucanase activity of the composition has been increased, e.g., by a factor of enrichment of at least 1.1.

[0600] The compositions can comprise a variant of the present application as the primary enzyme component, e.g., a single component composition. Alternatively, the compositions can comprise multiple enzyme activities, such as one or more enzymes selected from the group consisting of a hydrolase, isomerase, ligase, lyase, oxidoreductase, or transferase, e.g., a -galactosidase, a -glucosidase, an aminopeptidase, an amylase, a -galactosidase, a -glucosidase, a -xylosidase, a carbohydrase, a carboxypeptidase, a catalase, a cellobiohydrolase, a cellulase, a chitinase, a cutinase, a cyclodextrin glucanotransferase, a deoxyribonuclease, an endoglucanase, an esterase, a glucoamylase, a invertase, a laccase, a lipase, a mannosidase, a mutanase, a nuclease, an oxidase, a pectolytic enzyme, a peroxidase, a phytase, a polyphenoloxidase, a proteolytic enzyme, a ribonuclease, a transglutaminase, or a xylanase.

[0601] The compositions can be prepared according to methods known in the art, and the compositions can be in the form of a liquid or dry composition. The compositions can be stabilized according to methods known in the art.

[0602] Examples of preferred uses of the compositions of the present application are given below. The dosage of the composition, as well as other conditions for using the composition, can be determined based on methods known in the art.

[0603] For textile care, the selection of components can include the following considerations: the type of textile to be cleaned, the type and / or extent of soil, the temperature at which cleaning is performed, and the formulation of the detergent product. Although the components mentioned below are categorized by general headings according to a particular functionality, this is not to be construed as limiting, as the components can comprise additional functionalities as would be understood by the ordinary artisan.

[0604] In one embodiment of the present application, a composition (e.g., a detergent composition) comprises a xyloglucanase variant as disclosed herein and a detergent adjunct.

[0605] In one embodiment of the present application, the detergent adjunct ingredient is selected from the group consisting of surfactants, builders, flocculating aids, chelating agents, dye transfer inhibiting agents, enzymes, enzyme stabilizers, enzyme inhibitors, catalytic materials, bleach activators, hydrogen peroxide, sources of hydrogen peroxide, preformed peracids, polymeric dispersing agents, clay soil removal / anti-redeposition agents, brighteners, suds suppressors, dyes, perfumes, structure elasticizing agents, fabric softeners, carriers, hydrotropes, builders and co-builders, fabric hueing agents, antifoams, dispersants, processing aids, and / or pigments.

[0606] The detergent adjunct ingredient can be a surfactant. One advantage of including a surfactant in a detergent composition comprising a xyloglucanase variant is that wash performance is improved. In one embodiment, the detergent adjunct ingredient is a builder or clay soil removal / anti-redeposition agent.

[0607] In one embodiment, the detergent adjunct ingredient is an enzyme. The detergent composition can comprise one or more enzymes, as specified below. The one or more enzymes can be selected from the group consisting of proteases, lipases, cutinases, amylases, carbohydrases, cellulases, pectinases, mannanases, arabinosases, galactanases, xylanases, nucleases, and oxidases. Specific enzymes suitable for use in the detergent compositions of the present application are described below.

[0608] The detergent composition can be formulated as bars, uniform tablets, and tablets having two or more layers, pouches having one or more compartments, regular or compact powders, granulates, pastes, gels, or regular, compact or concentrated liquids. The detergent composition can be a liquid detergent, a powder detergent, or a granular detergent.

[0609] The xyloglucanases of the present application are suitable for use in cleaning, for example, laundry. The present application further relates to a method for laundering an article, the method comprising the steps of:

[0610] a. exposing the article to a wash liquor comprising a variant polypeptide having xyloglucanase activity or a detergent composition containing the polypeptide;

[0611] b. completing at least one wash cycle; and

[0612] c. optionally rinsing the article,

[0613] wherein the article is a textile.

[0614] The pH of the liquid solution is in the range of 1 to 11, such as in the range of 5.5 to 11, such as in the range of 7 to 9, in the range of 7 to 8, or in the range of 7 to 8.5.

[0615] The wash liquor can have a temperature in the range of 5°C to 95°C, or in the range of 10°C to 80°C, in the range of 10°C to 70°C, in the range of 10°C to 60°C, in the range of 10°C to 50°C, in the range of 15°C to 40°C, or in the range of 20°C to 30°C. In one embodiment, the temperature of the wash liquor is 30°C.

[0616] In one embodiment of the application, the method for washing an item further comprises draining the wash liquor or part of the wash liquor after completion of the wash cycle. The wash liquor can then be reused in a subsequent wash cycle or in a subsequent rinse cycle. The item can be exposed to the wash liquor during the first and optionally the second or third wash cycle. In one embodiment, the item is rinsed after exposure to the wash liquor. The item can be rinsed with water or with water comprising a conditioning agent. The present application further relates to an item washed according to the method of the present application. The xyloglucanase variant of the present application can be added to the wash liquor.

[0617] Thus, one embodiment of the present application relates to a detergent composition comprising one or more anionic surfactants; an enzyme selected from the group consisting of: protease, lipase, cutinase, amylase, carbohydrase, cellulase, pectinase, mannanase, arabinanase, galactanase, xylanase, nucleotide- dase, and oxidase; and a xyloglucanase variant of the present application.

[0618] One embodiment further relates to a method for washing a textile, the method comprising:

[0619] a. exposing the textile to a wash liquor comprising a xyloglucanase variant or a detergent composition containing at least one of these xyloglucanase variants,

[0620] b. completing at least one wash cycle; and

[0621] c. optionally rinsing the textile,

[0622] wherein the xyloglucanase variant comprises an alteration at one or more positions corresponding to positions selected from the group consisting of 111, 123, 159, 256, 294, 8, 18, 20, 41, 42, 76, 82, 83, 87, 94, 103, 104, 105, 121, 125, 126, 127, 136, 137, 146, 147, 148, 152, 153, 155, 165, 168, 169, 177, 184, 189, 203, 206, 210, 211, 214, 217, 219, 220, 226, 237, 238, 240, 243, 244, 248, 251, 252, 267, 271, 276, 289, 295, 298, 300, 302, 322, 329, 339, 347, 347, 353, 383, 384, 392, 394, 395, 402, 414, 427, 431, 445, 447, 459, 473, 474, 476, 482, 488, 489, 491, 492, 503, and 505 of the polypeptide of SEQ ID NO: 1.

[0623] Another embodiment relates to a textile washed according to the method of the application.

[0624] The concentration of the xyloglucanase variant in the wash liquor is typically in the range of 0.00004-100 ppm enzyme protein, such as in the range of 0.00008-100, in the range of 0.0001-100, in the range of 0.0002-100, in the range of 0.0004-100, in the range of 0.0008-100, in the range of 0.001-100 ppm enzyme protein, 0.01-100 ppm enzyme protein, preferably 0.05-50 ppm enzyme protein, more preferably 0.1-50 ppm enzyme protein, more preferably 0.1-30 ppm enzyme protein, more preferably 0.5-20 ppm enzyme protein, and most preferably 0.5-10 ppm enzyme protein.

[0625] The xyloglucanase variant of the application can be added to a detergent composition in an amount corresponding to at least 0.002 mg of xyloglucanase protein, such as at least 0.004 mg of xyloglucanase protein, at least 0.006 mg of xyloglucanase protein, at least 0.008 mg of xyloglucanase protein, at least 0.01 mg of xyloglucanase protein, at least 0.1 mg of protein, preferably at least 1 mg of protein, more preferably at least 10 mg of protein, even more preferably at least 15 mg of protein, most preferably at least 20 mg of protein, and even most preferably at least 25 mg of protein. Thus, the detergent composition can comprise at least 0.00008% xyloglucanase protein, preferably at least 0.002%, 0.003%, 0.004%, 0.005%, 0.006%, 0.008%, 0.01%, 0.02%, 0.03%, 0.05%, 0.1%, 0.2%, 0.3%, 0.4%, 0.6%, 0.7%, 0.8%, 0.9%, or 1.0% xyloglucanase protein.

[0626] Enzymes included in the detergent of the application can be stabilized using conventional stabilizing agents, e.g., a polyol such as propylene glycol or glycerol, a sugar or sugar alcohol, and a different salt such as NaCI and KCI. Proteases present in the detergent of the application can be stabilized using lactic acid, formic acid, boric acid, or a boronic acid derivative (e.g., an aromatic boronic ester), or a phenyl boronic acid derivative (e.g., 4-formylphenyl boronic acid), or a peptide aldehyde (e.g., a di-, tri-, or tetrapeptide aldehyde or aldehyde analog) (or having the form B1-B0-R, wherein R is H, CH3, CX3, CHX2, or CH2X (X = halogen), B0 is a single amino acid residue (preferably with an optionally substituted aliphatic or aromatic side chain); and B1 consists of one or more amino acid residues (preferably one, two, or three), optionally including an N-terminal protecting group, or a protease inhibitor of the protein type, such as RASI, BASI, WASI (rice, barley, and wheat bifunctional alpha-amylase / subtilisin inhibitors) or CI2 or SSI. The composition can be formulated as described in, e.g., WO 92 / 19709, WO 92 / 19708, and US 6,472,364. In some embodiments, the enzymes used herein are stabilized by water-soluble sources of zinc (II), calcium (II), and / or magnesium (II) ions, along with other metal ions (e.g., barium (II), scandium (II), iron (II), manganese (II), aluminum (III), tin (II), cobalt (II), copper (II), nickel (II), and vanadyl (IV)) present in the finished composition providing such ions for the enzymes.

[0627] In one embodiment, a peptide aldehyde or ketone is used to stabilize the polypeptide. Suitable peptide aldehydes are described in WO 94 / 04651, WO 95 / 25791, WO 98 / 13458, WO 98 / 13459, WO 98 / 13460, WO 98 / 13461, WO 98 / 13462, WO 07 / 141736, WO 07 / 145963, WO 09 / 118375, WO 10 / 055052, and WO 11 / 036153. The polypeptides of the present application can also be incorporated into the detergent formulations disclosed in WO 97 / 07202, which is hereby incorporated by reference.

[0628] In another embodiment, a phenyl boronic acid derivative is used to stabilize the polypeptide, which is 4-formylphenyl boronic acid (4-FPBA) having the following formula:

[0629]

[0630] The detergent composition can comprise two or more stabilizing agents, such as, for example, those selected from the group consisting of propylene glycol, glycerol, 4-formylphenyl boronic acid, and borate.

[0631] The detergent composition can comprise two or more stabilizing agents, such as, for example, those selected from the group consisting of propylene glycol, glycerol, 4-formylphenyl boronic acid, and borate.

[0632] The amount of the one or more stabilizing agents preferably present in the detergent composition is from 0.001 to about 5.0 wt%, from 0.01 to about 2.0 wt%, from 0.1 to about 3 wt%, or from 0.5% to about 1.5 wt%.

[0633] Surfactant

[0634] The detergent composition can comprise one or more surfactants, which can be anionic and / or cationic and / or nonionic and / or semi-polar and / or zwitterionic, or mixtures thereof. In particular embodiments, the detergent composition includes a mixture of one or more nonionic surfactants and one or more anionic surfactants. The one or more surfactants are typically present at a level of from about 5% to 60% by weight (e.g., from about 5% to about 50%, or from about 10% to about 50%, or from about 20% to about 50%). The one or more surfactants are selected based on the desired cleaning application, and can include any one or more conventional surfactants known in the art.

[0635] When included therein, the detergent will usually contain from about 5% to about 60%, such as from about 5% to about 40%, including from about 10% to about 25%, by weight of one or more anionic surfactants. Non-limiting examples of anionic surfactants include sulfates and sulfonates, specifically linear alkylbenzenesulfonates (LAS), isomers of LAS, branched alkylbenzenesulfonates (BABS), phenylalkanesulfonates, alpha-olefinsulfonates (AOS), olefin sulfonates, alkene sulfonates, alkane-2,3-diylbis(sulfates), hydroxyalkane sulfonates, and disulfonates, alkyl sulfates (AS) such as sodium dodecyl sulfate (SDS), fatty alcohol sulfates (FAS), primary alcohol sulfates (PAS), alcohol-ether sulfates (AES or AEOS or FES, also known as alcohol ethoxysulfates or fatty alcohol ether sulfates), secondary alkane sulfonates (SAS), paraffin sulfonates (PS), ester sulfonates, sulfonated fatty acid glycerol esters, alpha-sulfo fatty acid methyl esters (alpha-SFMe or SES) including methyl ester sulfonate (MES), alkyl or alkenyl succinic acid, dodecenyl / tetradecenyl succinic acid (DTSA), fatty acid derivatives of amino acids, diesters and monoesters of sulfosuccinic acid, or fatty acid salts (soaps) or fatty acids, and combinations thereof.

[0636] When included therein, the detergent will usually contain from about 0.1% to about 10%, for example from about 0,1% to about 5%, by weight of a cationic surfactant. Non-limiting examples of cationic surfactants include alkyl dimethyl ethylammonium ethosulfates (ADMEAQ), cetyltrimethylammonium bromide (CTAB), dimethyldistearylammonium chloride (DSDMAC), and alkylbenzyldimethylammonium, alkyl quaternary ammonium compounds, alkoxylated quaternary ammonium (AQA) compounds, ester quats, and combinations thereof.

[0637] When included, detergents will typically contain from about 0.2% to about 60% by weight (e.g. from about 1% to about 40%, particularly from about 5% to about 20%, from about 3% to about 15%) of nonionic surfactants. Non-limiting examples of nonionic surfactants include alcohol ethoxylates (AE or AEO), alcohol propoxylates, propoxylated fatty alcohols (PFA), alkoxylated fatty acid alkyl esters (e.g., ethoxylated and / or propoxylated fatty acid alkyl esters), alkylphenol ethoxylates (APE), nonylphenol ethoxylates (NPE), alkyl polysaccharides (APG), alkoxylated amines, fatty acid monoethanolamides (FAM), fatty acid diethanolamides (FADA), ethoxylated fatty acid monoethanolamides (EFAM), propoxylated fatty acid monoethanolamides (PFAM), polyhydroxyalkyl fatty acid amides, or N-acyl N-alkyl derivatives of glucosamine (glucosamide (GA) or fatty acid glucosamide (FAGA)), methyl ester ethoxylates (MEE), together with products available under the trade names SPAN and TWEEN, and combinations thereof.

[0638] When included therein, the detergent will typically contain from about 0.1% to about 10% by weight of a semi-polar surfactant. Non-limiting examples of semi-polar surfactants include amine oxides (AOs), such as alkyl dimethylamine oxides, N-(cocoylalkyl)-N,N-dimethylamine oxides, and N-(butter-alkyl)-N,N-bis(2-hydroxyethyl)amine oxides, and combinations thereof.

[0639] When included, detergents will typically contain from about 0.1% to about 10% by weight of a facultative zwitterionic surfactant. Non-limiting examples of facultative zwitterionic surfactants include betaines, such as alkyl dimethyl betaine, sulfobetaine, and combinations thereof.

[0640] Solvent System: A solvent system is required to dissolve surfactants and other detergent components. Solvents are typically water, alcohols, polyols, sugars, and / or mixtures thereof. Preferred solvents are water, glycerol, sorbitol, propylene glycol (MPG, 1,2-propanediol, or 1,3-propanediol), dipropylene glycol (DPG), polyethylene glycol family (PEG300-600), hexanediol, inositol, mannitol, ethanol, isopropanol, n-butoxypropoxypropanol, ethanolamines (monoethanolamine, diethanolamine, and triethanolamine), sucrose, dextrose, glucose, ribose, xylose, and related mono- and dipyranoside and furanoside.

[0641] The solvent system is typically present in total amounts of 5%-90%, 5%-60%, 5%-40%, and 10%-30% by weight.

[0642] The water content of the unit dose wrapped in PVA film is typically in the range of 1-15%, 2-12%, 3-10%, 5-10%.

[0643] The polyol content of the unit dose wrapped in PVA film is typically in the range of 5-50%, 10-40%, or 20-30%.

[0644] In embodiments, the surfactant is a non-naturally occurring surfactant.

[0645] Hydrotropes

[0646] Hydrotropes are compounds that solubilize hydrophobic compounds in aqueous solutions (or conversely, polar substances in non-polar environments). Typically, hydrotropes have both hydrophilic and hydrophobic character (as known from surfactants, the so-called amphiphilic character), however the molecular structure of hydrotropes generally does not favor spontaneous self-aggregation, see for example the review by Hodgdon and Kaler (2007), Current Opinion in Colloid & Interface Science 12: 121-128. Hydrotropes do not exhibit a critical concentration as seen in surfactants and lipids that form micelles, lamellar or other well-defined meso-phases above which self-aggregation occurs. Instead, many hydrotropes show a continuous type of aggregation process in which the size of the aggregates grows with increasing concentration. However, many hydrotropes alter the phase behavior, stability, and colloidal properties of systems containing polar and non-polar character, including mixtures of water, oil, surfactants, and polymers. Hydrotropes are routinely used in various industries from pharmaceutical, personal care, food to technical applications. The use of hydrotropes in detergent compositions allows for example more concentrated surfactant formulations (as in the process of compacting liquid detergents by removing water) without causing undesirable phenomena such as phase separation or high viscosity.

[0647] The detergent can contain 0-10% by weight, for example 0-5% by weight, for example about 0.5% to about 5%, or about 3% to about 5% of a hydrotrope. Any hydrotrope known in the art for use in detergents can be utilized. Non-limiting examples of hydrotropes include sodium benzenesulfonate, sodium p-toluenesulfonate (STS), sodium xylene sulfonate (SXS), sodium cumene sulfonate (SCS), sodium umbelliferone sulfonate, amine oxides, alcohols and polyglycol ethers, sodium hydroxynaphthalene carboxylate, sodium hydroxynaphthalene sulfonate, sodium ethylhexyl sulfonate, and combinations thereof.

[0648] Builders and co-builders

[0649] The detergent composition can contain from 0-65%, 0-20%, or 0.5%-5% of a detergent builder or co-builder, or mixtures thereof. In a dishwashing detergent, the level of builder is typically 10%-65%, especially 20%-40%. The builder and / or co-builder can be especially a chelating agent which forms water-soluble complexes with Ca and Mg. Any builder and / or co-builder known in the art for use in laundry detergents can be utilized. Non-limiting examples are citrate, sodium carbonate, sodium bicarbonate, and sodium citrate. Examples of phosphonate include 1-hydroxyethylidene-1,1-diphosphonic acid (HEDP, etidronic acid), diethylenetriamine pentakis(methylene phosphonic acid) (DTPMP), ethylenediamine tetra(methylene phosphonic acid) (EDTMPA), amino tri(methylene phosphonic acid) (ATMP), nitrilotri(methylene phosphonic acid) (NTMP), 2-aminoethyl phosphonic acid (AEPn), dimethyl methyl phosphonate (DMPP), tetramethylenediamine tetra(methylene phosphonic acid) (TDTMP), hexamethylenediamine tetra(methylene phosphonic acid) (HDTMP), phosphonobutanetris-carboxylic acid (PBTC), N-(phosphonomethyl)iminodiacetic acid (PMIDA), 2-carboxyethyl phosphonic acid (CEPA), 2-hydroxyphosphonyl carboxylic acid (HPAA), and amino-tris-(methylene-phosphonic acid) (AMP). L-glutamic acid N,N-diacetic acid tetrasodium salt (GLDA), methylglycinediacetic acid (MGDA). Non-limiting examples of builders include homopolymers of polyacrylates or copolymers thereof, such as poly(acrylic acid) (PAA) or co-poly(acrylic acid / maleic acid) (PAA / PMA). Further non-limiting examples include citrate, chelants (such as aminocarboxylates, aminopolycarboxylates, and phosphonates), and alkyl or alkenyl succinic acids.Further particular examples include 2,2',2"-nitrilotriacetic acid (NTA), ethylenediaminetetraacetic acid (EDTA), diethylenetriaminepentaacetic acid (DTPA), iminodisuccinic acid (IDS), ethylenediamine-N,N'-disuccinic acid (EDDS), methylglycinediacetic acid (MGDA), glutamic acid-N,N-diacetic acid (GLDA), 1 -hydroxyethane-1,1 -diphosphonic acid (HEDP), ethylenediaminetetra(methylene phosphonic acid) (EDTMPA), diethylenetriaminepenta(methylene phosphonic acid) (DTMPA or DTPMPA), N-(2-hydroxyethyl)iminodiacetic acid (EDG), aspartic acid-N-monocarboxylic acid (ASMA), aspartic acid-N,N-dicarboxylic acid (ASDA), aspartic acid-N-monopropionic acid (ASMP), iminodisuccinic acid (IDA), N-(2-sulfomethyl)-aspartic acid (SMAS), N-(2-sulfoethyl)-aspartic acid (SEAS), N-(2-sulfomethyl)-glutamic acid (SMGL), N-(2-sulfoethyl)-glutamic acid (SEGL), N-methyliminodiacetic acid (MIDA), a-alanine-N,N- diacetic acid (a-ALDA), serine-N,N-diacetic acid (SEDA), isoserine-N,N-diacetic acid (ISDA), phenylalanine-N,N-diacetic acid (PHDA), anthranilic acid-N,N-diacetic acid (ANDA), sulfanilic acid-N,N-diacetic acid (SLDA), taurine-N,N-diacetic acid (TUDA) and sulfomethyl-N,N-diacetic acid (SMDA), N-(2-hydroxyethyl)-ethylenediamine-N,N'- triacetic acid (HEDTA), diglycolic acid (DEG), diethylenetriaminepenta(methylene phosphonic acid) (DTPMP), amino tri(methylene phosphonic acid) (ATMP), and combinations and salts thereof. Further exemplary builders and / or co-builders are described in, for example, WO 09 / 102854, US 5977053.

[0650] In embodiments, the builder or co-builder is a non-naturally occurring builder or co-builder.

[0651] Bleaching system

[0652] The detergent can contain 0-30% by weight, for example from about 1% to about 20%, of a bleaching system. Any bleaching system known in the art for use in laundry detergents can be utilized. Suitable bleaching system components include bleach catalysts, photobleaches, bleach activators, sources of hydrogen peroxide such as sodium percarbonate, sodium perborate, and hydrogen peroxide-urea (1 :1), preformed peracids, and mixtures thereof. Suitable preformed peracids include, but are not limited to, peroxycarboxylic acids and salts, diperoxydicarboxylic acids, perimidic acids and salts, peroxymonosulfuric acid and salts (e.g., Oxone(R)), and mixtures thereof. Non-limiting examples of bleaching systems include peroxide-based bleaching systems in combination with a peracid-forming bleach activator, which can comprise, for example, inorganic salts, including alkali metal salts such as sodium salts of perborate (typically mono- or tetra-hydrate), percarbonate, persulfate, perphosphate, persilicate. The term bleach activator means herein a compound which reacts with hydrogen peroxide to form a peracid via perhydrolysis. The peracid thus formed constitutes the activated bleaching agent. Suitable bleach activators to be used herein include those belonging to the class of esters, amides, imides, or anhydrides. Suitable examples are tetraacetyl ethylene diamine (TAED), sodium 4-[(3,5,5-trimethylhexanoyl)oxy]benzene-1 -sulfonate (ISONOBS), sodium 4-(dodecanoyloxy)benzene-1 -sulfonate (LOBS), 4-(decanoyloxy)benzene-1 -sulfonate, 4-(decanoyloxy)benzoic acid salt (DOBS or DOBA), 4-(nonanoyloxy)benzene-1 -sulfonate (NOBS), and / or those disclosed in WO 98 / 17767. A particular family of bleach activators of interest is disclosed in EP 624154 and particularly preferred within this family is acetylcitric acid triethyl ester (ATC). ATC or short chain triglycerides like triacetin have the advantage that they are environmentally friendly. Furthermore, acetylcitric acid triethyl ester and triacetin have good hydrolytic stability in the product upon storage and are highly efficient bleach activators. Finally, ATC is multifunctional because the citrate salt released in the perhydrolysis reaction can act as a builder. Alternatively, the bleaching system can comprise a peroxy acid of the amide, imide, or sulfone type. The bleaching system can also comprise a peracid such as 6-(phthaloyl imino)peroxy hexanoic acid (PAP). The bleaching system can also include a bleach catalyst. In some embodiments, the bleach component can be an organic catalyst selected from the group consisting of organic catalysts having the formula:

[0653]

[0654] (iii) mixtures thereof,

[0655] wherein each R 1independently is a branched alkyl group containing from 9 to 24 carbons or a linear alkyl group containing from 11 to 24 carbons, preferably each R 1 independently is a branched alkyl group containing from 9 to 18 carbons or a linear alkyl group containing from 11 to 18 carbons, more preferably each R 1 is independently selected from the group consisting of 2-propylheptyl, 2-butyloctyl, 2-pentylnonyl, 2-hexyldecyl, dodecyl, tetradecyl, hexadecyl, octadecyl, isononyl, isodecyl, isotridecyl, and isopentadecyl. Other exemplary bleach systems are described, for example, in WO 2007 / 087258, WO 2007 / 087244, WO 2007 / 087259, EP 1867708 (vitamin K), and WO 2007 / 087242. A suitable photobleach can be, for example, sulfonated zinc phthalocyanine or aluminum phthalocyanine.

[0656] Preferably, in addition to the bleach catalyst, in particular the organic bleach catalyst, the bleach component further comprises a peracid source. The peracid source can be selected from (a) preformed peracid; (b) percarbonate, perborate or persulfate (hydrogen peroxide source), preferably in combination with a bleach activator; and (c) perhydrolytic enzyme and ester for in situ formation of peracid in the presence of water in a textile or hard surface treatment step.

[0657] In embodiments, the bleach system is a non-naturally occurring bleach system.

[0658] Polymer

[0659] The detergent can contain 0-10% by weight (e.g., 0.5-5%, 2-5%, 0.5-2%, or 0.2-1%) of a polymer. Any polymer known in the art for use in detergents can be utilized. The polymer can function as a co-builder as mentioned above, or can provide anti-redeposition, fiber protection, soil release, dye transfer inhibition, grease cleaning, and / or antifoam properties. Some polymers can have more than one of the above-mentioned properties and / or more than one of the below-mentioned motifs. Exemplary polymers include (carboxymethyl)cellulose (CMC), poly(vinyl alcohol) (PVA), poly(vinylpyrrolidone) (PVP), poly(ethylene glycol) or poly(ethylene oxide) (PEG), ethoxylated poly(ethyleneimine), carboxymethyl inulin (CMI), and polycarboxylates (such as PAA, PAA / PMA, poly-aspartic acid, and lauryl methacrylate / acrylic acid copolymer), hydrophobically modified CMC (HM-CMC) and silicone, copolymers of terephthalic acid and oligoethylene glycol, copolymers of poly(ethylene terephthalate) and poly(oxethylene terephthalate) (PET-POET), PVP, poly(vinylimidazole) (PVI), poly(vinylpyrrolidone-N-oxide) (PVPO or PVPNO), and polyvinylpyrrolidone-vinylimidazole (PVPVI). Additional exemplary polymers include sulfonated polycarboxylates, polyethylene oxide and polypropylene oxide (PEO-PPO), and diquaternary ammonium ethyl sulfate. Other exemplary polymers are disclosed in, e.g., WO 2006 / 130575. Salts of the above-mentioned polymers are also contemplated.

[0660] In embodiments, the polymer is a non-naturally occurring polymer.

[0661] Fabric hueing agents

[0662] The detergent composition of the present application can also include a fabric hueing agent, such as a dye or pigment, which when formulated in a detergent composition can deposit onto a fabric when said fabric is contacted with a wash liquor comprising said detergent composition, and thus alter the color of said fabric by absorption / reflection of visible light. Fluorescent whitening agents emit at least some visible light. In contrast, fabric hueing agents alter the color of a surface when they absorb at least a portion of the visible light spectrum. Suitable fabric hueing agents include dyes and dye-clay conjugates, and can also include pigments. Suitable dyes include small molecule dyes and polymeric dyes. Suitable small molecule dyes include small molecule dyes selected from the group consisting of dyes falling into the Colour Index (C.I.) classifications of Direct Blue, Direct Red, Direct Violet, Acid Blue, Acid Red, Acid Violet, Basic Blue, Basic Violet and Basic Red, or mixtures thereof, as described in WO 2005 / 03274, WO 2005 / 03275, WO 2005 / 03276 and EP 1876226 (hereby incorporated by reference). The detergent composition preferably comprises from about 0.00003 wt% to about 0.2 wt%, from about 0.00008 wt% to about 0.05 wt%, or even from about 0.0001 wt% to about 0.04 wt% of fabric hueing agent. The composition can comprise from 0.0001 wt% to 0.2 wt% of fabric hueing agent, which can be especially preferred when the composition is in the form of a unit dose pouch. Suitable hueing agents are also disclosed in, for example, WO 2007 / 087257 and WO 2007 / 087243.

[0663] Additional enzymes

[0664] The detergent additive as well as the detergent composition can comprise one or more [additional] enzymes such as hydrolases (EC 3.-.-.-), such as hydrolases acting on ester bonds (EC 3.1.-.-), glycosidases (EC 3.2.-.-), and hydrolases acting on peptide bonds (EC 3.4.-.-), oxidoreductases (EC 1.-.-.-), such as laccases (EC 1.10.-.-) or peroxidases (EC 1.11.-.-), or lyases (EC 4.-.-.-), such as carbon-oxygen lyases (EC 4.2.-.-). In particular embodiments, the detergent composition can comprise one or more [additional] enzymes such as proteases, lipases, cutinases, amylases, carbohydrases, cellulases, pectinases, mannanases, arabinosases, galactanases, xylanases, nucleases, oxidases (such as laccases), and / or peroxidases.

[0665] Generally, the properties of one or more enzymes selected should be compatible with the selected detergent (i.e., optimal pH, compatibility with other enzyme or non-enzyme components, etc.), and the one or more enzymes should be present in an effective amount.

[0666] Cellulase

[0667] Suitable cellulases include those of bacterial or fungal origin. This includes chemically modified mutants or protein-engineered mutants. Suitable cellulases include those from the genera *Bacillus*, *Pseudomonas*, *Pyrophyllus*, *Fusarium*, *Clostridium*, and *Cladosporium*, such as the fungal cellulases produced by *Pyrophyllus*, *Thermophyllus*, and *Fusarium* as disclosed in US 4,435,307, US 5,648,263, US 5,691,178, US 5,776,757, and WO 89 / 09259.

[0668] Particularly suitable cellulases are alkaline or neutral cellulases, which provide or maintain whiteness and prevent redeposition or have color-care benefits. Examples of such cellulases are those described in EP 0 495 257, EP0 531 372, WO 96 / 11262, WO 96 / 29397, and WO 98 / 08940. Other examples are, for instance, those cellulases described in WO 94 / 07998, EP 0 531 315, US 5,457,046, US 5,686,593, US 5,763,254, WO 95 / 24471, WO 98 / 12307, and WO 99 / 001544.

[0669] Other cellulases are endoglucanases having the following sequence, which has at least 97% identity with the amino acid sequence at positions 1 to 773 of SEQ ID NO:2 of WO 2002 / 099091, or family 44 xyloglucanases having the following sequence, which has at least 60% identity with positions 40-559 of SEQ ID NO:2 of WO 2001 / 062903.

[0670] Commercially available cellulases include Celluzyme TM and Carezyme TM (Novozymes A / S), Carezyme Premium) TM (Novozymes), Celluclean TM (Novozymes), Celluclean Classic TMCelluclean (Novozymes), Cellulolytic enzyme TM Whitezyme (Novozymes), Clazinase TM Clazinase (Novozymes), Clazinase TM Puradax HA (Novozymes), Purabrite TM KAC-500 (B) (Genencor International Inc.) and KAC-500 (B) TM Purabrite (Kao Corporation).

[0671] Mannanase

[0672] Suitable mannanases include those of bacterial or fungal origin. Mutants including chemical or genetic modifications are included. The mannanase can be an alkaline mannanase of family 5 or 26. It can be a wild type from Bacillus or Humicola, in particular from B. mucilaginosus, B. licheniformis, B. halodurans, B. clausii, or H. insolens. Suitable mannanases are described in WO 1999 / 064619. A commercially available mannanase is Mannaway (Novozymes).

[0673] Protease

[0674] Suitable proteases include those of bacterial, fungal, plant, viral, or animal origin, e.g. of vegetable or microbial origin. A microbial origin is preferred. Mutants including chemical modifications or protein engineered mutants are included. It can be an alkaline protease such as a serine protease or a metallo protease. The serine protease can e.g. be of S1 family (such as trypsin) or S8 family (such as subtilisin). The metallo protease can e.g. be a thermolysin from e.g. M4 family or other metallo proteases such as those from M5, M7 or M8 family.

[0675] The term "subtilase" refers to the sub-group of serine proteases according to Siezen et al., Protein Eng. 4 (1991) 719-737 and Siezen et al., Protein Science 6 (1997) 501-523. Serine proteases are a sub-group of the proteases characterized by a serine in the active site that forms a covalent adduct with the substrate during catalysis. Subtilases can be divided into six subfamilies, i.e. the subtilisin family, the thermitase family, the proteinase K family, the lanthionine antibiotic peptidase family, the Kexin family and the Pyrolysin family.

[0676] Examples of subtilases are those derived from Bacillus, such as Bacillus lentus, Bacillus alkalophilus, Bacillus subtilis, Bacillus amyloliquefaciens, Bacillus pumilus and Bacillus gibsonii described in US 7262042 and WO 09 / 021867; and Bacillus lentus protease, Bacillus subtilis protease Novo, Bacillus

[0677] Further preferred proteases are alkaline proteases from Bacillus lentus DSM 5483 (as described in e.g. WO 95 / 23221) and variants thereof (described in WO 92 / 21760, WO 95 / 23221, EP 1921147 and EP 1921148).

[0678] Examples of metalloproteases are the neutral metalloproteases as described in WO 07 / 044993 (Genencor International Inc.), such as those derived from Bacillus amyloliquefaciens.

[0679] Examples of useful proteases are the variants described in WO 92 / 19729, WO 96 / 034946, WO 98 / 20115, WO 98 / 20116, WO 99 / 011768, WO 01 / 44452, WO 03 / 006602, WO 04 / 03186, WO 04 / 041979, WO 07 / 006305, WO 11 / 036263, WO 11 / 036264, especially variants with substitutions in one or more of the following positions: 3, 4, 9, 15, 27, 36, 57, 68, 76, 87, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 106, 118, 120, 123, 128, 129, 130, 160, 167, 170, 194, 195, 199, 205, 206, 217, 218, 222, 224, 232, 235, 236, 245, 248, 252 and 274 using BPN' numbering. More preferred subtilase variants can comprise the following mutations: S3T, V4I, S9R, A15T, K27R, *36D, V68A, N76D, N87S,R, *97E, A98S, S99G,D,A, S99AD, S101G,M,R, S103A, V104I,Y,N, S106A, G118V,R, H120D,N, N123S, S128L, P129Q, S130A, G160D, Y167A, R170S, A194P, G195E, V199M, V205I, L217D, N218D, M222S, A232V, K235L, Q236H, Q245R, N252K, T274A (using BPN' numbering).

[0680] Suitable commercially available proteases include those sold under the trade names: Duralase Tm , Durazym Tm , Ultra, Ultra, Ultra, Ultra, Progress and Progress (Novozymes), those sold under the trade names: Purafect Preferenz Tm , Purafect Purafect Purafect Effectenz Tm , and (Proteolytic enzyme from Bacillus lentus, Danisco / DuPont), Axapem TM (Proteolytic enzyme from Bacillus lentus, Gist-Brocades N.V.), BLAP (sequence shown in Figure 29 of US 5352604) and variants thereof (Henkel AG) and KAP (Bacillus alkalophilus subtilisin) from Kao.

[0681] Lipase and cutinase

[0682] Suitable lipases and cutinases include those of bacterial or fungal origin. Both chemically modified mutant enzymes or protein engineered mutant enzymes are included. Examples include lipases from Thermomyces, e.g., from T. lanuginosus (formerly named Humicola lanuginosa) as described in EP 258068 and EP 305216; cutinases from Humicola, e.g., H. insolens (WO 96 / 13580); lipases from strains of Pseudomonas (some of which are now reclassified as Burkholderia), e.g., P. alcaligenes or P. pseudoalcaligenes (EP 218272), P. cepacia (EP 331376), Pseudomonas sp. strain SD705 (WO 95 / 06720 and WO 96 / 27002), P. wisconsinensis (WO 96 / 12012); GDSL-type Streptomyces lipases (WO 10 / 065455); cutinases from Magnaporthe grisea (WO 10 / 107560); cutinases from Pseudomonas mendocina (US 5,389,536); lipases from Thermobifida fusca (WO 11 / 084412); Geobacillus stearothermophilus lipase (WO 11 / 084417); lipases from Bacillus subtilis (WO 11 / 084599); and lipases from Streptomyces griseus (WO 11 / 150157) and S. pristinaespiralis (WO 12 / 137147).

[0683] Other examples are lipase variants such as those described in EP 407225, WO 92 / 05249, WO 94 / 01541, WO 94 / 25578, WO 95 / 14783, WO 95 / 30744, WO 95 / 35381, WO 95 / 22615, WO 96 / 00292, WO 97 / 04079, WO 97 / 07202, WO 00 / 34450, WO 00 / 60063, WO 01 / 92502, WO 07 / 87508, and WO 09 / 109500.

[0684] Preferred commercial lipase products include Lipolase TM , Lipex TM ; Lipolex TM and Lipoclean TM (Novozymes), Lumafast (originally from Genencor), and Lipomax (originally from Gist-Brocades).

[0685] Still other examples are lipases sometimes referred to as acyl transferases or perhydrolases, such as acyl transferases having homology to Candida antarctica lipase A (WO 10 / 111143), acyl transferases from Mycobacterium smegmatis (WO 05 / 56782), perhydrolases from the CE 7 family (WO 09 / 67279) and variants of Mycobacterium smegmatis perhydrolases, in particular the S54V variant used in the commercial product Gentle Power Bleach from Huntsman Textile Effects Pte Ltd (WO 10 / 100028).

[0686] Amylase

[0687] Suitable amylases which can be used with the variants of the present application can be alpha-amylases or glucoamylases and can be of bacterial or fungal origin. Included are chemically modified mutants or protein engineered mutants. Amylases include, for example, alpha-amylases obtained from certain strains of Bacillus, for example Bacillus licheniformis, described in greater detail in GB 1,296,839.

[0688] Suitable amylases include amylases having SEQ ID NO: 2 in WO 95 / 10603 or variants thereof having 90% sequence identity to SEQ ID NO: 3. Preferred variants are described in WO 94 / 02597, WO 94 / 18314, WO 97 / 43424 and in SEQ ID NO: 4 of WO 99 / 019467, such as variants having substitutions in one or more of the following positions: 15, 23, 105, 106, 124, 128, 133, 154, 156, 178, 179, 181, 188, 190, 197, 201, 202, 207, 208, 209, 211, 243, 264, 304, 305, 391, 408 and 444.

[0689] Different suitable amylases include amylases having SEQ ID NO: 6 in WO 02 / 010355 or a variant thereof having 90% sequence identity to SEQ ID NO: 6. Preferred variants of SEQ ID NO: 6 are those having a deletion in positions 181 and 182 and a substitution in position 193.

[0690] Other suitable amylases are hybrid alpha-amylases comprising residues 1-33 of the alpha-amylase derived from B. amyloliquefaciens shown in SEQ ID NO: 6 in WO 2006 / 066594 and residues 36-483 of the B. licheniformis alpha-amylase shown in SEQ ID NO: 4 in WO 2006 / 066594 or a variant thereof having 90% sequence identity. Preferred variants of this hybrid alpha-amylase are those having a substitution, a deletion or an insertion in one or more of the following positions: G48, T49, G107, H156, A181, N190, M197, I201, A209 and Q264. Most preferred variants of the hybrid alpha-amylase comprising residues 1-33 of the alpha-amylase derived from B. amyloliquefaciens shown in SEQ ID NO: 6 in WO 2006 / 066594 and residues 36-483 of SEQ ID NO: 4 are those having the following substitutions:

[0691] M197T;

[0692] H156Y+A181T+N190F+A209V+Q264S; or

[0693] G48A+T49I+G107A+H156Y+A181T+N190F+I201F+A209V+Q264S.

[0694] Further suitable amylases are amylases having SEQ ID NO: 6 in WO 99 / 019467 or a variant thereof having 90% sequence identity to SEQ ID NO: 6. Preferred variants of SEQ ID NO: 6 are those having a substitution, a deletion, or an insertion in one or more of the following positions: R181, G182, H183, G184, N195, I206, E212, E216 and K269. Particularly preferred amylases are those having a deletion in positions R181 and G182, or positions H183 and G184.

[0695] Further amylases that can be used are those having SEQ ID NO: 1, SEQ ID NO: 3, SEQ ID NO: 2, or SEQ ID NO: 7 of WO 96 / 023873, or a variant thereof having 90% sequence identity to SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, or SEQ ID NO: 7. Preferred variants of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, or SEQ ID NO: 7 are those having a substitution, deletion, or insertion at one or more of the following positions: 140, 181, 182, 183, 184, 195, 206, 212, 243, 260, 269, 304, and 476 (using SEQ ID 2 of WO 96 / 023873 for numbering). More preferred variants are those having a deletion in two positions selected from 181, 182, 183, and 184 (such as 181 and 182, 182 and 183, or positions 183 and 184). Most preferred amylase variants of SEQ ID NO: 1, SEQ ID NO: 2, or SEQ ID NO: 7 are those having a deletion in positions 183 and 184 and a substitution at one or more of positions 140, 195, 206, 243, 260, 304, and 476.

[0696] Other amylases that can be used are amylases having SEQ ID NO: 2 of WO 08 / 153815, SEQ ID NO: 10 in WO 01 / 66712, or a variant thereof having 90% sequence identity to SEQ ID NO: 2 of WO 08 / 153815, or a variant thereof having 90% sequence identity to SEQ ID NO: 10 in WO 01 / 66712. Preferred variants of SEQ ID NO: 10 in WO 01 / 66712 are those having a substitution, deletion, or insertion at one or more of the following positions: 176, 177, 178, 179, 190, 201, 207, 211, and 264.

[0697] Further suitable amylases are amylases having SEQ ID NO: 2 of WO 09 / 061380 or variants thereof having 90% sequence identity to SEQ ID NO: 2. Preferred variants of SEQ ID NO: 2 are those having a C-terminal truncation, and / or a substitution, a deletion or an insertion in one or more of the following positions: Q87, Q98, S125, N128, T131, T165, K178, R180, S181, T182, G183, M201, F202, N225, S243, N272, N282, Y305, R309, D319, Q320, Q359, K444 and G475. More preferred variants of SEQ ID NO: 2 are those having a substitution in one or more of the following positions: Q87E,R, Q98R, S125A, N128C, T131 I, T165I, K178L, T182G, M201 L, F202Y, N225E,R, N272E,R, S243Q,A,E,D, Y305R, R309A, Q320R, Q359E, K444E and G475K, and / or a deletion in position R180 and / or S181 or T182 and / or G183. Most preferred amylase variants of SEQ ID NO: 2 are those having the following substitutions:

[0698] N128C + K178L + T182G + Y305R + G475K;

[0699] N128C + K178L + T182G + F202Y + Y305R + D319T + G475K;

[0700] S125A + N128C + K178L + T182G + Y305R + G475K; or

[0701] S125A + N128C + T131 I + T165I + K178L + T182G + Y305R + G475K, wherein the variants are C-terminally truncated, and optionally further comprising a substitution in position 243 and / or a deletion in position 180 and / or position 181.

[0702] Other suitable amylases are alpha-amylases having SEQ ID NO: 12 in WO 01 / 66712 or variants thereof having at least 90% sequence identity to SEQ ID NO: 12. Preferred amylase variants are those having a substitution, deletion or insertion in one or more of the following positions of SEQ ID NO: 12 in WO 01 / 66712: R28, R118, N174; R181, G182, D183, G184, G186, W189, N195, M202, Y298, N299, K302, S303, N306, R310, N314; R320, H324, E345, Y396, R400, W439, R444, N445, K446, Q449, R458, N471, N484. Particularly preferred amylases include variants having a deletion of D183 and G184 and having the substitutions R118K, N195F, R320K and R458K, and variants additionally having a substitution at one or more positions selected from the group consisting of: M9, G149, G182, G186, M202, T257, Y295, N299, M323, E345 and A339, most preferably variants additionally having a substitution at all of these positions.

[0703] Other examples are amylase variants such as those described in WO 2011 / 098531, WO 2013 / 001078 and WO 2013 / 001087.

[0704] Commercially available amylases are Duramyl TM , Termamyl TM , Fungamyl TM , Stainzyme TM , Stainzyme Plus TM , Natalase TM and BAN TM from Novozymes), and Rapidase TM , Purastar TM / Effectenz TM , Powerase TM , Preferenz S1000 TM , Preferenz S1 10 TM and Preferenz S100 TM from Genencor / DuPont).

[0705] Peroxidase / oxidase

[0706] Peroxidases are peroxidases encompassed by the enzyme classification EC 1.11.1.7 as set forth by the Nomenclature Committee of the International Union of Biochemistry and Molecular Biology (IUBMB) or any fragment derived therefrom that exhibits peroxidase activity.

[0707] Suitable peroxidases include those of plant, bacterial or fungal origin. Chemically modified mutants or protein engineered mutants are included. Examples of useful peroxidases include peroxidases from Coprinopsis, for example from C. cinerea (EP 179,486), and variants thereof, such as those described in WO 93 / 24618, WO 95 / 10602 and WO 98 / 15257.

[0708] Peroxidases can also include haloperoxidases, such as chloroperoxidases, bromoperoxidases and compounds that exhibit chloroperoxidase or bromoperoxidase activity. Haloperoxidases are classified according to their specificity for halide ions. Chloroperoxidases (E.C. 1.11.1.10) catalyze the formation of hypochlorite from chloride ions.

[0709] In embodiments, the haloperoxidase is a chloroperoxidase. Preferably, the haloperoxidase is a vanadohaloperoxidase, i.e. a vanadate-containing haloperoxidase. In a preferred method of the application, a vanadate-containing haloperoxidase is combined with a source of chloride ions.

[0710] Haloperoxidases have been isolated from a number of different fungi, particularly from the group of dematiaceous hyphomycete fungi, such as Caldariomyces (e.g. C. fumago), Alternaria, Curvularia (e.g. C. verruculosa and C. inaequalis), Drechslera, Ulocladium and Botrytis.

[0711] Haloperoxidases have also been isolated from bacteria, such as Pseudomonas (e.g. P. pyrrocinia) and Streptomyces (e.g. S. aureofaciens).

[0712] In preferred embodiments, the haloperoxidase can be derived from a Curvularia species, in particular Curvularia verruculosa or Curvularia inaequalis, such as Curvularia inaequalis CBS 102.42 as described in WO 95 / 27046; or Curvularia verruculosa CBS 147.63 or Curvularia verruculosa CBS 444.70 as described in WO 97 / 04102; or from Drechslera hartlebii as described in WO 01 / 79459, Dendryphiella salina as described in WO 01 / 79458, Phaeotrichoconis crotalarie as described in WO 01 / 79461, or Geniculosporium sp. as described in WO 01 / 79460.

[0713] Oxidases can in particular include any laccase comprised by the enzyme classification EC 1.10.3.2 or any fragment derived therefrom that exhibits laccase activity, or compounds that exhibit similar activity, such as catechol oxidase (EC 1.10.3.1), ortho-aminophenol oxidase (EC 1.10.3.4) or biliverdin oxidase (EC 1.3.3.5).

[0714] Preferred laccases are enzymes of microbial origin. These enzymes can be derived from plants, bacteria or fungi, including filamentous fungi and yeasts.

[0715] Suitable examples from fungi include laccases derivable from strains of Aspergillus, Neurospora (e.g. N. crassa), Podospora, Botrytis, Collybia, Fomes, Lentinus, Pleurotus, Tricholoma (e.g. T. giganteum and T. borchii), Rhizoctonia (e.g. R. solani), Coprinus (e.g. C. cinereus, C. comatus, C. friesii and C. plicatilis), Psathyrella (e.g. P. condelleana), Panaeolus (e.g. P. papilionaceus), Myceliopthora (e.g. M. thermophilum), Schytalidium (e.g. S. thermophilum), Polyporus (e.g. P. pinsitus), Phlebia (e.g. P. radiata) (WO 92 / 01046) or Coriolus (e.g. C. hirsutus) (JP 2238885).

[0716] Suitable examples from bacteria include laccases derivable from strains of Bacillus.

[0717] Preferred are laccases derived from Coprinus or Myceliopthora; in particular laccases derived from C. cinereus, as disclosed in WO 97 / 08325, or M. thermophilum, as disclosed in WO 95 / 33836.

[0718] Nuclease

[0719] Suitable nucleases include deoxyribonucleases (DNases) and ribonucleases (RNases), which are any enzymes that catalyze the hydrolytic cleavage of phosphodiester bonds in DNA or RNA, respectively, thereby degrading DNA and RNA. There are two main classifications according to the site of activity. Exonucleases digest nucleic acids from the ends. Endonucleases act on regions in the middle of the target molecule. The nuclease is preferably a DNase, which is preferably obtainable from a microorganism, preferably a fungus or a bacterium. In particular, a DNase obtainable from a species of Bacillus is preferred; in particular, a DNase obtainable from Bacillus cibi, B. subtilis or B. licheniformis is preferred. Examples of such DNases are described in WO 2011 / 098579, WO 2014 / 087011 and WO 2017 / 060475. Also particularly preferred is a DNase obtainable from a species of Aspergillus; in particular, a DNase obtainable from A. oryzae, such as the DNase described in WO 2015 / 155350.

[0720] Licheninase

[0721] Suitable lichenase enzymes (licheninases) include enzymes that catal...

Claims

1. A xyloglucanase variant, which xyloglucanase variant comprises an alteration at a position selected from the group consisting of: A41L+P111Q+S123P+Q147K+V159M+V203T+A129T, A41L+P111Q+S123P+Q147K+V159M+K217R+A129T, P111Q+S123P+Q147K+V159M+I294Q+S402Q+A129T, A83E+P111Q+S123P+V159M+S256E+I294E+A129T, Q82E+P111Q+S123P+V159M+S256E+I294E+A129T, Q82E+P111Q+S123P+Q147K+V159M+I294E+A129T, K87E+P111Q+S123P+V159M+K217T+I294E+A129T, A83E+P111Q+S123P+V159M+K240F+K252E+A129T, K87E+P111Q+S123P+V159M+S256Q+I294E+A129T, and K87E+P111Q+S123P+V159M+K347E+N383E+A129T, wherein the variant has xyloglucanase activity and has at least 98% sequence identity to the polypeptide of SEQ ID NO:

2.

2. The variant of claim 1, wherein the variant has a half-life improvement factor of at least 1.1 compared to the reference polypeptide of SEQ ID NO:

2.

3. The variant of claim 1, wherein the variant has a half-life improvement factor of at least 1.2 compared to the reference polypeptide of SEQ ID NO:

2.

4. The variant of claim 1, wherein the variant has a half-life improvement factor of at least 1.3 compared to the reference polypeptide of SEQ ID NO:

2.

5. The variant of claim 1, wherein the variant has a half-life improvement factor of at least 1.4 compared to the reference polypeptide of SEQ ID NO:

2.

6. The variant of claim 1, wherein the variant has a half-life improvement factor of at least 1.5 compared to the reference polypeptide of SEQ ID NO:

2.

7. A detergent composition comprising the variant of any one of claims 1-6.

8. The detergent composition of claim 7, which is in the form of a bar, a uniform tablet, a unit dose product, a regular or compacted powder, a granule, a paste, a gel, or a regular or concentrated liquid.

9. An isolated polynucleotide encoding the variant of any one of claims 1-6.

10. A nucleic acid construct or expression vector comprising the polynucleotide of claim 9. ​ 11. A recombinant host cell of a non-animal or plant variety, transformed with the polynucleotide of claim 9.

12. A method of producing a variant, comprising: a. culturing the recombinant host cell of claim 11 under conditions suitable for expression of the variant; and b. recovering the variant.

13. Use of the variant of any one of claims 1-6 or the detergent composition of claim 7 or 8 for cleaning an article, for pretreating a stain on the article, and / or for preventing, reducing redeposition of soil during a wash cycle.

14. Use of the variant of any one of claims 1-6 or the detergent composition of claim 7 or 8 for removing redeposition of soil during a wash cycle.

15. Use of the variant of any one of claims 1-6 or the detergent composition of claim 7 or 8 for maintaining or improving whiteness of an article.

16. The use of claim 13 or 15, wherein the article is a textile or a hard surface.

17. A method of washing an article, comprising: c. exposing an article to a wash liquor comprising the variant of any one of claims 1-6 or the detergent composition of claim 7 or 8; d. completing at least one wash cycle; and e. optionally rinsing the article, wherein the article is a textile.

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