Cleaning compositions comprising polypeptides having fructan-degrading activity

Abstract

The present invention relates to a cleaning composition comprising a fructan-degrading enzyme and at least one cleaning component, and use of such a composition for cleaning an item, such as a textile or a surface.

Description

[0001] References to sequence lists

[0002] This application contains a sequence list in computer-readable form, which is incorporated herein by reference. Background of the Invention Technical Field

[0004] This invention relates to compositions comprising polypeptides with fructan-degrading activity, particularly cleaning compositions. The invention further relates to the use of novel fructan-degrading polypeptides, polynucleotides encoding these polypeptides, polypeptides with fructan-degrading activity in cleaning processes and / or for removing or reducing bacterial-derived fructans, and methods for removing or reducing fructans. The invention further relates to nucleic acid constructs, vectors, and host cells comprising polynucleotides encoding these polypeptides, and methods for producing and using these polypeptides. Background Technology

[0006] Enzymes have been used in detergents for decades. Often, mixtures of multiple enzymes are added to detergent compositions, containing two or more enzymes, each targeting a specific substrate; for example, amylase is active against starch stains, and protease is active against protein stains. Textiles and surfaces (such as clothing and tableware) can be soiled by many different types of dirt. Dirt can consist of proteins, fats, starches, etc. Complex stains composed of different organic substances (such as food stains, sebum, dead cell material, and extracellular polymeric matrix (EPS) from sources such as biological membranes) are difficult to remove completely with conventional automatic dishwashing (ADW) and laundry detergent compositions.

[0007] One component that can generate organic matter is fructans derived from bacteria, which are abundant in clothing. When bacteria dissolve, a component left behind on textiles or hard surfaces, such as the inner surface of a washing machine, is fructans derived from damaged cells. This fructan substrate can be sticky or adhesive; when present on textiles, it attracts dirt and can cause redeposition or re-dyeing, resulting in a graying of the textile. Furthermore, it is particularly difficult to remove odors from textiles, for example, from sweat, cigarette smoke, and pollution. Odor is a growing problem, especially with clothing that exhibits changing habits such as: washing at lower temperatures; front-loading washing machines, which save water but leave residual water between loads, leading to the proliferation of bacterial biofilms; hanging clothes to dry instead of using appliances to save energy; and the increasing popularity of synthetic fabrics, such as sportswear, which retain more odor than natural fabrics. In conventional detergent compositions, such as laundry detergents, these problems are often addressed by adding fragrances. However, this solution is not entirely effective because it is short-term and only masks the odor rather than addressing its root cause. Therefore, there is a need in this field for new solutions to overcome the problems of odor and redeposition. Summary of the Invention

[0008] This invention relates to a cleaning composition comprising a fructan-degrading enzyme and at least one cleaning component.

[0009] The present invention further relates to the use of such compositions in cleaning products, such as textiles or surfaces. The present invention further relates to a method of cleaning products, the method comprising contacting the product with a solution comprising the fructan-degrading enzyme of the present invention and at least one cleaning component. The present invention further relates to a composition comprising the fructan-degrading enzyme and at least one nuclease polypeptide having DNase or RNase activity, and its use in cleaning.

[0010] Sequence Overview

[0011] SEQ ID NO:1 encodes the DNA of a full-length polypeptide from Paenibacillus amylolyticus.

[0012] SEQ ID NO:2 is a polypeptide derived from SEQ ID NO:1

[0013] SEQ ID NO:3 Mature polypeptide obtained from Bacillus amyloliquefaciens

[0014] SEQ ID NO:4 encodes the DNA of a full-length polypeptide from Aspergillus deflectus.

[0015] SEQ ID NO:5 is a polypeptide derived from SEQ ID NO:4.

[0016] SEQ ID NO:6 Mature polypeptide obtained from Aspergillus curvature

[0017] SEQ ID NO:7 encodes the DNA of a full-length polypeptide from Aspergillus niger.

[0018] SEQ ID NO:8 is a polypeptide derived from SEQ ID NO:7.

[0019] SEQ ID NO:9 Mature polypeptide obtained from Aspergillus niger

[0020] SEQ ID NO:10 encodes the DNA of a full-length polypeptide from Penicillium ochrochloron.

[0021] SEQ ID NO:11 is a polypeptide derived from SEQ ID NO:10.

[0022] SEQ ID NO:12 Mature polypeptide obtained from Penicillium ochreae

[0023] SEQ ID NO:13 encodes the DNA of a full-length polypeptide from Penicillium murcianum.

[0024] SEQ ID NO:14 is a polypeptide derived from SEQ ID NO:13.

[0025] SEQ ID NO:15 Mature polypeptide obtained from Penicillium murcianum

[0026] SEQ ID NO:16 encodes the DNA of a full-length polypeptide from Bacillus licheniformis.

[0027] SEQ ID NO:17 is a polypeptide derived from SEQ ID NO:16.

[0028] SEQ ID NO:18 Mature polypeptide obtained from Bacillus licheniformis

[0029] SEQ ID NO:19 Mature DNAase polypeptide obtained from Bacillus cibi

[0030] SEQ ID NO:20 Mature DNAase polypeptide obtained from Bacillus licheniformis

[0031] SEQ ID NO:21 Mature DNAse polypeptide obtained from Bacillus subtilis

[0032] SEQ ID NO:22 Mature DNase polypeptide obtained from Aspergillus oryzae

[0033] SEQ ID NO:23 Mature DNase polypeptide obtained from Trichoderma harzianum

[0034] SEQ ID NO:24 motif [WPG][GMTH][NH][AEILDNMV]

[0035] SEQ ID NO:25 Motif A[YF]S[LN]D[QK]

[0036] SEQ ID NO:26 motif AYSNDK

[0037] SEQ ID NO:27 Motif [HS]WGH

[0038] SEQ ID NO:28 Motif [QLKEYMRVTANI][NSYHATRD][WF][MEVKTL][NGA][EVDLIH][PAEHDS][NATSKGVERQCD]

[0039] SEQ ID NO:29 motif KNWMNEPN

[0040] SEQ ID NO:30 Motif WMN[DE]

[0041] SEQ ID NO:31 motif WMN[DE]PNG

[0042] SEQ ID NO:32 Motif W[GMT]N[NDEI]

[0043] definition

[0044] Fructose-degrading enzyme: The term "fructose-degrading enzyme" refers to an enzyme with activity against peptidoglycan. In this context, the term may also refer to "fructanase" or a polypeptide or enzyme with fructan-degrading or fructanase activity.

[0045] Fructans are polymers of fructose molecules found in certain species of Gram-positive and Gram-negative bacteria, such as Bacillus, Streptococcus, Pseudomonas, Erwinia, and Actinomycetes, along with some fungi, such as Aspergillus and Penicillium. Bacterially produced fructan molecules are primarily composed of β(2,6)-linked fructose residues and some β(2,1)-linked branches. Such fructans are called levans and can reach a degree of polymerization (DP) exceeding 100,000 fructose units. (Vijn and Smeekens, Plant Physiology, 1999, Vol. 120: 351-359; Vanden Ende, J Exp Bot., 2018, Vol. 69(18): 4227-4231). Another major type of fructan, inulin, mainly consists of β-(2,1)-linked fructose residues and some β-(2,6)-linked branches.

[0046] The GH32 family of glycosyl hydrolases contains enzymes that hydrolyze fructose-containing polysaccharides. The GH32 family includes inulinase (EC 3.2.1.7), exoinulinase (fructan β-fructosylase) (EC 3.2.1.80), levofloxacinase (EC 3.2.1.65), 2,6-β-fructan 6-levofloxacin hydrolase (EC 3.2.1.64), fructan β-(2,1)-fructosylase (1-fructan exohydrolase) (EC 3.2.1.153), and fructan β-(2,6)-fructosylase (EC 3.2.1.154). Other enzymes in the GH32 family include those exhibiting transglycosylation activity, such as sucrose:sucrose 1-fructosyltransferase (EC 2.4.1.99), fructan:fructan 1-fructosyltransferase (2,1-fructan:2,1-fructan 1-fructosyltransferase) (EC 2.4.1.100), sucrose:fructan 6-fructosyltransferase (EC 2.4.1.10), fructan:fructan 6-G-fructosyltransferase (EC 2.4.1.243), and L-fructan glycosyltransferase (EC 2.4.1.-). (Mirjam Czjzek and Wim Van denEnde, “Glycoside Hydrolase Family 32” on CAZypedia, available at http: / / www.cazypedia.org / , accessed February 14, 2020).

[0047] Therefore, the enzymes used in the compositions of the present invention are derived from the GH32 family and may be referred to as GH32 fructanase, GH32 fructan degrading enzyme, or polypeptide. These enzymes may have both endo- and exo-activating activities.

[0048] In a preferred embodiment, these GH32 fructanases contain one or more of the aforementioned fructanase activities, such as one or more fructan β(2,1)-fructosylase activities (EC 3.2.1.153), fructan β(2,6)-fructosylase activities (EC 3.2.1.154), fructan β-fructosylase activities (EC 3.2.1.80), levofloxacinase activities (EC 3.2.1.65), 2,6-β-fructan 6-levofloxacin biohydrolase activities (EC 3.2.1.64), and inulinase activities (EC 3.2.1.7).

[0049] For the purposes of this invention, the fructan degradation activity was determined according to the method described in Example 2 below.

[0050] Biofilms are organic matter produced by any group of microorganisms in which cells adhere to one another or to a surface (such as textiles, tableware, or hard surfaces) or another surface. These adherent cells are typically embedded in an autotrophic matrix of extracellular polymeric substances (EPS). Biofilm EPS are polymeric aggregates generally composed of extracellular DNA, proteins, and polysaccharides. Biofilms can form on living or non-living surfaces. Microbial cells growing in biofilms are physiologically different from planktonic cells of the same organism (in contrast, planktonic cells are single cells that can float or float in a liquid medium). Bacteria living in biofilms often have significantly different characteristics from planktonic bacteria of the same species because the dense and protected environment of the membrane allows them to cooperate and interact in a variety of ways. One benefit of this environment for microorganisms is increased resistance to detergents and antibiotics, as the dense extracellular matrix and the outer layer of the cells protect the interior of the community. When the biofilm matrix is ​​damaged, bacteria living in biofilms do not lose their ability to live as planktonic cells. The bacteria that produce clothing, biofilms, or EPS can be found in the following species: Acinetobacter sp., Aeromicrobium sp., Brevundimonas sp., Microbacterium sp., Micrococcus luteus, Pseudomonas sp., Staphylococcus epidermidis, and Stenotrophomonas sp.

[0051] The term "catalytic domain" refers to the region of an enzyme that contains the catalytic mechanism of the enzyme.

[0052] The term "cDNA" refers to a DNA molecule that can be prepared by reverse transcription from mature, spliced ​​mRNA molecules obtained from eukaryotic or prokaryotic cells. cDNA lacks intron sequences that can be present in the corresponding genomic DNA. The initial primary RNA transcript is the precursor of mRNA, which is processed through a series of steps (including splicing) to become mature, spliced ​​mRNA.

[0053] The term "clade" refers to a group of polypeptides that are grouped together based on homology traits traced back to a common ancestor. A polypeptide clade can be viewed as a phylogenetic tree, and a clade is a group of polypeptides consisting of a common ancestor and all its direct descendants.

[0054] The term "coding sequence" refers to a polynucleotide that directly specifies the amino acid sequence of a polypeptide. The boundaries of a coding sequence are generally defined by an open reading frame (ORF), which begins with a start codon (such as ATG, GTG, or TTG) and ends with a stop codon (such as TAA, TAG, or TGA). Coding sequences can be genomic DNA, cDNA, synthetic DNA, or a combination thereof.

[0055] The term "control sequence" refers to the nucleic acid sequence necessary for the expression of the polynucleotide encoding the mature polypeptide of the present invention. Each control sequence may be native (i.e., from the same gene) or exogenous (i.e., from different genes) for the polynucleotide encoding the polypeptide, or native or exogenous relative to each other. Such control sequences include, but are not limited to, leader sequences, polyadenylated sequences, propeptide sequences, promoters, signal peptide sequences, and transcription terminators. At a minimum, these control sequences include promoters and transcription and translation termination signals. These control sequences may be provided with multiple linkers for the purpose of introducing specific restriction sites that facilitate the linking of control sequences to the coding region of the polynucleotide encoding the polypeptide.

[0056] The term "cleaning component" (or "detergent component") refers to a detergent adjuvant component that is different from the polypeptide of the present invention. The precise properties of these additional cleaning or adjuvant components, and their incorporation levels, will depend on the physical form of the composition and the nature of the operation in which the composition will be used. Suitable cleaning components include, but are not limited to, the components described below: for example, surfactants, builders and co-builders, flocculants, chelating agents, dye transfer inhibitors, enzymes (not of the present invention), enzyme stabilizers, enzyme inhibitors, catalytic materials, bleaching activators, hydrogen peroxide, hydrogen peroxide sources, pre-formed peracids, polymerizers, clay detergents / anti-redeposition agents, brighteners, defoamers, dyes, fragrances, structural elastic agents, fabric softeners, carriers, water-soluble auxiliaries, fabric colorants, antifoaming agents, dispersants, processing aids, and / or pigments. Cleaning compositions will typically contain at least one surfactant and additional components such as at least one builder and / or at least one bleaching component.

[0057] The term "cleaning composition" includes "detergent composition" and refers to a composition used to remove unwanted compounds from items to be cleaned, such as textiles. This detergent composition can be used, for example, to clean textiles, for both household and industrial cleaning. These terms encompass any material / compound selected for use in the form of a particular type of cleaning composition and product (e.g., liquid, gel, powder, granules, paste, or spray composition), and include, but are not limited to, detergent compositions such as liquid and / or solid laundry detergents and delicate fabric detergents; fabric fresheners; fabric softeners; and textile and garment pre-treatments / pre-treatments. In addition to the enzymes of the present invention, the detergent formulation may also contain one or more other enzymes (e.g., proteases, amylases, lipases, keratases, cellulases, endoglucanases, xyloglucanases, pectinases, pectin lyases, xanthan gumases, peroxidases, halogenated peroxidases, catalases, mannanases, nucleases, or any mixture thereof), and / or detergent adjuvant components such as surfactants, builders, chelating agents or chelating reagents, bleaching systems or bleaching components, polymers, fabric softeners, foaming agents, defoaming agents, dyes, fragrances, darkening inhibitors, optical brighteners, bactericides, fungicides, dirt suspending agents, corrosion inhibitors, enzyme inhibitors or stabilizers, enzyme activators, transferases, hydrolases, oxidoreductases, bluing agents and fluorescent dyes, antioxidants, and solubilizers.

[0058] The term “expression” includes any step involved in peptide production, including but not limited to: transcription, post-transcriptional modification, translation, post-translational modification, and secretion.

[0059] The term "expression vector" refers to a straight or circular DNA molecule that contains a polynucleotide encoding a polypeptide and is operatively linked to a control sequence that provides for its expression.

[0060] The term "fragment" refers to a polypeptide having one or more amino acids deleted from the amino and / or carboxyl terminus of a mature polypeptide or domain, wherein the fragment has fructan-degrading activity.

[0061] The term "host cell" refers to any cell type that is readily transformed, transfected, transduced, etc., using nucleic acid constructs or expression vectors containing the polynucleotides of the present invention. The term "host cell" also encompasses any parental cell progeny that differs from the parent cell due to mutations occurring during replication.

[0062] The term "isolated" means a substance in a form or environment not naturally occurring. Non-limiting examples of isolated substances include (1) any substance not naturally occurring, (2) any substance including, but not limited to, any enzyme, variant, nucleic acid, protein, peptide, or cofactor, which is at least partially removed from one or more naturally occurring components associated with its properties; (3) any substance artificially modified relative to a substance found in nature; or (4) any substance modified by increasing the amount of the substance relative to other components naturally associated with it (e.g., recombinant production in a host cell; multiple copies of the gene encoding the substance; and the use of a promoter stronger than the promoter naturally associated with the gene encoding the substance). Isolated substances may be present in fermentation broth samples; for example, host cells may be genetically modified to express the polypeptides of the present invention. Fermentation broth from such host cells will contain isolated polypeptides. It will be apparent to those skilled in the art that the polypeptides disclosed herein are preferably in an isolated form.

[0063] The term "laundry" encompasses both household and industrial laundry and refers to the process of treating textiles with a solution containing the cleaning or detergent composition of the present invention. The laundry process can be performed, for example, using a household or industrial washing machine or manually.

[0064] The term "foul odor" refers to an undesirable smell on cleaning products. Cleaning products should smell fresh and clean, without any foul odor adhering to them. One example of a foul odor is a compound with an unpleasant smell, which may be produced by microorganisms. Another example is an unpleasant odor, which may be the smell of sweat or body odor adhering to products that have come into contact with humans or animals. Yet another example of a foul odor can be the smell of fragrance ingredients that adhere to products, such as curry or other strongly scented fragrances.

[0065] The term "mature polypeptide" refers to a polypeptide in its mature form after N-terminal processing (e.g., removal of the signal peptide).

[0066] In one respect, the mature polypeptide is amino acids 1 to 892 of SEQ ID NO:2. Amino acids -31 to -1 of SEQ ID NO:2 are signal peptides.

[0067] In one respect, the mature polypeptide is amino acids 1 to 496 of SEQ ID NO:5. Amino acids -16 to -1 of SEQ ID NO:5 are signal peptides.

[0068] In one respect, the mature polypeptide is amino acids 1 to 491 of SEQ ID NO:8. Amino acids -25 to -1 of SEQ ID NO:8 are signal peptides.

[0069] In one respect, the mature polypeptide is amino acids 1 to 1411 of SEQ ID NO:11. Amino acids -21 to -1 of SEQ ID NO:11 are signal peptides.

[0070] In one respect, the mature polypeptide is amino acids 1 to 492 of SEQ ID NO:14. Amino acids -15 to -1 of SEQ ID NO:14 are signal peptides.

[0071] In one respect, the mature polypeptide is amino acids 1 to 653 of SEQ ID NO:17. Amino acids -24 to -1 of SEQ ID NO:17 are signal peptides.

[0072] It is known in the art that host cells can produce mixtures of two or more different mature polypeptides (i.e., with different C-terminal and / or N-terminal amino acids) expressed by the same polynucleotide. It is also known in the art that different host cells process polypeptides differently, and therefore a host cell expressing a polynucleotide can produce different mature polypeptides (e.g., with different C-terminal and / or N-terminal amino acids) compared to another host cell expressing the same polynucleotide.

[0073] The term "mature polypeptide coding sequence" refers to a polynucleotide that encodes a mature polypeptide with fructan degradation activity.

[0074] In one aspect, the mature polypeptide encoding sequence is nucleotides 94 to 2769 of SEQ ID NO:1, and nucleotides 1 to 93 of SEQ ID NO:1 encode a signal peptide.

[0075] In one aspect, the mature polypeptide encoding sequence is nucleotides 49 to 1536 of SEQ ID NO:4, and nucleotides 1 to 48 of SEQ ID NO:4 encode a signal peptide.

[0076] In one aspect, the mature polypeptide encoding sequence is nucleotides 76 to 1548 of SEQ ID NO:7, and nucleotides 1 to 75 of SEQ ID NO:7 encode a signal peptide.

[0077] In one aspect, the mature polypeptide encoding sequence is nucleotides 64 to 399 and 453 to 4349 of SEQ ID NO:10, and nucleotides 1 to 63 of SEQ ID NO:10 encode a signal peptide.

[0078] In one aspect, the mature polypeptide encoding sequence is nucleotides 46 to 1521 of SEQ ID NO:13, and nucleotides 1 to 45 of SEQ ID NO:13 encode a signal peptide.

[0079] In one aspect, the mature polypeptide encoding sequence is nucleotides 73 to 2031 of SEQ ID NO:16, and nucleotides 1 to 72 of SEQ ID NO:16 encode a signal peptide.

[0080] The term "nucleic acid construct" refers to a single-stranded or double-stranded nucleic acid molecule that is isolated from a naturally occurring gene, modified in a way that does not normally exist in nature to contain segments of nucleic acid, or is synthesized and contains one or more control sequences.

[0081] The term "operably linked" refers to a configuration in which a control sequence is placed at an appropriate position relative to the coding sequence of a polynucleotide, such that the control sequence directs the expression of the coding sequence.

[0082] The degree of association between two amino acid sequences or two nucleotide sequences is described by the parameter “sequence identity”. For the purposes of this invention, the Needleman-Wunsch algorithm (Needleman and Wunsch, 1970, J. Mol. Biol. 48:443-453) implemented by the Needleman program in the EMBOSS software package (EMBOSS: European Molecular Biology Open Software Suite, Rice et al., 2000, Trends Genet. 16:276-277) (preferably version 5.0.0 or later) is used to determine the sequence identity between two amino acid sequences. The parameters used are a vacancy opening penalty of 10, a vacancy extension penalty of 0.5, and an EBLOSUM62 (EMBOSS version of BLOSUM62) substitution matrix. The output of Needleman labeled “longest identity” (obtained using the non-simplification option) is used as the identity percentage and calculated as follows:

[0083] (identical residues × 100) / (alignment length - total number of vacancies in the alignment)

[0084] For the purposes of this invention, the Needleman-Wunsch algorithm (Needleman and Wunsch, 1970, ibid.) implemented by the Needleman program in the EMBOSS software package (EMBOSS: European Molecular Biology Open Software Suite, Rice et al., 2000, ibid.) (preferably version 5.0.0 or later) is used to determine sequence identity between two deoxynucleotide sequences. The parameters used are a vacancy opening penalty of 10, a vacancy extension penalty of 0.5, and an EDNAFULL (EMBOSS version of NCBI NUC4.4) substitution matrix. The output of Needleman labeled "Longest Identity" (obtained using the non-simplified option) is used as the identity percentage and calculated as follows:

[0085] (identical deoxyribonucleotides x 100) / (alignment length - total number of vacancies in the alignment)

[0086] The term "variant" refers to a polypeptide that exhibits fructan-degrading activity and includes alterations (i.e., substitution, insertion, and / or deletion) at one or more positions. Substitution means replacing an amino acid occupying a position with a different amino acid; deletion means removing an amino acid occupying a position; and insertion means adding an amino acid adjacent to and immediately following the amino acid occupying a position.

[0087] Nomenclature: For the purposes of this invention, the nomenclature [E / Q] or [EQ] indicates that the amino acid at this position can be glutamic acid (Glu, E) or glutamine (Gln, Q). Similarly, the nomenclature [V / G / A / I] or [VGAI] indicates that the amino acid at this position can be valine (Val, V), glycine (Gly, G), alanine (Ala, A), or isoleucine (Ile, I), and so on for other combinations as described herein. Unless further limited otherwise, amino acid X is defined such that it can be any of the 20 naturally occurring amino acids. Detailed Implementation

[0088] As mentioned in the background section above, textiles and surfaces (such as clothing and tableware) can be soiled by many different types of dirt. Single, complex stains, such as food stains, sebum, dead cell debris, EPS, or biofilm-related stains, are often composed of various organic materials such as proteins, polysaccharides, and oils, which are often difficult to remove completely with conventional detergent compositions. Furthermore, such stains can produce drawbacks such as redeposition or foul odor. The peptides of this invention address this problem, thereby providing excellent cleaning results for complex stains such as biofilms and EPS, and reducing redeposition and foul odor from, for example, textiles and tableware.

[0089] The polypeptides of the present invention are one or more fructan-degrading enzymes having the enzyme activities listed above, such as one or more fructan β(2,1)-fructosylase activities (EC 3.2.1.153), fructan β(2,6)-fructosylase activities (EC 3.2.1.154), fructan β-fructosylase activities (EC 3.2.1.80), levofloxacinase activities (EC 3.2.1.65), 2,6-β-fructan 6-levofloxacin biohydrolase activities (EC 3.2.1.64), and inulinase activities (EC 3.2.1.7). Preferred activities include β(2,6)-fructosylase activities (EC 3.2.1.154) and / or fructan β(2,1)-fructosylase activities (EC 3.2.1.153), and preferably at least β(2,6)-fructosylase activities.

[0090] The peptides of the present invention contain a GH32 domain, as defined in CAZY (Lombard, Henrissat et al., 2014. The carbohydrate-active enzymes database (CAZy), Nucleic Acids Research, 2013, 42, http: / / www.cazy.org / ). Preferably, these peptides also contain a 32C-terminal domain of a glycosyl hydrolase family (“GH32C domain”), as defined by Pfam domain number PF08244 (The Pfam protein families database: towards a more sustainable future, Finn et al., Nucleic Acids Research, 2016, Database Issue, 44: D279-D285).

[0091] Furthermore, this paper describes clusters or clades defined by specific motifs shared by polypeptides of a particular clade. The construction of the phylogenetic tree and the identification of clades and clusters are described in Example 3.

[0092] Peptides with fructan degradation activity

[0093] One embodiment of the present invention relates to a fructan-degrading polypeptide, namely GH32 fructanase having one or more of the above-described enzymatic activities, together with a cleaning composition comprising the fructan-degrading enzyme and its use for cleaning. In one embodiment, the fructan-degrading polypeptide is a polypeptide belonging to the WMNE clade and contains the motif [WPG][GMTH][NH][AEILDNMV] (SEQ ID NO:24), preferably the motif W[GMT]N[NDEI] (SEQ ID NO:32).

[0094] In another embodiment, the fructan-degrading enzyme belongs to the AYSN clade and contains the motif A[YF]S[LN]D[QK] (SEQ ID NO:25) and / or the motif [HS]WGH (SEQ ID NO:27).

[0095] In another embodiment, the fructan-degrading enzyme belongs to the WMNEP clade and contains the motif [QLKEYMRVTANI][NSYHATRD][WF][MEVKTL][NGA][EVDLIH][PAEHDS][NATSKGVERQCD](SEQID NO:28).

[0096] The motif of SEQ ID NO:24 preferably has the sequence W[GMT]N[NDEI] (SEQ ID NO:32), more particularly WMN[DE] (SEQ ID NO:30), for example WMN[DE]PNG (SEQ ID NO:31).

[0097] In one embodiment, the present invention relates to polypeptides having at least 60%, such as 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 with the mature polypeptide of SEQ ID NO:5 (these polypeptides have fructan-degrading activity), and their use in the compositions and cleaning methods of the present invention. In one aspect, these polypeptides differ from the mature polypeptide of SEQ ID NO:5 by up to 10 amino acids (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10). The polypeptide may also be a fragment of the mature polypeptide of SEQ ID NO:5 having fructan-degrading activity. The polypeptide preferably belongs to the WMNE clade and contains the motif [WPG][GMTH][NH][AEILDNMV] (SEQ ID NO:24), preferably the motif W[GMT]N[NDEI] (SEQ ID NO:32), and optionally also belongs to the AYSN clade and contains the motif A[YF]S[LN]D[QK] (SEQ ID NO:25) and / or the motif [HS]WGH (SEQ ID NO:27) and / or belongs to the WMNEP clade and contains the motif [QLKEYMRVTANI][NSYHATRD][WF][MEVKTL][NGA][EVDLIH][PAEHDS][NATSKGVERQCD] (SEQ ID NO:28).

[0098] In one embodiment, the present invention relates to polypeptides having at least 60%, such as 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 with the mature polypeptide of SEQ ID NO:8 (these polypeptides have fructan-degrading activity), and their use in the compositions and cleaning methods of the present invention. In one aspect, these polypeptides differ from the mature polypeptide of SEQ ID NO:8 by up to 10 amino acids (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10). The polypeptide may also be a fragment of the mature polypeptide of SEQ ID NO:8 having fructan-degrading activity. The polypeptide preferably belongs to the WMNE clade and contains the motif [WPG][GMTH][NH][AEILDNMV] (SEQ ID NO:24), preferably the motif W[GMT]N[NDEI] (SEQ ID NO:32), and optionally also belongs to the AYSN clade and contains the motif A[YF]S[LN]D[QK] (SEQ ID NO:25) and / or the motif [HS]WGH (SEQ ID NO:27) and / or belongs to the WMNEP clade and contains the motif [QLKEYMRVTANI][NSYHATRD][WF][MEVKTL][NGA][EVDLIH][PAEHDS][NATSKGVERQCD] (SEQ ID NO:28).

[0099] In one embodiment, the present invention relates to polypeptides having at least 60%, such as 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 with the mature polypeptide of SEQ ID NO:11 (these polypeptides have fructan-degrading activity), and their use in the compositions and cleaning methods of the present invention. In one aspect, these polypeptides differ from the mature polypeptide of SEQ ID NO:11 by up to 10 amino acids (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10). The polypeptide may also be a fragment of the mature polypeptide of SEQ ID NO:11 having fructan-degrading activity. The polypeptide preferably belongs to the WMNE clade and contains the motif [WPG][GMTH][NH][AEILDNMV] (SEQ ID NO:24), preferably the motif W[GMT]N[NDEI] (SEQ ID NO:32), and optionally also belongs to the AYSN clade and contains the motif A[YF]S[LN]D[QK] (SEQ ID NO:25) and / or the motif [HS]WGH (SEQ ID NO:27) and / or belongs to the WMNEP clade and contains the motif [QLKEYMRVTANI][NSYHATRD][WF][MEVKTL][NGA][EVDLIH][PAEHDS][NATSKGVERQCD] (SEQ ID NO:28).

[0100] In one embodiment, the present invention relates to polypeptides having at least 60%, such as 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 with the mature polypeptide of SEQ ID NO:14 (these polypeptides have fructan-degrading activity), and their use in the compositions and cleaning methods of the present invention. In one aspect, these polypeptides differ from the mature polypeptide of SEQ ID NO:14 by up to 10 amino acids (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10). The polypeptide may also be a fragment of the mature polypeptide of SEQ ID NO:14 having fructan-degrading activity. The polypeptide preferably belongs to the WMNE clade and contains the motif [WPG][GMTH][NH][AEILDNMV] (SEQ ID NO:24), preferably the motif W[GMT]N[NDEI] (SEQ ID NO:32), and optionally also belongs to the AYSN clade and contains the motif A[YF]S[LN]D[QK] (SEQ ID NO:25) and / or the motif [HS]WGH (SEQ ID NO:27) and / or belongs to the WMNEP clade and contains the motif [QLKEYMRVTANI][NSYHATRD][WF][MEVKTL][NGA][EVDLIH][PAEHDS][NATSKGVERQCD] (SEQ ID NO:28).

[0101] In one embodiment, the present invention relates to polypeptides having at least 60%, such as 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 with the mature polypeptide of SEQ ID NO:17 (these polypeptides have fructan-degrading activity), and their use in the compositions and cleaning methods of the present invention. In one aspect, these polypeptides differ from the mature polypeptide of SEQ ID NO:17 by up to 10 amino acids (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10). The polypeptide may also be a fragment of the mature polypeptide of SEQ ID NO:17 having fructan-degrading activity. The polypeptide preferably belongs to the WMNE clade and contains the motif [WPG][GMTH][NH][AEILDNMV] (SEQ ID NO:24), preferably the motif W[GMT]N[NDEI] (SEQ ID NO:32), and optionally also belongs to the AYSN clade and contains the motif A[YF]S[LN]D[QK] (SEQ ID NO:25) and / or the motif [HS]WGH (SEQ ID NO:27) and / or belongs to the WMNEP clade and contains the motif [QLKEYMRVTANI][NSYHATRD][WF][MEVKTL][NGA][EVDLIH][PAEHDS][NATSKGVERQCD] (SEQ ID NO:28).

[0102] In a particular embodiment, the present invention relates to a polypeptide having at least 60%, such as 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 with polypeptide SEQ ID NO:6 (and wherein the polypeptide has fructan-degrading activity), and its use in the compositions and cleaning methods of the present invention. The polypeptide may, for example, comprise or consist of the amino acid sequence of SEQ ID NO:6. The polypeptide may also be a fragment of SEQ ID NO:6 having fructan-degrading activity. The polypeptide preferably belongs to the WMNE clade and contains the motif [WPG][GMTH][NH][AEILDNMV] (SEQ ID NO:24), preferably the motif W[GMT]N[NDEI] (SEQ ID NO:32), and optionally also belongs to the AYSN clade and contains the motif A[YF]S[LN]D[QK] (SEQ ID NO:25) and / or the motif [HS]WGH (SEQ ID NO:27) and / or belongs to the WMNEP clade and contains the motif [QLKEYMRVTANI][NSYHATRD][WF][MEVKTL][NGA][EVDLIH][PAEHDS][NATSKGVERQCD] (SEQ ID NO:28).

[0103] In a particular embodiment, the present invention relates to a polypeptide having at least 60%, such as 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 with polypeptide SEQ ID NO:9 (and wherein the polypeptide has fructan-degrading activity), and its use in the compositions and cleaning methods of the present invention. The polypeptide may, for example, comprise or consist of the amino acid sequence of SEQ ID NO:9. The polypeptide may also be a fragment of SEQ ID NO:9 having fructan-degrading activity. The polypeptide preferably belongs to the WMNE clade and contains the motif [WPG][GMTH][NH][AEILDNMV] (SEQ ID NO:24), preferably the motif W[GMT]N[NDEI] (SEQ ID NO:32), and optionally also belongs to the AYSN clade and contains the motif A[YF]S[LN]D[QK] (SEQ ID NO:25) and / or the motif [HS]WGH (SEQ ID NO:27) and / or belongs to the WMNEP clade and contains the motif [QLKEYMRVTANI][NSYHATRD][WF][MEVKTL][NGA][EVDLIH][PAEHDS][NATSKGVERQCD] (SEQ ID NO:28).

[0104] In a particular embodiment, the present invention relates to a polypeptide having at least 60%, such as 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 with polypeptide SEQ ID NO:12 (and wherein the polypeptide has fructan-degrading activity), and its use in the compositions and cleaning methods of the present invention. The polypeptide may, for example, comprise or consist of the amino acid sequence of SEQ ID NO:12. The polypeptide may also be a fragment of SEQ ID NO:12 having fructan-degrading activity. The polypeptide preferably belongs to the WMNE clade and contains the motif [WPG][GMTH][NH][AEILDNMV] (SEQ ID NO:24), preferably the motif W[GMT]N[NDEI] (SEQ ID NO:32), and optionally also belongs to the AYSN clade and contains the motif A[YF]S[LN]D[QK] (SEQ ID NO:25) and / or the motif [HS]WGH (SEQ ID NO:27) and / or belongs to the WMNEP clade and contains the motif [QLKEYMRVTANI][NSYHATRD][WF][MEVKTL][NGA][EVDLIH][PAEHDS][NATSKGVERQCD] (SEQ ID NO:28).

[0105] In a particular embodiment, the present invention relates to a polypeptide having at least 60%, such as 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 with polypeptide SEQ ID NO:15 (and wherein the polypeptide has fructan-degrading activity), and its use in the compositions and cleaning methods of the present invention. The polypeptide may, for example, comprise or consist of the amino acid sequence of SEQ ID NO:15. The polypeptide may also be a fragment of SEQ ID NO:15 having fructan-degrading activity. The polypeptide preferably belongs to the WMNE clade and contains the motif [WPG][GMTH][NH][AEILDNMV] (SEQ ID NO:24), preferably the motif W[GMT]N[NDEI] (SEQ ID NO:32), and optionally also belongs to the AYSN clade and contains the motif A[YF]S[LN]D[QK] (SEQ ID NO:25) and / or the motif [HS]WGH (SEQ ID NO:27) and / or belongs to the WMNEP clade and contains the motif [QLKEYMRVTANI][NSYHATRD][WF][MEVKTL][NGA][EVDLIH][PAEHDS][NATSKGVERQCD] (SEQ ID NO:28).

[0106] In a particular embodiment, the present invention relates to a polypeptide having at least 60%, such as 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 with polypeptide SEQ ID NO:18 (and wherein the polypeptide has fructan-degrading activity), and its use in the compositions and cleaning methods of the present invention. The polypeptide may, for example, comprise or consist of the amino acid sequence of SEQ ID NO:18. The polypeptide may also be a fragment of SEQ ID NO:18 having fructan-degrading activity. The polypeptide preferably belongs to the WMNE clade and contains the motif [WPG][GMTH][NH][AEILDNMV] (SEQ ID NO:24), preferably the motif W[GMT]N[NDEI] (SEQ ID NO:32), and optionally also belongs to the AYSN clade and contains the motif A[YF]S[LN]D[QK] (SEQ ID NO:25) and / or the motif [HS]WGH (SEQ ID NO:27) and / or belongs to the WMNEP clade and contains the motif [QLKEYMRVTANI][NSYHATRD][WF][MEVKTL][NGA][EVDLIH][PAEHDS][NATSKGVERQCD] (SEQ ID NO:28).

[0107] In any of the embodiments disclosed herein, the polypeptide is preferably isolated, i.e., the polypeptide is in an "isolated" form as defined above or in an "isolated" environment as defined above.

[0108] Nucleic acid probes can be designed using polynucleotides of SEQ ID NO:4, SEQ ID NO:7, SEQ ID No:10, SEQ ID NO:13, or SEQ ID NO:16 or their subsequences, as well as fragments thereof, to identify and clone DNA encoding polypeptides with fructan-degrading activity from strains of different genera or species, according to methods well known in the art. In particular, such probes can be used to hybridize with genomic DNA or cDNA of target cells, following standard DNA blotting procedures, to identify and isolate the corresponding genes therein. Such probes may be significantly shorter than the complete sequence, but should be at least 15 nucleotides long, for example, at least 25, at least 35, or at least 70 nucleotides. Preferably, the nucleic acid probe is at least 100 nucleotides long, for example, at least 200, at least 300, at least 400, at least 500, at least 600, at least 700, at least 800, or at least 900 nucleotides long. Both DNA and RNA probes can be used. Probes are typically labeled (e.g., with...) 32 P, 3 H, 35 Probes (such as biotin or avidin) are used to detect corresponding genes. Such probes are covered in this invention.

[0109] DNA from genomic DNA or cDNA libraries prepared from other strains of this type that hybridizes to the probes described above and encodes polypeptides with fructan-degrading activity can be screened. Genomic DNA or other DNA from other strains of this type can be separated by agarose or polyacrylamide gel electrophoresis or other separation techniques. DNA from the library or separated DNA can be transferred and immobilized on nitrocellulose or another suitable carrier material. To identify clones or DNA that hybridize with SEQ ID NO:4, SEQ ID NO:7, SEQ ID NO:10, SEQ ID NO:13, or SEQ ID NO:16, the carrier material is used for DNA blotting.

[0110] For the purposes of this invention, hybridization refers to the hybridization of a polynucleotide with a nucleic acid probe labeled with the following under standard low to high stringency conditions: (i) SEQ ID NO:4, SEQ ID NO:7, SEQ ID NO:10, SEQ ID NO:13, or SEQ ID NO:16; (ii) the mature polypeptide coding sequence of SEQ ID NO:4, SEQ ID NO:7, SEQ ID NO:10, SEQ ID NO:13, or SEQ ID NO:16; (iii) its full-length complementary sequence; or (iv) its subsequence. Molecules hybridizing with the nucleic acid probe under these conditions can be detected using, for example, X-ray film or any other detection method known in the art.

[0111] In another embodiment, the present invention relates to a polypeptide having fructan-degrading activity, the polypeptide being encoded by a polynucleotide having 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%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity with the mature polypeptide encoding sequence of SEQ ID NO:4.

[0112] In another embodiment, the present invention relates to a polypeptide having fructan-degrading activity, the polypeptide being encoded by a polynucleotide having 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%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity with the mature polypeptide encoding sequence of SEQ ID NO: 7.

[0113] In another embodiment, the present invention relates to a polypeptide having fructan-degrading activity, the polypeptide being encoded by a polynucleotide having 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%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity with the mature polypeptide coding sequence of SEQ ID NO: 10.

[0114] In another embodiment, the present invention relates to a polypeptide having fructan-degrading activity, the polypeptide being encoded by a polynucleotide having 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%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity with the mature polypeptide encoding sequence of SEQ ID NO: 13.

[0115] In another embodiment, the present invention relates to a polypeptide having fructan-degrading activity, the polypeptide being encoded by a polynucleotide having 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%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity with the mature polypeptide encoding sequence of SEQ ID NO: 16.

[0116] In another embodiment, the present invention relates to a variant of the polypeptide represented by SEQ ID NO:3 comprising substitutions, deletions, and / or insertions at one or more sites. In one embodiment, up to 10 amino acid substitutions, deletions, and / or insertions are introduced into the polypeptide represented by SEQ ID NO:3, for example, 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10.

[0117] In another embodiment, the present invention relates to a variant of the polypeptide represented by SEQ ID NO:6 comprising substitutions, deletions, and / or insertions at one or more sites. In one embodiment, up to 10 amino acid substitutions, deletions, and / or insertions are introduced into the polypeptide represented by SEQ ID NO:6, for example, 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10.

[0118] In another embodiment, the present invention relates to a variant of the polypeptide represented by SEQ ID NO:9 comprising substitutions, deletions, and / or insertions at one or more sites. In one embodiment, up to 10 amino acid substitutions, deletions, and / or insertions are introduced into the polypeptide represented by SEQ ID NO:9, for example, 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10.

[0119] In another embodiment, the present invention relates to a variant of the polypeptide represented by SEQ ID NO:12 comprising substitutions, deletions, and / or insertions at one or more sites. In one embodiment, up to 10 amino acid substitutions, deletions, and / or insertions are introduced into the polypeptide represented by SEQ ID NO:12, such as 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10.

[0120] In another embodiment, the present invention relates to a variant of the polypeptide represented by SEQ ID NO:15 comprising substitutions, deletions, and / or insertions at one or more sites. In one embodiment, up to 10 amino acid substitutions, deletions, and / or insertions are introduced into the polypeptide represented by SEQ ID NO:15, for example, 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10.

[0121] In another embodiment, the present invention relates to a variant of the polypeptide represented by SEQ ID NO:18 comprising substitutions, deletions, and / or insertions at one or more sites. In one embodiment, up to 10 amino acid substitutions, deletions, and / or insertions are introduced into the polypeptide represented by SEQ ID NO:18, such as 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10.

[0122] Amino acid alterations in any of the above examples or elsewhere may be minor in nature, i.e., conserved amino acid substitutions or insertions that do not significantly affect protein folding and / or activity; typically small deletions of 1-30 amino acids; small N-terminal or C-terminal extensions, such as methionine residues at the N-terminus; small linker peptides of up to 20-25 residues; or small extensions that facilitate purification by altering net charge or another function (e.g., polyhistidine tags, antigenic epitopes, or binding domains).

[0123] Examples of conserved substitutions are found in the following group: 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 generally do not alter specific reactivity are known in the art and are described, for example, by H. Neurath and RL Hill, 1979, in *The Proteins*, Academic Press, New York. Common substitutes are Ala / Ser, Val / Ile, Asp / Glu, Thr / Ser, Ala / Gly, Ala / Thr, Ser / Asn, Ala / Val, Ser / Gly, Tyr / Phe, Ala / Pro, Lys / Arg, Asp / Asn, Leu / Ile, Leu / Val, Ala / Glu, and Asp / Gly.

[0124] Essential amino acids in peptides can be identified using 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, a single alanine mutation is introduced at each residue in the molecule, and the fructan-degrading activity of the resulting molecule is tested to identify amino acid residues critical to the activity of the molecule. See also Hilton et al., 1996, J. Biol. Chem. 271:4699-4708. Active sites of enzymes or other biological interactions can also be determined by physical analysis of the structure, such as by techniques including nuclear magnetic resonance, crystallography, electron diffraction, or photoaffinity labeling, along with mutagenesis of the amino acids at the putative contact sites. 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 essential amino acids can also be inferred from comparisons with related peptides.

[0125] Known mutagenesis, recombination, and / or hybridization methods can be used, followed by relevant screening procedures to perform single or multiple amino acid substitutions, deletions, and / or insertions and test them. These relevant screening procedures are, for example, those disclosed by 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; US 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).

[0126] Mutagenesis / co-coding methods can be combined with high-throughput, automated screening methods to detect the activity of cloned, mutagenesis-encoded peptides expressed by host cells (Ness et al., 1999, Nature Biotechnology 17:893-896). Mutagenesis-encoded DNA molecules encoding active peptides can be recovered from host cells and rapidly sequenced using standard methods in the art. These methods enable the rapid determination of the importance of individual amino acid residues within the peptide.

[0127] A polypeptide can be a hybrid polypeptide, in which a region of one polypeptide is fused to the N-terminus or C-terminus of a region of another polypeptide.

[0128] The polypeptide can be a fusion polypeptide or a cleavable fusion polypeptide, wherein another polypeptide is fused at the N-terminus or C-terminus of the polypeptide of the present invention. Fusion polypeptides are produced by fusing a polynucleotide encoding another polypeptide with the polynucleotide of the present invention. Techniques for producing fusion polypeptides are known in the art and include linking the coding sequences of the polypeptides such that they conform to reading frames, and that the expression of the fusion polypeptide is under the control of the same promoter and terminator. Fusion polypeptides can also be constructed using intron technology, wherein the fusion polypeptide is produced post-translationally (Cooper et al., 1993, EMBO J. [Journal of the European Society for Molecular Biology] 12:2575-2583; Dawson et al., 1994, Science 266:776-779).

[0129] Fusion peptides may further include a cleavage site between the two peptides. Upon secretion of the fusion protein, the site is cleaved, releasing both peptides. Examples of cleavage sites include, but are not limited to, those disclosed in the following literature: Martin et al., 2003, J. Ind. Microbiol. Biotechnol. [Journal of Industrial Microbiology and Biotechnology] 3:568-576; Svetina et al., 2000, J. Biotechnol. [Journal of Biotechnology] 76:245-251; Rasmussen-Wilson et al., 1997, Appl. Environ. Microbiol. [Applied and Environmental Microbiology] 63:3488-3493; Ward et al., 199 5. 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.

[0130] Sources of peptides with fructan degradation activity

[0131] The polypeptides possessing the fructan-degrading activity of the present invention can be obtained from any genus of microorganisms, particularly from bacteria or fungi. For the purposes of this invention, the term "obtained from," as used herein in conjunction with a given source, should mean that the polypeptide encoded by the polynucleotide is produced by that source or by a strain that has inserted a polynucleotide from that source. In one aspect, polypeptides obtained from a given source are secreted extracellularly, for example, obtained from bacteria.

[0132] The fructan-degrading polypeptide is preferably of microbial origin, such as bacterial or fungal origin. In this context, the term "origin" should be understood not only to polypeptides obtained from microorganisms (e.g., bacteria or fungi), but also to chemically modified mutants or protein-engineered variants. Those skilled in the art will recognize that protein engineering is commonly used to modify one or more amino acid residues in parental enzymes (e.g., wild-type enzymes) to improve one or more desired characteristics, such as enzyme activity, stability, expression level, etc.

[0133] In some respects, the polypeptide can be obtained from bacterial sources. In one respect, the polypeptide is a *Bacillus* spp. polypeptide. In another respect, the polypeptide is a *Bacillus* spp. polypeptide. In another respect, the polypeptide is a *Haloxymonas* spp. polypeptide. In another respect, the polypeptide is a *Korean* spp. polypeptide, for example, a polypeptide obtained from *Aluminum cristatum*. In another respect, the polypeptide is a *Streptomyces* spp. polypeptide, for example, a polypeptide obtained from *Streptomyces griseus*. In another respect, the polypeptide is a *Nonomura* spp. polypeptide, for example, a polypeptide obtained from *Nonomura coxsii*, *Nonomura disei*, or *Nonomura guangzhouensis*. In another respect, the polypeptide is a *Micromonospora* spp. polypeptide, for example, a polypeptide obtained from *Micromonospora apulia*, *Micromonospora crimsonii*, or *Micromonospora marinei*. In another respect, the polypeptide is a *Laceyella* spp. polypeptide, for example, a polypeptide obtained from *Laceyella sacchari*. In yet another respect, the polypeptide is a *Bacillus* spp. polypeptide, for example, a polypeptide obtained from *Bacillus thermoseptica* or *Bacillus coli*. In one aspect, the polypeptide is a Lysobacterium polypeptide, such as a polypeptide obtained from antibiotic Lysobacterium or capsicum Lysobacterium. In another aspect, the polypeptide is a Hamadaea polypeptide, such as a polypeptide obtained from Hamadaea dunnioni. In another aspect, the polypeptide is a Bacillus-like polypeptide, such as a polypeptide obtained from Bacillus pineae. In another aspect, the polypeptide is a Thermostaphylospora polypeptide, such as a polypeptide obtained from Thermostaphylospora chromogenica. In another aspect, the polypeptide is a Pseudomonas polypeptide, such as a polypeptide obtained from Pseudomonas sphaeroides or Pseudomonas alcaligenes.

[0134] In another respect, the polypeptide can be obtained from fungal sources, such as fungi of the Aspergillus family (e.g., Aspergillus or Penicillium).

[0135] It should be understood that, for the aforementioned species, this invention covers complete and incomplete stages, as well as other taxonomic equivalents, such as asexual forms, regardless of their known species names. Those skilled in the art will readily identify the appropriate equivalents.

[0136] Strains of these species are readily available to the public at many culture collections, such as the American Type Culture Collection (ATCC), the German Microbial Culture Collection (DSMZ), the Centralbureau Voor Schimmelcultures (CBS), and the Agricultural Research Service Patent Culture Collection, Northern Regional Research Center (NRRL).

[0137] The polypeptide can be identified and obtained from other sources, including microorganisms isolated from nature (e.g., soil, compost, water, etc.) or DNA samples obtained directly from natural materials (e.g., soil, compost, water, etc.), using the probes mentioned above. Techniques for directly isolating microorganisms and DNA from natural habitats are well known in the art. The polynucleotide encoding the polypeptide can then be obtained by similarly screening a library of genomic DNA or cDNA from another microorganism or a mixed DNA sample. Once the polynucleotide encoding the polypeptide has been detected with one or more probes, the polynucleotide can be isolated or cloned using techniques known to those skilled in the art (see, for example, Sambrook et al., 1989, ibid.).

[0138] Nucleic acid constructs

[0139] The present invention also relates to nucleic acid constructs comprising polynucleotides of the present invention operably linked to one or more control sequences, which, under conditions compatible with the control sequences, guide the expression of coding sequences in suitable host cells.

[0140] Polynucleotides can be manipulated in various ways to provide polypeptide expression. Depending on the expression vector, manipulating the polynucleotide before insertion into the vector may be desirable or necessary. Techniques for modifying polynucleotides using recombinant DNA methods are well known in the art.

[0141] The control sequence may be a promoter, i.e., a polynucleotide recognized by the host cell for expressing the polypeptide encoding the present invention. The promoter contains a transcriptional control sequence that mediates the expression of the polypeptide. The promoter can be any polynucleotide exhibiting transcriptional activity in the host cell, including variants, truncated and heterozygous promoters, and can be obtained from a gene encoding an extracellular or intracellular polypeptide that is homologous or heterologous to that of the host cell.

[0142] Examples of suitable promoters for guiding the transcription of the nucleic acid constructs of the present invention in bacterial host cells are those obtained from the following promoters: Bacillus amyloliquefaciens α-amylase gene (amyQ), Bacillus licheniformis α-amylase gene (amyL), Bacillus licheniformis penicillinase gene (penP), Bacillus thermophilus malt amylase gene (amyM), Bacillus subtilis fructan sucrase gene (sacB), Bacillus subtilis xylA and xylB genes, and Bacillus thuringiensis cryIIIA gene (Agaisse and Lereclus, 1994, Molecular). Microbiology 13:97-107), Escherichia coli lac operon, Escherichia coli trc promoter (Egon et al., 1988, Gene 69:301-315), Streptomyces agarase gene (dagA), and prokaryotic β-lactamase gene (Villa-Kamaroff et al., 1978, Proc. Natl. Acad. Sci. USA 75:3727-3731), along with tac promoter (DeBoer et al., 1983, Proc. Natl. Acad. Sci. USA 80:21-25). Other promoters are described in “Useful proteins from recombinant bacteria”, Gilbert et al., 1980, Scientific American 242:74-94; and Sambrook et al., 1989, ibid. Examples of tandem promoters are disclosed in WO 99 / 43835.

[0143] Examples of suitable promoters for guiding the transcription of the nucleic acid constructs of this invention in filamentous fungal host cells are promoters obtained from the following genes: Aspergillus nidulans acetamase, Aspergillus niger neutral α-amylase, Aspergillus niger acid-stable α-amylase, Aspergillus niger or Aspergillus awamori glucosylase (glaA), Aspergillus oryzae TAKA amylase, Aspergillus oryzae alkaline protease, Aspergillus oryzae triose phosphate isomerase, Fusarium oxysporum trypsin-like protease (WO 96 / 00787), Fusarium moniliforme amyloglucosidase (WO 00 / 56900), Fusarium moniliforme Daria (WO 00 / 56900), Fusarium moniliforme Quinn (WO 00 / 56900), and Rhizomucor miltiorrhiza. (miehei) Lipase, *Rhizopus miehei* aspartic protease, *Trichoderma reesei* β-glucosidase, *Trichoderma reesei* cellobiose hydrolase I, *Trichoderma reesei* cellobiose hydrolase II, *Trichoderma reesei* endoglucanase I, *Trichoderma reesei* endoglucanase II, *Trichoderma reesei* endoglucanase III, *Trichoderma reesei* endoglucanase V, *Trichoderma reesei* xylanase I, *Trichoderma reesei* xylanase II, *Trichoderma reesei* xylanase III, *Trichoderma reesei* β-xylosidase, and *Trichoderma reesei* translation elongation factor. And the NA2-tpi promoter (a modified promoter from the Aspergillus neutral α-amylase gene, wherein the untranslated leader sequence has been replaced with an untranslated leader sequence from the Aspergillus triose phosphate isomerase gene; non-limiting examples include a modified promoter from the Aspergillus niger neutral α-amylase gene, wherein the untranslated leader sequence has been replaced with an untranslated leader sequence from the Aspergillus nidus or Aspergillus oryzae triose phosphate isomerase gene); and its variants, truncated forms, and heterozygous promoters. Other promoters are described in U.S. Patent No. 6,011,147.

[0144] In the yeast host, useful promoters are derived from the following genes: *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-phosphate glycerate kinase. Other useful promoters in the yeast host cell are described by Romanos et al., 1992, Yeast [Yeast] 8:423-488.

[0145] The control sequence may also be a transcription terminator recognized by the host cell to terminate transcription. The terminator is operatively linked to the 3' end of a polynucleotide encoding a polypeptide. Any terminator that is functional in the host cell can be used in this invention.

[0146] The preferred terminator for bacterial host cells is obtained from the following genes: Bacillus clausti alkaline protease (aprH), Bacillus licheniformis α-amylase (amyL), and Escherichia coli ribosomal RNA (rrnB).

[0147] The preferred terminator for filamentous fungal host cells is derived from the following genes: Aspergillus nidulans acetamase, Aspergillus nidulans o-aminobenzoic acid synthase, Aspergillus niger glucosylamylase, Aspergillus niger α-glucosidase, Aspergillus oryzae TAKA amylase, Fusarium oxysporum trypsin-like protease, Trichoderma reesei β-glucosidase, Trichoderma reesei cellobiose hydrolase I, Trichoderma reesei cellobiose hydrolase II, Trichoderma reesei endoglucanase I, Trichoderma reesei endoglucanase II, Trichoderma reesei endoglucanase III, Trichoderma reesei endoglucanase V, Trichoderma reesei xylanase I, Trichoderma reesei xylanase II, Trichoderma reesei xylanase III, Trichoderma reesei β-xylosidase, and Trichoderma reesei translation elongation factor.

[0148] Preferred terminators for yeast host cells are derived from the following genes: *Saccharomyces cerevisiae* enolase, *Saccharomyces cerevisiae* cytochrome C (CYC1), and *Saccharomyces cerevisiae* glyceraldehyde-3-phosphate dehydrogenase. Other useful terminators for yeast host cells are described by Romanos et al. (1992, ibid.).

[0149] Control sequences can also be mRNA stabilizing regions downstream of the promoter and upstream of the gene's coding sequence, which enhance the expression of the gene.

[0150] Examples of suitable mRNA stable regions were obtained from the following genes: Bacillus thuringiensis cryIIIA gene (WO94 / 25612) and Bacillus subtilis SP82 gene (Hue et al., 1995, Journal of Bacteriology 177:3465-3471).

[0151] The control sequence can also be a leader sequence, which is a non-translated region of mRNA that is important for translation in the host cell. The leader sequence is operatively linked to the 5' end of a polynucleotide encoding a polypeptide. Any leader sequence that is functional in the host cell can be used.

[0152] The preferred leader sequence for filamentous fungal host cells is obtained from the following genes: Aspergillus oryzae TAKA amylase and Aspergillus nidulans triose phosphate isomerase.

[0153] The appropriate leader sequence for yeast host cells is obtained from the following genes: Saccharomyces cerevisiae enolase (ENO-1), Saccharomyces cerevisiae 3-phosphoglycerate kinase, Saccharomyces cerevisiae α-factor, and Saccharomyces cerevisiae alcohol dehydrogenase / glyceraldehyde-3-phosphate dehydrogenase (ADH2 / GAP).

[0154] The control sequence can also be a polyadenylation sequence, a sequence operatively linked to the 3' end of the polynucleotide and recognized by the host cell during transcription as a signal to add polyadenylation residues to the transcribed mRNA. Any polyadenylation sequence that is functional in the host cell can be used.

[0155] Preferred polyadenylated sequences for filamentous fungal host cells are derived from the following genes: Aspergillus nidulans o-aminobenzoic acid synthase, Aspergillus niger glucosyl amylase, Aspergillus niger α-glucosidase, Aspergillus oryzae TAKA amylase, and Fusarium oxysporum trypsin-like protease.

[0156] Useful polyadenylation sequences in yeast host cells are described by Guo and Sherman, 1995, Mol. Cellular Biol. [Molecular Cell Biology] 15: 5983-5990.

[0157] The control sequence can also be a signal peptide coding region encoding a signal peptide that is linked to the N-terminus of the polypeptide and guides the polypeptide into the secretory pathway of the cell. The 5' end of the polynucleotide coding sequence itself may contain a signal peptide coding sequence naturally linked to the coding sequence segment of the polypeptide within the translation reading frame. Alternatively, the 5' end of the coding sequence may contain a signal peptide coding sequence that is exogenous to the coding sequence. In cases where the coding sequence does not naturally contain a signal peptide coding sequence, an exogenous signal peptide coding sequence may be required. Alternatively, an exogenous signal peptide coding sequence may simply replace the natural signal peptide coding sequence to enhance polypeptide secretion. However, any signal peptide coding sequence that guides the secretory pathway of the expressed polypeptide into the host cell can be used.

[0158] The effective signal peptide coding sequences for bacterial host cells are derived from the following genes: maltose amylase produced by Bacillus NCIB 11837, subtilisin protease from Bacillus licheniformis, β-lactamase from Bacillus licheniformis, α-amylase from Bacillus thermophilus, neutral proteases (nprT, nprS, nprM) from Bacillus thermophilus, and prsA from Bacillus subtilis. Additional signal peptides are described by Simonen and Palva, 1993, Microbiological Reviews, 57:109-137.

[0159] The effective signal peptide coding sequences for filamentous fungal host cells are obtained from the following genes: Aspergillus niger neutral amylase, Aspergillus niger glucosylase, Aspergillus oryzae TAKA amylase, Aspergillus oryzae cellulase, Aspergillus oryzae endoglucanase V, Aspergillus sparsely cottony lipase, and Rhizopus oryzae aspartic protease.

[0160] Useful signal peptides in yeast host cells are obtained from genes such as *Saccharomyces cerevisiae* α-factor and *Saccharomyces cerevisiae* invertase. Sequences encoding other useful signal peptides are described by Romanos et al. (1992, ibid.).

[0161] The control sequence can also be a propeptide-coding sequence encoding the propeptide located at the N-terminus of the polypeptide. The resulting polypeptide is called a precursor enzyme or propeptide progenitor (or, in some cases, a zymogen). The propeptide progenitor is generally inactive and can be converted into an active polypeptide by catalytic cleavage or autocatalytic cleavage of the propeptide progenitor. Propeptide-coding sequences can be obtained from the following genes: Bacillus subtilis alkaline protease (aprE), Bacillus subtilis neutral protease (nprT), Myceliophthora thermophila laccase (WO 95 / 33836), Rhizopus miltiorrhiza aspartic protease, and Saccharomyces cerevisiae α-factor.

[0162] When both the signal peptide sequence and the propeptide sequence are present, the propeptide sequence is located immediately adjacent to the N-terminus of the polypeptide, and the signal peptide sequence is located immediately adjacent to the N-terminus of the propeptide sequence.

[0163] It is also desirable to add regulatory sequences that modulate the expression of host cell growth-related peptides. Examples of regulatory sequences are those that cause gene expression to turn on or off in response to chemical or physical stimuli, including the presence of regulatory compounds. Regulatory sequences in prokaryotic systems include the lac, tac, and trp operon systems. In yeast, the ADH2 or GAL1 system can be used. In filamentous fungi, the *Aspergillus niger* glucosylamylase promoter, the *Aspergillus oryzae* TAKA α-amylase promoter and *Aspergillus oryzae* glucosylamylase promoter, and the *Trichoderma reesei* cellobiose hydrolase I and cellobiose hydrolase II promoters can be used. Other examples of regulatory sequences are those that amplify genes. In eukaryotic systems, these regulatory sequences include dihydrofolate reductase genes amplified in the presence of methotrexate and metallothionein genes amplified with heavy metals. In these cases, the polynucleotide encoding the peptide is operatively linked to the regulatory sequence.

[0164] expression carrier

[0165] The present invention also relates to recombinant expression vectors comprising the polynucleotide of the present invention, a promoter, and transcription and translation termination signals. Multiple nucleotides and control sequences can be linked together to produce a recombinant expression vector, which may include one or more convenient restriction sites for the insertion or substitution of the polynucleotide encoding the polypeptide at such sites. Alternatively, the polynucleotide can be expressed by inserting the polynucleotide or a nucleic acid construct containing the polynucleotide into a suitable vector for expression. In producing the expression vector, the coding sequence is located in the vector such that the coding sequence is operatively linked to a suitable control sequence for expression.

[0166] Recombinant expression vectors can be any vector (e.g., plasmids or viruses) that can readily undergo recombinant DNA procedures and induce polynucleotide expression. The choice of vector will typically depend on its compatibility with the host cell to which it will be introduced. Vectors can be linear or closed circular plasmids.

[0167] The vector can be a self-replicating vector, that is, a vector that exists as an extrachromosomal entity and replicates independently of chromosome replication, such as a plasmid, extrachromosomal element, microchromosome, or artificial chromosome. The vector can contain any means to ensure self-replication. Alternatively, the vector can be one that integrates into the genome when introduced into a host cell and replicates along with one or more chromosomes in which it has already been integrated. Furthermore, a single vector or plasmid, or two or more vectors or plasmids collectively containing the total DNA of the host cell genome to be introduced, or transposons can be used.

[0168] The vector preferably contains one or more selective markers that allow for convenient selection of cells such as transformed cells, transfected cells, and transduced cells. A selective marker is a gene whose product provides resistance to biocides or viruses, resistance to heavy metals, or prototrophic auxotrophic traits, etc.

[0169] Examples of selective bacterial markers include the dal gene in Bacillus licheniformis or Bacillus subtilis, 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. Selective markers used in filamentous fungal host cells include, but are not limited to, adeA (phosphogluconoylaminoimidazolium-succinate synthase), adeB (phosphogluconoylaminoimidazolium synthase), amdS (acetamipase), argB (ornithine carbamoyltransferase), bar (glufosinate acetyltransferase), hph (hygromycin phosphotransferase), niaD (nitrate reductase), niaA (nitrite reductase), pyrG (orotate nucleoside-5'-phosphate decarboxylase), sC (adenosyl sulfate transtransferase), and trpC (o-aminobenzoic acid synthase), and their equivalents. Preferred genes used in Aspergillus cells include the amdS and pyrG genes from Aspergillus nidus or Aspergillus oryzae, and the bar gene from Streptomyces hygroscopicus. Preferred genes used in Trichoderma cells include the adeA, adeB, amdS, hph, and pyrG genes.

[0170] Selective labeling can be a biselective labeling system as described in WO 2010 / 039889. In one aspect, biselective labeling is the hph-tk biselective labeling system.

[0171] The vector preferably contains elements that allow the vector to integrate into the host cell genome or to replicate autonomously in the cell independently of the genome.

[0172] For integration into the host cell genome, the vector can rely on a polynucleotide sequence encoding the polypeptide or any other element of the vector for integration into the genome via homologous or non-homologous recombination. Alternatively, the vector can contain additional polynucleotides to guide integration into the host cell genome at a precise location on the chromosome via homologous recombination. To increase the likelihood of integration at a precise location, the integrative element should contain a sufficient number of nucleic acids, such as 100 to 10,000 base pairs, 400 to 10,000 base pairs, and 800 to 10,000 base pairs, that have high sequence identity with the corresponding target sequence to enhance the probability of homologous recombination. The integrative element can be any sequence homologous to the target sequence within the host cell genome. Moreover, the integrative element can be a non-coding or coding polynucleotide. On the other hand, the vector can integrate into the host cell genome via non-homologous recombination.

[0173] For autonomous replication, the vector may further include an origin of replication, which enables the vector to replicate autonomously within the host cell discussed in this context. The origin of replication can be any plasmid replicon that mediates autonomous replication and functions within the cell. The terms "origin of replication" or "plasmid replicon" refer to the polynucleotide that enables a plasmid or vector to replicate in vivo.

[0174] Examples of bacterial origins of replication are the origins of replication of plasmids pBR322, pUC19, pACYC177, and pACYC184, which allow replication in Escherichia coli, and the origins of replication of plasmids pUB110, pE194, pTA1060, and pAMβ1, which allow replication in Bacillus.

[0175] Examples of replication origins used in yeast host cells are 2-micron replication origins, ARS1, ARS4, combinations of ARS1 and CEN3, and combinations of ARS4 and CEN6.

[0176] Examples of useful origins of replication in filamentous fungal cells are AMA1 and ANS1 (Gems et al., 1991, Gene 98:61-67; Cullen et al., 1987, Nucleic Acids Res. 15:9163-9175; WO00 / 24883). The AMA1 gene can be isolated and plasmids or vectors containing the gene can be constructed according to the methods disclosed in WO 00 / 24883.

[0177] More than one copy of the polynucleotide of the present invention can be inserted into host cells to enhance polypeptide production. An increased 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 selective marker gene along with the polynucleotide, wherein cells containing the amplified copy of the selective marker gene and thus additional copies of the polynucleotide can be selected by culturing cells in the presence of a suitable selective reagent.

[0178] The procedures for connecting the above-described elements to construct the recombinant expression vector of the present invention are well known to those skilled in the art (see, for example, Sambrook et al., 1989, ibid.).

[0179] host cells

[0180] This invention also relates to recombinant host cells containing polynucleotides of the invention operably linked to one or more control sequences that direct the production of polypeptides of the invention. A construct or vector containing the polynucleotide is introduced into the host cell such that the construct or vector is maintained as a chromosomal integrase or as an autonomously replicating extrachromosomal vector, as previously described. The term "host cell" encompasses any parental cell progeny that is not identical to the parental cell due to mutations occurring during replication. The selection of the host cell will depend largely on the gene encoding the polypeptide and its origin.

[0181] The host cell can be any cell useful in the recombinant production of the polypeptides of the present invention, such as prokaryotes or eukaryotes.

[0182] Prokaryotic host cells can be any Gram-positive or Gram-negative bacteria. Gram-positive bacteria include, but are not limited to: Bacillus, Clostridium, Enterococcus, Bacillus aeruginosa, Lactobacillus, Lactococcus, Bacillus cereus, Staphylococcus, Streptococcus, and Streptomyces. Gram-negative bacteria include, but are not limited to: Campylobacter, Escherichia coli, Flavobacterium, Fusobacterium, Helicobacter, Staphylococcus, Neisseria, Pseudomonas, Salmonella, and Ureaplasma.

[0183] The bacterial host cell can be any Bacillus genus cell, including but not limited to: Bacillus alkalophilus, Bacillus altitudinis, Bacillus amyloliquefaciens subsp. plantarum, Bacillus brevis, Bacillus circulans, Bacillus croceae, Bacillus scoagulans, Bacillus firmus, Bacillus lautus, Bacillus lentus, Bacillus licheniformis, Bacillus megaterium, Bacillus methylotrophicus, Bacillus pumilus, and Bacillus safranin. (Safensis), thermophilic steatobacterium, Bacillus subtilis and Bacillus thuringiensis cells.

[0184] The bacterial host cell can also be any streptococcus cell, including but not limited to Streptococcus equisimilis, Streptococcus pyogenes, Streptococcus uberis, and Streptococcus equisubsp. Zooepidemicus.

[0185] 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.

[0186] DNA can be introduced into Bacillus cells via protoplast transformation (see, for example, Chang and Cohen, 1979, Mol. Gen. Genet. [Molecular Genetics and Genomics] 168:111-115), competent cell transformation (see, for example, Young and Spizizen, 1961, J. Bacteriol. [Journal of Bacteriology] 81:823-829; or Dubnau and Davidoff-Abelson, 1971, J. Mol. Biol. [Journal of Molecular Biology] 56:209-221), electroporation (see, for example, Shigekawa and Dower, 1988, Biotechniques [Biotechnology] 6:742-751), or conjugation (see, for example, Koehler and Thorne, 1987, J. Bacteriol. [Journal of Bacteriology] 169:5271-5278). DNA can be introduced into E. coli cells by protoplast transformation (see, for example, Hanahan, 1983, J. Mol. Biol. [Journal of Molecular Biology] 166:557-580) or electroporation (see, for example, Dower et al., 1988, Nucleic Acids Res. [Nucleic Acid Research] 16:6127-6145). DNA can be introduced into Streptomyces cells via protoplast transformation, electroporation (see, for example, Gong et al., 2004, Folia Microbiol. (Praha) 49:399-405), conjugation (see, for example, Mazodier et al., 1989, J. Bacteriol. 171:3583-3585), or transduction (see, for example, Burke et al., 2001, Proc. Natl. Acad. Sci. USA 98:6289-6294). DNA can be introduced into Pseudomonas cells by electroporation (see, for example, Choi et al., 2006, J. Microbiol. Methods 64:391-397) or conjugation (see, for example, Pinedo and Smets, 2005, Appl. Environ. Microbiol. 71:51-57).DNA can be introduced into Streptococcus cells via: native competent cells (see, for example, Perry and Kuramitsu, 1981, Infect. Immun. 32:1295-1297), protoplast transformation (see, for example, Catt and Jollick, 1991, Microbios 68:189-207), electroporation (see, for example, Buckley et al., 1999, Appl. Environ. Microbiol. 65:3800-3804), or conjugation (see, for example, Clewell, 1981, Microbiol. Rev. 45:409-436). However, any method known in the art for introducing DNA into host cells can be used.

[0187] The host cell can also be a eukaryotic cell, such as a mammalian, insect, plant, or fungal cell.

[0188] The host cell can be a fungal cell. As used herein, “fungus” includes Ascomycota, Basidiomycota, Chytridiomycota, Zygomycota, Oomycota, and all mitotic fungi (as defined by Hawksworth et al. in Ainsworth and Bisby’s Dictionary of The Fungi, 8th edition, 1995, CAB International, University Press, Cambridge, UK).

[0189] The host cell for fungi can be a yeast cell. As used herein, “yeast” includes ascosporogenous yeast (Endomycetales), basidiosporogenous yeast, and yeasts belonging to the class Fungi Imperfecti (Blastomycetes). Since yeast classification may change in the future, for the purposes of this invention, yeast should be defined as described in *Biology and Activities of Yeast* (edited by Skinner, Passmore, and Davenport, Soc. App. Bacteriol. Symposium Series No. 9, 1980).

[0190] Yeast host cells can be cells from the genera *Candida*, *Hansenula*, *Kluyveromyces*, *Pichia*, *Saccharomyces*, *Schizosaccharomyces*, or *Yarrowia*, such as *Kluyveromyces lactis*, *Saccharomyces carlsbergensis*, *Saccharomyces diastaticus*, *Saccharomyces douglasii*, *Saccharomyces kluyveri*, *Saccharomyces norbensis*, *Saccharomyces oviformis*, or *Yarrowia lipolytica*.

[0191] The host cell for fungi can be a filamentous fungal cell. "Filamentous fungi" includes all filamentous forms within the phylum Eumycota and subphyla Oomycota (as defined by Hawksworth et al., 1995, ibid.). Filamentous fungi are generally characterized by a hyphal wall composed of chitin, cellulose, glucan, chitosan, mannan, and other complex polysaccharides. Vegetative growth occurs through hyphal extension, and carbon metabolism is obligate aerobic. In contrast, the vegetative growth of yeasts (such as Saccharomyces cerevisiae) occurs through budding of single-celled cells, and carbon metabolism can be fermentative.

[0192] The host cells for this filamentous fungus can be *Aureobasidium*, *Bjerkandera*, *Ceriporiopsis*, *Chrysosporium*, *Coprinus*, *Coriolus*, *Cryptococcus*, *Filibasidium*, *Fusarium*, *Humicola*, *Magnaporthe*, *Mucor*, and *Mycelium*. Cells of the genera *iophthora*, *Neocallimastix*, *Neurospora*, *Paecilomyces*, *Penicillium*, *Phanerochaete*, *Phlebia*, *Piromyces*, *Pleurotus*, *Schizophyllum*, *Basilaria*, *Thermoascus*, *Thielavia*, *Tolypocladium*, *Trametes*, or *Trichoderma*.

[0193] For example, the host cells of filamentous fungi can be *Aspergillus bubomori*, *Aspergillus sulphureus*, *Aspergillus fumigatus*, *Aspergillus japonicus*, *Aspergillus nidus*, *Aspergillus oryzae*, *Bjerkandera adusta*, *Ceriporiopsis aneirina*, *Ceriporiopsis caregiea*, *Ceriporiopsis gilvescens*, *Ceriporiopsis pannocinta*, *Ceriporiopsis rivulosa*, *Ceriporiopsis subrufa*, *Ceriporiopsis subvermispora*, *Chrysosporium inops*, *Chrysosporium lucknowense*, and *Chrysosporium lucknowense*. *Chrysosporium pannicola*, *Chrysosporium queenslandicum*, *Chrysosporium zonatum*, *Coprinus cinereus*, *Coriolushirsutus*, *Fusarium graminearum ... Chrysosporium), Phlebiaradiata, Pleurotus eryngii, Clostridium terrestris, Trametes villosa, Trametes versicolor, Trichoderma harzianum, Trichoderma corningensis, Trichoderma longifolia, Trichoderma reesei, or green Trichoderma cells.

[0194] Fungal cells can be transformed in a manner known per se through processes involving protoplast formation, protoplast transformation, and cell wall regeneration. Suitable procedures for transforming Aspergillus and Trichoderma host cells are described in EP 238023 and Yelton et al., 1984, Proc. Natl. Acad. Sci. USA [Proceedings of the National Academy of Sciences] 81:1470-1474, and Christensen et al., 1988, Bio / Technology [Biology / Technology] 6:1419-1422. Suitable methods for transforming Fusarium species are described by Malardier et al., 1989, Gene [Gene] 78:147-156 and WO 96 / 00787. Yeast can be transformed using procedures described in the following literature: Becker and Guarente, in Abelson, JN and Simon, MI (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.

[0195] Production methods

[0196] The present invention also relates to methods for producing the polypeptides of the present invention, the methods comprising (a) culturing cells under conditions favorable for the production of the polypeptide, the cells producing the polypeptide in their wild-type form; and optionally (b) recovering the polypeptide.

[0197] The present invention also relates to recombinant methods for producing the polypeptides of the present invention, the methods comprising (a) culturing recombinant host cells of the present invention capable of expressing the polypeptide under conditions favorable for the production of the polypeptide; and optionally (b) recovering the polypeptide.

[0198] One embodiment of the present invention relates to a method for producing a polypeptide selected from the group consisting of SEQ ID NO:3, SEQ ID NO:6, SEQ ID NO:9, SEQ ID NO:12, SEQ ID NO:15 and SEQ ID NO:18, and polypeptides having at least 60% sequence identity with these polypeptides, such as 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, the method comprising (a) culturing a recombinant host cell of the present invention capable of expressing one of these polypeptides under conditions conducive to the production of the polypeptide; and optionally (b) recovering the polypeptide.

[0199] The host cells are cultured in a suitable nutrient medium for producing the peptide using methods known in the art. For example, cells can be cultured by shake flask culture or by small-scale or large-scale fermentation (including continuous, batch, fed-batch, or solid-state fermentation) in a laboratory or industrial fermenter, in a suitable medium and under conditions that allow for peptide expression and / or isolation. Using procedures known in the art, the culture occurs in a suitable nutrient medium containing carbon and nitrogen sources and inorganic salts. Suitable media are available from commercial suppliers or can be prepared according to publicly available compositions (e.g., in the catalogue of the U.S. Center for Type Culture Collection). If the peptide is secreted into the nutrient medium, it can be recovered directly from the medium. If the peptide is not secreted, it can be recovered from cell lysates.

[0200] Peptides can be detected using methods known in the art that are specific to peptides. These detection methods include, but are not limited to, the use of specific antibodies, the formation of enzyme products, or the disappearance of enzyme substrates. For example, enzyme assays can be used to determine the activity of a peptide.

[0201] Peptides can be recovered using methods known in the art. For example, peptides can be recovered from nutrient media through conventional procedures, including but not limited to collection, centrifugation, filtration, extraction, spray drying, evaporation, or precipitation. In one aspect, fermentation broth containing peptides can be recovered.

[0202] Peptides can be purified using a variety of procedures known in the art, including but not limited to chromatography (e.g., ion exchange chromatography, affinity chromatography, hydrophobic chromatography, focusing chromatography, and size exclusion chromatography), electrophoresis procedures (e.g., preparative isoelectric focusing electrophoresis), differential dissolution (e.g., ammonium sulfate precipitation), SDS-PAGE, or extraction (see, for example, Protein Purification, edited by Janson and Ryden, VCH Publishers, New York, 1989) to obtain substantially pure peptides.

[0203] In terms of alternatives, instead of recovering the peptide, the host cells of the present invention expressing the peptide are used as the peptide source. Another option is to use the supernatant from peptide expression as the peptide source.

[0204] Fermentation broth preparations or cell compositions

[0205] The present invention also relates to fermentation broth formulations or cell compositions comprising the polypeptides of the present invention. The fermentation broth product further comprises additional components used in the fermentation process, such as cells (including host cells containing genes encoding the polypeptides of the present invention, these host cells being used to produce the target polypeptide), cell debris, biomass, fermentation medium, and / or fermentation products. In some embodiments, the composition is a cell-killing whole culture medium containing organic acids, killed cells and / or cell debris, and a culture medium.

[0206] As used herein, the term "fermentation broth" refers to a preparation produced by cell fermentation that undergoes little or no recovery and / or purification. For example, fermentation broth is produced when a microbial culture is incubated to saturation under carbon-limited conditions that enable protein synthesis (e.g., expression of enzymes by the host cell) and secretion of proteins into the cell culture medium. Fermentation broth may contain unfractionated or fractionated contents of fermentation material derived at the end of fermentation. Typically, fermentation broth is unfractionated and contains used culture medium and cell debris remaining after, for example, removal of microbial cells (e.g., filamentous fungal cells) by centrifugation. In some embodiments, fermentation broth contains used cell culture medium, extracellular enzymes, and viable and / or non-viable microbial cells.

[0207] In the embodiments, the fermentation broth formulation and cell composition comprise a first organic acid component (containing at least one organic acid with 1-5 carbons and / or its salt) and a second organic acid component (containing at least one organic acid with 6 or more carbons and / or its salt). In specific embodiments, the first organic acid component is acetic acid, formic acid, propionic acid, its salt, or a mixture of two or more of the foregoing; and the second organic acid component is benzoic acid, cyclohexanecarboxylic acid, 4-methylvaleric acid, phenylacetic acid, its salt, or a mixture of two or more of the foregoing.

[0208] In one aspect, the composition contains at least one organic acid and optionally further contains killed cells and / or cell debris. In one embodiment, these killed cells and / or cell debris are removed from the cell-killing whole culture medium to provide a composition free of these components.

[0209] These fermentation broth formulations or cell compositions may further contain preservatives and / or antimicrobial (e.g., bacteriostatic) agents, including but not limited to sorbitol, sodium chloride, potassium sorbate, and other reagents known in the art.

[0210] The cell-killing whole culture or composition may contain ungraded contents of fermentation material derived at the end of fermentation. Typically, the cell-killing whole culture or composition contains used culture medium and cell debris present after microbial cells (e.g., filamentous fungal cells) have grown to saturation under carbon-limited conditions for protein synthesis. In some embodiments, the cell-killing whole culture or composition contains used cell culture medium, extracellular enzymes, and killed filamentous fungal cells. In some embodiments, methods known in the art can be used to permeate and / or lyse the microbial cells present in the cell-killing whole culture or composition.

[0211] The whole culture medium or cell composition described herein is typically a liquid, but may contain insoluble components, such as killed cells, cell debris, culture medium components, and / or insoluble enzymes. In some embodiments, insoluble components may be removed to provide a clear liquid composition.

[0212] The whole culture medium formulations and cell compositions of the present invention can be produced by the methods described in WO 90 / 15861 or WO 2010 / 096673.

[0213] Composition

[0214] The present invention also relates to compositions comprising the polypeptides of the present invention, particularly cleaning compositions.

[0215] These compositions may contain the polypeptides of the present invention as the main enzyme component, for example, as a single-component composition. Alternatively, these compositions may contain a variety of enzyme activities, such as one or more enzymes selected from the group consisting of: hydrolases, isomerases, ligases, lyases, oxidoreductases, or transferases, for example, α-galactosidase, α-glucosidase, aminopeptidase, amylase, β-galactosidase, β-glucosidase, β-xylosidase, glycosylase, carboxypeptidase, catalase, cellobiose hydrolase, cellulase, chitinase, keratinase, cyclodextrin glucosyltransferase, deoxyribonuclease, endoglucanase, esterase, glucosylamylase, invertase, laccase, lipase, mannosidase, polysaccharidase, oxidase, pectinase, peroxidase, phytase, polyphenol oxidase, proteolytic enzyme, ribonuclease, transglutaminase, or xylanase.

[0216] This invention relates to cleaning compositions or detergent compositions comprising a fructan-degrading enzyme and one or more other cleaning composition components. The selection of the other components is within the capabilities of a person skilled in the art and includes conventional components, including exemplary non-limiting components set forth below.

[0217] One aspect of the present invention relates to a cleaning composition comprising a polypeptide having fructan-degrading activity, particularly a polypeptide comprising a GH32 domain and preferably also comprising a GH32C domain, and at least one cleaning component.

[0218] The polypeptide with fructan-degrading activity in the cleaning composition of the present invention preferably belongs to the WMNE clade and contains the motif [WPG][GMTH][NH][AEILDNMV] (SEQ ID NO:24), preferably the motif W[GMT]N[NDEI] (SEQ ID NO:32), and optionally also belongs to the AYSN clade and contains the motif A[YF]S[LN]D[QK] (SEQ ID NO:25) and / or the motif [HS]WGH (SEQ ID NO:27) and / or belongs to the WMNEP clade and contains the motif [QLKEYMRVTANI][NSYHATRD][WF][MEVKTL][NGA][EVDLIH][PAEHDS][NATSKGVERQCD] (SEQ ID NO:28).

[0219] One aspect of the present invention relates to a cleaning composition comprising:

[0220] a) A polypeptide with fructan degradation activity, wherein the polypeptide is selected from the group consisting of:

[0221] i. A polypeptide having at least 60%, such as 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 with SEQ ID NO:6.

[0222] ii. A polypeptide having 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%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity with SEQ ID NO:9.

[0223] iii. A polypeptide having 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%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity with SEQ ID NO:12.

[0224] iv. A polypeptide having at least 60%, such as 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 with SEQ ID NO:15, and

[0225] v. A polypeptide having at least 60%, such as 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 with SEQ ID NO:18; and

[0226] b) At least one cleaning component, preferably selected from surfactants, detergents, bleaching agents, polymers, dispersants and other enzymes.

[0227] The fructan-degrading enzyme may be included in the cleaning composition of the present invention at levels of at least 0.0001 to at least 100, at least 0.001 to at least 100, at least 0.01 to at least 100, at least 0.02 to at least 100, at least 0.01 to at least 100, at least 0.1 to at least 100, at least 0.2 to at least 100, or at least 0.5 to at least 100 mg / mL. Preferably, the concentration of the fructan-degrading enzyme in the cleaning composition, such as a detergent, is in the range of 0.01 to 100, 0.1 to 50, or 1 to 10 mg / mL. Therefore, the detergent composition may contain at least 0.00008%, 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% of fructan-degrading enzyme protein.

[0228] Polypeptides with DNA enzyme activity

[0229] A particular aspect of the invention relates to cleaning compositions as described elsewhere herein, which, in addition to at least one fructan-degrading enzyme containing a GH32 domain and preferably also containing a GH32C domain, also contain at least one nuclease, such as a DNase or RNase, and preferably an enzyme having DNase activity.

[0230] The term "DNase" refers to a nuclease polypeptide with DNase activity, which catalyzes the hydrolytic cleavage of phosphodiester bonds in the DNA backbone, thereby degrading DNA. The terms "DNase" and "polypeptide with / having DNase activity" are used interchangeably herein. DNase activity can be determined according to the procedures described in Assay 1 or Assay 2 of PCT / EP 2019 / 076825.

[0231] Preferably, the DNase is selected from any of the following enzyme categories: EC3.1.21.X, where X = 1, 2, 3, 4, 5, 6, 7, 8 or 9, such as deoxyribonuclease I, deoxyribonuclease IV, type I site-specific deoxyribonuclease, type II site-specific deoxyribonuclease, type III site-specific deoxyribonuclease, CC-preferred endonuclease, deoxyribonuclease V, T(4) deoxyribonuclease II, T(4) deoxyribonuclease IV or EC3.1.22.Y, where Y = 1, 2, 4 or 5, such as deoxyribonuclease II, Aspergillus deoxyribonuclease K(1), cross-linked endonuclease, or deoxyribonuclease X.

[0232] Preferably, the DNase activity is obtained from microorganisms, and the DNase is a microbial enzyme. The DNase is preferably of fungal or bacterial origin.

[0233] DNase can be obtained from bacteria, such as Bacillus, like Bacillus licheniformis, Bacillus subtilis, Bacillus sp., Bacillus horikoshii, Bacillus horneckiae, Bacillus cibi, Bacillus idriensis, Bacillus algicola, Bacillus vietnamensis, Bacillus hwajinpoensis, Bacillus indicus, Bacillus marisflavi, Bacillus luciferensis, or Bacillus species SA2-6.

[0234] DNase can also be obtained from any of the following: *Pyrenochaetopsis* sp., *Vibrissea flavovirens*, *Setosphaeria rostrate*, *Endophragmiella valdina*, *Corynespora cassiicola*, *Paraphoma* sp., *Monilinia fructicola*, *Curvularialunata*, *Penicillium reticulisporum*, *Penicillium quercetorum*, *Setophaeosphaeria* sp., *Alternaria* sp., *Trichoderma reesei*, *Chaetomium thermophilum*, *Scytalidium thermophilum*, *Metapochonia*. suchlasporia, Daldinia fissa, Acremonium sp., Acremonium dichromosporum, Sarocladium sp., Metarhizium sp. HNA15-2, Isaria tenuipes, Scytalidium circinatum, Metarhizium lepidiotae, Thermobisporabispora, Spormia fimetaria, Pycnidiophora cf. dispera, Clavicipitaceae sp., Westerdykella sp., Humicolopsis cephalosporioides, Neosartorya massa, Roussoella Intermedia, Pleosporales, Phaeosphaeria, or Didymosphaeria futilis.

[0235] In some embodiments, these polypeptides with DNase activity are polypeptides containing the PFAM domain DUF1524 (http: / / pfam.xfam.org / , “The Pfam protein families database: towards a more sustainable future”, R.D. Finn et al., Nucleic Acids Research (2016) Database Issue 44: D279-D285). The DUF1524 domain contains a conserved HXXP sequence (where H is histidine, P is proline, and X is any amino acid motif) commonly found in nucleases (MA.Machnicka et al., Phylogenomics and sequence-structure-function relationships in the GmrSD family of Type IV restriction enzymes, BMC). Bioinformatics [BMC Bioinformatics], 2015, 16, 336. DUF represents a domain with unknown function and a family of peptides containing DUFs that have been collected together in the Pfam database, which provides sequence alignments and hidden Markov models that define the domains of the collected proteins.

[0236] Therefore, in some embodiments, these polypeptides having DNase activity in the compositions of the present invention comprise the DUF1524 domain. For further information regarding DNases comprising the DUF1524 domain, see WO 2017 / 060475, which is incorporated herein by reference.

[0237] In some embodiments, the DNase is a NUC1 or NUC1_A DNase. A NUC1 DNase is a DNase containing a domain called the NUC1 domain, and polypeptides containing this domain, in addition to DNase activity, contain features of certain motifs. Similarly, a NUC1 subdomain, called the NUC1_A domain, has been identified, which also contains features of specific motifs. NUC1 and NUC1_A DNases are described in WO 2017 / 060475 and WO 2018 / 184873, which are incorporated herein by reference.

[0238] The preparation of polypeptides having the DNA enzyme activity described herein can be carried out, for example, as described in WO 2017 / 059802, particularly in the sections on nucleic acid constructs, expression vectors, host cells, production and fermentation broth formulations.

[0239] In another embodiment, the cleaning composition of the present invention comprises, in addition to a fructan-degrading enzyme, a polypeptide having DNase activity, and the polypeptide 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 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity with the polypeptide shown in SEQ ID NO:19. In one embodiment, the polypeptide has an amino acid sequence comprising or consisting of SEQ ID NO:19. The polypeptide may also be a fragment of SEQ ID NO:19 having DNase activity.

[0240] In another embodiment, the cleaning composition of the present invention comprises, in addition to a fructan-degrading enzyme, a polypeptide having DNase activity, and the polypeptide 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 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity with the polypeptide shown in SEQ ID NO:20. In one embodiment, the DNase has an amino acid sequence comprising or consisting of SEQ ID NO:20. The polypeptide may also be a fragment of SEQ ID NO:20 having DNase activity.

[0241] In another embodiment, the cleaning composition of the present invention comprises, in addition to a fructan-degrading enzyme, a polypeptide having DNase activity, and the polypeptide 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 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity with the polypeptide shown in SEQ ID NO:21. In one embodiment, the DNase has an amino acid sequence comprising or consisting of SEQ ID NO:21. The polypeptide may also be a fragment of SEQ ID NO:21 having DNase activity.

[0242] In another embodiment, the cleaning composition of the present invention comprises, in addition to a fructan-degrading enzyme, a polypeptide having DNase activity, and the polypeptide and the polypeptide shown in SEQ ID NO:22 have 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 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity. In one embodiment, the DNase has an amino acid sequence comprising or consisting of SEQ ID NO:22.

[0243] The polypeptide with DNase activity can also be a fragment of SEQ ID NO:22 with DNase activity. The polypeptide expressing SEQ ID NO:22 can be expressed by different N-terminal truncations, for example as a polypeptide of 206 amino acid residues corresponding to amino acids 16 to 221 of SEQ ID NO:22, or as a polypeptide of 204 amino acid residues corresponding to amino acids 18 to 221 of SEQ ID NO:22, or as a mixture of two or more such polypeptides.

[0244] In another embodiment, the cleaning composition of the present invention comprises, in addition to a fructan-degrading enzyme, a polypeptide having DNase activity, and the polypeptide 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 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity with the polypeptide shown in SEQ ID NO:23. In one embodiment, the DNase has an amino acid sequence comprising or consisting of SEQ ID NO:23. The polypeptide may also be a fragment of SEQ ID NO:23 having DNase activity.

[0245] In one particular embodiment, the cleaning composition comprises:

[0246] • A fructan-degrading enzyme comprising or consisting of SEQ ID NO:3, or a variant thereof, the variant having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity with SEQ ID NO:3; and preferably comprising the motif of SEQ ID NO:24, more preferably the motif of SEQ ID NO:32, and optionally also comprising the motifs of SEQ ID NO:25, SEQ ID NO:27, and / or SEQ ID NO:28; and

[0247] • A DNA enzyme comprising or consisting of SEQ ID NO:19, or a variant thereof, having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity with SEQ ID NO:19.

[0248] In another particular embodiment, the cleaning composition comprises:

[0249] • A fructan-degrading enzyme comprising or consisting of SEQ ID NO:3, or a variant thereof, the variant having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity with SEQ ID NO:3; and preferably comprising the motif of SEQ ID NO:24, more preferably the motif of SEQ ID NO:32, and optionally also comprising the motifs of SEQ ID NO:25, SEQ ID NO:27, and / or SEQ ID NO:28; and

[0250] • A DNA enzyme comprising or consisting of SEQ ID NO:20, or a variant thereof, having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity with SEQ ID NO:20.

[0251] In another particular embodiment, the cleaning composition comprises:

[0252] • A fructan-degrading enzyme comprising or consisting of SEQ ID NO:3, or a variant thereof, the variant having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity with SEQ ID NO:3; and preferably comprising the motif of SEQ ID NO:24, more preferably the motif of SEQ ID NO:32, and optionally also comprising the motifs of SEQ ID NO:25, SEQ ID NO:27, and / or SEQ ID NO:28; and

[0253] • A DNA enzyme comprising or consisting of SEQ ID NO:21, or a variant thereof, having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity with SEQ ID NO:21.

[0254] In another particular embodiment, the cleaning composition comprises:

[0255] • A fructan-degrading enzyme comprising or consisting of SEQ ID NO:3, or a variant thereof, the variant having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity with SEQ ID NO:3; and preferably comprising the motif of SEQ ID NO:24, more preferably the motif of SEQ ID NO:32, and optionally also comprising the motifs of SEQ ID NO:25, SEQ ID NO:27, and / or SEQ ID NO:28; and

[0256] • A DNA enzyme comprising or consisting of SEQ ID NO:22, or a variant thereof, having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity with SEQ ID NO:22.

[0257] In another particular embodiment, the cleaning composition comprises:

[0258] • A fructan-degrading enzyme comprising or consisting of SEQ ID NO:3, or a variant thereof, the variant having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity with SEQ ID NO:3; and preferably comprising the motif of SEQ ID NO:24, more preferably the motif of SEQ ID NO:32, and optionally also comprising the motifs of SEQ ID NO:25, SEQ ID NO:27, and / or SEQ ID NO:28; and

[0259] • A DNA enzyme comprising or consisting of SEQ ID NO:23, or a variant thereof, having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity with SEQ ID NO:23.

[0260] In another particular embodiment, the cleaning composition comprises:

[0261] • A fructan-degrading enzyme comprising or consisting of SEQ ID NO:6, or a variant thereof, the variant having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity with SEQ ID NO:6; and preferably comprising the motif of SEQ ID NO:24, more preferably the motif of SEQ ID NO:32, and optionally also comprising the motifs of SEQ ID NO:25, SEQ ID NO:27, and / or SEQ ID NO:28; and

[0262] • A DNA enzyme comprising or consisting of SEQ ID NO:19, or a variant thereof, having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity with SEQ ID NO:19.

[0263] In another particular embodiment, the cleaning composition comprises:

[0264] • A fructan-degrading enzyme comprising or consisting of SEQ ID NO:6, or a variant thereof, the variant having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity with SEQ ID NO:6; and preferably comprising the motif of SEQ ID NO:24, more preferably the motif of SEQ ID NO:32, and optionally also comprising the motifs of SEQ ID NO:25, SEQ ID NO:27, and / or SEQ ID NO:28; and

[0265] • A DNA enzyme comprising or consisting of SEQ ID NO:20, or a variant thereof, having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity with SEQ ID NO:20.

[0266] In another particular embodiment, the cleaning composition comprises:

[0267] • A fructan-degrading enzyme comprising or consisting of SEQ ID NO:6, or a variant thereof, the variant having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity with SEQ ID NO:6; and preferably comprising the motif of SEQ ID NO:24, more preferably the motif of SEQ ID NO:32, and optionally also comprising the motifs of SEQ ID NO:25, SEQ ID NO:27, and / or SEQ ID NO:28; and

[0268] • A DNA enzyme comprising or consisting of SEQ ID NO:21, or a variant thereof, having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity with SEQ ID NO:21.

[0269] In another particular embodiment, the cleaning composition comprises:

[0270] • A fructan-degrading enzyme comprising or consisting of SEQ ID NO:6, or a variant thereof, the variant having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity with SEQ ID NO:6; and preferably comprising the motif of SEQ ID NO:24, more preferably the motif of SEQ ID NO:32, and optionally also comprising the motifs of SEQ ID NO:25, SEQ ID NO:27, and / or SEQ ID NO:28; and

[0271] • A DNA enzyme comprising or consisting of SEQ ID NO:22, or a variant thereof, having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity with SEQ ID NO:22.

[0272] In another particular embodiment, the cleaning composition comprises:

[0273] • A fructan-degrading enzyme comprising or consisting of SEQ ID NO:6, or a variant thereof, the variant having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity with SEQ ID NO:6; and preferably comprising the motif of SEQ ID NO:24, more preferably the motif of SEQ ID NO:32, and optionally also comprising the motifs of SEQ ID NO:25, SEQ ID NO:27, and / or SEQ ID NO:28; and

[0274] • A DNA enzyme comprising or consisting of SEQ ID NO:23, or a variant thereof, having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity with SEQ ID NO:23.

[0275] In one particular embodiment, the cleaning composition comprises:

[0276] • A fructan-degrading enzyme comprising or consisting of SEQ ID NO:9, or a variant thereof, the variant having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity with SEQ ID NO:9; and preferably comprising the motif of SEQ ID NO:24, more preferably the motif of SEQ ID NO:32, and optionally also comprising the motifs of SEQ ID NO:25, SEQ ID NO:27, and / or SEQ ID NO:28; and

[0277] • A DNA enzyme comprising or consisting of SEQ ID NO:19, or a variant thereof, having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity with SEQ ID NO:19.

[0278] In another particular embodiment, the cleaning composition comprises:

[0279] • A fructan-degrading enzyme comprising or consisting of SEQ ID NO:9, or a variant thereof, the variant having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity with SEQ ID NO:9; and preferably comprising the motif of SEQ ID NO:24, more preferably the motif of SEQ ID NO:32, and optionally also comprising the motifs of SEQ ID NO:25, SEQ ID NO:27, and / or SEQ ID NO:28; and

[0280] • A DNA enzyme comprising or consisting of SEQ ID NO:20, or a variant thereof, having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity with SEQ ID NO:20.

[0281] In another particular embodiment, the cleaning composition comprises:

[0282] • A fructan-degrading enzyme comprising or consisting of SEQ ID NO:9, or a variant thereof, the variant having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity with SEQ ID NO:9; and preferably comprising the motif of SEQ ID NO:24, more preferably the motif of SEQ ID NO:32, and optionally also comprising the motifs of SEQ ID NO:25, SEQ ID NO:27, and / or SEQ ID NO:28; and

[0283] • A DNA enzyme comprising or consisting of SEQ ID NO:21, or a variant thereof, having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity with SEQ ID NO:21.

[0284] In another particular embodiment, the cleaning composition comprises:

[0285] • A fructan-degrading enzyme comprising or consisting of SEQ ID NO:9, or a variant thereof, the variant having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity with SEQ ID NO:9; and preferably comprising the motif of SEQ ID NO:24, more preferably the motif of SEQ ID NO:32, and optionally also comprising the motifs of SEQ ID NO:25, SEQ ID NO:27, and / or SEQ ID NO:28; and

[0286] • A DNA enzyme comprising or consisting of SEQ ID NO:22, or a variant thereof, having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity with SEQ ID NO:22.

[0287] In another particular embodiment, the cleaning composition comprises:

[0288] • A fructan-degrading enzyme comprising or consisting of SEQ ID NO:9, or a variant thereof, the variant having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity with SEQ ID NO:9; and preferably comprising the motif of SEQ ID NO:24, more preferably the motif of SEQ ID NO:32, and optionally also comprising the motifs of SEQ ID NO:25, SEQ ID NO:27, and / or SEQ ID NO:28; and

[0289] • A DNA enzyme comprising or consisting of SEQ ID NO:23, or a variant thereof, having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity with SEQ ID NO:23.

[0290] In one particular embodiment, the cleaning composition comprises:

[0291] • A fructan-degrading enzyme comprising or consisting of SEQ ID NO:12, or a variant thereof, the variant having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity with SEQ ID NO:12; and preferably comprising the motif of SEQ ID NO:24, more preferably the motif of SEQ ID NO:32, and optionally also comprising the motifs of SEQ ID NO:25, SEQ ID NO:27, and / or SEQ ID NO:28; and

[0292] • A DNA enzyme comprising or consisting of SEQ ID NO:19, or a variant thereof, having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity with SEQ ID NO:19.

[0293] In another particular embodiment, the cleaning composition comprises:

[0294] • A fructan-degrading enzyme comprising or consisting of SEQ ID NO:12, or a variant thereof, the variant having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity with SEQ ID NO:12; and preferably comprising the motif of SEQ ID NO:24, more preferably the motif of SEQ ID NO:32, and optionally also comprising the motifs of SEQ ID NO:25, SEQ ID NO:27, and / or SEQ ID NO:28; and

[0295] • A DNA enzyme comprising or consisting of SEQ ID NO:20, or a variant thereof, having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity with SEQ ID NO:20.

[0296] In another particular embodiment, the cleaning composition comprises:

[0297] • A fructan-degrading enzyme comprising or consisting of SEQ ID NO:12, or a variant thereof, the variant having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity with SEQ ID NO:12; and preferably comprising the motif of SEQ ID NO:24, more preferably the motif of SEQ ID NO:32, and optionally also comprising the motifs of SEQ ID NO:25, SEQ ID NO:27, and / or SEQ ID NO:28; and

[0298] • A DNA enzyme comprising or consisting of SEQ ID NO:21, or a variant thereof, having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity with SEQ ID NO:21.

[0299] In another particular embodiment, the cleaning composition comprises:

[0300] • A fructan-degrading enzyme comprising or consisting of SEQ ID NO:12, or a variant thereof, the variant having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity with SEQ ID NO:12; and preferably comprising the motif of SEQ ID NO:24, more preferably the motif of SEQ ID NO:32, and optionally also comprising the motifs of SEQ ID NO:25, SEQ ID NO:27, and / or SEQ ID NO:28; and

[0301] • A DNA enzyme comprising or consisting of SEQ ID NO:22, or a variant thereof, having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity with SEQ ID NO:22.

[0302] In another particular embodiment, the cleaning composition comprises:

[0303] • A fructan-degrading enzyme comprising or consisting of SEQ ID NO:12, or a variant thereof, the variant having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity with SEQ ID NO:12; and preferably comprising the motif of SEQ ID NO:24, more preferably the motif of SEQ ID NO:32, and optionally also comprising the motifs of SEQ ID NO:25, SEQ ID NO:27, and / or SEQ ID NO:28; and

[0304] • A DNA enzyme comprising or consisting of SEQ ID NO:23, or a variant thereof, having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity with SEQ ID NO:23.

[0305] In one particular embodiment, the cleaning composition comprises:

[0306] • A fructan-degrading enzyme comprising or consisting of SEQ ID NO:15, or a variant thereof, having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity with SEQ ID NO:15; and preferably comprising the motif of SEQ ID NO:24, more preferably the motif of SEQ ID NO:32, and optionally also comprising the motifs of SEQ ID NO:25, SEQ ID NO:27, and / or SEQ ID NO:28; and

[0307] • A DNA enzyme comprising or consisting of SEQ ID NO:19, or a variant thereof, having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity with SEQ ID NO:19.

[0308] In another particular embodiment, the cleaning composition comprises:

[0309] • A fructan-degrading enzyme comprising or consisting of SEQ ID NO:15, or a variant thereof, having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity with SEQ ID NO:15; and preferably comprising the motif of SEQ ID NO:24, more preferably the motif of SEQ ID NO:32, and optionally also comprising the motifs of SEQ ID NO:25, SEQ ID NO:27, and / or SEQ ID NO:28; and

[0310] • A DNA enzyme comprising or consisting of SEQ ID NO:20, or a variant thereof, having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity with SEQ ID NO:20.

[0311] In another particular embodiment, the cleaning composition comprises:

[0312] • A fructan-degrading enzyme comprising or consisting of SEQ ID NO:15, or a variant thereof, having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity with SEQ ID NO:15; and preferably comprising the motif of SEQ ID NO:24, more preferably the motif of SEQ ID NO:32, and optionally also comprising the motifs of SEQ ID NO:25, SEQ ID NO:27, and / or SEQ ID NO:28; and

[0313] • A DNA enzyme comprising or consisting of SEQ ID NO:21, or a variant thereof, having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity with SEQ ID NO:21.

[0314] In another particular embodiment, the cleaning composition comprises:

[0315] • A fructan-degrading enzyme comprising or consisting of SEQ ID NO:15, or a variant thereof, having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity with SEQ ID NO:15; and preferably comprising the motif of SEQ ID NO:24, more preferably the motif of SEQ ID NO:32, and optionally also comprising the motifs of SEQ ID NO:25, SEQ ID NO:27, and / or SEQ ID NO:28; and

[0316] • A DNA enzyme comprising or consisting of SEQ ID NO:22, or a variant thereof, having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity with SEQ ID NO:22.

[0317] In another particular embodiment, the cleaning composition comprises:

[0318] • A fructan-degrading enzyme comprising or consisting of SEQ ID NO:15, or a variant thereof, having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity with SEQ ID NO:15; and preferably comprising the motif of SEQ ID NO:24, more preferably the motif of SEQ ID NO:32, and optionally also comprising the motifs of SEQ ID NO:25, SEQ ID NO:27, and / or SEQ ID NO:28; and

[0319] • A DNA enzyme comprising or consisting of SEQ ID NO:23, or a variant thereof, having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity with SEQ ID NO:23.

[0320] In one particular embodiment, the cleaning composition comprises:

[0321] • A fructan-degrading enzyme comprising or consisting of SEQ ID NO:18, or a variant thereof, the variant having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity with SEQ ID NO:18; and preferably comprising the motif of SEQ ID NO:24, more preferably the motif of SEQ ID NO:32, and optionally also comprising the motifs of SEQ ID NO:25, SEQ ID NO:27, and / or SEQ ID NO:28; and

[0322] • A DNA enzyme comprising or consisting of SEQ ID NO:19, or a variant thereof, having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity with SEQ ID NO:19.

[0323] In another particular embodiment, the cleaning composition comprises:

[0324] • A fructan-degrading enzyme comprising or consisting of SEQ ID NO:18, or a variant thereof, the variant having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity with SEQ ID NO:18; and preferably comprising the motif of SEQ ID NO:24, more preferably the motif of SEQ ID NO:32, and optionally also comprising the motifs of SEQ ID NO:25, SEQ ID NO:27, and / or SEQ ID NO:28; and

[0325] • A DNA enzyme comprising or consisting of SEQ ID NO:20, or a variant thereof, having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity with SEQ ID NO:20.

[0326] In another particular embodiment, the cleaning composition comprises:

[0327] • A fructan-degrading enzyme comprising or consisting of SEQ ID NO:18, or a variant thereof, the variant having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity with SEQ ID NO:18; and preferably comprising the motif of SEQ ID NO:24, more preferably the motif of SEQ ID NO:32, and optionally also comprising the motifs of SEQ ID NO:25, SEQ ID NO:27, and / or SEQ ID NO:28; and

[0328] • A DNA enzyme comprising or consisting of SEQ ID NO:21, or a variant thereof, having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity with SEQ ID NO:21.

[0329] In another particular embodiment, the cleaning composition comprises:

[0330] • A fructan-degrading enzyme comprising or consisting of SEQ ID NO:18, or a variant thereof, the variant having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity with SEQ ID NO:18; and preferably comprising the motif of SEQ ID NO:24, more preferably the motif of SEQ ID NO:32, and optionally also comprising the motifs of SEQ ID NO:25, SEQ ID NO:27, and / or SEQ ID NO:28; and

[0331] • A DNA enzyme comprising or consisting of SEQ ID NO:22, or a variant thereof, having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity with SEQ ID NO:22.

[0332] In another particular embodiment, the cleaning composition comprises:

[0333] • A fructan-degrading enzyme comprising or consisting of SEQ ID NO:18, or a variant thereof, the variant having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity with SEQ ID NO:18; and preferably comprising the motif of SEQ ID NO:24, more preferably the motif of SEQ ID NO:32, and optionally also comprising the motifs of SEQ ID NO:25, SEQ ID NO:27, and / or SEQ ID NO:28; and

[0334] • A DNA enzyme comprising or consisting of SEQ ID NO:23, or a variant thereof, having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity with SEQ ID NO:23.

[0335] In another embodiment, the cleaning composition of the present invention may include any DNase polypeptide disclosed in WO 2015 / 155350, WO 2017 / 059802, WO 2017 / 060475, WO 2017 / 060505, WO 2017 / 064269, WO 2018 / 011277, WO 2018 / 177938, WO 2018 / 185285, WO 2018 / 184873, WO 2019 / 081721 or WO 2019 / 081724, the contents of which are incorporated herein by reference.

[0336] When present in the cleaning compositions of the present invention, the DNase polypeptide will typically be present in amounts ranging from 0.01 to 1000 ppm, from 0.1 to 1000 ppm, from 1 ppm to 1000 ppm, from 10 ppm to 1000 ppm, from 50 ppm to 1000 ppm, from 100 ppm to 1000 ppm, from 150 ppm to 1000 ppm, from 200 ppm to 1000 ppm, from 250 ppm to 1000 ppm, from 250 ppm to 750 ppm, or from 250 ppm to 500 ppm, based on the activated protein.

[0337] Cleaning components

[0338] The selection of cleaning components may include (for textile care) the type of textile to be cleaned, the type and / or extent of soiling, the temperature at which cleaning is performed, and considerations for the formulation of the detergent product. Although the components mentioned below are classified under a general heading according to specific functionality, this is not to be construed as limiting, as components may contain additional functionality as would be understood by a person skilled in the art.

[0339] surfactants

[0340] Cleaning compositions may contain one or more surfactants, which may be anionic and / or cationic and / or nonionic and / or semi-polar and / or zwitterionic, or mixtures thereof. In certain embodiments, the detergent composition comprises a mixture of one or more nonionic surfactants and one or more anionic surfactants. The surfactant is typically present at levels ranging from about 1% to 70% by weight, for example from about 1 wt% to about 40 wt%, or from about 3 wt% to about 20 wt%, or from about 3 wt% to about 10 wt%.

[0341] The one or more surfactants are selected based on the desired cleaning application, and the one or more surfactants may include any conventional surfactants known in the art.

[0342] When included therein, the detergent will typically contain about 1% to about 70% by weight of anionic surfactant, such as about 5 wt% to about 50 wt%, including about 5 wt% to about 20 wt%, or about 15 wt% to about 20 wt%, or about 20 wt% to about 25 wt%, or at least 30 wt%, at least 40 wt%, or at least 50 wt% of anionic surfactant. Non-limiting examples of anionic surfactants include sulfates and sulfonates, particularly alkylbenzene sulfonates, such as linear alkylbenzene sulfonates (LAS), isomers of LAS, branched alkylbenzene sulfonates (BABS), phenyl alkyl sulfonates, α-olefin sulfonates (AOS), olefin sulfonates, chain olefin sulfonates, alkyl-2,3-dimethylbis(sulfate), hydroxyalkyl sulfonates, and disulfonates, alkyl sulfates (AS) (such as sodium dodecyl sulfate (SDS)), fatty alcohol sulfates (FAS), primary alcohol sulfates (PA). S), alcohol ether sulfates (AES or AEOS or FES, also known as alcohol ethoxy sulfates or fatty alcohol ether sulfates), secondary alkyl sulfonates (SAS), paraffin sulfonates (PS), ester sulfonates, sulfonated fatty acid glycerides, α-sulfo fatty acid methyl esters (α-SFMe or SES) (including methyl ester sulfonates (MES)), alkyl succinic acids or alkenyl succinic acids, dodecenyl / tetradecenyl succinic acids (DTSA), fatty acid derivatives of amino acids, diesters and monoesters or fatty acid salts (soaps) of sulfosuccinic acids, and combinations thereof.

[0343] When included therein, the detergent will typically contain about 1% to about 40% by weight of a cationic surfactant, for example about 0.5% to about 30%, particularly about 1% to about 20%, about 3% to about 10%, such as about 3% to about 5%, about 8% to about 12%, or about 10% to about 12%. Non-limiting examples of cationic surfactants include alkyl dimethyl ethanol quaternary ammonium (ADMEAQ), hexadecyl trimethyl ammonium bromide (CTAB), dimethyl distearate ammonium chloride (DSDMAC), and alkyl benzyl dimethyl ammonium, alkyl quaternary ammonium compounds, alkoxylated quaternary ammonium (AQA) compounds, ester quaternary ammonium compounds, and combinations thereof.

[0344] When included therein, the detergent will typically contain from about 0.2% to about 40% by weight of a nonionic surfactant, for example from about 0.5 wt% to about 30 wt%, particularly from about 1 wt% to about 20 wt%, from about 3 wt% to about 10 wt%, such as from about 3 wt% to about 5 wt%, from about 8 wt% to about 12 wt%, or from about 10 wt% to about 12 wt%. Non-limiting examples of nonionic surfactants include alcohol ethoxylates (AE or AEO), alcohol propoxylates, propoxylated fatty alcohols (PFA), alkoxylated fatty acid alkyl esters (such as 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)), as well as products available under the trade names SPAN and TWEEN, and combinations thereof.

[0345] When included therein, the detergent will typically contain about 0.01% 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.

[0346] When included therein, the detergent will typically contain about 0.01% to about 10% by weight of a zwitterionic surfactant. Non-limiting examples of zwitterionic surfactants include betaines, such as alkyldimethyl betaine, sulfobetaine, and combinations thereof.

[0347] Typically, the cleaning composition contains more than one surfactant, for example, at least one anionic and at least one nonionic surfactant. Preferably, the total amount of all surfactants present, i.e., the amount of anionic, nonionic, amphoteric, and cationic surfactants present, is preferably from about 1 wt% to 80 wt% by weight, for example from about 1 wt% to 70 wt%, for example from about 1 wt% to 50 wt%, for example from about 1 wt% to about 40 wt%, or from about 5 wt% to about 40 wt%, or from about 10 wt% to about 60 wt%. The ratio between the present surfactants depends on the specific composition, but when the composition includes anionic and nonionic surfactants, the weight ratio can be 30:1 to 10:1, 20:1 to 1:10, 25:1 to 1:2, or 20:1 to 1:5 (anionic to nonionic surfactant weight ratio).

[0348] One embodiment relates to a cleaning composition comprising a fructan-degrading enzyme, wherein the cleaning composition is at least one surfactant, preferably anionic and / or nonionic surfactant, preferably, wherein the composition comprises 1 to 70 wt%, more preferably 5 to 40 wt%, of a surfactant, wherein the surfactant is preferably selected from alkylbenzene sulfonates such as LAS, alkyl sulfates (AS) and mixtures thereof, preferably, the cleaning composition comprises at least 20 wt% of an alkylbenzene sulfonate surfactant.

[0349] One embodiment relates to a cleaning composition comprising a fructan-degrading enzyme, wherein the cleaning composition comprises at least one anionic surfactant, and wherein the cleaning composition further comprises a nonionic surfactant, and preferably wherein the weight ratio of the anionic to the nonionic surfactant is 25:1 to 1:2 or 1.5:1 to 1:10.

[0350] Builders and co-builders

[0351] The cleaning composition may contain, by weight, about 0-65%, such as about 5% to about 50%, such as about 0.5% to about 20%, of a detergent builder or co-builder, or a mixture thereof. In dishwashing detergents, the level of the builder is typically 40%-65%, particularly 50%-65%. The builder and / or co-builder may be chelating agents, particularly those that form water-soluble complexes with Ca and Mg. Any builder and / or co-builder known in the art for use in cleaning detergents may be used. Non-limiting examples of detergent builders include zeolites, diphosphates (pyrophosphates), triphosphates such as sodium triphosphate (STP or STPP), carbonates such as sodium carbonate, soluble silicates such as sodium metasilicate, layered silicates (e.g., SKS-6 from Hoechst), ethanolamines such as 2-aminoethanol (MEA), diethanolamine (DEA, also known as 2,2'-iminodiethanol), triethanolamine (TEA, also known as 2,2',2”-nitrotriethanolamine), and (carboxymethyl)inulin (CMI), and combinations thereof.

[0352] The detergent composition may also contain 0-50% by weight, such as about 5% to about 30%, of a detergent co-agent. The detergent composition may include a co-agent alone or in combination with a builder (e.g., a zeolite builder). Non-limiting examples of co-agents include homopolymers of polyacrylates or copolymers thereof, such as poly(acrylic acid) (PAA) or copolymers of (acrylic acid / maleic acid) (PAA / PMA). Other non-limiting examples include citrates, chelating agents (such as aminocarboxylates, aminopolycarboxylates, and phosphonates), and alkyl succinic acids or alkenyl succinic acids. Other specific examples include 2,2',2”-N-aminotriacetic acid (NTA), ethylenediaminetetraacetic acid (EDTA), diethylenetriaminepentaacetic acid (DTPA), iminodisuccinic acid (IDS), ethylenediamine-N,N'-dibutanoic acid (EDDS), methylglycine diacetic acid (MGDA), glutamic acid-N,N-diacetic acid (GLDA), 1-hydroxyethane-1,1-diphosphonic acid (HEDP), ethylenediaminetetra(methylenephosphonic acid) (EDTMPA), diethylenetriaminepenta(methylenephosphonic acid) (DTMPA or DTPMPA), N-(2-hydroxyethyl)iminodiacetic acid (EDG), aspartic-N-monoacetic acid (ASMA), aspartic-N,N-diacetic acid (ASDA), aspartic-N-monopropionic acid (ASMP), iminodisuccinic acid (IDA), N-(2-sulfomethyl)-aspartic acid (SMAS), N-(2-sulfoethyl)-aspartic acid... Aspartic acid (SEAS), N-(2-sulfomethyl)-glutamic acid (SMGL), N-(2-sulfoethyl)-glutamic acid (SEGL), N-methyliminodiacetic acid (MIDA), α-alanine-N,N-diacetic acid (α-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), sulfomethyl-N,N-diacetic acid (SMDA), N-(2-hydroxyethyl)ethylenediamine-N,N',N”-triacetic acid (HEDTA), diethanolglycine (DEG), diethylenetriaminepenta (methylenephosphonic acid) (DTPMP), aminotris (methylenephosphonic acid) (ATMP), and combinations thereof and their salts. Other exemplary builders and / or co-builders are described, for example, in WO 09 / 102854 and US 5977053.

[0353] bleaching system

[0354] The cleaning composition may contain 0-30% by weight, such as from about 1% to about 20%, or from about 0.01% to about 10%, of a bleaching system. Any bleaching system containing components known in the art for use in cleaning detergents may be used. Suitable bleaching system components include a hydrogen peroxide source; a peracid source; and a bleaching catalyst or synergist.

[0355] Suitable sources of hydrogen peroxide are inorganic peroxide salts, including alkali metal salts (such as sodium percarbonate and sodium perborate (usually monohydrate or tetrahydrate)) and hydrogen peroxide-urea (1 / 1).

[0356] The peracid can be (a) directly incorporated as a pre-formed peracid, or (b) formed in situ in the washing solution from hydrogen peroxide and a bleaching activator (perhydrolysis), or (c) formed in situ in the washing solution from hydrogen peroxide and a perhydrolase and a suitable substrate (e.g., an ester).

[0357] Suitable preformed peracids include, but are not limited to, peroxycarboxylic acids (such as peroxybenzoic acid) and their cyclically substituted derivatives, peroxy-α-naphthoic acid, peroxyphthalic acid, peroxylauric acid, peroxystearic acid, ε-phthaliminoperoxyhexanoic acid [phthaliminoperoxyhexanoic acid (PAP)], and o-carboxybenzoiminoperoxyhexanoic acid; aliphatic and aromatic disperoxydicarboxylic acids, such as disperoxydodecanoic acid, disperoxyazelaic acid, disperoxysedicarboxylic acid, disperoxybasilicic acid, 2-decyldisperoxysuccinic acid, and disperoxyphthalic acid, -isophthalic acid and -terephthalic acid; perimidic acid; peroxymonosulfate; peroxydisulfate; peroxyphosphoric acid; peroxysilicic acid; and mixtures of said compounds. It should be understood that in some cases, the mentioned peracids may preferably be added as suitable salts, such as alkali metal salts (e.g., ) or alkaline earth metal salts.

[0358] Suitable bleaching activators include those belonging to the classes of esters, amides, imides, nitriles, or acid anhydrides, and, where applicable, their salts. Suitable examples are tetraacetyl ethylenediamine (TAED), sodium 4-[(3,5,5-trimethylhexanoyl)oxy]benzene-1-sulfonate (ISONOBS), sodium 4-(dodecanoyloxy)benzene-1-sulfonate (LOBS), sodium 4-(decanoyloxy)benzene-1-sulfonate, 4-(decanoyloxy)benzoic acid (DOBA), sodium 4-(nonanoyloxy)benzene-1-sulfonate (NOBS), and / or those disclosed in WO 98 / 17767. A particular family of bleaching activators for this purpose is disclosed in EP 624154, and is particularly preferred in this family is triethyl acetyl citrate (ATC). ATC or short-chain triglycerides (like triacetin) have the advantage that they are environmentally friendly. Furthermore, triethyl acetylglucosyl citrate and triacetin exhibit good hydrolytic stability in the product during storage and are effective bleaching activators. Finally, ATC is versatile because the citrate released during the hydrolysis reaction can act as a detergent builder.

[0359] Bleaching catalysts and synergists

[0360] The bleaching system may also include a bleaching catalyst or synergist. Some non-limiting examples of bleaching catalysts that can be used in the compositions of the present invention include manganese oxalate, manganese acetate, manganese collagen, cobalt-amine catalysts, and manganese triazacyclononane (MnTACN) catalysts; particularly preferred are complexes of manganese with 1,4,7-trimethyl-1,4,7-triazacyclononane (Me3-TACN) or 1,2,4,7-tetramethyl-1,4,7-triazacyclononane (Me4-TACN), especially Me3-TACN, such as the binuclear manganese complex [(Me3-TACN)Mn(O)3Mn(Me3-TACN)](PF6)2, and [2,2',2”-nitrotris(ethane-1,2-diylazalkylidene-κN-methylidene)triphenol-κ3O]manganese(III). These bleaching catalysts may also be other metal compounds, such as iron or cobalt complexes.

[0361] In some embodiments, including those containing a peracid source, an organic bleaching catalyst or bleaching enhancer having one of the following formulas may be used:

[0362]

[0363] (iii) and mixtures thereof; wherein each R1 is independently a branched alkyl group containing 9 to 24 carbons or a straight-chain alkyl group containing 11 to 24 carbons, preferably each R1 is independently a branched alkyl group containing 9 to 18 carbons or a straight-chain alkyl group containing 11 to 18 carbons, more preferably each R1 is independently selected from the group consisting of: 2-propylheptyl, 2-butyloctyl, 2-pentylnonyl, 2-hexyldecyl, dodecyl, tetradecyl, hexadecyl, octadecyl, isononyl, isodecyl, isothidecyl, isothidecyl and isopentadecyl.

[0364] Other exemplary bleaching systems are described, for example, in WO 2007 / 087258, WO 2007 / 087244, WO 2007 / 087259, EP 1867708 (Vitamin K) and WO 2007 / 087242. Suitable photobleaching agents may be, for example, sulfonated zinc phthalocyanine or aluminum phthalocyanine.

[0365] Metal care products

[0366] Metal care products can prevent or reduce rust, corrosion, or oxidation of metals, including aluminum, stainless steel, and non-ferrous metals such as silver and copper. Suitable examples include one or more of the following:

[0367] (a) Benzotriazoles, including benzotriazoles or bis-benzotriazoles and their substituted derivatives. Benzotriazole derivatives are those compounds in which the available substitution sites on the aromatic ring are partially or completely substituted. Suitable substituents include straight-chain or branched Ci-C20-alkyl groups (e.g., C1-C20-alkyl groups) and hydroxyl, thio, phenyl, or halogen (e.g., fluorine, chlorine, bromine, and iodine).

[0368] (b) Metal salts and complexes selected from the group consisting of: zinc, manganese, titanium, zirconium, hafnium, vanadium, cobalt, gallium, and cesium salts and / or complexes, wherein these metals are in one of oxidation states II, III, IV, V, or VI. In one aspect, suitable metal salts and / or metal complexes may be selected from the group consisting of: Mn(II) sulfates, Mn(II) citrates, Mn(II) stearates, Mn(II) acetylacetonates, K^TiF6 (e.g., K2TiF6), K^ZrF6 (e.g., K2ZrF6), CoSO4, Co(NOs)2, and Ce(NOs)3, zinc salts, such as zinc sulfate, zinc hydroxide, or zinc acetate;

[0369] (c) Silicates, including sodium silicate or potassium silicate, disodium silicate, sodium metasilicate, crystalline silicates and mixtures thereof.

[0370] WO 94 / 26860 and WO 94 / 26859 disclose other suitable organic and inorganic redox active substances for use as silver / copper corrosion inhibitors. Preferably, the compositions of the present invention comprise 0.1% to 5% by weight of a metal care agent, preferably a zinc salt.

[0371] Water-soluble additives

[0372] The cleaning composition may contain 0-10% by weight, for example 0-5% by weight, such as about 0.5% to about 5%, or about 3% to about 5% of a water-soluble adjuvant. Any water-soluble adjuvant known in the art for use in detergents may be used. Non-limiting examples of water-soluble adjuvants include sodium benzenesulfonate, sodium p-toluenesulfonate (STS), sodium xylenesulfonate (SXS), sodium cumenesulfonate (SCS), sodium cymene sulfonate, amine oxides, alcohols and polyethylene glycol ethers, sodium hydroxynaphthoate, sodium hydroxynaphthoate, sodium ethylhexyl sulfonate, sodium ethylhexyl sulfonate, and combinations thereof.

[0373] polymer

[0374] The cleaning composition may contain 0-10% by weight, such as 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 may be used. The polymer may function as a co-cleaning agent as mentioned above, or may provide anti-redeposition, fiber protection, dirt release, dye transfer inhibition, grease removal, and / or defoaming properties. Some polymers may have more than one of the properties mentioned above and / or more than one of the motifs mentioned below. 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 polycarboxylic acid esters such as PAA, PAA / PMA, poly-aspartic acid, and lauryl methacrylate / acrylic acid copolymers, hydrophobically modified CMC (HM-CMC) and silicone, copolymers of terephthalic acid and oligomeric polyethylene glycol, copolymers of poly(ethylene terephthalate) and poly(ethylene oxyterephthalate) (PET-POET), PVP, poly(vinylimidazolium) (PVI), poly(vinylpyridine-N-oxide) (PVPO or PVPNO), and polyvinylpyrrolidone-vinylimidazolium (PVPVI). Suitable examples include the PVP-K15, PVP-K30, ChromaBond S-400, ChromaBond S-403E, and ChromaBond S-100 from Ashland's Aqualon, and BASF's... HP 165 HP 50 (dispersant) HP 53 (dispersant) HP 59 (dispersion reagent), HP 56 (dye transfer inhibitor), HP 66K (dye transfer inhibitor). Other exemplary polymers include sulfonated polycarboxylate esters, polyethylene oxide and polypropylene oxide (PEO-PPO), and diquaternary ammonium ethoxysulfate. Other exemplary polymers are disclosed, for example, in WO 2006 / 130575. Salts of the polymers mentioned above are also considered. Particularly preferred polymers are ethoxylated homopolymers from BASF. HP 20 helps prevent the redeposition of dirt in the detergent solution.

[0375] Fabric colorant

[0376] The cleaning compositions of the present invention may also include fabric colorants, such as dyes or pigments, which, when formulated in a detergent composition, can deposit on the fabric upon contact with a detergent solution containing the detergent composition, and thus alter the color of the fabric through the absorption / reflection of visible light. Fluorescent whitening agents emit at least some visible light. Conversely, fabric colorants alter the color of a surface when they absorb at least a portion of the visible spectrum. Suitable fabric colorants include dyes and dye-clay conjugates, and may also include pigments. Suitable dyes include small molecule dyes and polymer dyes. Suitable small molecule dyes include those selected from the following dyes falling into the Colour Index (CI) classification: Direct Blue, Direct Red, Direct Violet, Acid Blue, Acid Red, Acid Violet, Basic Blue, Basic Violet, and Basic Red, or mixtures thereof, such as those described in WO 2005 / 03274, WO 2005 / 03275, WO 2005 / 03276 and EP 1876226 (incorporated herein by reference). The detergent composition preferably contains 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 a fabric toner. The composition may contain from 0.0001 wt% to 0.2 wt% of a fabric toner, which may be particularly preferred when the composition is in the form of a unit-dosage pouch. Suitable toners are also disclosed, for example, in WO 2007 / 087257 and WO2007 / 087243.

[0377] dispersant

[0378] The cleaning compositions of the present invention may also contain dispersants. In particular, powdered detergents may contain dispersants. Suitable water-soluble organic materials include homopolymeric or copolymeric acids or salts thereof, wherein the polycarboxylic acid contains at least two carboxyl groups separated from each other by no more than two carbon atoms. Suitable dispersants are described, for example, in Powdered Detergents, Surfactant Science Series, Volume 71, Marcel Dekker, Inc.

[0379] Dye transfer inhibitors

[0380] The cleaning compositions of the present invention may further include one or more dye transfer inhibitors. Suitable polymeric dye transfer inhibitors include, but are not limited to, polyvinylpyrrolidone polymers, polyamine N-oxide polymers, copolymers of N-vinylpyrrolidone and N-vinylimidazole, polyvinyloxazolidinone, and polyvinylimidazole or mixtures thereof. When present in the subject composition, the dye transfer inhibitor may be present at a level from about 0.0001% to about 10%, from about 0.01% to about 5%, or even from about 0.1% to about 3% by weight of the composition.

[0381] Fluorescent whitening agent

[0382] The cleaning compositions of the present invention will also preferably contain additional components that can color the items being cleaned, such as optical brighteners or fluorescent whitening agents. When present, the level of the brightening agent is preferably from about 0.01% to about 0.5%. Any suitable optical brightener used in laundry detergent compositions can be used in the compositions of the present invention. The most commonly used optical brighteners are those belonging to the following categories: diaminostilbene-sulfonic acid derivatives, diarylpyrazoline derivatives, and diphenyl-bistyryl derivatives. Examples of diaminostilbene-sulfonic acid derivatives of fluorescent whitening agents include the following sodium salts: 4,4'-bis-(2-diethanolamino-4-anilino-s-triazin-6-ylamino)stilbene-2,2'-disulfonate, 4,4'-bis-(2,4-diphenylamino-s-triazin-6-ylamino)stilbene-2,2'-disulfonate, 4,4'-bis-(2-anilino-4-(N-methyl-N-2-hydroxy-ethylamino)-s-triazin-6-ylamino)stilbene-2,2'-disulfonate, 4,4'-bis-(4-phenyl-1,2,3-triazol-2-yl)stilbene-2,2'-disulfonate, and sodium 5-(2H-naphtho[1,2-d][1,2,3]triazol-2-yl)-2-[(E)-2-phenylvinyl]benzenesulfonate. Preferred fluorescent whitening agents are Tinopal DMS and Tinopal CBS, available from Ciba-Geigy AG (Basel, Switzerland). Tinopal DMS is the disodium salt of 4,4'-bis-(2-morpholino-4-anilino-s-triazine-6-ylamino)stilbene-2,2'-disulfonate. Tinopal CBS is the disodium salt of 2,2'-bis-(phenyl-styryl)-disulfonate. Also preferred is Parawhite KX, commercially available and supplied by Paramount Minerals and Chemicals in Mumbai, India. Other suitable fluorescent agents for use in this invention include 1,3-diarylpyrazoline and 7-aminoalkylcoumarin. Suitable levels of fluorescent brighteners range from a low level of about 0.01 wt%, from 0.05 wt%, from about 0.1 wt%, or even from a low level of about 0.2 wt% to a high level of 0.5 wt% or even 0.75 wt%.

[0383] Fouling release polymers

[0384] The cleaning compositions of the present invention may also include one or more dirt-releasing polymers that help remove dirt from fabrics (e.g., cotton and polyester-based fabrics), particularly hydrophobic dirt from polyester-based fabrics. Dirt-releasing polymers may be, for example, polymers based on nonionic or anionic terephthalic acid, polyvinylcaprolactam and related copolymers, vinyl graft copolymers, polyester polyamides, see, for example, Powdered Detergents, Surfactant Science Series, Volume 71, Chapter 7, Marcel Decker. Another type of dirt-releasing polymer is an amphiphilic alkoxylated grease-cleaning polymer comprising a core structure and a plurality of alkoxylated groups attached to that core structure. The core structure may comprise a polyalkylimide structure or a polyalkanolamine structure, as detailed in WO 2009 / 087523 (incorporated herein by reference). Moreover, random graft copolymers are suitable dirt-releasing polymers. Suitable graft copolymers are described in more detail in WO 2007 / 138054, WO 2006 / 108856 and WO 2006 / 113314 (incorporated herein by reference). Suitable polyethylene glycol polymers include randomly grafted copolymers comprising: (i) a hydrophilic backbone containing polyethylene glycol; and (ii) one or more side chains selected from the group consisting of: C4-C25 alkyl groups, polypropylene, polybutene, vinyl esters of saturated C1-C6 monocarboxylic acids, C1-C6 alkyl esters of acrylic acid or methacrylic acid, and mixtures thereof. Suitable polyethylene glycol polymers have a polyethylene glycol backbone (with randomly grafted polyvinyl acetate side chains). The average molecular weight of the polyethylene glycol backbone can range from 2,000 Da to 20,000 Da, or from 4,000 Da to 8,000 Da. The molecular weight ratio of the polyethylene glycol backbone to the polyvinyl acetate side chain can range from 1:1 to 1:5, or from 1:1.2 to 1:2. The average number of grafting sites per ethylene oxide unit can be less than 1 or less than 0.8, and the average number of grafting sites per ethylene oxide unit can range from 0.5 to 0.9, or from 0.1 to 0.5, or from 0.2 to 0.4. A suitable polyethylene glycol polymer is Sokalan HP22. Other fouling-releasing polymers are substituted polysaccharide structures, especially substituted cellulose structures, such as modified cellulose derivatives, as described in EP 1867808 or WO 2003 / 040279 (both of which are incorporated herein by reference). Suitable cellulose polymers include cellulose, cellulose ethers, cellulose esters, cellulose amides, and mixtures thereof.Suitable cellulose polymers include anionicly modified cellulose, nonionicly modified cellulose, cationicly modified cellulose, zwitterionicly modified cellulose, and mixtures thereof. Suitable cellulose polymers include methylcellulose, carboxymethylcellulose, ethylcellulose, hydroxyethylcellulose, hydroxypropylmethylcellulose, ester carboxymethylcellulose, and mixtures thereof.

[0385] Anti-redeposition agent

[0386] The cleaning compositions of the present invention may further include one or more anti-redeposition agents, such as carboxymethyl cellulose (CMC), polyvinyl alcohol (PVA), polyvinylpyrrolidone (PVP), polyethylene oxide and / or polyethylene glycol (PEG), homopolymers of acrylic acid, copolymers of acrylic acid and maleic acid, and ethoxylated polyethyleneimine. The cellulose-based polymers described above under the category of dirt-releasing polymers may also function as anti-redeposition agents.

[0387] rheology modifiers

[0388] The cleaning compositions of the present invention may further include one or more rheology modifiers, structural agents, or thickeners, unlike viscosity reducers. Rheology modifiers are selected from the group consisting of non-polymeric crystals, hydroxyl functional materials, and polymeric rheology modifiers, which impart shear-thinning characteristics to the aqueous liquid matrix of the liquid detergent composition. The rheology and viscosity of the detergent can be modified and adjusted by methods known in the art, for example, as shown in EP 2169040.

[0389] Other suitable cleaning composition components include, but are not limited to, anti-shrinkage agents, anti-wrinkle agents, bactericides, adhesives, carriers, dyes, enzyme stabilizers, fabric softeners, fillers, foam conditioners, water-soluble additives, fragrances, pigments, defoamers, solvents, and structural agents and / or structural elastic agents for liquid detergents.

[0390] polymer

[0391] The cleaning composition may contain 0-10% by weight, such as 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 may be used. The polymer may function as a co-cleaning agent as mentioned above, or may provide anti-redeposition, fiber protection, dirt release, dye transfer inhibition, grease removal, and / or defoaming properties. Some polymers may have more than one of the properties mentioned above and / or more than one of the motifs mentioned below. 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 polycarboxylic acid esters such as PAA, PAA / PMA, poly-aspartic acid, and lauryl methacrylate / acrylic acid copolymers, hydrophobically modified CMC (HM-CMC) and silicone, copolymers of terephthalic acid and oligomeric polyethylene glycol, copolymers of poly(ethylene terephthalate) and poly(ethylene oxyterephthalate) (PET-POET), PVP, poly(vinylimidazolium) (PVI), poly(vinylpyridine-N-oxide) (PVPO or PVPNO), and polyvinylpyrrolidone-vinylimidazolium (PVPVI). Suitable examples include the PVP-K15, PVP-K30, ChromaBond S-400, ChromaBond S-403E, and ChromaBond S-100 from Ashland's Aqualon, and BASF's... HP 165 HP 50 (dispersant) HP 53 (dispersant) HP 59 (dispersion reagent), HP 56 (dye transfer inhibitor), HP 66K (dye transfer inhibitor). Other exemplary polymers include sulfonated polycarboxylate esters, polyethylene oxide and polypropylene oxide (PEO-PPO), and diquaternary ammonium ethoxysulfate. Other exemplary polymers are disclosed, for example, in WO 2006 / 130575. Salts of the polymers mentioned above are also considered. Particularly preferred polymers are ethoxylated homopolymers from BASF. HP 20 helps prevent the redeposition of dirt in the detergent solution.

[0392] Other enzymes

[0393] In addition to at least one fructan-degrading enzyme and optionally at least one DNase, the cleaning composition may contain one or more other enzymes, such as one or more lipases, keratases, amylases, glycoses, cellulases, pectinsases, mannanases, arabinases, galactanases, xylanases, oxidases (e.g., laccase), and / or peroxidases. Generally, the selected enzyme should be compatible with the selected detergent (i.e., optimal pH, compatibility with other enzymes and non-enzyme components, etc.), and the enzyme should be present in an effective amount.

[0394] Mannanase

[0395] Suitable mannanases include those of bacterial or fungal origin, including chemically or genetically modified mutants. Mannanases can be basic mannanases of families 5 or 26. They can be wild-type from the genera *Bacillus* or *Pythium*, particularly from *Bacillus agaricus*, *Bacillus licheniformis*, *Bacillus halodurans*, *Bacillus clausii*, or specific *Pythium* species. Suitable mannanases are described in WO 1999 / 064619. Commercially available mannanases are from Mannaway (Novozymes).

[0396] Cellulase

[0397] Suitable cellulases include intact cellulases or single-component endoglucanases of bacterial or fungal origin. This includes chemically or genetically modified mutants. Cellulases can be, for example, single-component endo-1,4-β-glucanases (often simply referred to as endoglucans) or mixtures thereof. Suitable cellulases include fungal cellulases derived from specific *Pyrophyte* species (US 4,435,307) or from the *Trichoderma* genus, such as *Trichoderma reesei* or *Trichoderma viride*. Examples of cellulases are described in EP 0495257. Other suitable cellulases are derived from the *Clostridium* genus, such as *Clostridium terrestrialum* as described in WO 96 / 29397 or *Fusarium oxysporum* as described in WO 91 / 17244, or from the *Bacillus* genus as described in WO 02 / 099091 and JP2000210081. Other examples are cellulase variants, such as those described in WO 94 / 07998, EP0531315, US 5,457,046, US 5,686,593, US 5,763,254, WO 95 / 24471, and WO 98 / 12307. Commercially available cellulases include... and (Novozymes A / S) Puradax HA and Puradax EG (from Genencor).

[0398] protease

[0399] Suitable proteases can be of any origin, but are preferably of bacterial or fungal origin, optionally in the form of protein-engineered or chemically modified mutants. Proteases can be basic proteases, such as serine proteases or metalloproteinases. Serine proteases can be, for example, members of the S1 family (such as trypsin) or the S8 family (such as subtilisin). Metalloproteinases can be, for example, thermophilic proteases, such as thermophilic proteases from the M4 family, or other metalloproteinases, such as those from the M5, M7, or M35 families.

[0400] The term "subtilase" refers to a subgroup of serine proteases, according to Siezen et al., Protein Eng. 4 (1991) 719-737 and Siezen et al., Protein Sci. 6 (1997) 501-523. Serine proteases are a subgroup of proteases characterized by the presence of a serine residue at its active site that forms a covalent adduct with the substrate. Subtilases can be classified into six subclasses: the subtilisin family, the thermophilic protease family, the proteinase K family, the lanathionine antibiotic peptidase family, the Kexin family, and the Pyrolysin family.

[0401] While suitable proteases for detergent applications can be obtained from a variety of organisms, including fungi such as Aspergillus, detergent proteases are generally derived from bacteria, particularly Bacillus species. Examples of Bacillus species that derive subtilisases include Bacillus lentus, Bacillus alkalophilus, Bacillus subtilis, Bacillus amyloliquefaciens, Bacillus licheniformis, Bacillus pumilus, and Bacillus gibsonii. Specific subtilisin proteases include subtilisin lentus, subtilisin Novo, subtilisin Carlsberg, subtilisin BPN', subtilisin 309, subtilisin 147, and subtilisin 168, as well as protease PD138 (described in WO 93 / 18140). Other useful proteases are those described, for example, in WO 01 / 16285 and WO 02 / 16547.

[0402] Examples of trypsin-like proteases include Fusarium proteases (described in WO 94 / 25583 and WO2005 / 040372) and chymotrypsins derived from Cellumonas (described in WO 2005 / 052161 and WO 2005 / 052146).

[0403] Examples of metalloproteinases include neutral metalloproteinases (e.g., those derived from Bacillus amylolyticus) described in WO 2007 / 044993, and metalloproteinases described, for example, in WO 2015 / 158723 and WO 2016 / 075078.

[0404] Examples of useful proteases are protease variants described in WO 89 / 06279, WO 92 / 19729, WO 96 / 34946, WO 98 / 20115, WO 98 / 20116, WO 99 / 11768, WO 01 / 44452, WO 03 / 006602, WO 2004 / 003186, WO2004 / 041979, WO 2007 / 006305, WO 2011 / 036263, WO 2014 / 207227, WO 2016 / 087617 and WO2016 / 174234. Preferred protease variants may, for example, contain one or more mutations selected from the group consisting of: S3T, V4I, S9R, S9E, A15T, S24G, S24R, K27R, N42R, S55P, G59E, G59D, N60D, N60E, V66A, N74D, S85R, A96S, S97G, S97D, S97A, S97SD, S99E, S99D, S99G, S99M, S99N, S99R, S99H, S101A, V102I, V102Y, V102N, S104A, G116V, G116R, H118D, H118N, A120S, S126L, P1 27Q, S128A, S154D, A156E, G157D, G157P, S158E, Y161A, R164S, Q176E, N179E, S182E, Q185N, A188P, G189E, V193M, N198D, V199I, Q200L, Y203W, S206G, L211Q, L211D, N212D, N212S, M216S, A226V, K229L, Q230H, Q239R, N246K, S253D, N255W, N255D, N255E, L256E, L256DT268A and R269H, where the position number corresponds to WO The location of the Bacillus retarded protease shown in SEQ ID NO:1 of WO 2016 / 001449. Protease variants having one or more of these mutations are preferably the Bacillus retarded protease shown in SEQ ID NO:1 of WO 2016 / 001449. Also known as variants of Bacillus subtilis protease 309 or variants of Bacillus amyloliquefaciens protease (BPN') shown in SEQ ID NO:2 of WO2016 / 001449. Such protease variants preferably have at least 80% sequence identity with SEQ ID NO:1 or SEQ ID NO:2 of WO 2016 / 001449.

[0405] Another target protease is an alkaline protease from Bacillus tarda DSM 5483 (as described, for example, in WO 91 / 02792) and its variants (which are described, for example, in WO 92 / 21760, WO 95 / 23221, EP 1921147, EP1921148 and WO 2016 / 096711).

[0406] Alternatively, the protease may be a variant of the TY145 protease having SEQ ID NO:1 of WO 2004 / 067737, for example, containing a substituted variant at one or more positions corresponding to positions 27, 109, 111, 171, 173, 174, 175, 180, 182, 184, 198, 199 and 297 of SEQ ID NO:1 of WO 2004 / 067737, wherein the protease variant has at least 75% but less than 100% sequence identity with SEQ ID NO:1 of WO 2004 / 067737. The target TY145 variants are described in, for example, WO 2015 / 014790, WO 2015 / 014803, WO 2015 / 014804, WO 2016 / 097350, WO2016 / 097352, WO 2016 / 097357 and WO 2016 / 097354.

[0407] Examples of preferred proteases include:

[0408] (a) A variant of SEQ ID NO:1 of WO 2016 / 001449 comprising two or more substitutions selected from the group consisting of: S9E, N43R, N76D, Q206L, Y209W, S259D and L262E, for example having substitutions for S9E, N43R, N76D, V205I, Q206L, Y209W, S259D, N261W and L262E, or having a variant having substitutions for S9E, N43R, N76D, N185E, S188E, Q191N, A194P, Q206L, Y209W, S259D and L262E, wherein the position number is based on the number of SEQ ID NO:2 of WO 2016 / 001449;

[0409] (b) A variant of the polypeptide of SEQ ID NO:1 of WO 2016 / 001449 with the mutation S99SE, wherein the position number is based on the number of SEQ ID NO:2 of WO 2016 / 001449;

[0410] (c) A variant of the polypeptide of SEQ ID NO:1 of WO 2016 / 001449 with the mutation S99AD, wherein the position number is based on the number of SEQ ID NO:2 of WO 2016 / 001449;

[0411] (d) A variant of the polypeptide of SEQ ID NO:1 of WO 2016 / 001449 having a substitution Y167A+R170S+A194P, wherein the position number is based on the number of SEQ ID NO:2 of WO 2016 / 001449;

[0412] (e) A variant of the polypeptide of SEQ ID NO:1 of WO 2016 / 001449 having a substitution S9R+A15T+V68A+N218D+Q245R, wherein the position number is based on the number of SEQ ID NO:2 of WO 2016 / 001449;

[0413] (f) A variant of the polypeptide of SEQ ID NO:1 of WO 2016 / 001449 having a substitution S9R+A15T+G61E+V68A+A194P+V205I+Q245R+N261D, wherein the position number is based on the number of SEQ ID NO:2 of WO 2016 / 001449;

[0414] (g) A variant of the polypeptide of SEQ ID NO:1 of WO 2016 / 001449 having a substitution for S99D+S101R / E+S103A+V104I+G160S; for example, a variant of SEQ ID NO:1 of WO 2016 / 001449 having a substitution for S3T+V4I+S99D+S101E+S103A+V104I+G160S+V205I, wherein the position number is based on the number of SEQ ID NO:2 of WO 2016 / 001449;

[0415] (h) A variant of the polypeptide of SEQ ID NO:2 of WO 2016 / 001449 having the substitution S24G+S53G+S78N+S101N+G128A / S+Y217Q, wherein the position number is based on the number of SEQ ID NO:2 of WO 2016 / 001449;

[0416] (i) The polypeptide disclosed in GENESEQP with accession number BER84782, which corresponds to SEQ ID NO:302 in WO 2017 / 210295;

[0417] (j) A variant of the polypeptide of SEQ ID NO:1 of WO 2016 / 001449 having the substitution S99D+S101E+S103A+V104I+S156D+G160S+L262E, wherein the position number is based on the number of SEQ ID NO:2 of WO 2016 / 001449;

[0418] (k) A variant of the polypeptide of SEQ ID NO:1 of WO 2016 / 001449 having a substitution of S9R+A15T+G61E+V68A+N76D+S99G+N218D+Q245R, wherein the position number is based on the number of SEQ ID NO:2 of WO 2016 / 001449;

[0419] (l) A variant of the polypeptide of SEQ ID NO:1 of WO 2016 / 001449 having a substitution for V68A+S106A, wherein the position number is based on the number of SEQ ID NO:2 of WO 2016 / 001449; and

[0420] (m) A variant of the polypeptide of SEQ ID NO:1 of WO 2004 / 067737 having a substitution of S27K+N109K+S111E+S171E+S173P+G174K+S175P+F180Y+G182A+L184F+Q198E+N199+T297P, wherein the position number is based on the number of SEQ ID NO:1 of WO2004 / 067737.

[0421] Suitable commercially available proteases include those sold under the following trade names: Duralase TM Durazym TM , Ultra Ultra, Prime TM , Ultra Ultra Blaze 100T, Blaze 125T, Blaze 150T, Blaze 200T Uno、 In and Excel (Novozymes), those sold under the following trade name: Maxatase TM Maxacal TM , Ox, OxP, FN2 TM FN3 TM FN4 exTM , Excellenz TM P1000, Excellenz TM P1250, Eraser TM , P100, Purafect Prime P110, P300, Effectiveenz P1000 TM , Effectenz P1050 TM , Ox、Effectenz TM P2000, Purafast TM , Opticlean TM and (Danisco / DuPont), BLAP (sequence shown in Figure 29 of US 5352604) and its variants (Henkel AG), and KAP (Bacillus subtilis protease) from Kao Corporation.

[0422] Lipase and keratinase

[0423] Suitable lipases and keratins include those derived from bacteria or fungi. This includes chemically modified mutant enzymes or protein-engineered mutant enzymes. Examples include lipases from the genus *Thermophilic*, such as those from *Thermophilic Hypotherium* (formerly named *Pyrophyte*) as described in EP 258068 and EP 305216; cutinases from the genus *Pyrophyte*, such as *Pyrophyte Specific* (WO 96 / 13580); lipases from strains of the genus *Pseudomonas* (some of which are now renamed *Burkholderia*), such as *Alcaligenes* or *Alcaligenes-like* (EP 218272), *Pseudomonas cepacia* (EP 331376), *Pseudomonas* strains SD705 (WO 95 / 06720 and WO 96 / 27002), *Pseudomonas wisconsinensis* (WO 96 / 12012); and GDSL-type *Streptomyces* lipases (WO 96 / 12012). 10 / 065455); cutinase from rice blast fungus (WO 10 / 107560); cutinase from Pseudomonas mendoza (US 5,389,536); lipase from Thermobifida fusca (WO 11 / 084412); lipase from Bacillus stearothermophilus (WO 11 / 084417); lipase from Bacillus subtilis (WO 11 / 084599); and lipase from Streptomyces griseus (WO11 / 150157) and Streptomyces pristinaespiralis (WO 12 / 137147).

[0424] 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.

[0425] Preferred commercially available lipase products include Lipolase TM Lipex TM Lipolex TM and Lipoclean TM(Novozymes A / S), Lumafast (from Genencor), and Lipomax (from Gist-Brocades). Other examples are lipases sometimes called acyltransferases or perhydrolases, such as an acyltransferase homologous to Candida antarctica lipase A (WO10 / 111143), an acyltransferase from Mycobacterium smegmatis (WO 05 / 56782), a perhydrolases from the CE 7 family (WO 09 / 67279), and variants of Mycobacterium smegmatis perhydrolases (particularly the S54V variant used in the commercial product Gentle Power Bleach from Huntsman Textile Effects Pte Ltd) (WO 10 / 100028).

[0426] amylase

[0427] Suitable amylases can be α-amylases or glucosylamylases and can be of bacterial or fungal origin. This includes chemically modified mutants or protein-engineered mutants. Amylases include, for example, α-amylases obtained from specific strains of Bacillus, such as Bacillus licheniformis (described in more detail in GB 1,296,839).

[0428] Suitable amylases include the amylase having SEQ ID NO:2 in WO 95 / 10603 or a variant thereof having 90% sequence identity with SEQ ID NO:3. Preferred variants are described in SEQ ID NO:4 of WO 94 / 02597, WO 94 / 18314, WO 97 / 43424, and WO 99 / 019467, such as variants having substitutions at 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.

[0429] Suitable amylases include the amylase having SEQ ID NO:6 in WO 02 / 010355 or its variants having 90% sequence identity with SEQ ID NO:6. Preferred variants of SEQ ID NO:6 are those with deletions at positions 181 and 182 and substitutions at position 193.

[0430] Other suitable amylases are hybrid α-amylases comprising residues 1-33 of the α-amylase derived from *Bacillus amylolyticus* as shown in SEQ ID NO:6 of WO 2006 / 066594 and residues 36-483 of the α-amylase from *Bacillus licheniformis* as shown in SEQ ID NO:4 of WO 2006 / 066594, or variants thereof having 90% sequence identity. Preferred variants of this hybrid α-amylase are those having substitutions, deletions, or insertions at one or more of the following positions: G48, T49, G107, H156, A181, N190, M197, I201, A209, and Q264. The most preferred hybrid α-amylase comprising residues 1-33 of the α-amylase derived from Bacillus amylolyticus as shown in SEQ ID NO:6 of WO 2006 / 066594 and residues 36-483 of SEQ ID NO:4 is a variant having the following substitutions:

[0431] M197T;

[0432] H156Y+A181T+N190F+A209V+Q264S; or

[0433] G48A+T49I+G107A+H156Y+A181T+N190F+I201F+A209V+Q264S.

[0434] Other suitable amylases are those having SEQ ID NO:6 in WO 99 / 019467 or variants thereof having 90% sequence identity with SEQ ID NO:6. Preferred variants of SEQ ID NO:6 are those having substitutions, deletions, or insertions at one or more of the following positions: R181, G182, H183, G184, N195, I206, E212, E216, and K269. Particularly preferred amylases are those having deletions at positions R181 and G182, or positions H183 and G184.

[0435] Other amylases that may be used are those of SEQ ID NO:1, SEQ ID NO:3, SEQ ID NO:2, or SEQ ID NO:7 of WO 96 / 023873, or variants thereof that have 90% sequence identity with 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 with substitutions, deletions, or insertions at one or more of the following positions: 140, 181, 182, 183, 184, 195, 206, 212, 243, 260, 269, 304, and 476 (numbered using SEQ ID 2 of WO 96 / 023873). More preferred variants are those with deletions at two positions selected from 181, 182, 183, and 184 (such as 181 and 182, 182 and 183, or positions 183 and 184). The most preferred amylase variants of SEQ ID NO:1, SEQ ID NO:2, or SEQ ID NO:7 are those with deletions at positions 183 and 184 and substitutions at one or more positions 140, 195, 206, 243, 260, 304, and 476.

[0436] Other amylases that may be used are those having SEQ ID NO:2 of WO 08 / 153815, SEQ ID NO:10 of WO 01 / 66712, or variants thereof having 90% sequence identity with SEQ ID NO:2 of WO 08 / 153815, or variants thereof having 90% sequence identity with SEQ ID NO:10 of WO 01 / 66712. Preferred variants of SEQ ID NO:10 of WO 01 / 66712 are those having substitutions, deletions, or insertions at one or more of the following positions: 176, 177, 178, 179, 190, 201, 207, 211, and 264.

[0437] Other suitable amylases are the amylase of SEQ ID NO:2 having WO 09 / 061380 or its variants having 90% sequence identity with SEQ ID NO:2. Preferred variants of SEQ ID NO:2 are those with C-terminal truncation, and / or substitution, deletion, or insertion at 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 substitutions at one or more of the following positions: Q87E,R, Q98R, S125A, N128C, T131I, T165I, K178L, T182G, M201L, F202Y, N225E,R, N272E,R, S243Q,A,E,D, Y305R, R309A, Q320R, Q359E, K444E, and G475K, and / or those having deletions at positions R180 and / or S181 or T182 and / or G183. The most preferred amylase variant of SEQ ID NO:2 is those having the following substitutions:

[0438] N128C+K178L+T182G+Y305R+G475K;

[0439] N128C+K178L+T182G+F202Y+Y305R+D319T+G475K;

[0440] S125A+N128C+K178L+T182G+Y305R+G475K; or

[0441] S125A+N128C+T131I+T165I+K178L+T182G+Y305R+G475K, wherein these variants are C-terminated and optionally further include a substitution at position 243 and / or a deletion at positions 180 and / or 181.

[0442] Other suitable amylases are the amylase of SEQ ID NO:1 having WO 13184577 or its variants having 90% sequence identity with SEQ ID NO:1. Preferred variants of SEQ ID NO:1 are those with substitutions, deletions, or insertions at one or more of the following positions: K176, R178, G179, T180, G181, E187, N192, M199, I203, S241, R458, T459, D460, G476, and G477. More preferred variants of SEQ ID NO:1 are those having substitutions at one or more of the following positions: K176L, E187P, N192FYH, M199L, I203YF, S241QADN, R458N, T459S, D460T, G476K, and G477K, and / or those having deletions at positions R178 and / or S179 or T180 and / or G181. Most preferred amylase variants of SEQ ID NO:1 are those having the following substitutions:

[0443] E187P+I203Y+G476K

[0444] E187P+I203Y+R458N+T459S+D460T+G476K

[0445] These variants optionally include a substitution at position 241 and / or a deletion at positions 178 and / or 179.

[0446] Other suitable amylases are the amylase of SEQ ID NO:1 having WO 10104675 or its variants having 90% sequence identity with SEQ ID NO:1. Preferred variants of SEQ ID NO:1 are those with substitutions, deletions, or insertions at one or more of the following positions: N21, D97, V128, K177, R179, S180, I181, G182, M200, L204, E242, G477, and G478. More preferred variants of SEQ ID NO:1 are those with substitutions at one or more of the following positions: N21D, D97N, V128I, K177L, M200L, L204YF, E242QA, G477K, and G478K, and / or those with deletions at positions R179 and / or S180 or I181 and / or G182. The most preferred amylase variant of SEQ ID NO:1 is those with the following substitutions:

[0447] N21D+D97N+V128I,

[0448] These variants optionally include a substitution at position 200 and / or a deletion at positions 180 and / or 181.

[0449] Other suitable amylases are α-amylases having SEQ ID NO:12 in WO 01 / 66712 or variants having at least 90% sequence identity with SEQ ID NO:12. Preferred amylase variants are those having substitutions, deletions, or insertions at one or more of the following positions in 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 deletions of D183 and G184 and having substitutions for R118K, N195F, R320K, and R458K, as well as variants having substitutions at one or more positions selected from the group consisting of M9, G149, G182, G186, M202, T257, Y295, N299, M323, E345, and A339, with the most preferred being variants having substitutions at all of these positions.

[0450] Other examples are amylase variants, such as those described in WO 2011 / 098531, WO 2013 / 001078 and WO 2013 / 001087.

[0451] Commercially available amylase is Duramyl TM Terminyl TM Fungayl TM Stainzyme TM StainzymePlus TM Natalase TM Liquozyme X and BAN TM (From Novozymes) and Rapidase TM Purastar TM / Effectenz TM Powerase, Preferenz TM S1000, Preferenz TM S100, Preferenz TM S110 and PreferenzTM S210 (from Genencor International Inc. / DuPont).

[0452] Peroxidase / oxidase

[0453] Peroxidases can be any enzyme classifying EC 1.11.1.7 as stated by the Nomenclature Committee of the International Union of Biochemistry and Molecular Biology (IUBMB), or any fragment derived therefrom that exhibits peroxidase activity. Suitable peroxidases include those of plant, bacterial, or fungal origin. This includes chemically modified mutants or protein-engineered mutants. Examples of useful peroxidases include peroxidases from the genus *Coprinus*, such as those from *C. cinerea* (EP 179,486), and their variants, such as those described in WO 93 / 24618, WO 95 / 10602, and WO 98 / 15257. Peroxidases can also include halogenated peroxidases, such as chloride peroxidase, bromoperoxidase, and compounds exhibiting chloride or bromoperoxidase activity. Halogenated peroxidases are classified according to their specificity for halide ions. Chloride peroxidase (EC 1.11.1.10) catalyzes the formation of hypochlorite from chloride ions. Halogenated peroxidases can be chloride peroxidases. Preferably, the halogenated peroxidase is a vanadium haloperoxidase, i.e., a vanadate-containing halogenated peroxidase. In a preferred method, the vanadate-containing halogenated peroxidase is combined with a chloride ion source. Halogenated peroxidases have been isolated from many different fungi, particularly from the dematiaceous hyphomycete fungal group, such as *Caldariomyces* (e.g., *C. fumago*), *Alternaria*, *Curvularia* (e.g., *C. verruculosa* and *C. inaequalis*), *Hymenopterus*, *C. spp.*, and *Botrytis*. Halogenated peroxidases have also been isolated from bacteria such as *Pseudomonas* (e.g., *P. pyrrocinia*) and *Streptomyces* (e.g., *Streptomyces aureofaciens*). Halogenated peroxidases can be derived from species of the genus Curvularia, particularly Curvularia vulgaris or Curvularia anisotropis, such as Curvularia anisotropis CBS102.42 as described in WO 95 / 27046; or Curvularia vulgaris CBS 147.63 or Curvularia vulgaris CBS 444.70 as described in WO 97 / 04102; or from species of the genus Geniculosporium as described in WO 01 / 79459, Dendryphiella salina as described in WO 01 / 79458, Phaeochrichoconis crotalarie as described in WO 01 / 79461, or Geniculosporium as described in WO 01 / 79460.

[0454] Oxidases include any laccase included in enzyme classification EC 1.10.3.2 or any fragment derived therefrom exhibiting laccase activity, or compounds exhibiting similar activity, such as catechol oxidase (EC 1.10.3.1), o-aminophenol oxidase (EC 1.10.3.4), or bilirubin oxidase (EC 1.3.3.5). Preferred laccases are microbial enzymes. These enzymes may be derived from plants, bacteria, or fungi (including filamentous fungi and yeasts). Suitable examples of fungi include laccases derived from strains of the following: *Aspergillus*, *Neurospora* (e.g., *Neurospora crassa*), *Stenocystis*, *Botrytis*, *Collybia*, *Fomes*, *Lentinula*, *Pleurotus*, *Coralus* (e.g., *Coralus longifolius* and *Coralus discolor*), *Rhizoctonia* (e.g., *Rhizoctonia solani*), *Coprinus* (e.g., *Coprinus comatus*, *Coprinus friesii*, and *Coprinus plicarius*). *Psathyrella* (e.g., *P. condelleana*), *P. papilionaceus*, *P. thermophilicus*, *Schytalidium* (e.g., *S. thermophilum*), *P. pinsitus*, *P. radiata* (WO 92 / 01046), or *C. hirsutus* (JP 2238885). Suitable examples of bacteria include laccases that may be derived from strains of the genus *Bacillus*. The preferred laccases are those derived from the genera *Coprinus* or *Desmodium*; particularly those derived from *Coprinus gracilistylus*, as disclosed in WO 97 / 08325; or those derived from *Desmodium thermophilum*, as disclosed in WO 95 / 33836.

[0455] microorganism

[0456] The detergent additives and detergent compositions may also contain one or more microorganisms, such as one or more fungi, yeasts, or bacteria. In an embodiment, the one or more microorganisms are dehydrated (e.g., by freeze-drying) bacteria or yeasts, such as strains of Lactobacillus. In another embodiment, the microorganisms are one or more microbial spores / buds (as opposed to vegetative cells), such as bacterial spores; or fungal spores, conidia, hyphae. Preferably, the one or more spores / buds are Bacillus spores; even more preferably, the one or more spores / buds are spores of Bacillus subtilis, Bacillus licheniformis, Bacillus amyloliquefaciens, or Bacillus megaterium. Microorganisms may be included in the detergent composition or additive in the same manner as enzymes (see above).

[0457] Preparation of detergent products

[0458] For example, the cleaning compositions of the present invention can be formulated as hand-wash or machine-washing detergent compositions, including suitable laundry detergent additive compositions for pre-treating stained fabrics and rinse-additive fabric softener compositions, or formulated as detergent compositions for general household hard surface cleaning operations, or formulated for hand-wash or machine-washing dishwashing operations. In a particular aspect, the present invention provides detergent additives comprising one or more enzymes as described herein. The cleaning compositions of the present invention can be in any conventional form, such as strips, homogeneous tablets, tablets having two or more layers, bags having one or more chambers, regular or compressed powders, granules, pastes, gels, or regular, compressed or concentrated liquids.

[0459] The bag can be configured as a single chamber or multiple chambers. It can have any form, shape, and material suitable for containing the composition, for example, preventing the composition from being released from the bag before contact with water. The bag is made of a water-soluble membrane containing an internal volume. This internal volume can be divided into chambers of the bag. Preferred membranes are polymeric materials, preferably polymers forming films or sheets. Preferred polymers, copolymers, or derivatives thereof are selected from polyacrylates and water-soluble acrylate copolymers, methylcellulose, carboxymethylcellulose, sodium dextrin, ethylcellulose, hydroxyethylcellulose, hydroxypropyl methylcellulose, maltodextrin, polymethyl acrylates, and most preferably polyvinyl alcohol copolymers and hydroxypropyl methylcellulose (HPMC). Preferably, the polymer level in the membrane, such as PVA, is at least about 60%. Preferred average molecular weights will typically be from about 20,000 to about 150,000. The membrane can also be a blend composition comprising a hydrolyzable and water-soluble polymer blend, such as polylactic acid and polyvinyl alcohol (known under Trade Reference No. M8630, as sold by MonoSol LLC, Indiana, USA), plus plasticizers such as glycerin, ethylene glycol, propylene glycol, sorbitol, and mixtures thereof. The bag may contain a solid laundry detergent composition or portions thereof and / or a liquid detergent composition or portions thereof separated by a water-soluble membrane. Chambers for liquid components may differ in composition from those containing solids: US2009 / 0011970 A1.

[0460] Detergent components can be physically separated from each other by compartments in different layers of water-soluble pouches or tablets. This avoids undesirable storage interactions between components. Furthermore, the different dissolution profiles of each compartment in the detergent solution can cause delayed dissolution of the selected components.

[0461] Non-unit-volume liquid or gel detergents can be aqueous, typically containing at least 20% and up to 95% water by weight, such as up to about 70%, up to about 65%, up to about 55%, up to about 45%, or up to about 35%. Other types of liquids, including but not limited to alkanols, amines, glycols, ethers, and polyols, can be included in aqueous liquids or gels. Aqueous liquid or gel detergents can contain from 0 to 30% organic solvents. Liquid or gel detergents can also be non-aqueous.

[0462] Preparation of enzymes in granules

[0463] Dust-free particles can be produced, for example, as disclosed in US 4,106,991 and 4,661,452, and can optionally be coated by methods known in the art. Examples of waxy coating materials are poly(ethylene oxide) products (polyethylene glycol, PEG) with an average molecular weight of 1,000 to 20,000; ethoxylated nonylphenol having 16 to 50 ethylene oxide units; ethoxylated fatty alcohols containing 12 to 20 carbon atoms and having 15 to 80 ethylene oxide units present; fatty alcohols; fatty acids; and monoglycerides, diglycerides, and triglycerides of fatty acids. Examples of suitable film-forming coating materials for applications via fluidized bed technology are given in GB 1483591. Liquid enzyme preparations can be stabilized, for example, by adding polyols (such as propylene glycol), sugars or sugar alcohols, lactic acid, or boric acid according to established methods. Protected enzymes can be prepared according to the methods disclosed in EP 238,216.

[0464] The compositions of the present invention can be formulated as granules, for example, as co-granules incorporating one or more enzymes. Each enzyme will then be present in multiple granules, ensuring a more uniform distribution of the enzyme in the detergent. This also reduces the physical isolation of different enzymes due to varying particle sizes. A method for producing multi-enzyme co-granules for the detergent industry is disclosed in IP.com disclosure IPCOM000200739D.

[0465] Another example of a formulation using co-particle enzymes is disclosed in WO 2013 / 188331, which relates to a detergent composition comprising: (a) multi-enzyme co-particles; (b) less than 10 wt% zeolite (anhydrous); and (c) less than 10 wt% phosphate (anhydrous), wherein the enzyme co-particles comprise a sink component of 10 wt% to 98 wt%, and the composition further comprises a detergent sink component of 20 wt% to 80 wt%. WO 2013 / 188331 also relates to a method of treating and / or cleaning a surface (preferably a fabric surface), the method comprising the steps of: (i) contacting the surface with a detergent composition in an aqueous detergent solution, and (ii) rinsing and / or drying the surface.

[0466] The multi-enzyme co-particle may contain the enzymes of the present invention and one or more enzymes selected from the group consisting of: proteases, lipases, cellulases, xyloglucans, hydrolases, peroxidases, lipoxygenases, laccases, hemicellulases, proteases, cellulases, cellobiose dehydrogenases, xylanases, phospholipases, esterases, keratinases, pectinases, mannanases, pectin lyases, keratinases, reductases, oxidases, phenol oxidases, ligninases, amylopectinases, tanninases, pentosanases, lichen polysaccharides, glucans, arabinosidases, hyaluronidases, chondroitinases, amylases, nucleases, aminohexosidases, and mixtures thereof.

[0467] One embodiment of the invention relates to enzyme particles comprising the enzyme of the invention. The particle consists of a core and optionally one or more coatings (outer layers) surrounding the core. Typically, the particle size (measured as equivalent sphere diameter (volume-based average particle size)) is 20-2000 μm, particularly 50-1500 μm, 100-1500 μm, or 250-1200 μm.

[0468] The core may include additional materials such as fillers, fibrous materials (cellulose or synthetic fibers), stabilizers, solubilizers, suspending agents, viscosity modifiers, light spheres, plasticizers, salts, lubricants, and fragrances.

[0469] The core may include adhesives such as synthetic polymers, waxes, fats, or carbohydrates.

[0470] The core, typically a homogeneous blend, can contain salts of polyvalent cations, reducing agents, antioxidants, peroxide decomposition catalysts, and / or acidic buffer components.

[0471] The core can consist of inert particles into which the enzyme is adsorbed or applied (e.g., via fluidized bed coating) to the surface of the inert particles.

[0472] The diameter of the core can be 20-2000μm, especially 50-1500μm, 100-1500μm or 250-1200μm.

[0473] The core can be prepared by blends of granulated components, for example by methods including granulation techniques such as crystallization, precipitation, pan-coating, fluidized bed coating, fluidized bed agglomeration, rotary atomization, extrusion, granulation, spheronization, particle size reduction, drum granulation, and / or high-shear granulation.

[0474] The method for preparing the core can be found in Handbook of Powder Technology; Particle size enlargement by CECapes; Volume 1, 1980, Elsevier.

[0475] The core of the enzyme particle / particle may be surrounded by at least one coating, for example, to improve storage stability, reduce dust formation during processing, or for coloring the particle. One or more optional coatings may include salt coatings or other suitable coating materials such as polyethylene glycol (PEG), methyl hydroxypropyl cellulose (MHPC), and polyvinyl alcohol (PVA). Examples of enzyme particles with multiple coatings are shown in WO 93 / 07263 and WO 97 / 23606.

[0476] The coating may be applied in an amount of at least 0.1% (e.g., at least 0.5%, 1%, or 5%) of the core weight. This amount may be at most 100%, 70%, 50%, 40%, or 30%.

[0477] The coating is preferably at least 0.1 μm thick, particularly at least 0.5 μm, at least 1 μm, or at least 5 μm thick. In a particular embodiment, the coating thickness is less than 100 μm. In a more particular embodiment, the coating thickness is less than 60 μm. In even more particular embodiments, the total coating thickness is less than 40 μm.

[0478] The coating should seal the core unit by forming a substantially continuous layer. A substantially continuous layer should be understood as a coating with very few or no pores, such that the sealed / enclosed core unit has very few or no uncoated areas. This layer or coating should be uniform in thickness.

[0479] The coating may further contain other materials known in the art, such as fillers, anti-sticking agents, pigments, dyes, plasticizers and / or binders, such as titanium dioxide, kaolin, calcium carbonate or talc.

[0480] Salt coatings may contain at least 60% salt by weight (w / w), for example, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% salt by weight (w / w).

[0481] The salt can be added from a salt solution (where the salt is completely dissolved) or from a salt suspension (where the fine particles are less than 50 μm, such as less than 10 μm or less than 5 μm).

[0482] The salt coating may contain a single salt or a mixture of two or more salts. The salt may be water-soluble, particularly having a solubility of at least 0.1 g in 100 g of water at 20°C, preferably at least 0.5 g / 100 g of water, for example, at least 1 g / 100 g of water, for example, at least 5 g / 100 g of water.

[0483] Salts can be inorganic salts, such as sulfates, sulfites, phosphates, phosphonates, nitrates, chlorides, or carbonates, or salts of simple organic acids (less than 10 carbon atoms, such as 6 or fewer carbon atoms), such as citrates, malonates, or acetates. Examples of cations in these salts are alkali or alkaline earth metal ions, ammonium ions, or first transition metal ions, such as sodium, potassium, magnesium, calcium, zinc, or aluminum. Examples of anions include chloride, bromine, iodine, sulfate, sulfite, bisulfite, thiosulfate, phosphate, dihydrogen phosphate, dibasic phosphate, hypophosphite, dihydrogen pyrophosphate, tetraborate, borate, carbonate, bicarbonate, silicate, citrate, malate, maleate, malonic acid, succinate, lactate, formate, acetate, butyrate, propionate, benzoate, tartrate, ascorbate, or gluconate. In particular, alkali or alkaline earth metal salts of sulfate, sulfite, phosphate, phosphonate, nitrate, chloride or carbonate, or salts of simple organic acids such as citrate, malonate or acetate can be used.

[0484] The salt in the coating may have a constant humidity of more than 60% at 20°C, particularly more than 70%, more than 80% or more than 85%, or it may be another hydrated form of the salt (e.g., anhydrous). Salt coating may be as described in WO00 / 01793 or WO 2006 / 034710.

[0485] A specific example of a suitable salt is NaCl (CH3) 20℃ =76%), Na2CO3(CH 20℃ =92%), NaNO3(CH 20℃ =73%), Na2HPO4(CH 20℃ =95%), Na3PO4(CH 25℃ =92%), NH4Cl(CH 20℃ =79.5%), (NH4)2HPO4(CH 20℃ =93.0%), NH4H2PO4(CH 20℃ =93.1%), (NH4)2SO4(CH 20℃ =81.1%), KCl(CH 20℃ =85%), K2HPO4(CH 20℃ =92%), KH2PO4(CH 20℃=96.5%), KNO3(CH 20℃ =93.5%), Na2SO4(CH 20℃ =93%), K2SO4(CH 20℃ =98%), KHSO4(CH 20℃ =86%), MgSO4(CH 20℃ =90%), ZnSO4(CH 20℃ =90%) and sodium citrate (CH 25℃ =86%). Other examples include NaH2PO4, (NH4)H2PO4, CuSO4, Mg(NO3)2, and magnesium acetate.

[0486] The salt can be in anhydrous form, or it can be a hydrated salt, i.e., a crystalline salt hydrate with bound water, such as that described in WO 99 / 32595. Specific examples include anhydrous sodium sulfate (Na₂SO₄), anhydrous magnesium sulfate (MgSO₄), magnesium sulfate heptahydrate (MgSO₄·7H₂O), zinc sulfate heptahydrate (ZnSO₄·7H₂O), disodium hydrogen phosphate heptahydrate (Na₂HPO₄·7H₂O), magnesium nitrate hexahydrate (Mg(NO₃)₂(6H₂O)), sodium citrate dihydrate, and magnesium acetate tetrahydrate. Preferably, the salt is used as a salt solution, for example, using a fluidized bed.

[0487] Therefore, in another aspect, the present invention provides particles comprising:

[0488] (a) Contains the core of the enzyme according to the invention.

[0489] (b) Optionally a coating consisting of one or more layers surrounding the core; and

[0490] (c) wherein the particle is preferably a co-particle containing one or more additional enzymes, wherein preferably at least one additional enzyme is selected from proteases, amylases and cellulases.

[0491] In one embodiment, the present invention provides particles comprising:

[0492] (a) Contains a core of peptides with fructan degradation activity.

[0493] (b) Optionally a coating consisting of one or more layers surrounding the core; and

[0494] (c) wherein the particle is preferably a co-particle containing one or more additional enzymes, wherein preferably at least one additional enzyme is selected from proteases, amylases and cellulases.

[0495] In one embodiment, the present invention provides particles comprising:

[0496] (a) A core comprising a polypeptide having fructan-degrading activity, wherein the polypeptide is selected from the group consisting of SEQ ID NO:3, SEQ ID NO:6, SEQ ID NO:9, SEQ ID NO:12, SEQ ID NO:15 and SEQ ID NO:18, and polypeptides having at least 60% sequence identity with these polypeptides, such as 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% sequence identity.

[0497] (b) Optionally a coating consisting of one or more layers surrounding the core; and

[0498] (c) wherein the particle is preferably a co-particle containing one or more additional enzymes, wherein preferably at least one additional enzyme is selected from proteases, amylases and cellulases.

[0499] use

[0500] The present invention also relates to methods for using the compositions thereof for cleaning clothing / textiles / fabrics (including household and industrial clothing) and for cleaning hard surfaces (including automated dishwashing (ADW), car washes and industrial surface cleaning).

[0501] Uses of cleaning compositions

[0502] For example, the detergent compositions of the present invention can be formulated as laundry detergent compositions for hand or machine washing, including as suitable laundry detergent additive compositions or rinse-additive fabric softener compositions for pretreatment of stained fabrics, or as detergent compositions for general household hard surface cleaning operations, or as formulations for hand or machine washing of dishes. Also provided herein are detergent additives comprising one or more of the fructan-degrading enzymes described herein, and optionally nucleases, such as DNases.

[0503] In one embodiment, the present invention relates to the use of a cleaning composition comprising a fructan-degrading enzyme as described herein (and optionally a nuclease, such as a DNase for cleaning products, wherein the product is a textile or surface).

[0504] In another embodiment, the present invention relates to the use of the fructan-degrading enzymes described herein (optionally in combination with nucleases such as DNases) for removing biofilms from articles (e.g., textiles or surfaces).

[0505] method

[0506] The present invention further relates to a method for treating fabrics, such as for removing biofilms from fabrics, the method comprising:

[0507] a) Contacting the fabric with an aqueous solution of a polysaccharide-degrading enzyme, and optionally with at least one nuclease polypeptide having DNase or RNase activity; and optionally...

[0508] b) Rinse and dry the fabric.

[0509] The present invention further relates to a method for using cleaning or washing products, the method comprising the following steps:

[0510] a) The product is exposed to a detergent containing at least one fructan-degrading enzyme, or a detergent composition containing such enzyme, and optionally to a nuclease polypeptide containing at least one DNAse or RNAse activity.

[0511] b) Complete at least one washing cycle; and optionally

[0512] c) Rinse the product.

[0513] The item in question is made of fabric.

[0514] The present invention further relates to a method for using cleaning or washing products, the method comprising the following steps:

[0515] a) Exposing the article to a detergent containing a polypeptide or a detergent composition containing a polypeptide, wherein the polypeptide is selected from the group consisting of: SEQ ID NO:3, SEQ ID NO:6, SEQ ID NO:9, SEQ ID NO:12, SEQ ID NO:15 and SEQ ID NO:18, and polypeptides having at least 60% sequence identity with these polypeptides, such as 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% sequence identity, and optionally at least one nuclease polypeptide having DNase or RNase activity;

[0516] b) Complete at least one washing cycle; and optionally

[0517] c) Rinse the product.

[0518] The item in question is made of fabric.

[0519] The pH of liquid solutions typically ranges from about 5.5 to about 12, for example from about 7 to about 11, for example from about 7 to about 10, for example from about 7 to about 9.

[0520] The washing liquid can have a temperature in the range of 5°C to 95°C, for example, in the range of 10°C to 80°C, 10°C to 70°C, 10°C to 60°C, 10°C to 50°C, 15°C to 40°C, or in the range of 20°C to 30°C. In one aspect, the temperature of the washing liquid is approximately 30°C.

[0521] The concentration of fructan-degrading enzyme in the washing solution is typically in the following ranges: at least 0.00001 ppm to at least 10 ppm, at least 0.00002 ppm to at least 10 ppm, at least 0.0001 ppm to at least 10 ppm, at least 0.0002 ppm to at least 10 ppm, at least 0.001 ppm to at least 10 ppm, at least 0.002 ppm to at least 10 ppm, at least 0.01 ppm to at least 10 ppm, at least 0.02 ppm to at least 10 ppm, at least 0.1 ppm to at least 10 ppm, at least 0.2 ppm to at least 10 ppm, at least 0.5 ppm to at least 5 ppm.

[0522] The invention is further described by the following examples, which should not be construed as limiting the scope of the invention.

[0523] Example

[0524] Example 1: Washing test

[0525] Preparation of small biofilm samples

[0526] Biofilm samples (10cm x 10cm) were prepared by growing *Syntrophus* species on polyester fabric samples for three days. The biofilm samples were rinsed twice in water and dried under flow in a laboratory bench for 2 hours. They were then pressed into circular samples (0.6cm diameter) and placed into the wells of an MTP96 microtiter plate, and stored at 4°C for further use.

[0527] Washing experiment

[0528] The washing performance of the fructanase of this invention was screened separately and also screened together with DNase. Small pieces of cloth were transferred from MTP96 plates to deep-well MTP96 plates for washing assays. The deep-well plates were placed in a Hamilton robot and a washing simulation program was performed under the following conditions: oscillation speed: 1000 rpm for 30 seconds; washing cycle duration: 30 minutes with oscillation; temperature: 30°C; total washing solution volume: 500 μl / well (490 μl of standard detergent A + 10 μl of sample).

[0529] A standard detergent solution (A) was prepared by dissolving 3.3 g / L of detergent in water with a hardness of 15°dH. Subsequently, dirt (WFK 09V pigment dirt; Testgewebe GmbH, Germany) was added to achieve a concentration of 0.7 g dirt / L.

[0530] Table 1: Standard Detergent A Composition

[0531]

[0532]

[0533] A 96-well plate was filled with each enzyme sample, and the program was started on a robot. Fructanase was tested at a concentration of 0.5 ppm, with or without a low dose (0.00001 ppm) of DNase from Bacillus foodae (SEQ ID NO: 19). Enzyme samples were tested against a blank consisting of small pieces of biofilm fabric without any enzyme. After completing a simulated washing cycle, the small pieces of fabric were removed from the washing solution and dried on filter paper. The dried small pieces of fabric were fixed onto a sheet of white paper for scanning, and the scanned images were analyzed using colorimeter software. Each sample was measured for intensity (from colorimeter software analysis), which was used to calculate the Δ intensity (reflectance) compared to the blank by subtracting the intensity value of the blank without enzyme from the intensity value of the sample with enzyme. Values ​​above 20 are visible to the human eye.

[0534] The reflectance value in this experiment is 168.

[0535] Table 2 below shows the washing performance of the seven fructan-degrading enzymes of the present invention, including the measured reflectance intensity and Δ intensity compared to the blank for a single enzyme, as well as the measured intensity and Δ intensity of the combination of two enzymes (0.5 ppm fructanase + 0.00001 ppm DNase) compared to the blank. The intensity and Δ intensity of the sample containing only DNase are also given.

[0536] It can be seen that all fructanases are able to remove biofilm from small cloth samples, and further, a significant synergistic effect can be obtained by using a combination of fructanase and low-dose DNase.

[0537] Table 2. Washing performance of fructanases with and without DNase on biofilms

[0538]

[0539]

[0540] Example 2: Fructanase Activity Assay

[0541] Fructanase and insoluble fructan substrate K-FRUCHK (Megazyme) were incubated as follows: 3 mg of fructan (Megazyme K-FRUCHK) was weighed into a centrifuge tube for each sample. Each sample was diluted 10-fold in 100 mM sodium acetate, pH 4.5 (except for sample SEQ ID NO: 12, which was already diluted and therefore pipetted directly). The sample was added to a final concentration of 100 ppm (0.1 mg / mL).

[0542]

[0543] The sample containing the enzyme and substrate was incubated at 37°C and 1400 rpm for 60 min, with a sample without added enzyme used as a control. After incubation, the sample was centrifuged at 16100 x G at room temperature for 5 min. The supernatant was analyzed by 1) liquid chromatography and 2) reduction end determination, as follows:

[0544] In Dionex with PD-10 column (Thermo Fisher Scientific) TM - Perform liquid chromatography on the IC300 system using the following gradient:

[0545]

[0546]

[0547] Fructose and glucose (Sigma) were used as standard references.

[0548] For the determination of reducing sugars, a working buffer was prepared by weighing 50 g of potassium sodium tartrate (K-Na-tartaric acid, Merck 8087) and 20 g of NaOH (Merck 1.06498) into 1 L of water.

[0549] The reducing agent is also called PAHBAH reagent, which is prepared by weighing 225 mg of PAHBAH into 15 ml of buffer solution to prepare PAHBAH (4-hydroxybenzoyl hydrazide, Sigma H-9882) solution.

[0550] The colorimetric reaction was performed by transferring 75 μL of the supernatant of each sample to a PCR plate and adding 150 μL of PAHBAH reagent.

[0551] After incubating at 95°C for 10 minutes on a PCR instrument, 150 μl of each sample was transferred to a microtiter plate to read the absorbance at 405 nm.

[0552] result

[0553] Liquid chromatography showed that fructanase was active on fructans. However, two enzymes bearing SEQ ID NO:12 and SEQ ID NO:18 produced hydrolysates different from the other enzymes. These enzymes produced monosaccharides, fructose, and glucose, indicating exoglycosidic activity. Other enzymes tested here produced oligosaccharide distributions of varying sizes, indicating endoglycosidic activity.

[0554] The table below describes the area of ​​the monosaccharides released by liquid chromatography, measured in nanocoulombs per minute (nC*min). The two sugars identified are glucose and fructose. For other fructooligosaccharides, the degree of polymerization (dp) is in the range of 1–10.

[0555]

[0556]

[0557] The levels of reducing sugars with SEQ ID NO:12 and SEQ ID NO:18 are consistent with the results in the table above, and these two enzymes have higher levels of activity.

[0558] The absorbance at 405 nm reflects the activity of fructan as follows:

[0559] 6 1.83 9 1.94 12 2.70 15 2.02 18 2.55 No enzymes 0.21

[0560] Example 3: Construction of clades and phylogenetic trees

[0561] GH32 evolutionary tree

[0562] Phylogenetic trees were constructed from polypeptide sequences of the present invention containing the GH32 domain, as defined in CAZY (Lombard, Henrissat et al., 2014. The carbohydrate-active enzymes database (CAZy), Nucleic Acids Res., 2013, http: / / www.cazy.org / ). Phylogenetic trees were also constructed from multiple alignments of mature polypeptide sequences containing at least one GH32 domain. Sequences were aligned using MUSCLE algorithm version 3.8.31 (Edgar, 2004. Nucleic Acids Research, 32(5):1792-1797), and the tree was constructed using FastTree version 2.1.8 (Price et al., 2010. PloS one, 5(3)). The tree was visualized using iTOL (Letunic and Bork, 2007. Bioinformatics, 23(1):127-128).

[0563] The peptide subtype containing the GH32 domain also contains the 32C-terminal domain (GH32C) of the glycosyl hydrolase family, as defined by Pfam domain ID PF08244 (The Pfam protein families database: towards a more sustainable future, Finn et al., Nucleic Acids Research (2016) database Issue 44: D279-D285). The peptides of this invention contain both the GH32 domain and the 32C-terminal domain of the glycosyl hydrolase family. The 32C-terminal domain of the glycosyl hydrolase family will be denoted as the GH32C domain. As an example, in SEQ ID NO:6 from *Aspergillus brevis*, the GH32C domain is located at positions 351 to 484.

[0564] Clade formation

[0565] In addition to containing the GH32 domain and the GH32C domain, the polypeptide of the present invention also contains one or more of several motifs.

[0566] One example is the motif [WPG][GMTH][NH][AEILDNMV] (SEQ ID NO:24), which is located at positions 15 to 18 in Aspergillus lanceolata (SEQ ID NO:6). Preferably, the sequence of the motif at these positions is W[GMT]N[NDEI] (SEQ ID NO:32).

[0567] Peptides containing the GH32 domain can be separated into different subclusters or clades, defined by one or more short sequence motifs, and containing both the GH32 and GH32C domains.

[0568] We denote the subcluster containing the motif [WPG][GMTH][NH][AEILDNMV] (SEQ ID NO:24) as the WMNE clade. Preferably, this motif has the sequence W[GMT]N[NDEI] (SEQ ID NO:32). Peptide sequences containing the GH32 domain, the GH32C domain, and the motif of SEQ ID NO:24 (preferably the motif of SEQ ID NO:32) will be represented as belonging to the WMNE clade.

[0569] The polypeptides in the WMNE clade can be further separated into several distinct subclusters or clades. The different motifs of each of these clades are described in detail below.

[0570] The formation of the WMNE clade

[0571] Using the MUSCLE algorithm and FastTree, phylogenetic trees were constructed from multiple alignments of peptide sequences containing the GH32 domain as defined above, and from mature peptide sequences containing at least one GH32 domain, and visualized using iTOL. The phylogenetic trees allowed peptides within GH32 to be separated into distinct subclusters, which we denoted as WMNE.

[0572] The characteristic motif of this subcluster is the motif [WPG][GMTH][NH][AEILDNMV] (SEQ ID NO:24), which corresponds to amino acids WMNE at positions 15 to 18 in Aspergillus brevis (SEQ ID NO:6). Preferably, the sequence of the motif at these positions is W[GMT]N[NDEI] (SEQ ID NO:32).

[0573] Peptides containing the GH32 domain, the GH32C domain, and this motif belong to the WMNE clade.

[0574] The motif of SEQ ID NO:24 is preferably W[GMT]N[NDEI] (SEQ ID NO:32), and more particularly in some embodiments is WMN[DE] (SEQ ID NO:30), such as WMN[DE]PNG (SEQ ID NO:31). Examples of polypeptides having these specific motifs in the WMNE clade are SEQ ID NO:6, 9, 12, 15, and 18.

[0575] WMNE can be divided into subclusters, one represented by the AYSN clade and the other by the WMNEP clade.

[0576] The generation of the AYSN clade

[0577] The polypeptide of the AYSN clade contains the GH32 domain (belonging to the WMNE clade) and includes the motif A[YF]S[LN]D[QK] (SEQ ID NO:25), which corresponds to amino acids AYSNDK (SEQ ID NO:26) at positions 120 to 125 in Bacillus licheniformis (SEQ ID NO:18), wherein the D corresponding to position 124 is completely conserved in the AYSN clade.

[0578] Another motif contained in the polypeptide of the AYSN clade is [HS]WGH (SEQ ID NO:27), located at positions 53 to 56 in Bacillus licheniformis (SEQ ID NO:18).

[0579] Examples of peptides from the AYSN clade include SEQ ID NO:12 and SEQ ID NO:18.

[0580] Generation of the WMNEP clade

[0581] The WMNE clade can be divided into another, separate subcluster, denoted as the WMNEP clade. The polypeptide of the WMNEP clade contains the motif [QLKEYMRVTANI][NSYHATRD][WF][MEVKTL][NGA][EVDLIH][PAEHDS][NATSKGVERQCD] (SEQ ID NO:28), corresponding to amino acids KNWMNEPN (SEQ ID NO:29) at positions 13 to 20 of Aspergillus flexus (SEQ ID NO:6).

[0582] Examples of peptides in the WMNEP clade include SEQ ID NO:6, SEQ ID NO:9, and SEQ ID NO:15. sequence list <110> Novozymes A / S <120> Contains peptides with fructan degradation activity Cleaning composition <130> 14962-WO-PCT <160> 32 <170> PatentIn version 3.5 <210> 1 <211> 2769 <212> DNA <213> Amylolytic Bacillus <220> <221> CDS <222> (1)..(2769) <220> <221> signal peptide <222> (1)..(93) <220> <221> Mature peptides <222> (94)..(2769) <400> 1 atg aaa gat aaa tct aaa ttc gca ata tgt ctt ctg atg gca acg gga 48 Met Lys Asp Lys Ser Lys Phe Ala Ile Cys Leu Leu Met Ala Thr Gly -30 -25 -20 tta act ctg act tca aac ggt att ttt aca tct aaa tca att gcg gct 96 Leu Thr Leu Thr Ser Asn Gly Ile Phe Thr Ser Lys Ser Ile Ala Ala -15 -10 -5 -1 1 agt gct gta ctt gct gat cag gaa aca aat aaa cag gct aag gag gag 144 Ser Ala Val Leu Ala Asp Gln Glu Thr Asn Lys Gln Ala Lys Glu Glu 5 10 15 ctt gat att ttt gaa agc gca tcc aag tca aat acc aat ctg ata ggc 192 Leu Asp Ile Phe Glu Ser Ala Ser Lys Ser Asn Thr Asn Leu Ile Gly 20 25 30 tgg caa gtg aaa gga aaa gga gga tta gag gat act tca gaa gga atc ​​240 Trp Gln Val Lys Gly Lys Gly Gly Leu Glu Asp Thr Ser Glu Gly Ile 35 40 45 ttg cta act tca cag cct aaa gaa aat gta atg gcc atc tca gag aca 288 Leu Leu Thr Ser Gln Pro Lys Glu Asn Val Met Ala Ile Ser Glu Thr 50 55 60 65 gtg tcc gat gat ttt atc tat gaa gct gat gtc atg atc agg gat acg 336 Val Ser Asp Asp Phe Ile Tyr Glu Ala Asp Val Met Ile Arg Asp Thr 70 75 80 aag gca gac gca aca ttg ttg ttt cgt tct aat gag gat ggc ttt agc 384 Lys Ala Asp Ala Thr Leu Leu Phe Arg Ser Asn Glu Asp Gly Phe Ser 85 90 95 tcg tac atg ctg caa atc ​​gtt ccg gac gca ggt ctg att cgg tta aga 432 Ser Tyr Met Leu Gln Ile Val Pro Asp Ala Gly Leu Ile Arg Leu Arg 100 105 110 gat gca aga act gga gat ggg aaa tta aag gag gag cgt aaa gtt tct 480 Asp Ala Arg Thr Gly Asp Gly Lys Leu Lys Glu Glu Arg Lys Val Ser 115 120 125 gtt gaa atg ggg caa atc ​​tat cat ctc aaa gtg aag gca gtg ggt tcc 528 Val Glu Met Gly Gln Ile Tyr His Leu Lys Val Lys Ala Val Gly Ser 130 135 140 145 tcg cta aaa gta tat tgg ggc aat caa tat aaa cca tta att gat gtt 576 Ser Leu Lys Val Tyr Trp Gly Asn Gln Tyr Lys Pro Leu Ile Asp Val 150 155 160 caa gac att tcg tat caa agc gga aag ctt gga ctc cat gta tgg gat 624 Gln Asp Ile Ser Tyr Gln Ser Gly Lys Leu Gly Leu His Val Trp Asp 165 170 175 gga tcc gcc ttg ttt tcg aac atc gtg gtg agc gat ctg aaa ggt aat 672 Gly Ser Ala Leu Phe Ser Asn Ile Val Val Ser Asp Leu Lys Gly Asn 180 185 190 ttg ggt acg gtg ctt tct agc aag gga aaa tgg cag cct gat atc aac 720 Leu Gly Thr Val Leu Ser Ser Lys Gly Lys Trp Gln Pro Asp Ile Asn 195 200 205 ggc aaa aga ggg acg gta gaa cag ggg agc ata gcr cag caa atc ​​tat 768 Gly Lys Arg Gly Thr Val Glu Gln Gly Ser Ile Ala Gln Gln Ile Tyr 210 215 220 225 aac aaa gag gcg act gat atg gtc tat gaa ggg aat att acc ctt cgt 816 Asn Lys Glu Ala Thr Asp Met Val Tyr Glu Gly Asn Ile Thr Leu Arg 230 235 240 cct gat tct att gca gct ctc gca ttt cga tct tca acc gat gga gct 864 Pro Asp Ser Ile Ala Ala Leu Ala Phe Arg Ser Ser Thr Asp Gly Ala 245 250 255 gaa gga tat gaa gct act ctt acg aag gaa gga gat cag gtc cgt gta 912 Glu Gly Tyr Glu Ala Thr Leu Thr Lys Glu Gly Asp Gln Val Arg Val 260 265 270 agt ttg acg aat aca aaa gga aca gta att gca agt tcg caa cgt act 960 Ser Leu Thr Asn Thr Lys Gly Thr Val Ile Ala Ser Ser Gln Arg Thr 275 280 285 tat ccg agt cag atg gga gcc aaa cat cat gtg gaa atc aag gca aag 1008 Tyr Pro Ser Gln Met Gly Ala Lys His His Val Glu Ile Lys Ala Lys 290 295 300 305 gga gat cga att cag gtc ttc ctg gac ggg tac act acg gcc gcg att 1056 Gly Asp Arg Ile Gln Val Phe Leu Asp Gly Tyr Thr Thr Ala Ala Ile 310 315 320 gac gtg aaa gat tca acc tac aag agt gga agt acc gga atc gtc gta 1104 Asp Val Lys Asp Ser Thr Tyr Lys Ser Gly Ser Thr Gly Ile Val Val 325 330 335 aaa aag ggg acg gct tac ttt cag gat act tat gtg acg gaa ctg agc 1152 Lys Lys Gly Thr Ala Tyr Phe Gln Asp Thr Tyr Val Thr Glu Leu Ser 340 345 350 caa tat tac aat gag ata tat cgt cca caa tat cac tac acg cca ata 1200 Gln Tyr Tyr Asn Glu Ile Tyr Arg Pro Gln Tyr His Tyr Thr Pro Ile 355 360 365 cgt ggt tca gcg agt gat ccg aat gga ctt gtc tac ttc gaa gga gaa 1248 Arg Gly Ser Ala Ser Asp Pro Asn Gly Leu Val Tyr Phe Glu Gly Glu 370 375 380 385 tat cat ctc ttc cat cag gat gga gga aca tgg gcg cat gcg gta agt 1296 Tyr His Leu Phe His Gln Asp Gly Gly Thr Trp Ala His Ala Val Ser 390 395 400 aaa gat atg ctg aac tgg aaa cgg ctt ccg att gca ctt ccc tgg aat 1344 Lys Asp Met Leu Asn Trp Lys Arg Leu Pro Ile Ala Leu Pro Trp Asn 405 410 415 gat cac gga cat gtc tgg tct ggg tcg gct gtt gcg gat atg aca aat 1392 Asp His Gly His Val Trp Ser Gly Ser Ala Val Ala Asp Met Thr Asn 420 425 430 gca tct ggt tta ttc gga gat tca gga ggc aag ggc ctt att gca tac 1440 Ala Ser Gly Leu Phe Gly Asp Ser Gly Gly Lys Gly Leu Ile Ala Tyr 435 440 445 tat act tcc ttt aat ccg gat agc ccg aat ggg aac cag cgt ata ggt 1488 Tyr Thr Ser Phe Asn Pro Asp Ser Pro Asn Gly Asn Gln Arg Ile Gly 450 455 460 465 cta gct tac agt aaa gac caa ggt cgt act tgg gag tat tcg aag gag 1536 Leu Ala Tyr Ser Lys Asp Gln Gly Arg Thr Trp Glu Tyr Ser Lys Glu 470 475 480 cgc ccg att gtg atc gag aac ccg ggt aag agc gga aat gaa gct ggg 1584 Arg Pro Ile Val Ile Glu Asn Pro Gly Lys Ser Gly Asn Glu Ala Gly 485 490 495 aat tgg gat ttc cgc gat ccg aaa gta atc ​​cgt gat gat gag aat aac 1632 Asn Trp Asp Phe Arg Asp Pro Lys Val Ile Arg Asp Asp Glu Asn Asn 500 505 510 cga tgg gtt atg gtt gtg tcc gga gga gat cat att cgt ttc tat act 1680 Arg Trp Val Met Val Val Ser Gly Gly Asp His Ile Arg Phe Tyr Thr 515 520 525 tca aca aat tta ctt gac tgg aca ttg aca gat aat tgg gga tat ggg 1728 Ser Thr Asn Leu Leu Asp Trp Thr Leu Thr Asp Asn Trp Gly Tyr Gly 530 535 540 545 gat tac gtc cgc gga gga gta tgg gaa tgc cct gat ttg ttc cag ctt 1776 Asp Tyr Val Arg Gly Gly Val Trp Glu Cys Pro Asp Leu Phe Gln Leu 550 555 560 ccg gta gac gga acg tca cag aaa aag tgg gtt atg atg atc agt aca 1824 Pro Val Asp Gly Thr Ser Gln Lys Lys Trp Val Met Met Ile Ser Thr 565 570 575 gga gcg aat ccc aaa aca ggc gga tca gat gcc gag tat ttt atc ggt 1872 Gly Ala Asn Pro Lys Thr Gly Gly Ser Asp Ala Glu Tyr Phe Ile Gly 580 585 590 cat tta aca gct gat ggt aaa ttc gta aac gat aat ccg gca ggt aag 1920 His Leu Thr Ala Asp Gly Lys Phe Val Asn Asp Asn Pro Ala Gly Lys 595 600 605 gtg tta aga aca gat ttt ggt aaa gaa ttt tac gct tcg atg tct ttt 1968 Val Leu Arg Thr Asp Phe Gly Lys Glu Phe Tyr Ala Ser Met Ser Phe 610 615 620 625 gct aac atg cct gat cat cgc aca gtg atg atg gcg tgg atg acg aat 2016 Ala Asn Met Pro Asp His Arg Thr Val Met Met Ala Trp Met Thr Asn 630 635 640 tgg gat tat ccg ttc gct ttc cca acg tcc aat tgg aaa ggt gaa cta 2064 Trp Asp Tyr Pro Phe Ala Phe Pro Thr Ser Asn Trp Lys Gly Glu Leu 645 650 655 acc att ccg aga gaa gta tca ttg gta acg acc gaa gat gga att cgg 2112 Thr Ile Pro Arg Glu Val Ser Leu Val Thr Thr Glu Asp Gly Ile Arg 660 665 670 atg gtg caa agt cca atc ​​aaa gaa ttg gaa tca ctg cgt aaa cct ttg 2160 Met Val Gln Ser Pro Ile Lys Glu Leu Glu Ser Leu Arg Lys Pro Leu 675 680 685 tat tct gct tct aac aag tcg gtg agc cct tct tcc ggg aat ctg cta 2208 Tyr Ser Ala Ser Asn Lys Ser Val Ser Pro Ser Ser Gly Asn Leu Leu 690 695 700 705 aaa ggt att att tca ggt gct tac gaa att gaa gct gaa att gaa atc ​​2256 Lys Gly Ile Ile Ser Gly Ala Tyr Glu Ile Glu Ala Glu Ile Glu Ile 710 715 720 cct gaa acc agc aca gtg acc gag ttt ggc ttt aac att cgt gag ggt 2304 Pro Glu Thr Ser Thr Val Thr Glu Phe Gly Phe Asn Ile Arg Glu Gly 725 730 735 gca aat cag aag acg gtc gtg ggt tat aag gca agc gat agt cgt atg 2352 Only Asn Gln Lys Thr Val Val Gly Tyr Lys Ala Ser Asp Ser Arg Met 740,745,750 ttt gtg gac cga act gca tcc ggt gaa acg gat ttt tct aac ctt ttc 2400 Phe Val Asp Arg Thr Ala Ser Gly Glu Thr Asp Phe Ser Asn Leu Phe 755,760,765 agt aag aaa cat gaa gcg cct acg caa atg gag at at cgg atc aag 2448 Serving Lys Lys His Glu Ala Pro Thr Gln Met Glu Asn Asn Arg Ile Lys 770 775 780 785 atg cgt att ctt gtg gat gag tct tct gtt gaa gcc ttt ggt aac gac 2496 Met Arg Ile Leu Val Asp Glu Ser Ser Val Glu Ala Phe Gly Asn Asp 790,795,800 ggc aaa gtc gtc ttt tca gat gtt ata ttt ccg gat cct gct agt aga 2544 Gly Lys Val Val Phe Ser Asp Val Ile Phe Pro Asp Pro Ala Ser Arg 805 810 815 gcg atg agt ttt tac gta aaa ggc ggg aat gtg aat gtt gtt tcc ttg 2592 Ala Met Ser Phe Tyr Val Lys Gly Gly Asn Val Asn Val Val Ser Leu 820 825 830 aaa gtg cat caa ctt caa tct gtc tgg at gcg gac att cct tca aaa 2640 Lys Will His Gln Leu Gln Ser Val Trp Asn Ala Asp Ile Pro Ser Lys 835 840 845 gtt caa ata aag atg gat acg agc gtt cgg gaa ctg ggg gta ggc gag 2688 Val Gln Ile Lys Met Asp Thr Served Val Arg Glu Leu Gly Val Gly Glu 850 855 860 865 tcg gat aca ctg cag gcg atg gtc gag tat ggc ccg ggt cta ggc gtt 2736 Ser Asp Thr Leu Gln Ala Met Val Glu Tyr Gly Pro Gly Leu Gly Val 870 875 880 CA CCG CTT AAG Tgg AT CC GTA AT ACG ACG 2769 Gln Pro Leu Lys Trp Asn Pro Val Ile Thr Thr 885,890 <210> 2 <211> 923 <212> PRT <213> Paenibacillus amyloliquefaciens <400> 2 Met Lys Asp Lys Ser Lys Phe Ala Ile Cys Leu Leu Met Ala Thr Gly -30 -25 -20 Leu Thr Leu Thr Ser Asn Gly Ile Phe Thr Ser Lys Ser Ile Ala Ala -15 -10 -5 -1 1 Ser Ala Val Leu Ala Asp Gln Glu Thr Asn Lys Gln Ala Lys Glu Glu 5 10 15 Leu Asp Ile Phe Glu Ser Ala Ser Lys Ser Asn Thr Asn Leu Ile Gly 20 25 30 Trp Gln Val Lys Gly Lys Gly Gly Leu Glu Asp Thr Ser Glu Gly Ile 35 40 45 Leu Leu Thr Ser Gln Pro Lys Glu Asn Val Met Ala Ile Ser Glu Thr 50 55 60 65 Val Ser Asp Asp Phe Ile Tyr Glu Ala Asp Val Met Ile Arg Asp Thr 70 75 80 Lys Ala Asp Ala Thr Leu Leu Phe Arg Ser Asn Glu Asp Gly Phe Ser 85 90 95 Ser Tyr Met Leu Gln Ile Val Pro Asp Ala Gly Leu Ile Arg Leu Arg 100 105 110 Asp Ala Arg Thr Gly Asp Gly Lys Leu Lys Glu Glu Arg Lys Val Ser 115 120 125 Val Glu Met Gly Gln Ile Tyr His Leu Lys Val Lys Ala Val Gly Ser 130 135 140 145 Ser Leu Lys Val Tyr Trp Gly Asn Gln Tyr Lys Pro Leu Ile Asp Val 150 155 160 Gln Asp Ile Ser Tyr Gln Ser Gly Lys Leu Gly Leu His Val Trp Asp 165 170 175 Gly Ser Ala Leu Phe Ser Asn Ile Val Val Ser Asp Leu Lys Gly Asn 180 185 190 Leu Gly Thr Val Leu Ser Ser Lys Gly Lys Trp Gln Pro Asp Ile Asn 195 200 205 Gly Lys Arg Gly Thr Val Glu Gln Gly Ser Ile Ala Gln Gln Ile Tyr 210 215 220 225 Asn Lys Glu Ala Thr Asp Met Val Tyr Glu Gly Asn Ile Thr Leu Arg 230 235 240 Pro Asp Ser Ile Ala Ala Leu Ala Phe Arg Ser Ser Thr Asp Gly Ala 245 250 255 Glu Gly Tyr Glu Ala Thr Leu Thr Lys Glu Gly Asp Gln Val Arg Val 260 265 270 Ser Leu Thr Asn Thr Lys Gly Thr Val Ile Ala Ser Ser Gln Arg Thr 275 280 285 Tyr Pro Ser Gln Met Gly Ala Lys His His Val Glu Ile Lys Ala Lys 290 295 300 305 Gly Asp Arg Ile Gln Val Phe Leu Asp Gly Tyr Thr Thr Ala Ala Ile 310 315 320 Asp Val Lys Asp Ser Thr Tyr Lys Ser Gly Ser Thr Gly Ile Val Val 325 330 335 Lys Lys Gly Thr Ala Tyr Phe Gln Asp Thr Tyr Val Thr Glu Leu Ser 340 345 350 Gln Tyr Tyr Asn Glu Ile Tyr Arg Pro Gln Tyr His Tyr Thr Pro Ile 355 360 365 Arg Gly Ser Ala Ser Asp Pro Asn Gly Leu Val Tyr Phe Glu Gly Glu 370 375 380 385 Tyr His Leu Phe His Gln Asp Gly Gly Thr Trp Ala His Ala Val Ser 390 395 400 Lys Asp Met Leu Asn Trp Lys Arg Leu Pro Ile Ala Leu Pro Trp Asn 405 410 415 Asp His Gly His Val Trp Ser Gly Ser Ala Val Ala Asp Met Thr Asn 420 425 430 Ala Ser Gly Leu Phe Gly Asp Ser Gly Gly Lys Gly Leu Ile Ala Tyr 435 440 445 Tyr Thr Ser Phe Asn Pro Asp Ser Pro Asn Gly Asn Gln Arg Ile Gly 450 455 460 465 Leu Ala Tyr Ser Lys Asp Gln Gly Arg Thr Trp Glu Tyr Ser Lys Glu 470 475 480 Arg Pro Ile Val Ile Glu Asn Pro Gly Lys Ser Gly Asn Glu Ala Gly 485 490 495 Asn Trp Asp Phe Arg Asp Pro Lys Val Ile Arg Asp Asp Glu Asn Asn 500 505 510 Arg Trp Val Met Val Val Ser Gly Gly Asp His Ile Arg Phe Tyr Thr 515 520 525 Ser Thr Asn Leu Leu Asp Trp Thr Leu Thr Asp Asn Trp Gly Tyr Gly 530 535 540 545 Asp Tyr Val Arg Gly Gly Val Trp Glu Cys Pro Asp Leu Phe Gln Leu 550 555 560 Pro Val Asp Gly Thr Ser Gln Lys Lys Trp Val Met Met Ile Ser Thr 565 570 575 Gly Ala Asn Pro Lys Thr Gly Gly Ser Asp Ala Glu Tyr Phe Ile Gly 580 585 590 His Leu Thr Ala Asp Gly Lys Phe Val Asn Asp Asn Pro Ala Gly Lys 595 600 605 Val Leu Arg Thr Asp Phe Gly Lys Glu Phe Tyr Ala Ser Met Ser Phe 610 615 620 625 Ala Asn Met Pro Asp His Arg Thr Val Met Met Ala Trp Met Thr Asn 630 635 640 Trp Asp Tyr Pro Phe Ala Phe Pro Thr Ser Asn Trp Lys Gly Glu Leu 645 650 655 Thr Ile Pro Arg Glu Val Ser Leu Val Thr Thr Glu Asp Gly Ile Arg 660 665 670 Met Val Gln Ser Pro Ile Lys Glu Leu Glu Ser Leu Arg Lys Pro Leu 675 680 685 Tyr Ser Ala Ser Asn Lys Ser Val Ser Pro Ser Ser Gly Asn Leu Leu 690 695 700 705 Lys Gly Ile Ile Ser Gly Ala Tyr Glu Ile Glu Ala Glu Ile Glu Ile 710 715 720 Pro Glu Thr Ser Thr Val Thr Glu Phe Gly Phe Asn Ile Arg Glu Gly 725 730 735 Ala Asn Gln Lys Thr Val Val Gly Tyr Lys Ala Ser Asp Ser Arg Met 740 745 750 Phe Val Asp Arg Thr Ala Ser Gly Glu Thr Asp Phe Ser Asn Leu Phe 755 760 765 Ser Lys Lys His Glu Ala Pro Thr Gln Met Glu Asn Asn Arg Ile Lys 770 775 780 785 Met Arg Ile Leu Val Asp Glu Ser Ser Val Glu Ala Phe Gly Asn Asp 790 795 800 Gly Lys Val Val Phe Ser Asp Val Ile Phe Pro Asp Pro Ala Ser Arg 805 810 815 Ala Met Ser Phe Tyr Val Lys Gly Gly Asn Val Asn Val Val Ser Leu 820 825 830 Lys Val His Gln Leu Gln Ser Val Trp Asn Ala Asp Ile Pro Ser Lys 835 840 845 Val Gln Ile Lys Met Asp Thr Ser Val Arg Glu Leu Gly Val Gly Glu 850 855 860 865 Ser Asp Thr Leu Gln Ala Met Val Glu Tyr Gly Pro Gly Leu Gly Val 870 875 880 Gln Pro Leu Lys Trp Asn Pro Val Ile Thr Thr 885 890 <210> 3 <211> 892 <212> PRT <213> Paenibacillus amylolyticus <400> 3 Ala Ser Ala Val Leu Ala Asp Gln Glu Thr Asn Lys Gln Ala Lys Glu 1 5 10 15 Glu Leu Asp Ile Phe Glu Ser Ala Ser Lys Ser Asn Thr Asn Leu Ile 20 25 30 Gly Trp Gln Val Lys Gly Lys Gly Gly Leu Glu Asp Thr Ser Glu Gly 35 40 45 Ile Leu Leu Thr Ser Gln Pro Lys Glu Asn Val Met Ala Ile Ser Glu 50 55 60 Thr Val Ser Asp Asp Phe Ile Tyr Glu Ala Asp Val Met Ile Arg Asp 65 70 75 80 Thr Lys Ala Asp Ala Thr Leu Leu Phe Arg Ser Asn Glu Asp Gly Phe 85 90 95 Ser Ser Tyr Met Leu Gln Ile Val Pro Asp Ala Gly Leu Ile Arg Leu 100 105 110 Arg Asp Ala Arg Thr Gly Asp Gly Lys Leu Lys Glu Glu Arg Lys Val 115 120 125 Ser Val Glu Met Gly Gln Ile Tyr His Leu Lys Val Lys Ala Val Gly 130 135 140 Ser Ser Leu Lys Val Tyr Trp Gly Asn Gln Tyr Lys Pro Leu Ile Asp 145 150 155 160 Val Gln Asp Ile Ser Tyr Gln Ser Gly Lys Leu Gly Leu His Val Trp 165 170 175 Asp Gly Ser Ala Leu Phe Ser Asn Ile Val Val Ser Asp Leu Lys Gly 180 185 190 Asn Leu Gly Thr Val Leu Ser Ser Lys Gly Lys Trp Gln Pro Asp Ile 195 200 205 Asn Gly Lys Arg Gly Thr Val Glu Gln Gly Ser Ile Ala Gln Gln Ile 210 215 220 Tyr Asn Lys Glu Ala Thr Asp Met Val Tyr Glu Gly Asn Ile Thr Leu 225 230 235 240 Arg Pro Asp Ser Ile Ala Ala Leu Ala Phe Arg Ser Ser Thr Asp Gly 245 250 255 Ala Glu Gly Tyr Glu Ala Thr Leu Thr Lys Glu Gly Asp Gln Val Arg 260 265 270 Val Ser Leu Thr Asn Thr Lys Gly Thr Val Ile Ala Ser Ser Gln Arg 275 280 285 Thr Tyr Pro Ser Gln Met Gly Ala Lys His His Val Glu Ile Lys Ala 290 295 300 Lys Gly Asp Arg Ile Gln Val Phe Leu Asp Gly Tyr Thr Thr Ala Ala 305 310 315 320 Ile Asp Val Lys Asp Ser Thr Tyr Lys Ser Gly Ser Thr Gly Ile Val 325 330 335 Val Lys Lys Gly Thr Ala Tyr Phe Gln Asp Thr Tyr Val Thr Glu Leu 340 345 350 Ser Gln Tyr Tyr Asn Glu Ile Tyr Arg Pro Gln Tyr His Tyr Thr Pro 355 360 365 Ile Arg Gly Ser Ala Ser Asp Pro Asn Gly Leu Val Tyr Phe Glu Gly 370 375 380 Glu Tyr His Leu Phe His Gln Asp Gly Gly Thr Trp Ala His Ala Val 385 390 395 400 Ser Lys Asp Met Leu Asn Trp Lys Arg Leu Pro Ile Ala Leu Pro Trp 405 410 415 Asn Asp His Gly His Val Trp Ser Gly Ser Ala Val Ala Asp Met Thr 420 425 430 Asn Ala Ser Gly Leu Phe Gly Asp Ser Gly Gly Lys Gly Leu Ile Ala 435 440 445 Tyr Tyr Thr Ser Phe Asn Pro Asp Ser Pro Asn Gly Asn Gln Arg Ile 450 455 460 Gly Leu Ala Tyr Ser Lys Asp Gln Gly Arg Thr Trp Glu Tyr Ser Lys 465 470 475 480 Glu Arg Pro Ile Val Ile Glu Asn Pro Gly Lys Ser Gly Asn Glu Ala 485 490 495 Gly Asn Trp Asp Phe Arg Asp Pro Lys Val Ile Arg Asp Asp Glu Asn 500 505 510 Asn Arg Trp Val Met Val Val Ser Gly Gly Asp His Ile Arg Phe Tyr 515 520 525 Thr Ser Thr Asn Leu Leu Asp Trp Thr Leu Thr Asp Asn Trp Gly Tyr 530 535 540 Gly Asp Tyr Val Arg Gly Gly Val Trp Glu Cys Pro Asp Leu Phe Gln 545 550 555 560 Leu Pro Val Asp Gly Thr Ser Gln Lys Lys Trp Val Met Met Ile Ser 565 570 575 Thr Gly Ala Asn Pro Lys Thr Gly Gly Ser Asp Ala Glu Tyr Phe Ile 580 585 590 Gly His Leu Thr Ala Asp Gly Lys Phe Val Asn Asp Asn Pro Ala Gly 595 600 605 Lys Val Leu Arg Thr Asp Phe Gly Lys Glu Phe Tyr Ala Ser Met Ser 610 615 620 Phe Ala Asn Met Pro Asp His Arg Thr Val Met Met Ala Trp Met Thr 625 630 635 640 Asn Trp Asp Tyr Pro Phe Ala Phe Pro Thr Ser Asn Trp Lys Gly Glu 645 650 655 Leu Thr Ile Pro Arg Glu Val Ser Leu Val Thr Thr Glu Asp Gly Ile 660 665 670 Arg Met Val Gln Ser Pro Ile Lys Glu Leu Glu Ser Leu Arg Lys Pro 675 680 685 Leu Tyr Ser Ala Ser Asn Lys Ser Val Ser Pro Ser Ser Gly Asn Leu 690 695 700 Leu Lys Gly Ile Ile Ser Gly Ala Tyr Glu Ile Glu Ala Glu Ile Glu 705 710 715 720 Ile Pro Glu Thr Ser Thr Val Thr Glu Phe Gly Phe Asn Ile Arg Glu 725 730 735 Gly Ala Asn Gln Lys Thr Val Val Gly Tyr Lys Ala Ser Asp Ser Arg 740 745 750 Met Phe Val Asp Arg Thr Ala Ser Gly Glu Thr Asp Phe Ser Asn Leu 755 760 765 Phe Ser Lys Lys His Glu Ala Pro Thr Gln Met Glu Asn Asn Arg Ile 770 775 780 Lys Met Arg Ile Leu Val Asp Glu Ser Ser Val Glu Ala Phe Gly Asn 785 790 795 800 Asp Gly Lys Val Val Phe Ser Asp Val Ile Phe Pro Asp Pro Ala Ser 805 810 815 Arg Ala Met Ser Phe Tyr Val Lys Gly Gly Asn Val Asn Val Val Ser 820 825 830 Leu Lys Val His Gln Leu Gln Ser Val Trp Asn Ala Asp Ile Pro Ser 835 840 845 Lys Val Gln Ile Lys Met Asp Thr Ser Val Arg Glu Leu Gly Val Gly 850 855 860 Glu Ser Asp Thr Leu Gln Ala Met Val Glu Tyr Gly Pro Gly Leu Gly 865 870 875 880 Val Gln Pro Leu Lys Trp Asn Pro Val Ile Thr Thr 885 890 <210> 4 <211> 1539 <212> DNA <213> Aspergillus effusus <220> <221> CDS <222> (1)..(1536) <220> <221> signal peptide <222> (1)..(48) <220> <221> mature peptide <222> (49)..(1536) <400> 4 atg ctc ctg tct ctc tca ttg att gca ctt gcc acg caa gct gtg gcc 48 Met Leu Leu Ser Leu Ser Leu Ile Ala Leu Ala Thr Gln Ala Val Ala -15 -10 -5 -1 gac gac ttc cgc ccc gtc ttc cac ttc gtc ccc gag aag aac tgg atg 96 Asp Asp Phe Arg Pro Val Phe His Phe Val Pro Glu Lys Asn Trp Met 1 5 10 15 aac gag ccc aac ggc ttg atc cag atc gac tcc gtc tgg cac ctg ttc 144 Asn Glu Pro Asn Gly Leu Ile Gln Ile Asp Ser Val Trp His Leu Phe 20 25 30 tac cag cac aac cca acc gcg aac gtc tgg ggc aat ctg aac tgg ggc 192 Tyr Gln His Asn Pro Thr Ala Asn Val Trp Gly Asn Leu Asn Trp Gly 35 40 45 cac gca acc agc agt gac cta gtc cac tgg act cac gag ccc ctc gca 240 His Ala Thr Ser Ser Asp Leu Val His Trp Thr His Glu Pro Leu Ala 50 55 60 atc acc agc gag aac ggc gtc gaa gcc ttc acc ggc acc gcc ttt tac 288 Ile Thr Ser Glu Asn Gly Val Glu Ala Phe Thr Gly Thr Ala Phe Tyr 65 70 75 80 gac gcc gag aat acc tct ggt ctc gga tct gcc gcc agt ccg ccg tac 336 Asp Ala Glu Asn Thr Ser Gly Leu Gly Ser Ala Ala Ser Pro Pro Tyr 85 90 95 ctg gcg tgg tac act ggg tat ctc ccg tcg aat ggg acc cag gac cag 384 Leu Ala Trp Tyr Thr Gly Tyr Leu Pro Ser Asn Gly Thr Gln Asp Gln 100 105 110 cgt ctt gcc ttt agt atc gat gac ggt gta acc tgg acc aag ttc cag 432 Arg Leu Ala Phe Ser Ile Asp Asp Gly Val Thr Trp Thr Lys Phe Gln 115 120 125 ggc aat ccc atc atc tca gcc gcg cag gag aca cca cat gac gag acg 480 Gly Asn Pro Ile Ile Ser Ala Ala Gln Glu Thr Pro His Asp Glu Thr 130 135 140 gcc ggg ctg gaa agc cgc gat ccc aag gtc ttc ttc cat gaa ccg tcc 528 Ala Gly Leu Glu Ser Arg Asp Pro Lys Val Phe Phe His Glu Pro Ser 145 150 155 160 ggg aaa tgg gtc atg gtg ctc gcg cat gga gga cag gac aag ttg acc 576 Gly Lys Trp Val Met Val Leu Ala His Gly Gly Gln Asp Lys Leu Thr 165 170 175 ttc tgg acg tca acg gac gcc ata tcc tgg acg tgg aag agc gat ctt 624 Phe Trp Thr Ser Thr Asp Ala Ile Ser Trp Thr Trp Lys Ser Asp Leu 180 185 190 tcg gcg gac cag gtc gat ggt ctg cct tct gac atc acc ggg tgg gag 672 Ser Ala Asp Gln Val Asp Gly Leu Pro Ser Asp Ile Thr Gly Trp Glu 195 200 205 gtg ccg gat atg ttc gag ctc ccc atc gag ggc acc gac gag acc acc 720 Val Pro Asp Met Phe Glu Leu Pro Ile Glu Gly Thr Asp Glu Thr Thr 210 215 220 tgg gtg ctg att atg act ccc gca gag ggg tcc cca gcc ggt ggg aat 768 Trp Val Leu Ile Met Thr Pro Ala Glu Gly Ser Pro Ala Gly Gly Asn 225 230 235 240 ggc gtc ctc gcc ttg acg ggc tcc ttc gat ggg tct gtc ttc acg cct 816 Gly Val Leu Ala Leu Thr Gly Ser Phe Asp Gly Ser Val Phe Thr Pro 245 250 255 gac cct gtc gac cct gcg tct tct ggc ctt tgg ctc gac tac ggt cgt 864 Asp Pro Val Asp Pro Ala Ser Ser Gly Leu Trp Leu Asp Tyr Gly Arg 260 265 270 gac ttc gac ggc gcc atg agc tgg gag aat gtg cct gcc gca gat gga 912 Asp Phe Asp Gly Ala Met Ser Trp Glu Asn Val Pro Ala Ala Asp Gly 275 280 285 cgg cgg atc ctg gcc tcc gtc atg aat agc tac ggg gca agc ccg ccg 960 Arg Arg Ile Leu Ala Ser Val Met Asn Ser Tyr Gly Ala Ser Pro Pro 290 295 300 acc aac acc tgg aaa ggg atg ctg tcg ttc ccg cgc acg atc gca ttg 1008 Thr Asn Thr Trp Lys Gly Met Leu Ser Phe Pro Arg Thr Ile Ala Leu 305 310 315 320 agg gaa aca gac acc aag aag aga tac ttc ctt caa cag cct gtc acc 1056 Arg Glu Thr Asp Thr Lys Lys Arg Tyr Phe Leu Gln Gln Pro Val Thr 325 330 335 gaa ctt agc tcc gtc agc tcc tct cct ctc gcg aca atc aag aat cag 1104 Glu Leu Ser Ser Val Ser Ser Ser Pro Leu Ala Thr Ile Lys Asn Gln 340 345 350 acc gtc acg ccc ggc caa acc ttg ctg tcg tcg gtt cac ggc acc gcg 1152 Thr Val Thr Pro Gly Gln Thr Leu Leu Ser Ser Val His Gly Thr Ala 355 360 365 ctg gat atc aaa ctg gcc ttc tcc atg aac gaa ggc gcc acc ctc gcc 1200 Leu Asp Ile Lys Leu Ala Phe Ser Met Asn Glu Gly Ala Thr Leu Ala 370 375 380 ctg gca gtg cgc aaa gca gcc tcg gag cag acg gtc atc tcg tac agc 1248 Leu Ala Val Arg Lys Ala Ala Ser Glu Gln Thr Val Ile Ser Tyr Ser 385 390 395 400 cag tcg aac ggg acc ttg tcc gtg gac cgc acg gcg agt ggg aat acc 1296 Gln Ser Asn Gly Thr Leu Ser Val Asp Arg Thr Ala Ser Gly Asn Thr 405 410 415 tcg tac gac cct gcg gcc ggc gga gtc cac agc acg tcg ctg cag ccc 1344 Ser Tyr Asp Pro Ala Ala Gly Gly Val His Ser Thr Ser Leu Gln Pro 420 425 430 gac gac tct gga gtg gtt cac atc cgg gtt ctg gtt gac act tgc tcg 1392 Asp Asp Ser Gly Val Val His Ile Arg Val Leu Val Asp Thr Cys Ser 435 440 445 gtg gag gtc ttt gga ggg gac ggc gag gtg gtg att tca gac ctg gtc 1440 Val Glu Val Phe Gly Gly Asp Gly Glu Val Val Ile Ser Asp Leu Val 450 455 460 ttc ccc agc gag acc tct gat ggg ctg tcc ctg cag gtg tct ggg ggg 1488 Phe Pro Ser Glu Thr Ser Asp Gly Leu Ser Leu Gln Val Ser Gly Gly 465 470 475 480 gtg gcc aat cta cac ttg gtc gag gtg cgc gga att tcg ctc cag gag 1536 Val Ala Asn Leu His Leu Val Glu Val Arg Gly Ile Ser Leu Gln Glu 485 490 495 taa 1539 <210> 5 <211> 512 <212> PRT <213> *Aspergillus deflectus* <400> 5 Met Leu Leu Ser Leu Ser Leu Ile Ala Leu Ala Thr Gln Ala Val Ala -15 -10 -5 -1 Asp Asp Phe Arg Pro Val Phe His Phe Val Pro Glu Lys Asn Trp Met 1 5 10 15 Asn Glu Pro Asn Gly Leu Ile Gln Ile Asp Ser Val Trp His Leu Phe 20 25 30 Tyr Gln His Asn Pro Thr Ala Asn Val Trp Gly Asn Leu Asn Trp Gly 35 40 45 His Ala Thr Ser Ser Asp Leu Val His Trp Thr His Glu Pro Leu Ala 50 55 60 Ile Thr Ser Glu Asn Gly Val Glu Ala Phe Thr Gly Thr Ala Phe Tyr 65 70 75 80 Asp Ala Glu Asn Thr Ser Gly Leu Gly Ser Ala Ala Ser Pro Pro Tyr 85 90 95 Leu Ala Trp Tyr Thr Gly Tyr Leu Pro Ser Asn Gly Thr Gln Asp Gln 100 105 110 Arg Leu Ala Phe Ser Ile Asp Asp Gly Val Thr Trp Thr Lys Phe Gln 115 120 125 Gly Asn Pro Ile Ile Ser Ala Ala Gln Glu Thr Pro His Asp Glu Thr 130 135 140 Ala Gly Leu Glu Ser Arg Asp Pro Lys Val Phe Phe His Glu Pro Ser 145 150 155 160 Gly Lys Trp Val Met Val Leu Ala His Gly Gly Gln Asp Lys Leu Thr 165 170 175 Phe Trp Thr Ser Thr Asp Ala Ile Ser Trp Thr Trp Lys Ser Asp Leu 180 185 190 Ser Ala Asp Gln Val Asp Gly Leu Pro Ser Asp Ile Thr Gly Trp Glu 195 200 205 Val Pro Asp Met Phe Glu Leu Pro Ile Glu Gly Thr Asp Glu Thr Thr 210 215 220 Trp Val Leu Ile Met Thr Pro Ala Glu Gly Ser Pro Ala Gly Gly Asn 225 230 235 240 Gly Val Leu Ala Leu Thr Gly Ser Phe Asp Gly Ser Val Phe Thr Pro 245 250 255 Asp Pro Val Asp Pro Ala Ser Ser Gly Leu Trp Leu Asp Tyr Gly Arg 260 265 270 Asp Phe Asp Gly Ala Met Ser Trp Glu Asn Val Pro Ala Ala Asp Gly 275 280 285 Arg Arg Ile Leu Ala Ser Val Met Asn Ser Tyr Gly Ala Ser Pro Pro 290 295 300 Thr Asn Thr Trp Lys Gly Met Leu Ser Phe Pro Arg Thr Ile Ala Leu 305 310 315 320 Arg Glu Thr Asp Thr Lys Lys Arg Tyr Phe Leu Gln Gln Pro Val Thr 325 330 335 Glu Leu Ser Ser Val Ser Ser Ser Pro Leu Ala Thr Ile Lys Asn Gln 340 345 350 Thr Val Thr Pro Gly Gln Thr Leu Leu Ser Ser Val His Gly Thr Ala 355 360 365 Leu Asp Ile Lys Leu Ala Phe Ser Met Asn Glu Gly Ala Thr Leu Ala 370 375 380 Leu Ala Val Arg Lys Ala Ala Ser Glu Gln Thr Val Ile Ser Tyr Ser 385 390 395 400 Gln Ser Asn Gly Thr Leu Ser Val Asp Arg Thr Ala Ser Gly Asn Thr 405 410 415 Ser Tyr Asp Pro Ala Ala Gly Gly Val His Ser Thr Ser Leu Gln Pro 420 425 430 Asp Asp Ser Gly Val Val His Ile Arg Val Leu Val Asp Thr Cys Ser 435 440 445 Val Glu Val Phe Gly Gly Asp Gly Glu Val Val Ile Ser Asp Leu Val 450 455 460 Phe Pro Ser Glu Thr Ser Asp Gly Leu Ser Leu Gln Val Ser Gly Gly 465 470 475 480 Val Ala Asn Leu His Leu Val Glu Val Arg Gly Ile Ser Leu Gln Glu 485 490 495 <210> 6 <211> 496 <212> PRT <213> Aspergillus effusus <400> 6 Asp Asp Phe Arg Pro Val Phe His Phe Val Pro Glu Lys Asn Trp Met 1 5 10 15 Asn Glu Pro Asn Gly Leu Ile Gln Ile Asp Ser Val Trp His Leu Phe 20 25 30 Tyr Gln His Asn Pro Thr Ala Asn Val Trp Gly Asn Leu Asn Trp Gly 35 40 45 His Ala Thr Ser Ser Asp Leu Val His Trp Thr His Glu Pro Leu Ala 50 55 60 Ile Thr Ser Glu Asn Gly Val Glu Ala Phe Thr Gly Thr Ala Phe Tyr 65 70 75 80 Asp Ala Glu Asn Thr Ser Gly Leu Gly Ser Ala Ala Ser Pro Pro Tyr 85 90 95 Leu Ala Trp Tyr Thr Gly Tyr Leu Pro Ser Asn Gly Thr Gln Asp Gln 100 105 110 Arg Leu Ala Phe Ser Ile Asp Asp Gly Val Thr Trp Thr Lys Phe Gln 115 120 125 Gly Asn Pro Ile Ile Ser Ala Ala Gln Glu Thr Pro His Asp Glu Thr 130 135 140 Ala Gly Leu Glu Ser Arg Asp Pro Lys Val Phe Phe His Glu Pro Ser 145 150 155 160 Gly Lys Trp Val Met Val Leu Ala His Gly Gly Gln Asp Lys Leu Thr 165 170 175 Phe Trp Thr Ser Thr Asp Ala Ile Ser Trp Thr Trp Lys Ser Asp Leu 180 185 190 Ser Ala Asp Gln Val Asp Gly Leu Pro Ser Asp Ile Thr Gly Trp Glu 195 200 205 Val Pro Asp Met Phe Glu Leu Pro Ile Glu Gly Thr Asp Glu Thr Thr 210 215 220 Trp Val Leu Ile Met Thr Pro Ala Glu Gly Ser Pro Ala Gly Gly Asn 225 230 235 240 Gly Val Leu Ala Leu Thr Gly Ser Phe Asp Gly Ser Val Phe Thr Pro 245 250 255 Asp Pro Val Asp Pro Ala Ser Ser Gly Leu Trp Leu Asp Tyr Gly Arg 260 265 270 Asp Phe Asp Gly Ala Met Ser Trp Glu Asn Val Pro Ala Ala Asp Gly 275 280 285 Arg Arg Ile Leu Ala Ser Val Met Asn Ser Tyr Gly Ala Ser Pro Pro 290 295 300 Thr Asn Thr Trp Lys Gly Met Leu Ser Phe Pro Arg Thr Ile Ala Leu 305 310 315 320 Arg Glu Thr Asp Thr Lys Lys Arg Tyr Phe Leu Gln Gln Pro Val Thr 325 330 335 Glu Leu Ser Ser Val Ser Ser Ser Pro Leu Ala Thr Ile Lys Asn Gln 340 345 350 Thr Val Thr Pro Gly Gln Thr Leu Leu Ser Ser Val His Gly Thr Ala 355 360 365 Leu Asp Ile Lys Leu Ala Phe Ser Met Asn Glu Gly Ala Thr Leu Ala 370 375 380 Leu Ala Val Arg Lys Ala Ala Ser Glu Gln Thr Val Ile Ser Tyr Ser 385 390 395 400 Gln Ser Asn Gly Thr Leu Ser Val Asp Arg Thr Ala Ser Gly Asn Thr 405 410 415 Ser Tyr Asp Pro Ala Ala Gly Gly Val His Ser Thr Ser Leu Gln Pro 420 425 430 Asp Asp Ser Gly Val Val His Ile Arg Val Leu Val Asp Thr Cys Ser 435 440 445 Val Glu Val Phe Gly Gly Asp Gly Glu Val Val Ile Ser Asp Leu Val 450 455 460 Phe Pro Ser Glu Thr Ser Asp Gly Leu Ser Leu Gln Val Ser Gly Gly 465 470 475 480 Val Ala Asn Leu His Leu Val Glu Val Arg Gly Ile Ser Leu Gln Glu 485 490 495 <210> 7 <211> 1551 <212> DNA <213> Aspergillus niger <220> <221> CDS <222> (1)..(1548) <220> <221> signal peptide <222> (1)..(75) <220> <221> mature peptide <222> (76)..(1548) <400> 7 atg ttg aat ccg aag gtt gcc tac atg gtc tgg atg acg tgc ctg ggt 48 Met Leu Asn Pro Lys Val Ala Tyr Met Val Trp Met Thr Cys Leu Gly -25 -20 -15 -10 tta acg ttg ccc agc cag gca cag tct aat gat tac cgt cct tca tac 96 Leu Thr Leu Pro Ser Gln Ala Gln Ser Asn Asp Tyr Arg Pro Ser Tyr -5 -1 1 5 cac ttc aca ccg gac cag tac tgg atg aac gag cca aac ggc ctg att 144 His Phe Thr Pro Asp Gln Tyr Trp Met Asn Glu Pro Asn Gly Leu Ile 10 15 20 aaa atc gga tcc acc tgg cac ctg ttc ttt caa cac aat ccg acg gcc 192 Lys Ile Gly Ser Thr Trp His Leu Phe Phe Gln His Asn Pro Thr Ala 25 30 35 aat gta tgg ggc aac ata tgc tgg ggg cac gct acg agc acc gat ctg 240 Asn Val Trp Gly Asn Ile Cys Trp Gly His Ala Thr Ser Thr Asp Leu 40 45 50 55 atg cac tgg gca cac aaa ccc act gcc att gcg gat gag aac gga gtc 288 Met His Trp Ala His Lys Pro Thr Ala Ile Ala Asp Glu Asn Gly Val 60 65 70 gaa gcg ttt acc ggt aca gcc tat tat gat cca aac aat acc tct ggc 336 Glu Ala Phe Thr Gly Thr Ala Tyr Tyr Asp Pro Asn Asn Thr Ser Gly 75 80 85 ctt ggg gat tcg gca aac cca ccc tat ctg gcc tgg ttc aca ggt tat 384 Leu Gly Asp Ser Ala Asn Pro Pro Tyr Leu Ala Trp Phe Thr Gly Tyr 90 95 100 acc act tca agc caa aca cag gac cag cgc ctg gct ttc agt gtg gat 432 Thr Thr Ser Ser Gln Thr Gln Asp Gln Arg Leu Ala Phe Ser Val Asp 105 110 115 aac ggg gcg acg tgg acc aaa ttt caa ggc aat ccc atc ata tca act 480 Asn Gly Ala Thr Trp Thr Lys Phe Gln Gly Asn Pro Ile Ile Ser Thr 120 125 130 135 agc cag gaa gca cca cat gat ata acg ggc ggc ctc gag agt agg gat 528 Ser Gln Glu Ala Pro His Asp Ile Thr Gly Gly Leu Glu Ser Arg Asp 140 145 150 cca aag gta ttc ttc cat cgc caa tcg ggg aac tgg atc atg gtt ctc 576 Pro Lys Val Phe Phe His Arg Gln Ser Gly Asn Trp Ile Met Val Leu 155 160 165 gcc cat ggc ggg cag gac aag ctg tct ttc tgg acg tct gca gac acc 624 Ala His Gly Gly Gln Asp Lys Leu Ser Phe Trp Thr Ser Ala Asp Thr 170 175 180 ata aac tgg aca tgg cag agt gac ctg aag tcc acc tcg atc aat ggc 672 Ile Asn Trp Thr Trp Gln Ser Asp Leu Lys Ser Thr Ser Ile Asn Gly 185 190 195 cta tcg tcc gat att aca ggg tgg gaa gtc ccc gac atg ttt gaa ctc 720 Leu Ser Ser Asp Ile Thr Gly Trp Glu Val Pro Asp Met Phe Glu Leu 200 205 210 215 ccg gtt gaa ggc act gag gag acc acg tgg gtg gtg atg atg acg ccg 768 Pro Val Glu Gly Thr Glu Glu Thr Thr Trp Val Val Met Met Thr Pro 220 225 230 gct gaa gga tcc cct gcc ggt ggt aac ggg gtc tta gct atc acc ggt 816 Ala Glu Gly Ser Pro Ala Gly Gly Asn Gly Val Leu Ala Ile Thr Gly 235 240 245 tct ttt gac ggg aaa agt ttt acg gca gat ccc gtc gat gct tcg acc 864 Ser Phe Asp Gly Lys Ser Phe Thr Ala Asp Pro Val Asp Ala Ser Thr 250 255 260 atg tgg ctg gac aat ggg cgt gat ttc gat ggc gct ctg agc tgg gtg 912 Met Trp Leu Asp Asn Gly Arg Asp Phe Asp Gly Ala Leu Ser Trp Val 265 270 275 aac gtg cct gcg tcc gat gga cgg cgg att atc gcc gcc gtc atg aat 960 Asn Val Pro Ala Ser Asp Gly Arg Arg Ile Ile Ala Ala Val Met Asn 280 285 290 295 agc tac ggt tcc aac ccg cct aca acc acc tgg aaa ggg atg ctc tcc 1008 Ser Tyr Gly Ser Asn Pro Pro Thr Thr Thr Trp Lys Gly Met Leu Ser 300 305 310 ttt ccc cgg acg ctg tcg ctc aag aaa gtt ggc acg cag cag cac ttt 1056 Phe Pro Arg Thr Leu Ser Leu Lys Lys Val Gly Thr Gln Gln His Phe 315 320 325 gtt caa cag ccg atc aca gag ttg gat aca atc agt acc agt ctg caa 1104 Val Gln Gln Pro Ile Thr Glu Leu Asp Thr Ile Ser Thr Ser Leu Gln 330 335 340 ata cta gca aac cag acc att acc cct ggc caa aca ttg ctg tca tcg 1152 Ile Leu Ala Asn Gln Thr Ile Thr Pro Gly Gln Thr Leu Leu Ser Ser 345 350 355 att cgg gga act gct ctc gat gtt cga gtt gct ttt tac cct gat gct 1200 Ile Arg Gly Thr Ala Leu Asp Val Arg Val Ala Phe Tyr Pro Asp Ala 360 365 370 375 ggc tcg gtt ctg tcc ctc gcc gtc cga aag ggt gct tcg gag caa aca 1248 Gly Ser Val Leu Ser Leu Ala Val Arg Lys Gly Ala Ser Glu Gln Thr 380 385 390 gtc att aag tac acc cag tca gat gcc aca ttg tcg gtt gat cga aca 1296 Val Ile Lys Tyr Thr Gln Ser Asp Ala Thr Leu Ser Val Asp Arg Thr 395 400 405 gag agt gga gat atc tcg tat gac ccg gcc gca ggt ggc gtc cat acc 1344 Glu Ser Gly Asp Ile Ser Tyr Asp Pro Ala Ala Gly Gly Val His Thr 410 415 420 gcc aag ttg gaa gag gac ggc acc gga ctg gtt tcc atc cgg gtg ttg 1392 Ala Lys Leu Glu Glu Asp Gly Thr Gly Leu Val Ser Ile Arg Val Leu 425 430 435 gtg gat acg tgt tct gta gag gtt ttt ggc gga caa gga gaa gcc gtc 1440 Val Asp Thr Cys Ser Val Glu Val Phe Gly Gly Gln Gly Glu Ala Val 440 445 450 455 att tcc gac ctc atc ttc ccg agt gac agt tct gac ggc ctg gcc ttg 1488 Ile Ser Asp Leu Ile Phe Pro Ser Asp Ser Ser Asp Gly Leu Ala Leu 460 465 470 gag gta act ggc gga aat gca gtg ctg cag tcg gtg gac gtg cgg agt 1536 Glu Val Thr Gly Gly Asn Ala Val Leu Gln Ser Val Asp Val Arg Ser 475 480 485 gtt tca ctt gaa tga 1551 Val Ser Leu Glu 490 <210> 8 <211> 516 <212> PRT <213> Aspergillus niger <400> 8 Met Leu Asn Pro Lys Val Ala Tyr Met Val Trp Met Thr Cys Leu Gly -25 -20 -15 -10 Leu Thr Leu Pro Ser Gln Ala Gln Ser Asn Asp Tyr Arg Pro Ser Tyr -5 -1 1 5 His Phe Thr Pro Asp Gln Tyr Trp Met Asn Glu Pro Asn Gly Leu Ile 10 15 20 Lys Ile Gly Ser Thr Trp His Leu Phe Phe Gln His Asn Pro Thr Ala 25 30 35 Asn Val Trp Gly Asn Ile Cys Trp Gly His Ala Thr Ser Thr Asp Leu 40 45 50 55 Met His Trp Ala His Lys Pro Thr Ala Ile Ala Asp Glu Asn Gly Val 60 65 70 Glu Ala Phe Thr Gly Thr Ala Tyr Tyr Asp Pro Asn Asn Thr Ser Gly 75 80 85 Leu Gly Asp Ser Ala Asn Pro Pro Tyr Leu Ala Trp Phe Thr Gly Tyr 90 95 100 Thr Thr Ser Ser Gln Thr Gln Asp Gln Arg Leu Ala Phe Ser Val Asp 105 110 115 Asn Gly Ala Thr Trp Thr Lys Phe Gln Gly Asn Pro Ile Ile Ser Thr 120 125 130 135 Ser Gln Glu Ala Pro His Asp Ile Thr Gly Gly Leu Glu Ser Arg Asp 140 145 150 Pro Lys Val Phe Phe His Arg Gln Ser Gly Asn Trp Ile Met Val Leu 155 160 165 Ala His Gly Gly Gln Asp Lys Leu Ser Phe Trp Thr Ser Ala Asp Thr 170 175 180 Ile Asn Trp Thr Trp Gln Ser Asp Leu Lys Ser Thr Ser Ile Asn Gly 185 190 195 Leu Ser Ser Asp Ile Thr Gly Trp Glu Val Pro Asp Met Phe Glu Leu 200 205 210 215 Pro Val Glu Gly Thr Glu Glu Thr Thr Trp Val Val Met Met Thr Pro 220 225 230 Ala Glu Gly Ser Pro Ala Gly Gly Asn Gly Val Leu Ala Ile Thr Gly 235 240 245 Ser Phe Asp Gly Lys Ser Phe Thr Ala Asp Pro Val Asp Ala Ser Thr 250 255 260 Met Trp Leu Asp Asn Gly Arg Asp Phe Asp Gly Ala Leu Ser Trp Val 265 270 275 Asn Val Pro Ala Ser Asp Gly Arg Arg Ile Ile Ala Ala Val Met Asn 280 285 290 295 Ser Tyr Gly Ser Asn Pro Pro Thr Thr Thr Trp Lys Gly Met Leu Ser 300 305 310 Phe Pro Arg Thr Leu Ser Leu Lys Lys Val Gly Thr Gln Gln His Phe 315 320 325 Val Gln Gln Pro Ile Thr Glu Leu Asp Thr Ile Ser Thr Ser Leu Gln 330 335 340 Ile Leu Ala Asn Gln Thr Ile Thr Pro Gly Gln Thr Leu Leu Ser Ser 345 350 355 Ile Arg Gly Thr Ala Leu Asp Val Arg Val Ala Phe Tyr Pro Asp Ala 360 365 370 375 Gly Ser Val Leu Ser Leu Ala Val Arg Lys Gly Ala Ser Glu Gln Thr 380 385 390 Val Ile Lys Tyr Thr Gln Ser Asp Ala Thr Leu Ser Val Asp Arg Thr 395 400 405 Glu Ser Gly Asp Ile Ser Tyr Asp Pro Ala Ala Gly Gly Val His Thr 410 415 420 Ala Lys Leu Glu Glu Asp Gly Thr Gly Leu Val Ser Ile Arg Val Leu 425 430 435 Val Asp Thr Cys Ser Val Glu Val Phe Gly Gly Gln Gly Glu Ala Val 440 445 450 455 Ile Ser Asp Leu Ile Phe Pro Ser Asp Ser Ser Asp Gly Leu Ala Leu 460 465 470 Glu Val Thr Gly Gly Asn Ala Val Leu Gln Ser Val Asp Val Arg Ser 475 480 485 Val Ser Leu Glu 490 <210> 9 <211> 491 <212> PRT <213> Aspergillus niger <400> 9 Asn Asp Tyr Arg Pro Ser Tyr His Phe Thr Pro Asp Gln Tyr Trp Met 1 5 10 15 Asn Glu Pro Asn Gly Leu Ile Lys Ile Gly Ser Thr Trp His Leu Phe 20 25 30 Phe Gln His Asn Pro Thr Ala Asn Val Trp Gly Asn Ile Cys Trp Gly 35 40 45 His Ala Thr Ser Thr Asp Leu Met His Trp Ala His Lys Pro Thr Ala 50 55 60 Ile Ala Asp Glu Asn Gly Val Glu Ala Phe Thr Gly Thr Ala Tyr Tyr 65 70 75 80 Asp Pro Asn Asn Thr Ser Gly Leu Gly Asp Ser Ala Asn Pro Pro Tyr 85 90 95 Leu Ala Trp Phe Thr Gly Tyr Thr Thr Ser Ser Gln Thr Gln Asp Gln 100 105 110 Arg Leu Ala Phe Ser Val Asp Asn Gly Ala Thr Trp Thr Lys Phe Gln 115 120 125 Gly Asn Pro Ile Ile Ser Thr Ser Gln Glu Ala Pro His Asp Ile Thr 130 135 140 Gly Gly Leu Glu Ser Arg Asp Pro Lys Val Phe Phe His Arg Gln Ser 145 150 155 160 Gly Asn Trp Ile Met Val Leu Ala His Gly Gly Gln Asp Lys Leu Ser 165 170 175 Phe Trp Thr Ser Ala Asp Thr Ile Asn Trp Thr Trp Gln Ser Asp Leu 180 185 190 Lys Ser Thr Ser Ile Asn Gly Leu Ser Ser Asp Ile Thr Gly Trp Glu 195 200 205 Val Pro Asp Met Phe Glu Leu Pro Val Glu Gly Thr Glu Glu Thr Thr 210 215 220 Trp Val Val Met Met Thr Pro Ala Glu Gly Ser Pro Ala Gly Gly Asn 225 230 235 240 Gly Val Leu Ala Ile Thr Gly Ser Phe Asp Gly Lys Ser Phe Thr Ala 245 250 255 Asp Pro Val Asp Ala Ser Thr Met Trp Leu Asp Asn Gly Arg Asp Phe 260 265 270 Asp Gly Ala Leu Ser Trp Val Asn Val Pro Ala Ser Asp Gly Arg Arg 275 280 285 Ile Ile Ala Ala Val Met Asn Ser Tyr Gly Ser Asn Pro Pro Thr Thr 290 295 300 Thr Trp Lys Gly Met Leu Ser Phe Pro Arg Thr Leu Ser Leu Lys Lys 305 310 315 320 Val Gly Thr Gln Gln His Phe Val Gln Gln Pro Ile Thr Glu Leu Asp 325 330 335 Thr Ile Ser Thr Ser Leu Gln Ile Leu Ala Asn Gln Thr Ile Thr Pro 340 345 350 Gly Gln Thr Leu Leu Ser Ser Ile Arg Gly Thr Ala Leu Asp Val Arg 355 360 365 Val Ala Phe Tyr Pro Asp Ala Gly Ser Val Leu Ser Leu Ala Val Arg 370 375 380 Lys Gly Ala Ser Glu Gln Thr Val Ile Lys Tyr Thr Gln Ser Asp Ala 385 390 395 400 Thr Leu Ser Val Asp Arg Thr Glu Ser Gly Asp Ile Ser Tyr Asp Pro 405 410 415 Ala Ala Gly Gly Val His Thr Ala Lys Leu Glu Glu Asp Gly Thr Gly 420 425 430 Leu Val Ser Ile Arg Val Leu Val Asp Thr Cys Ser Val Glu Val Phe 435 440 445 Gly Gly Gln Gly Glu Ala Val Ile Ser Asp Leu Ile Phe Pro Ser Asp 450 455 460 Ser Ser Asp Gly Leu Ala Leu Glu Val Thr Gly Gly Asn Ala Val Leu 465 470 475 480 Gln Ser Val Asp Val Arg Ser Val Ser Leu Glu 485 490 <210> 10 <211> 4352 <212> DNA <213> Ochre-green Penicillium <220> <221> CDS <222> (1)..(399) <220> <221> signal peptide <222> (1)..(63) <220> <221> Mature peptides <222> (64)..(399) <220> <221> CDS <222> (453)..(4349) <400> 10 atg gct tct cgg ctt gtg aca gcc ttg act ctg ggt ctt cat tta ttc 48 Met Ala Ser Arg Leu Val Thr Ala Leu Thr Leu Gly Leu His Leu Phe -20 -15 -10 caa gca gca ttt gct caa acc tac gat gag ctc tac cgc cct cag tat 96 Gln Ala Ala Phe Ala Gln Thr Tyr Asp Glu Leu Tyr Arg Pro Gln Tyr -5 -1 1 5 10 cac ttt acc ccc cct gtg aac tgg atg aat gat ccc aac gga ctt tta 144 His Phe Thr Pro Pro Val Asn Trp Met Asn Asp Pro Asn Gly Leu Leu 15 20 25 tat cac aat ggc ctc tat cat ctg tac tat caa tac aac cca gga ggg 192 Tyr His Asn Gly Leu Tyr His Leu Tyr Tyr Gln Tyr Asn Pro Gly Gly 30 35 40 aac acc tgg gga gct atg tct tgg ggc cat gcc act agc act gat ctg 240 Asn Thr Trp Gly Ala Met Ser Trp Gly His Ala Thr Ser Thr Asp Leu 45 50 55 act cac tgg aat cac cag cct att gcc ctc ctc gct cgt ggc tac cca 288 Thr His Trp Asn His Gln Pro Ile Ala Leu Leu Ala Arg Gly Tyr Pro 60 65 70 75 ggt gat atc act gag atg ttc ttc tct ggc tct gct gtc gcc gat act 336 Gly Asp Ile Thr Glu Met Phe Phe Ser Gly Ser Ala Val Ala Asp Thr 80 85 90 cag aac acg agt gga ttt ggt act agt gga aag gtg cca ttt gtt gcg 384 Gln Asn Thr Ser Gly Phe Gly Thr Ser Gly Lys Val Pro Phe Val Ala 95 100 105 atg tat act tct tat gtgagtattt gaaaccttta aagtcggcta ctctaaccct 439 Met Tyr Thr Ser Tyr 110 ttttcggcct tag tac cct gca tcc cag aac ctc ccc agt ggg aag tta 488 Tyr Pro Ala Ser Gln Asn Leu Pro Ser Gly Lys Leu 115 120 gtc aac ggg ggc cag caa gcg cag tca att gcc tac agt ttg gac gaa 536 Val Asn Gly Gly Gln Gln Ala Gln Ser Ile Ala Tyr Ser Leu Asp Glu 125 130 135 140 ggc atg aca tgg aca acc tac gac gcc gcc aat cct gtc att ctc aat 584 Gly Met Thr Trp Thr Thr Tyr Asp Ala Ala Asn Pro Val Ile Leu Asn 145 150 155 ccc cct acg cca tat aca gac caa tgg cgg gac ttc cga gat cca ttc 632 Pro Pro Thr Pro Tyr Thr Asp Gln Trp Arg Asp Phe Arg Asp Pro Phe 160 165 170 ctg ttc tgg cac gaa gcc agc cag aag tgg atc tct gtg gtt tca ctg 680 Leu Phe Trp His Glu Ala Ser Gln Lys Trp Ile Ser Val Val Ser Leu 175 180 185 gcc caa ctc cac aaa tta ctt att tac acc tcc ccg aat ctt aag gat 728 Ala Gln Leu His Lys Leu Leu Ile Tyr Thr Ser Pro Asn Leu Lys Asp 190 195 200 tgg act tat tca agc gaa ttt gga cct tgg aat gca gtc ggg ggt gtt 776 Trp Thr Tyr Ser Ser Glu Phe Gly Pro Trp Asn Ala Val Gly Gly Val 205 210 215 220 tgg gag tgt cct agc ctc ttt cca ctt gct gtt gac gga gat gac gcc 824 Trp Glu Cys Pro Ser Leu Phe Pro Leu Ala Val Asp Gly Asp Asp Ala 225 230 235 aat aca aaa tgg att atg caa atc ggg ctg aac cct ggc ggg ccc cct 872 Asn Thr Lys Trp Ile Met Gln Ile Gly Leu Asn Pro Gly Gly Pro Pro 240 245 250 gga gtg act ggc tca gga acg cag tat att gtg gga acc ttt gat gga 920 Gly Val Thr Gly Ser Gly Thr Gln Tyr Ile Val Gly Thr Phe Asp Gly 255 260 265 aca aaa ttt gtt gcg gat ccc aac tct cca ctt tcg gca cat aca tcc 968 Thr Lys Phe Val Ala Asp Pro Asn Ser Pro Leu Ser Ala His Thr Ser 270 275 280 acc agt acc act acc tca gca tcc gtc act acc tca gca tcc atc act 1016 Thr Ser Thr Thr Thr Ser Ala Ser Val Thr Thr Ser Ala Ser Ile Thr 285 290 295 300 acc tca gca tcc atc act ccc aca acc aca aaa gca act gcg act gct 1064 Thr Ser Ala Ser Ile Thr Pro Thr Thr Thr Lys Ala Thr Ala Thr Ala 305 310 315 act gca act gga gac atc atc ttt cag gat ttt gaa ggc gcc ggt gac 1112 Thr Ala Thr Gly Asp Ile Ile Phe Gln Asp Phe Glu Gly Ala Gly Asp 320 325 330 ttc gcc tct cgt ggc tgg gtt gcc acc ggt ggt ttg ctt ggg gct gct 1160 Phe Ala Ser Arg Gly Trp Val Ala Thr Gly Gly Leu Leu Gly Ala Ala 335 340 345 cca gct aaa ggg act ctt gcg gga cag caa acc gtc act gga tat gct 1208 Pro Ala Lys Gly Thr Leu Ala Gly Gln Gln Thr Val Thr Gly Tyr Ala 350 355 360 gga agc cag ctg gtc aat acc ttc cta agc gga gac tct aca act ggc 1256 Gly Ser Gln Leu Val Asn Thr Phe Leu Ser Gly Asp Ser Thr Thr Gly 365 370 375 380 acg ctc aca tcc ccg gct ttc act ata tca ctt cca tat gtc aat ttc 1304 Thr Leu Thr Ser Pro Ala Phe Thr Ile Ser Leu Pro Tyr Val Asn Phe 385 390 395 cta att ggc ggc ggg aat gct cca ggg acg gaa tgc atc aac ctg aag 1352 Leu Ile Gly Gly Gly Asn Ala Pro Gly Thr Glu Cys Ile Asn Leu Lys 400 405 410 gtg caa ggc cag gct gtc cgg acg gcg aca ggc gcg aac gtg gaa caa 1400 Val Gln Gly Gln Ala Val Arg Thr Ala Thr Gly Ala Asn Val Glu Gln 415 420 425 ctc gca cca aca acc tgg aat gtc acg gac ctg atg ggt caa acc gct 1448 Leu Ala Pro Thr Thr Trp Asn Val Thr Asp Leu Met Gly Gln Thr Ala 430 435 440 gtg ctt gag atc gtg gat cag caa aca ggt ggc tgg gga cac ata ttg 1496 Val Leu Glu Ile Val Asp Gln Gln Thr Gly Gly Trp Gly His Ile Leu 445 450 455 460 att gat cag att acc ttc acc agc agt acc agc acc aac aac ctc cac 1544 Ile Asp Gln Ile Thr Phe Thr Ser Ser Thr Ser Thr Asn Asn Leu His 465 470 475 aaa cgc agc gtt tct agc gtt act tgg gat ttc aat ggt act agc act 1592 Lys Arg Ser Val Ser Ser Val Thr Trp Asp Phe Asn Gly Thr Ser Thr 480 485 490 ttt gca gac tat ggc tgg act ccg tct gga gat ttt gtt gga aaa ggg 1640 Phe Ala Asp Tyr Gly Trp Thr Pro Ser Gly Asp Phe Val Gly Lys Gly 495 500 505 cca gcc cag ggc aca ctt gcg gga caa cag gtt gtg act ggg aat atg 1688 Pro Ala Gln Gly Thr Leu Ala Gly Gln Gln Val Val Thr Gly Asn Met 510 515 520 gga aac ttc gtc aac aca ttt ttg agc gga gat ggt acc act gga aca 1736 Gly Asn Phe Val Asn Thr Phe Leu Ser Gly Asp Gly Thr Thr Gly Thr 525 530 535 540 ctc acg tcc ccc gcc ttc acc ata act cag aag aag att aac ttt ctc 1784 Leu Thr Ser Pro Ala Phe Thr Ile Thr Gln Lys Lys Ile Asn Phe Leu 545 550 555 atc ggt gga ggc aat tct ccg ggc atg gaa tgc atc aat ctc aag atc 1832 Ile Gly Gly Gly Asn Ser Pro Gly Met Glu Cys Ile Asn Leu Lys Ile 560 565 570 caa ggc cag gtt gta cga acg gct aca ggc gct aat gca gaa caa ctc 1880 Gln Gly Gln Val Val Arg Thr Ala Thr Gly Ala Asn Ala Glu Gln Leu 575 580 585 gta cca acc acc tgg gat gtc acc gac ctg ata ggc cag tct gcg gtc 1928 Val Pro Thr Thr Trp Asp Val Thr Asp Leu Ile Gly Gln Ser Ala Val 590 595 600 atc gag att gtt gat ctc agt acg acc ggc tgg ggg cac att ctg atc 1976 Ile Glu Ile Val Asp Leu Ser Thr Thr Gly Trp Gly His Ile Leu Ile 605 610 615 620 gat gag att tcc ttc tca aac ata tca att gtg cca tat ggg ccc aac 2024 Asp Glu Ile Ser Phe Ser Asn Ile Ser Ile Val Pro Tyr Gly Pro Asn 625 630 635 tgg ttg gac tat ggg ccg gac ttc tac gct gca aca aca ttc aat gga 2072 Trp Leu Asp Tyr Gly Pro Asp Phe Tyr Ala Ala Thr Thr Phe Asn Gly 640 645 650 tta tca tcc aca aat cga atc ​​gat att gcg tgg atg aac aat tgg caa 2120 Leu Ser Ser Thr Asn Arg Ile Asp Ile Ala Trp Met Asn Asn Trp Gln 655 660 665 tac gct agc gca atc ​​cct act tct cca tgg cgc agc atg ctt tca gtt 2168 Tyr Ala Ser Ala Ile Pro Thr Ser Pro Trp Arg Ser Met Leu Ser Val 670 675 680 gcc cgg aag ctt tca ctc aag acg atc gac gac agg cca aga ctg atc 2216 Ala Arg Lys Leu Ser Leu Lys Thr Ile Asp Asp Arg Pro Arg Leu Ile 685 690 695 700 cag caa cct gag gca aac tgg acg agt cta cag aca aca acg tat tct 2264 Gln Gln Pro Glu Ala Asn Trp Thr Ser Leu Gln Thr Thr Thr Tyr Ser 705 710 715 acc agc ttt gat aca gtt gct gaa ggg aat cag ctc gta caa ctc tcc 2312 Thr Ser Phe Asp Thr Val Ala Glu Gly Asn Gln Leu Val Gln Leu Ser 720 725 730 gga aaa ttg ctt gac atc aca gtg gcc ttt tca gac aga gtt gca gcg 2360 Gly Lys Leu Leu Asp Ile Thr Val Ala Phe Ser Asp Arg Val Ala Ala 735 740 745 tcg tcc tca tcg caa ttc ggc ata att ctc aga gca aca tct gac cta 2408 Ser Ser Ser Ser Gln Phe Gly Ile Ile Leu Arg Ala Thr Ser Asp Leu 750 755 760 gca cag caa aca cgg att ggc tat gat ttt agc aca aag agg ctt ttt 2456 Ala Gln Gln Thr Arg Ile Gly Tyr Asp Phe Ser Thr Lys Arg Leu Phe 765 770 775 780 gtt gat cga acg aaa tct gga aat gtt ggc ttc gac ggg act ttt ccc 2504 Val Asp Arg Thr Lys Ser Gly Asn Val Gly Phe Asp Gly Thr Phe Pro 785 790 795 agc acc tat ttt gct cct ttg gca ccc tcc ggt gat ggc cga gtc aca 2552 Ser Thr Tyr Phe Ala Pro Leu Ala Pro Ser Gly Asp Gly Arg Val Thr 800 805 810 atg cgc att ctt ctt gac agg tcc tcc gtt gaa gtc ttt gga ggc cag 2600 Met Arg Ile Leu Leu Asp Arg Ser Ser Val Glu Val Phe Gly Gly Gln 815 820 825 ggt gag gtc aca ata tcc gct caa atc ​​ttt ccg cag gac aat ggt gtt 2648 Gly Glu Val Thr Ile Ser Ala Gln Ile Phe Pro Gln Asp Asn Gly Val 830 835 840 gat gtc cgg ctt ttc tcc aga ggc ggg agt aca aat gct atc acg att 2696 Asp Val Arg Leu Phe Ser Arg Gly Gly Ser Thr Asn Ala Ile Thr Ile 845 850 855 860 gat gca att gtg ctt gat tca gtt tat gat tca tcc acc cca agt gct 2744 Asp Ala Ile Val Leu Asp Ser Val Tyr Asp Ser Ser Thr Pro Ser Ala 865 870 875 tct ctc agt acg agc tca acc tcc atc ggt act gtc acc acc acc ttc 2792 Ser Leu Ser Thr Ser Ser Thr Ser Ile Gly Thr Val Thr Thr Thr Phe 880 885 890 agt aac acc gtg act acc tct gtc ggt aac acc ctg acc acc agt aca 2840 Ser Asn Thr Val Thr Thr Ser Val Gly Asn Thr Leu Thr Thr Ser Thr 895 900 905 aga acc gtc act agc gct agc cct acg tct aca gcg ccc tac gat ttt 2888 Arg Thr Val Thr Ser Ala Ser Pro Thr Ser Thr Ala Pro Tyr Asp Phe 910 915 920 cgt cct gtt ttt cat ttt gtg cca gaa gag aac tgg atg aat gag cct 2936 Arg Pro Val Phe His Phe Val Pro Glu Glu Asn Trp Met Asn Glu Pro 925 930 935 940 aac gga ctg atc aaa att gac tct acc tgg cac ctt ttc ttt caa cac 2984 Asn Gly Leu Ile Lys Ile Asp Ser Thr Trp His Leu Phe Phe Gln His 945 950 955 aac cca act gga aac ttt tgg ggc aac tta agt tgg ggt cat gct act 3032 Asn Pro Thr Gly Asn Phe Trp Gly Asn Leu Ser Trp Gly His Ala Thr 960 965 970 agc att gac ctt gtg tcc tgg gat tat aaa ccg gtt gcg att tca agc 3080 Ser Ile Asp Leu Val Ser Trp Asp Tyr Lys Pro Val Ala Ile Ser Ser 975 980 985 gca gat ggg att cag gct ttc aca gga acc tca tac ttt gat gcg atg 3128 Ala Asp Gly Ile Gln Ala Phe Thr Gly Thr Ser Tyr Phe Asp Ala Met 990 995 1000 aat ctc tcg ggg ctt ggg aca tca tca aac cag ccg tac ctt gcc 3173 Asn Leu Ser Gly Leu Gly Thr Ser Ser Asn Gln Pro Tyr Leu Ala 1005 1010 1015 ttt tat act ggc tac gca cct tca agt ggc ata cag gat caa agg 3218 Phe Tyr Thr Gly Tyr Ala Pro Ser Ser Gly Ile Gln Asp Gln Arg 1020 1025 1030 ctt gcg tat agc ttg gac cag gga gcg aca tgg atg aaa tac cag 3263 Leu Ala Tyr Ser Leu Asp Gln Gly Ala Thr Trp Met Lys Tyr Gln 1035 1040 1045 ggt aat cca att ata tcc caa agc caa gaa gcg cca cac gat ata 3308 Gly Asn Pro Ile Ile Ser Gln Ser Gln Glu Ala Pro His Asp Ile 1050 1055 1060 acc cag ggt ttg gag ata cgt gat cca aag gtg ttc tac cac agt 3353 Thr Gln Gly Leu Glu Ile Arg Asp Pro Lys Val Phe Tyr His Ser 1065 1070 1075 ccg acg agc aga tgg gtt atg atc ctg gcg cac gga gga caa aac 3398 Pro Thr Ser Arg Trp Val Met Ile Leu Ala His Gly Gly Gln Asn 1080 1085 1090 aaa cta aca ttc tgg acg tct acc gat gca aag aac tgg acc tgg 3443 Lys Leu Thr Phe Trp Thr Ser Thr Asp Ala Lys Asn Trp Thr Trp 1095 1100 1105 aga agt aat ttc gct gca agc aat att ctt gga ttc cct agt ggg 3488 Arg Ser Asn Phe Ala Ala Ser Asn Ile Leu Gly Phe Pro Ser Gly 1110 1115 1120 gtc aac ggt tgg gaa gtg cca gac ttt ttc gaa ctt cgg att caa 3533 Val Asn Gly Trp Glu Val Pro Asp Phe Phe Glu Leu Arg Ile Gln 1125 1130 1135 ggt act acg caa tat aaa tgg gtg ttg att gtg acc ccc gcc gct 3578 Gly Thr Thr Gln Tyr Lys Trp Val Leu Ile Val Thr Pro Ala Ala 1140 1145 1150 gga tcg cct gct ggt ggt aat ggg gtc ttt gca ctc acc gga tct 3623 Gly Ser Pro Ala Gly Gly Asn Gly Val Phe Ala Leu Thr Gly Ser 1155 1160 1165 ttc gac ggc gag gta ttt aca gca gac gcg gtt gac cct acc act 3668 Phe Asp Gly Glu Val Phe Thr Ala Asp Ala Val Asp Pro Thr Thr 1170 1175 1180 cta tgg ctc gat tat ggt cgc gat tgg gat ggg gta atg agt tgg 3713 Leu Trp Leu Asp Tyr Gly Arg Asp Trp Asp Gly Val Met Ser Trp 1185 1190 1195 gaa aac gtg cct gct tcg gac ggg cgc agg gtt ctt gct gca gta 3758 Glu Asn Val Pro Ala Ser Asp Gly Arg Arg Val Leu Ala Ala Val 1200 1205 1210 atg aac agt tat ggt ggc aac ccc cca act aat acg tgg aag gga 3803 Met Asn Ser Tyr Gly Gly Asn Pro Pro Thr Asn Thr Trp Lys Gly 1215 1220 1225 atg ctt tca ttc cct cgt act ctg gaa ctc aag caa ctt aat ggc 3848 Met Leu Ser Phe Pro Arg Thr Leu Glu Leu Lys Gln Leu Asn Gly 1230 1235 1240 aaa ctc cga ttc ctc caa ttg ccc gtg agc gaa cta gac gcg gtc 3893 Lys Leu Arg Phe Leu Gln Leu Pro Val Ser Glu Leu Asp Ala Val 1245 1250 1255 ggt gct tca gtt gcg act atc acg aat cag act ctt gca cct gga 3938 Gly Ala Ser Val Ala Thr Ile Thr Asn Gln Thr Leu Ala Pro Gly 1260 1265 1270 caa aca ttg ctc tct aac atc cat tcc cgg tca ttg gac atc ctt 3983 Gln Thr Leu Leu Ser Asn Ile His Ser Arg Ser Leu Asp Ile Leu 1275 1280 1285 att acg ttt gtt cca gcc cag ggc tct aca ctg tct ctc tcc gtt 4028 Ile Thr Phe Val Pro Ala Gln Gly Ser Thr Leu Ser Leu Ser Val 1290 1295 1300 cgg aag gga ggg tct caa caa acc gtg att gaa tat gtc cag tcc 4073 Arg Lys Gly Gly Ser Gln Gln Thr Val Ile Glu Tyr Val Gln Ser 1305 1310 1315 aac aac caa ctt tcg gtc gat cgc aat gaa agt gga aac acc tca 4118 Asn Asn Gln Leu Ser Val Asp Arg Asn Glu Ser Gly Asn Thr Ser 1320 1325 1330 tac gat cct gca gct ggc ggt gtc cac agc gcc gct ctc cac gct 4163 Tyr Asp Pro Ala Ala Gly Gly Val His Ser Ala Ala Leu His Ala 1335 1340 1345 gat gag aat ggg aag gtg caa ttg cga gta ttg gtt gat gaa tgc 4208 Asp Glu Asn Gly Lys Val Gln Leu Arg Val Leu Val Asp Glu Cys 1350 1355 1360 tcc att gag gtc tat ggc ggg caa ggg gag gcg gtg atc tct gat 4253 Ser Ile Glu Val Tyr Gly Gly Gln Gly Glu Ala Val Ile Ser Asp 1365 1370 1375 ttg ata ttc ccc gat ttt tcc tcg gat ggc ctg gct ttg tcc act 4298 Leu Ile Phe Pro Asp Phe Ser Ser Asp Gly Leu Ala Leu Ser Thr 1380 1385 1390 agt caa ggg aac gtg gta ttg gaa tca gtc gac gtg cgg tcg att 4343 Ser Gln Gly Asn Val Val Leu Glu Ser Val Asp Val Arg Ser Ile 1395 1400 1405 ttg ctc tga 4352 Leu Leu 1410 <210> 11 <211> 1432 <212> PRT <213> Penicillium ochrochloron <400> 11 Met Ala Ser Arg Leu Val Thr Ala Leu Thr Leu Gly Leu His Leu Phe -20 -15 -10 Gln Ala Ala Phe Ala Gln Thr Tyr Asp Glu Leu Tyr Arg Pro Gln Tyr -5 -1 1 5 10 His Phe Thr Pro Pro Val Asn Trp Met Asn Asp Pro Asn Gly Leu Leu 15 20 25 Tyr His Asn Gly Leu Tyr His Leu Tyr Tyr Gln Tyr Asn Pro Gly Gly 30 35 40 Asn Thr Trp Gly Ala Met Ser Trp Gly His Ala Thr Ser Thr Asp Leu 45 50 55 Thr His Trp Asn His Gln Pro Ile Ala Leu Leu Ala Arg Gly Tyr Pro 60 65 70 75 Gly Asp Ile Thr Glu Met Phe Phe Ser Gly Ser Ala Val Ala Asp Thr 80 85 90 Gln Asn Thr Ser Gly Phe Gly Thr Ser Gly Lys Val Pro Phe Val Ala 95 100 105 Met Tyr Thr Ser Tyr Tyr Pro Ala Ser Gln Asn Leu Pro Ser Gly Lys 110 115 120 Leu Val Asn Gly Gly Gln Gln Ala Gln Ser Ile Ala Tyr Ser Leu Asp 125 130 135 Glu Gly Met Thr Trp Thr Thr Tyr Asp Ala Ala Asn Pro Val Ile Leu 140 145 150 155 Asn Pro Pro Thr Pro Tyr Thr Asp Gln Trp Arg Asp Phe Arg Asp Pro 160 165 170 Phe Leu Phe Trp His Glu Ala Ser Gln Lys Trp Ile Ser Val Val Ser 175 180 185 Leu Ala Gln Leu His Lys Leu Leu Ile Tyr Thr Ser Pro Asn Leu Lys 190 195 200 Asp Trp Thr Tyr Ser Ser Glu Phe Gly Pro Trp Asn Ala Val Gly Gly 205 210 215 Val Trp Glu Cys Pro Ser Leu Phe Pro Leu Ala Val Asp Gly Asp Asp 220 225 230 235 Ala Asn Thr Lys Trp Ile Met Gln Ile Gly Leu Asn Pro Gly Gly Pro 240 245 250 Pro Gly Val Thr Gly Ser Gly Thr Gln Tyr Ile Val Gly Thr Phe Asp 255 260 265 Gly Thr Lys Phe Val Ala Asp Pro Asn Ser Pro Leu Ser Ala His Thr 270 275 280 Ser Thr Ser Thr Thr Thr Ser Ala Ser Val Thr Thr Ser Ala Ser Ile 285 290 295 Thr Thr Ser Ala Ser Ile Thr Pro Thr Thr Thr Lys Ala Thr Ala Thr 300 305 310 315 Ala Thr Ala Thr Gly Asp Ile Ile Phe Gln Asp Phe Glu Gly Ala Gly 320 325 330 Asp Phe Ala Ser Arg Gly Trp Val Ala Thr Gly Gly Leu Leu Gly Ala 335 340 345 Ala Pro Ala Lys Gly Thr Leu Ala Gly Gln Gln Thr Val Thr Gly Tyr 350 355 360 Ala Gly Ser Gln Leu Val Asn Thr Phe Leu Ser Gly Asp Ser Thr Thr 365 370 375 Gly Thr Leu Thr Ser Pro Ala Phe Thr Ile Ser Leu Pro Tyr Val Asn 380 385 390 395 Phe Leu Ile Gly Gly Gly Asn Ala Pro Gly Thr Glu Cys Ile Asn Leu 400 405 410 Lys Val Gln Gly Gln Ala Val Arg Thr Ala Thr Gly Ala Asn Val Glu 415 420 425 Gln Leu Ala Pro Thr Thr Trp Asn Val Thr Asp Leu Met Gly Gln Thr 430 435 440 Ala Val Leu Glu Ile Val Asp Gln Gln Thr Gly Gly Trp Gly His Ile 445 450 455 Leu Ile Asp Gln Ile Thr Phe Thr Ser Ser Thr Ser Thr Asn Asn Leu 460 465 470 475 His Lys Arg Ser Val Ser Ser Val Thr Trp Asp Phe Asn Gly Thr Ser 480 485 490 Thr Phe Ala Asp Tyr Gly Trp Thr Pro Ser Gly Asp Phe Val Gly Lys 495 500 505 Gly Pro Ala Gln Gly Thr Leu Ala Gly Gln Gln Val Val Thr Gly Asn 510 515 520 Met Gly Asn Phe Val Asn Thr Phe Leu Ser Gly Asp Gly Thr Thr Gly 525 530 535 Thr Leu Thr Ser Pro Ala Phe Thr Ile Thr Gln Lys Lys Ile Asn Phe 540 545 550 555 Leu Ile Gly Gly Gly Asn Ser Pro Gly Met Glu Cys Ile Asn Leu Lys 560 565 570 Ile Gln Gly Gln Val Val Arg Thr Ala Thr Gly Ala Asn Ala Glu Gln 575 580 585 Leu Val Pro Thr Thr Trp Asp Val Thr Asp Leu Ile Gly Gln Ser Ala 590 595 600 Val Ile Glu Ile Val Asp Leu Ser Thr Thr Gly Trp Gly His Ile Leu 605 610 615 Ile Asp Glu Ile Ser Phe Ser Asn Ile Ser Ile Val Pro Tyr Gly Pro 620 625 630 635 Asn Trp Leu Asp Tyr Gly Pro Asp Phe Tyr Ala Ala Thr Thr Phe Asn 640 645 650 Gly Leu Ser Ser Thr Asn Arg Ile Asp Ile Ala Trp Met Asn Asn Trp 655 660 665 Gln Tyr Ala Ser Ala Ile Pro Thr Ser Pro Trp Arg Ser Met Leu Ser 670 675 680 Val Ala Arg Lys Leu Ser Leu Lys Thr Ile Asp Asp Arg Pro Arg Leu 685 690 695 Ile Gln Gln Pro Glu Ala Asn Trp Thr Ser Leu Gln Thr Thr Thr Tyr 700 705 710 715 Ser Thr Ser Phe Asp Thr Val Ala Glu Gly Asn Gln Leu Val Gln Leu 720 725 730 Ser Gly Lys Leu Leu Asp Ile Thr Val Ala Phe Ser Asp Arg Val Ala 735 740 745 Ala Ser Ser Ser Ser Gln Phe Gly Ile Ile Leu Arg Ala Thr Ser Asp 750 755 760 Leu Ala Gln Gln Thr Arg Ile Gly Tyr Asp Phe Ser Thr Lys Arg Leu 765 770 775 Phe Val Asp Arg Thr Lys Ser Gly Asn Val Gly Phe Asp Gly Thr Phe 780 785 790 795 Pro Ser Thr Tyr Phe Ala Pro Leu Ala Pro Ser Gly Asp Gly Arg Val 800 805 810 Thr Met Arg Ile Leu Leu Asp Arg Ser Ser Val Glu Val Phe Gly Gly 815 820 825 Gln Gly Glu Val Thr Ile Ser Ala Gln Ile Phe Pro Gln Asp Asn Gly 830 835 840 Val Asp Val Arg Leu Phe Ser Arg Gly Gly Ser Thr Asn Ala Ile Thr 845 850 855 Ile Asp Ala Ile Val Leu Asp Ser Val Tyr Asp Ser Ser Thr Pro Ser 860 865 870 875 Ala Ser Leu Ser Thr Ser Ser Thr Ser Ile Gly Thr Val Thr Thr Thr 880 885 890 Phe Ser Asn Thr Val Thr Thr Ser Val Gly Asn Thr Leu Thr Thr Ser 895 900 905 Thr Arg Thr Val Thr Ser Ala Ser Pro Thr Ser Thr Ala Pro Tyr Asp 910 915 920 Phe Arg Pro Val Phe His Phe Val Pro Glu Glu Asn Trp Met Asn Glu 925 930 935 Pro Asn Gly Leu Ile Lys Ile Asp Ser Thr Trp His Leu Phe Phe Gln 940 945 950 955 His Asn Pro Thr Gly Asn Phe Trp Gly Asn Leu Ser Trp Gly His Ala 960 965 970 Thr Ser Ile Asp Leu Val Ser Trp Asp Tyr Lys Pro Val Ala Ile Ser 975 980 985 Ser Ala Asp Gly Ile Gln Ala Phe Thr Gly Thr Ser Tyr Phe Asp Ala 990 995 1000 Met Asn Leu Ser Gly Leu Gly Thr Ser Ser Asn Gln Pro Tyr Leu 1005 1010 1015 Ala Phe Tyr Thr Gly Tyr Ala Pro Ser Ser Gly Ile Gln Asp Gln 1020 1025 1030 Arg Leu Ala Tyr Ser Leu Asp Gln Gly Ala Thr Trp Met Lys Tyr 1035 1040 1045 Gln Gly Asn Pro Ile Ile Ser Gln Ser Gln Glu Ala Pro His Asp 1050 1055 1060 Ile Thr Gln Gly Leu Glu Ile Arg Asp Pro Lys Val Phe Tyr His 1065 1070 1075 Ser Pro Thr Ser Arg Trp Val Met Ile Leu Ala His Gly Gly Gln 1080 1085 1090 Asn Lys Leu Thr Phe Trp Thr Ser Thr Asp Ala Lys Asn Trp Thr 1095 1100 1105 Trp Arg Ser Asn Phe Ala Ala Ser Asn Ile Leu Gly Phe Pro Ser 1110 1115 1120 Gly Val Asn Gly Trp Glu Val Pro Asp Phe Phe Glu Leu Arg Ile 1125 1130 1135 Gln Gly Thr Thr Gln Tyr Lys Trp Val Leu Ile Val Thr Pro Ala 1140 1145 1150 Ala Gly Ser Pro Ala Gly Gly Asn Gly Val Phe Ala Leu Thr Gly 1155 1160 1165 Ser Phe Asp Gly Glu Val Phe Thr Ala Asp Ala Val Asp Pro Thr 1170 1175 1180 Thr Leu Trp Leu Asp Tyr Gly Arg Asp Trp Asp Gly Val Met Ser 1185 1190 1195 Trp Glu Asn Val Pro Ala Ser Asp Gly Arg Arg Val Leu Ala Ala 1200 1205 1210 Val Met Asn Ser Tyr Gly Gly Asn Pro Pro Thr Asn Thr Trp Lys 1215 1220 1225 Gly Met Leu Ser Phe Pro Arg Thr Leu Glu Leu Lys Gln Leu Asn 1230 1235 1240 Gly Lys Leu Arg Phe Leu Gln Leu Pro Val Ser Glu Leu Asp Ala 1245 1250 1255 Val Gly Ala Ser Val Ala Thr Ile Thr Asn Gln Thr Leu Ala Pro 1260 1265 1270 Gly Gln Thr Leu Leu Ser Asn Ile His Ser Arg Ser Leu Asp Ile 1275 1280 1285 Leu Ile Thr Phe Val Pro Ala Gln Gly Ser Thr Leu Ser Leu Ser 1290 1295 1300 Val Arg Lys Gly Gly Ser Gln Gln Thr Val Ile Glu Tyr Val Gln 1305 1310 1315 Ser Asn Asn Gln Leu Ser Val Asp Arg Asn Glu Ser Gly Asn Thr 1320 1325 1330 Ser Tyr Asp Pro Ala Ala Gly Gly Val His Ser Ala Ala Leu His 1335 1340 1345 Ala Asp Glu Asn Gly Lys Val Gln Leu Arg Val Leu Val Asp Glu 1350 1355 1360 Cys Ser Ile Glu Val Tyr Gly Gly Gln Gly Glu Ala Val Ile Ser 1365 1370 1375 Asp Leu Ile Phe Pro Asp Phe Ser Ser Asp Gly Leu Ala Leu Ser 1380 1385 1390 Thr Ser Gln Gly Asn Val Val Leu Glu Ser Val Asp Val Arg Ser 1395 1400 1405 Ile Leu Leu 1410 <210> 12 <211> 1411 <212> PRT <213> Penicillium ochrochloron <400> 12 Gln Thr Tyr Asp Glu Leu Tyr Arg Pro Gln Tyr His Phe Thr Pro Pro 1 5 10 15 Val Asn Trp Met Asn Asp Pro Asn Gly Leu Leu Tyr His Asn Gly Leu 20 25 30 Tyr His Leu Tyr Tyr Gln Tyr Asn Pro Gly Gly Asn Thr Trp Gly Ala 35 40 45 Met Ser Trp Gly His Ala Thr Ser Thr Asp Leu Thr His Trp Asn His 50 55 60 Gln Pro Ile Ala Leu Leu Ala Arg Gly Tyr Pro Gly Asp Ile Thr Glu 65 70 75 80 Met Phe Phe Ser Gly Ser Ala Val Ala Asp Thr Gln Asn Thr Ser Gly 85 90 95 Phe Gly Thr Ser Gly Lys Val Pro Phe Val Ala Met Tyr Thr Ser Tyr 100 105 110 Tyr Pro Ala Ser Gln Asn Leu Pro Ser Gly Lys Leu Val Asn Gly Gly 115 120 125 Gln Gln Ala Gln Ser Ile Ala Tyr Ser Leu Asp Glu Gly Met Thr Trp 130 135 140 Thr Thr Tyr Asp Ala Ala Asn Pro Val Ile Leu Asn Pro Pro Thr Pro 145 150 155 160 Tyr Thr Asp Gln Trp Arg Asp Phe Arg Asp Pro Phe Leu Phe Trp His 165 170 175 Glu Ala Ser Gln Lys Trp Ile Ser Val Val Ser Leu Ala Gln Leu His 180 185 190 Lys Leu Leu Ile Tyr Thr Ser Pro Asn Leu Lys Asp Trp Thr Tyr Ser 195 200 205 Ser Glu Phe Gly Pro Trp Asn Ala Val Gly Gly Val Trp Glu Cys Pro 210 215 220 Ser Leu Phe Pro Leu Ala Val Asp Gly Asp Asp Ala Asn Thr Lys Trp 225 230 235 240 Ile Met Gln Ile Gly Leu Asn Pro Gly Gly Pro Pro Gly Val Thr Gly 245 250 255 Ser Gly Thr Gln Tyr Ile Val Gly Thr Phe Asp Gly Thr Lys Phe Val 260 265 270 Ala Asp Pro Asn Ser Pro Leu Ser Ala His Thr Ser Thr Ser Thr Thr 275 280 285 Thr Ser Ala Ser Val Thr Thr Ser Ala Ser Ile Thr Thr Ser Ala Ser 290 295 300 Ile Thr Pro Thr Thr Thr Lys Ala Thr Ala Thr Ala Thr Ala Thr Gly 305 310 315 320 Asp Ile Ile Phe Gln Asp Phe Glu Gly Ala Gly Asp Phe Ala Ser Arg 325 330 335 Gly Trp Val Ala Thr Gly Gly Leu Leu Gly Ala Ala Pro Ala Lys Gly 340 345 350 Thr Leu Ala Gly Gln Gln Thr Val Thr Gly Tyr Ala Gly Ser Gln Leu 355 360 365 Val Asn Thr Phe Leu Ser Gly Asp Ser Thr Thr Gly Thr Leu Thr Ser 370 375 380 Pro Ala Phe Thr Ile Ser Leu Pro Tyr Val Asn Phe Leu Ile Gly Gly 385 390 395 400 Gly Asn Ala Pro Gly Thr Glu Cys Ile Asn Leu Lys Val Gln Gly Gln 405 410 415 Ala Val Arg Thr Ala Thr Gly Ala Asn Val Glu Gln Leu Ala Pro Thr 420 425 430 Thr Trp Asn Val Thr Asp Leu Met Gly Gln Thr Ala Val Leu Glu Ile 435 440 445 Val Asp Gln Gln Thr Gly Gly Trp Gly His Ile Leu Ile Asp Gln Ile 450 455 460 Thr Phe Thr Ser Ser Thr Ser Thr Asn Asn Leu His Lys Arg Ser Val 465 470 475 480 Ser Ser Val Thr Trp Asp Phe Asn Gly Thr Ser Thr Phe Ala Asp Tyr 485 490 495 Gly Trp Thr Pro Ser Gly Asp Phe Val Gly Lys Gly Pro Ala Gln Gly 500 505 510 Thr Leu Ala Gly Gln Gln Val Val Thr Gly Asn Met Gly Asn Phe Val 515 520 525 Asn Thr Phe Leu Ser Gly Asp Gly Thr Thr Gly Thr Leu Thr Ser Pro 530 535 540 Ala Phe Thr Ile Thr Gln Lys Lys Ile Asn Phe Leu Ile Gly Gly Gly 545 550 555 560 Asn Ser Pro Gly Met Glu Cys Ile Asn Leu Lys Ile Gln Gly Gln Val 565 570 575 Val Arg Thr Ala Thr Gly Ala Asn Ala Glu Gln Leu Val Pro Thr Thr 580 585 590 Trp Asp Val Thr Asp Leu Ile Gly Gln Ser Ala Val Ile Glu Ile Val 595 600 605 Asp Leu Ser Thr Thr Gly Trp Gly His Ile Leu Ile Asp Glu Ile Ser 610 615 620 Phe Ser Asn Ile Ser Ile Val Pro Tyr Gly Pro Asn Trp Leu Asp Tyr 625 630 635 640 Gly Pro Asp Phe Tyr Ala Ala Thr Thr Phe Asn Gly Leu Ser Ser Thr 645 650 655 Asn Arg Ile Asp Ile Ala Trp Met Asn Asn Trp Gln Tyr Ala Ser Ala 660 665 670 Ile Pro Thr Ser Pro Trp Arg Ser Met Leu Ser Val Ala Arg Lys Leu 675 680 685 Ser Leu Lys Thr Ile Asp Asp Arg Pro Arg Leu Ile Gln Gln Pro Glu 690 695 700 Ala Asn Trp Thr Ser Leu Gln Thr Thr Thr Tyr Ser Thr Ser Phe Asp 705 710 715 720 Thr Val Ala Glu Gly Asn Gln Leu Val Gln Leu Ser Gly Lys Leu Leu 725 730 735 Asp Ile Thr Val Ala Phe Ser Asp Arg Val Ala Ala Ser Ser Ser Ser 740 745 750 Gln Phe Gly Ile Ile Leu Arg Ala Thr Ser Asp Leu Ala Gln Gln Thr 755 760 765 Arg Ile Gly Tyr Asp Phe Ser Thr Lys Arg Leu Phe Val Asp Arg Thr 770 775 780 Lys Ser Gly Asn Val Gly Phe Asp Gly Thr Phe Pro Ser Thr Tyr Phe 785 790 795 800 Ala Pro Leu Ala Pro Ser Gly Asp Gly Arg Val Thr Met Arg Ile Leu 805 810 815 Leu Asp Arg Ser Ser Val Glu Val Phe Gly Gly Gln Gly Glu Val Thr 820 825 830 Ile Ser Ala Gln Ile Phe Pro Gln Asp Asn Gly Val Asp Val Arg Leu 835 840 845 Phe Ser Arg Gly Gly Ser Thr Asn Ala Ile Thr Ile Asp Ala Ile Val 850 855 860 Leu Asp Ser Val Tyr Asp Ser Ser Thr Pro Ser Ala Ser Leu Ser Thr 865 870 875 880 Ser Ser Thr Ser Ile Gly Thr Val Thr Thr Thr Phe Ser Asn Thr Val 885 890 895 Thr Thr Ser Val Gly Asn Thr Leu Thr Thr Ser Thr Arg Thr Val Thr 900 905 910 Ser Ala Ser Pro Thr Ser Thr Ala Pro Tyr Asp Phe Arg Pro Val Phe 915 920 925 His Phe Val Pro Glu Glu Asn Trp Met Asn Glu Pro Asn Gly Leu Ile 930 935 940 Lys Ile Asp Ser Thr Trp His Leu Phe Phe Gln His Asn Pro Thr Gly 945 950 955 960 Asn Phe Trp Gly Asn Leu Ser Trp Gly His Ala Thr Ser Ile Asp Leu 965 970 975 Val Ser Trp Asp Tyr Lys Pro Val Ala Ile Ser Ser Ala Asp Gly Ile 980 985 990 Gln Ala Phe Thr Gly Thr Ser Tyr Phe Asp Ala Met Asn Leu Ser Gly 995 1000 1005 Leu Gly Thr Ser Ser Asn Gln Pro Tyr Leu Ala Phe Tyr Thr Gly 1010 1015 1020 Tyr Ala Pro Ser Ser Gly Ile Gln Asp Gln Arg Leu Ala Tyr Ser 1025 1030 1035 Leu Asp Gln Gly Ala Thr Trp Met Lys Tyr Gln Gly Asn Pro Ile 1040 1045 1050 Ile Ser Gln Ser Gln Glu Ala Pro His Asp Ile Thr Gln Gly Leu 1055 1060 1065 Glu Ile Arg Asp Pro Lys Val Phe Tyr His Ser Pro Thr Ser Arg 1070 1075 1080 Trp Val Met Ile Leu Ala His Gly Gly Gln Asn Lys Leu Thr Phe 1085 1090 1095 Trp Thr Ser Thr Asp Ala Lys Asn Trp Thr Trp Arg Ser Asn Phe 1100 1105 1110 Ala Ala Ser Asn Ile Leu Gly Phe Pro Ser Gly Val Asn Gly Trp 1115 1120 1125 Glu Val Pro Asp Phe Phe Glu Leu Arg Ile Gln Gly Thr Thr Gln 1130 1135 1140 Tyr Lys Trp Val Leu Ile Val Thr Pro Ala Ala Gly Ser Pro Ala 1145 1150 1155 Gly Gly Asn Gly Val Phe Ala Leu Thr Gly Ser Phe Asp Gly Glu 1160 1165 1170 Val Phe Thr Ala Asp Ala Val Asp Pro Thr Thr Leu Trp Leu Asp 1175 1180 1185 Tyr Gly Arg Asp Trp Asp Gly Val Met Ser Trp Glu Asn Val Pro 1190 1195 1200 Ala Ser Asp Gly Arg Arg Val Leu Ala Ala Val Met Asn Ser Tyr 1205 1210 1215 Gly Gly Asn Pro Pro Thr Asn Thr Trp Lys Gly Met Leu Ser Phe 1220 1225 1230 Pro Arg Thr Leu Glu Leu Lys Gln Leu Asn Gly Lys Leu Arg Phe 1235 1240 1245 Leu Gln Leu Pro Val Ser Glu Leu Asp Ala Val Gly Ala Ser Val 1250 1255 1260 Ala Thr Ile Thr Asn Gln Thr Leu Ala Pro Gly Gln Thr Leu Leu 1265 1270 1275 Ser Asn Ile His Ser Arg Ser Leu Asp Ile Leu Ile Thr Phe Val 1280 1285 1290 Pro Ala Gln Gly Ser Thr Leu Ser Leu Ser Val Arg Lys Gly Gly 1295 1300 1305 Ser Gln Gln Thr Val Ile Glu Tyr Val Gln Ser Asn Asn Gln Leu 1310 1315 1320 Ser Val Asp Arg Asn Glu Ser Gly Asn Thr Ser Tyr Asp Pro Ala 1325 1330 1335 Ala Gly Gly Val His Ser Ala Ala Leu His Ala Asp Glu Asn Gly 1340 1345 1350 Lys Val Gln Leu Arg Val Leu Val Asp Glu Cys Ser Ile Glu Val 1355 1360 1365 Tyr Gly Gly Gln Gly Glu Ala Val Ile Ser Asp Leu Ile Phe Pro 1370 1375 1380 Asp Phe Ser Ser Asp Gly Leu Ala Leu Ser Thr Ser Gln Gly Asn 1385 1390 1395 Val Val Leu Glu Ser Val Asp Val Arg Ser Ile Leu Leu 1400 1405 1410 <210> 13 <211> 1524 <212> DNA <213> Murcia penicillium <220> <221> CDS <222> (1)..(1521) <220> <221> signal peptide <222> (1)..(45) <220> <221> Mature peptides <222> (46) (1521) <400> 13 atg ttc ctc gcc tta gca ttc gcg ctc gcc att cgg gta atg gcg gat 48 Met Phe Leu Ala Leu Ala Phe Ala Leu Ala Ile Arg Val Met Ala Asp -15 -10 -5 -1 1 gac ttc cgc cca gtc tac cat ttt gtc cct aag gag aac tgg atg aac 96 Asp Phe Arg Pro Val Tyr His Phe Val Pro Lys Glu Asn Trp Met Asn 5 10 15 gaa ccc aat ggg ttg att aag atc aaa tca acc tgg cac ctg ttc tat 144 Glu Pro Asn Gly Leu Ile Lys Ile Lys Ser Thr Trp His Leu Phe Tyr 20 25 30 caa cac aac cca acg gca aat gtc tgg ggc aat ctg aac tgg ggc cac 192 Gln His Asn Pro Thr Ala Asn Val Trp Gly Asn Leu Asn Trp Gly His 35 40 45 gca acc agt agt gac ctg gtc cac tgg act cac gag ccc ctt gcc ata 240 Ala Thr Ser Ser Asp Leu Val His Trp Thr His Glu Pro Leu Ala Ile 50 55 60 65 ata agc gag aac ggt gtc gaa gcg ttc acc ggc act agc tat tat gat 288 Ile Ser Glu Asn Gly Val Glu Ala Phe Thr Gly Thr Ser Tyr Tyr Asp 70 75 80 gcc gac aac acc tcc gct cta gga tct tct acc agc cca ccc tat ctg 336 Ala Asp Asn Thr Ser Ala Leu Gly Ser Ser Thr Ser Pro Pro Tyr Leu 85 90 95 gcc tac tac act ggg tat ttt cct tct aac ggg acc cag gac cag cgt 384 Ala Tyr Tyr Thr Gly Tyr Phe Pro Ser Asn Gly Thr Gln Asp Gln Arg 100 105 110 ctt gcc ttt agt gtc gac gat ggc gtg acc tgg acg aag tac cag ggc 432 Leu Ala Phe Ser Val Asp Asp Gly Val Thr Trp Thr Lys Tyr Gln Gly 115 120 125 aat cct att atc tca gcc gct cag gag tcg ccg cat gat aag acg gcg 480 Asn Pro Ile Ile Ser Ala Ala Gln Glu Ser Pro His Asp Lys Thr Ala 130 135 140 145 ggg ctg gag acc cgc gac cct aag gtc ttt ttt cac cag gca tct ggg 528 Gly Leu Glu Thr Arg Asp Pro Lys Val Phe Phe His Gln Ala Ser Gly 150 155 160 aaa tgg gtc atg gtc ctt gcc cat ggc gga cag gac aag cta acg ttt 576 Lys Trp Val Met Val Leu Ala His Gly Gly Gln Asp Lys Leu Thr Phe 165 170 175 tgg acg tcg tcg aac gca aag tcc tgg acg tgg aag agc gac ctc tct 624 Trp Thr Ser Ser Asn Ala Lys Ser Trp Thr Trp Lys Ser Asp Leu Ser 180 185 190 gct agc cag atc gag ggt cta ccc tct gga att act ggg tgg gaa gtg 672 Ala Ser Gln Ile Glu Gly Leu Pro Ser Gly Ile Thr Gly Trp Glu Val 195 200 205 cct gat atg ttc gaa ctc cct gtc cag gac act aat gcg acc acc tgg 720 Pro Asp Met Phe Glu Leu Pro Val Gln Asp Thr Asn Ala Thr Thr Trp 210 215 220 225 gtg tta att ata acc cct gca aac gga tct ccc gct ggt ggt aat ggt 768 Val Leu Ile Ile Thr Pro Ala Asn Gly Ser Pro Ala Gly Gly Asn Gly 230 235 240 gtt cta gcc ctg aca ggc tcc ttt gat gga tca aca ttc aaa cct gac 816 Val Leu Ala Leu Thr Gly Ser Phe Asp Gly Ser Thr Phe Lys Pro Asp 245 250 255 cct gtc gac cct tct gct ctt tgg ctc gac tat ggt cgc gat ttc gac 864 Pro Val Asp Pro Ser Ala Leu Trp Leu Asp Tyr Gly Arg Asp Phe Asp 260 265 270 ggt gct ttg agt tgg gaa aat gtc cct gct gca gat agt cgg cgg cgg 912 Gly Ala Leu Ser Trp Glu Asn Val Pro Ala Ala Asp Ser Arg Arg Arg 275 280 285 atc ctt gcc tct gtc atg aac agc tac ggc gct agc cca cca acc aat 960 Ile Leu Ala Ser Val Met Asn Ser Tyr Gly Ala Ser Pro Pro Thr Asn 290 295 300 305 acc tgg aaa ggg atg ctg tct ttc cct cgg acc att acc cta aag gaa 1008 Thr Trp Lys Gly Met Leu Ser Phe Pro Arg Thr Ile Thr Leu Lys Glu 310 315 320 aca gcc tct aag cga tac ttc ctc cag cag cct gtc act gag ctc tcc 1056 Thr Ala Ser Lys Arg Tyr Phe Leu Gln Gln Pro Val Thr Glu Leu Ser 325 330 335 ctc gtt ggt tcc tcg tta gtg aca atc ​​cag aat caa aca att acc acc 1104 Leu Val Gly Ser Ser Leu Val Thr Ile Gln Asn Gln Thr Ile Thr Thr 340 345 350 ggc caa acg ttg ctg tca tcc att cac gga acc gcg tta gat atc aaa 1152 Gly Gln Thr Leu Leu Ser Ser Ile His Gly Thr Ala Leu Asp Ile Lys 355 360 365 att gcc ttt acc att aac gca ggt gcc aaa ctt tcg ctg gca gtg cgc 1200 Ile Ala Phe Thr Ile Asn Ala Gly Ala Lys Leu Ser Leu Ala Val Arg 370 375 380 385 aag gcc gcg tca gag cag aca atc ​​atc cag tat gac cag tca aat tcg 1248 Lys Ala Ala Ser Glu Gln Thr Ile Ile Gln Tyr Asp Gln Ser Asn Ser 390 395 400 tcg ata tct gtt gat cga acg acg agt gga gac atc tcc tac gac ccc 1296 Ser Ile Ser Val Asp Arg Thr Thr Ser Gly Asp Ile Ser Tyr Asp Pro 405 410 415 gct gca ggc gga gtc cac agt gct tcc ctg aaa cca gat ggt tct ggg 1344 Ala Ala Gly Gly Val His Ser Ala Ser Leu Lys Pro Asp Gly Ser Gly 420 425 430 gtt att cat ctc cgc gtc cta gtc gac acc tgc tcg gtg gag gtc ttt 1392 Val Ile His Leu Arg Val Leu Val Asp Thr Cys Ser Val Glu Val Phe 435 440 445 ggc ggg gaa gga gaa gtt gtg att tca gac ctg att ttt ccc agc gag 1440 Gly Gly Glu Gly Glu Val Val Ile Ser Asp Leu Ile Phe Pro Ser Glu 450 455 460 465 acc tcc gac ggg ctg tcc ttg gag gta act ggg gga tcg gcc aat ttg 1488 Thr Ser Asp Gly Leu Ser Leu Glu Val Thr Gly Gly Ser Ala Asn Leu 470 475 480 cac tct gtc gag gtg cgc agc att tcg ctc gaa taa 1524 His Ser Val Glu Val Arg Ser Ile Ser Leu Glu 485 490 <210> 14 <211> 507 <212> PRT <213> Penicillium murcianum <400> 14 Met Phe Leu Ala Leu Ala Phe Ala Leu Ala Ile Arg Val Met Ala Asp -15 -10 -5 -1 1 Asp Phe Arg Pro Val Tyr His Phe Val Pro Lys Glu Asn Trp Met Asn 5 10 15 Glu Pro Asn Gly Leu Ile Lys Ile Lys Ser Thr Trp His Leu Phe Tyr 20 25 30 Gln His Asn Pro Thr Ala Asn Val Trp Gly Asn Leu Asn Trp Gly His 35 40 45 Ala Thr Ser Ser Asp Leu Val His Trp Thr His Glu Pro Leu Ala Ile 50 55 60 65 Ile Ser Glu Asn Gly Val Glu Ala Phe Thr Gly Thr Ser Tyr Tyr Asp 70 75 80 Ala Asp Asn Thr Ser Ala Leu Gly Ser Ser Thr Ser Pro Pro Tyr Leu 85 90 95 Ala Tyr Tyr Thr Gly Tyr Phe Pro Ser Asn Gly Thr Gln Asp Gln Arg 100 105 110 Leu Ala Phe Ser Val Asp Asp Gly Val Thr Trp Thr Lys Tyr Gln Gly 115 120 125 Asn Pro Ile Ile Ser Ala Ala Gln Glu Ser Pro His Asp Lys Thr Ala 130 135 140 145 Gly Leu Glu Thr Arg Asp Pro Lys Val Phe Phe His Gln Ala Ser Gly 150 155 160 Lys Trp Val Met Val Leu Ala His Gly Gly Gln Asp Lys Leu Thr Phe 165 170 175 Trp Thr Ser Ser Asn Ala Lys Ser Trp Thr Trp Lys Ser Asp Leu Ser 180 185 190 Ala Ser Gln Ile Glu Gly Leu Pro Ser Gly Ile Thr Gly Trp Glu Val 195 200 205 Pro Asp Met Phe Glu Leu Pro Val Gln Asp Thr Asn Ala Thr Thr Trp 210 215 220 225 Val Leu Ile Ile Thr Pro Ala Asn Gly Ser Pro Ala Gly Gly Asn Gly 230 235 240 Val Leu Ala Leu Thr Gly Ser Phe Asp Gly Ser Thr Phe Lys Pro Asp 245 250 255 Pro Val Asp Pro Ser Ala Leu Trp Leu Asp Tyr Gly Arg Asp Phe Asp 260 265 270 Gly Ala Leu Ser Trp Glu Asn Val Pro Ala Ala Asp Ser Arg Arg Arg 275 280 285 Ile Leu Ala Ser Val Met Asn Ser Tyr Gly Ala Ser Pro Pro Thr Asn 290 295 300 305 Thr Trp Lys Gly Met Leu Ser Phe Pro Arg Thr Ile Thr Leu Lys Glu 310 315 320 Thr Ala Ser Lys Arg Tyr Phe Leu Gln Gln Pro Val Thr Glu Leu Ser 325 330 335 Leu Val Gly Ser Ser Leu Val Thr Ile Gln Asn Gln Thr Ile Thr Thr 340 345 350 Gly Gln Thr Leu Leu Ser Ser Ile His Gly Thr Ala Leu Asp Ile Lys 355 360 365 Ile Ala Phe Thr Ile Asn Ala Gly Ala Lys Leu Ser Leu Ala Val Arg 370 375 380 385 Lys Ala Ala Ser Glu Gln Thr Ile Ile Gln Tyr Asp Gln Ser Asn Ser 390 395 400 Ser Ile Ser Val Asp Arg Thr Thr Ser Gly Asp Ile Ser Tyr Asp Pro 405 410 415 Ala Ala Gly Gly Val His Ser Ala Ser Leu Lys Pro Asp Gly Ser Gly 420 425 430 Val Ile His Leu Arg Val Leu Val Asp Thr Cys Ser Val Glu Val Phe 435 440 445 Gly Gly Glu Gly Glu Val Val Ile Ser Asp Leu Ile Phe Pro Ser Glu 450 455 460 465 Thr Ser Asp Gly Leu Ser Leu Glu Val Thr Gly Gly Ser Ala Asn Leu 470 475 480 His Ser Val Glu Val Arg Ser Ile Ser Leu Glu 485 490 <210> 15 <211> 492 <212> PRT <213> Penicillium murcianum <400> 15 Asp Asp Phe Arg Pro Val Tyr His Phe Val Pro Lys Glu Asn Trp Met 1 5 10 15 Asn Glu Pro Asn Gly Leu Ile Lys Ile Lys Ser Thr Trp His Leu Phe 20 25 30 Tyr Gln His Asn Pro Thr Ala Asn Val Trp Gly Asn Leu Asn Trp Gly 35 40 45 His Ala Thr Ser Ser Asp Leu Val His Trp Thr His Glu Pro Leu Ala 50 55 60 Ile Ile Ser Glu Asn Gly Val Glu Ala Phe Thr Gly Thr Ser Tyr Tyr 65 70 75 80 Asp Ala Asp Asn Thr Ser Ala Leu Gly Ser Ser Thr Ser Pro Pro Tyr 85 90 95 Leu Ala Tyr Tyr Thr Gly Tyr Phe Pro Ser Asn Gly Thr Gln Asp Gln 100 105 110 Arg Leu Ala Phe Ser Val Asp Asp Gly Val Thr Trp Thr Lys Tyr Gln 115 120 125 Gly Asn Pro Ile Ile Ser Ala Ala Gln Glu Ser Pro His Asp Lys Thr 130 135 140 Ala Gly Leu Glu Thr Arg Asp Pro Lys Val Phe Phe His Gln Ala Ser 145 150 155 160 Gly Lys Trp Val Met Val Leu Ala His Gly Gly Gln Asp Lys Leu Thr 165 170 175 Phe Trp Thr Ser Ser Asn Ala Lys Ser Trp Thr Trp Lys Ser Asp Leu 180 185 190 Ser Ala Ser Gln Ile Glu Gly Leu Pro Ser Gly Ile Thr Gly Trp Glu 195 200 205 Val Pro Asp Met Phe Glu Leu Pro Val Gln Asp Thr Asn Ala Thr Thr 210 215 220 Trp Val Leu Ile Ile Thr Pro Ala Asn Gly Ser Pro Ala Gly Gly Asn 225 230 235 240 Gly Val Leu Ala Leu Thr Gly Ser Phe Asp Gly Ser Thr Phe Lys Pro 245 250 255 Asp Pro Val Asp Pro Ser Ala Leu Trp Leu Asp Tyr Gly Arg Asp Phe 260 265 270 Asp Gly Ala Leu Ser Trp Glu Asn Val Pro Ala Ala Asp Ser Arg Arg 275 280 285 Arg Ile Leu Ala Ser Val Met Asn Ser Tyr Gly Ala Ser Pro Pro Thr 290 295 300 Asn Thr Trp Lys Gly Met Leu Ser Phe Pro Arg Thr Ile Thr Leu Lys 305 310 315 320 Glu Thr Ala Ser Lys Arg Tyr Phe Leu Gln Gln Pro Val Thr Glu Leu 325 330 335 Ser Leu Val Gly Ser Ser Leu Val Thr Ile Gln Asn Gln Thr Ile Thr 340 345 350 Thr Gly Gln Thr Leu Leu Ser Ser Ile His Gly Thr Ala Leu Asp Ile 355 360 365 Lys Ile Ala Phe Thr Ile Asn Ala Gly Ala Lys Leu Ser Leu Ala Val 370 375 380 Arg Lys Ala Ala Ser Glu Gln Thr Ile Ile Gln Tyr Asp Gln Ser Asn 385 390 395 400 Ser Ser Ile Ser Val Asp Arg Thr Thr Ser Gly Asp Ile Ser Tyr Asp 405 410 415 Pro Ala Ala Gly Gly Val His Ser Ala Ser Leu Lys Pro Asp Gly Ser 420 425 430 Gly Val Ile His Leu Arg Val Leu Val Asp Thr Cys Ser Val Glu Val 435 440 445 Phe Gly Gly Glu Gly Glu Val Val Ile Ser Asp Leu Ile Phe Pro Ser 450 455 460 Glu Thr Ser Asp Gly Leu Ser Leu Glu Val Thr Gly Gly Ser Ala Asn 465 470 475 480 Leu His Ser Val Glu Val Arg Ser Ile Ser Leu Glu 485 490 <210> 16 <211> 2034 <212> DNA <213> Bacillus licheniformis <220> <221> CDS <222> (1)..(2031) <220> <221> signal peptide <222> (1)..(72) <220> <221> mature peptide <222> (73)..(2031) <400> 16 atg aaa aag aga atg att cag atg ggg atc ata ggg gcc atg atg ttt 48 Met Lys Lys Arg Met Ile Gln Met Gly Ile Ile Gly Ala Met Met Phe -20 -15 -10 ccg gaa gcg ttt tct gca gct gcg gcc gac ccg gac tac tat aac gag 96 Pro Glu Ala Phe Ser Ala Ala Ala Ala Asp Pro Asp Tyr Tyr Asn Glu -5 -1 1 5 gat cat cgg ccc aag tat cat ttc act ccg gaa gca aac tgg atg aat 14...

Claims

1. A cleaning composition comprising GH32 fructan-degrading enzyme and at least one cleaning component, wherein the fructan-degrading enzyme comprises a GH32 domain and optionally a GH32C domain, and is a polypeptide having at least 99% sequence identity with SEQ ID NO: 18, wherein the fructan-degrading enzyme comprises the motif WMND; and further comprises the motifs AYSNDK and HWGH, wherein the fructan-degrading enzyme is derived from Bacillus licheniformis.

2. The cleaning composition of claim 1, wherein the fructan-degrading enzyme is a polypeptide having 100% sequence identity with SEQ ID NO:

18.

3. The cleaning composition of claim 1 or 2, wherein the fructan-degrading enzyme has one or more enzyme activities selected from the group consisting of: fructan β(2,1)-fructosylase activity, fructan β(2,6)-fructosylase activity, fructan β-fructosylase activity, levanase activity, 2,6-β-fructan 6-levanbiohydrolase activity, and inulinase activity.

4. The cleaning composition of claim 1 or 2, wherein the cleaning composition comprises at least one surfactant, and optionally: (a) at least one additional cleaning component selected from the self-cleaning and bleaching components, and / or (b) at least one additional enzyme.

5. The cleaning composition of claim 1 or 2, wherein the cleaning composition further comprises at least one nuclease polypeptide having DNAse or RNAse activity.

6. Use of the cleaning composition according to any one of claims 1-5 for cleaning products, wherein the products are textiles.

7. The use of the cleaning composition according to any one of claims 1-5 for cleaning articles, wherein the article is a surface.

8. Use of the fructan-degrading enzyme as described in any one of claims 1-3 for removing biofilms from articles, optionally wherein the fructan-degrading enzyme is used in combination with at least one nuclease polypeptide having DNAase or RNAase activity.

9. The use as claimed in claim 8, wherein the article is a textile.

10. The use as claimed in claim 8, wherein the article is a surface.

11. A method for producing a cleaning product, the method comprising contacting the product with a solution comprising a fructan-degrading enzyme as described in any one of claims 1-3 and at least one cleaning component, and optionally contacting it with at least one nuclease polypeptide having DNase or RNase activity.

12. A polypeptide having fructan-degrading enzyme activity, wherein the polypeptide has at least 99% sequence identity with SEQ ID NO: 18, wherein the fructan-degrading enzyme comprises the motif WMND; and further comprises the motifs AYSNDK and HWGH, wherein the fructan-degrading enzyme is derived from Bacillus licheniformis.

13. The polypeptide of claim 12, wherein the polypeptide comprises a GH32 domain and optionally a GH32C domain.

14. The polypeptide of claim 12 or 13, wherein the polypeptide has 100% sequence identity with SEQ ID NO:

18.

15. The polypeptide of claim 12 or 13, wherein the polypeptide has one or more enzymatic activities selected from the group consisting of: fructan β(2,1)-fructosylase activity, fructan β(2,6)-fructosylase activity, fructan β-fructosylase activity, levanase activity, 2,6-β-fructan 6-levanbiohydrolase activity, and inulinase activity.

16. The polypeptide of claim 14, wherein the polypeptide has one or more enzymatic activities selected from the group consisting of: fructan β(2,1)-fructosylase activity, fructan β(2,6)-fructosylase activity, fructan β-fructosylase activity, levanase activity, 2,6-β-fructan 6-levanbiohydrolase activity, and inulinase activity.

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