CLEANING COMPOSITIONS COMPRISING AMYLASE VARIANTS - Patent application

Abstract

To provide cleaning compositions comprising variants of an α-amylase which exhibits an improved property, such as specific activity when compared to the parent polypeptide, and can be used in washing, dishwashing and / or cleaning processes at low temperature.SOLUTION: There is provided cleaning compositions comprising: a) variants of an α-amylase of a parent α-amylase having α-amylase activity which comprises one of the following substitutions at positions N3E, N3F, N3H, N3L, N3Q, N3T and N3V in a specific sequence, wherein the variant has at least 90%, but less than 100% sequence identity to the polypeptide of the specific sequence, and wherein the variant has α-amylase activity and wherein the α-amylase variant has an improved property relative to the parent polypeptide of the specific sequence; and b) a cleaning adjunct.SELECTED DRAWING: None

Description

[Technical Field]

[0001] (Reference to array list) The present specification contains a Sequence Listing in computer readable form, which is incorporated herein by reference.

[0002] FIELD OF THE INVENTION The present invention relates to cleaning compositions containing amylase enzymes, methods for making them, and methods for using them. The compositions and methods of the present invention are suitable for use in household cleaning or treatment compositions, particularly laundry and dishwashing compositions, including manual and automatic laundry compositions and / or manual and automatic dishwashing compositions. The present invention is particularly useful for cleaning laundry. [Background technology]

[0003] Alpha-amylases (α-1,4-glucan-4-glucanohydrolases, EC 3.2.1.1) constitute a group of enzymes that catalyze the hydrolysis of starch and other linear and branched 1,4-glucoside oligosaccharides and polysaccharides. They have been known for use in household cleaning compositions for many years, including the first bacterial α-amylases (also known as Termamyl or alkaline amylases) obtained from Bacillus licheniformis, such as AA560 (WO 2000 / 060060) and SP707 (described in Tsukamoto et al., 1988, Biochem. Biophys. Res. Comm. 151:25-31). Modifications of these and other amylases are known to improve their stability in detergents. For example, WO 96 / 23873 discloses deleting amino acids 181+182 or amino acids 183+184 of SP707 (SEQ ID NO: 7 of WO 96 / 23873) to improve the stability of this amylase. WO 96 / 23873 further discloses modifying SP707 amylase by substituting M202, for example, with leucine, to stabilize the molecule against oxidation.

[0004] For detergent formulators, it is always a challenge to improve the effectiveness of cleaning compositions, especially in an environmentally friendly way.For environmental reasons, it is also becoming increasingly important to provide detergent compositions that are effective even in quick wash cycles, which often use less water and / or lower temperatures.Therefore, it is desirable to have amylolytic enzymes that exhibit improved properties, such as specific activity, compared to their parent polypeptides.In particular, it is desirable for preferred amylolytic enzymes to be able to function at low temperatures while retaining or increasing other desirable properties, such as specific activity (amylolytic activity), stability, and / or cleaning performance, to enable good cleaning at lower temperatures and / or shorter wash cycles. [Prior art documents] [Patent documents]

[0005] [Patent Document 1] International Publication No. 2000 / 060060 [Patent Document 2] International Publication No. 96 / 23873 [Non-patent literature]

[0006] [Non-Patent Document 1] Tsukamoto et al.,1988,Biochem.Biophys.Res.Comm.151:25-31 Summary of the Invention [Problem to be solved by the invention]

[0007] It is therefore an object of the present invention to provide cleaning compositions comprising α-amylase variants that exhibit improved properties, such as specific activity, when compared to the parent polypeptide, which can be used in washing, dishwashing and / or cleaning processes at low temperatures. It is a further object of the present invention to provide cleaning compositions containing α-amylase variants that have improved cleaning performance at low temperatures compared to the parent α-amylase or compared to cleaning compositions containing the α-amylase of any of SEQ ID NOs: 1, 2, 3, 4, 5, 6, 7, or 8. [Means for solving the problem]

[0008] The present invention provides a cleaning composition comprising: (a) an α-amylase variant of a parent α-amylase polypeptide, wherein the variant is selected from the group consisting of a nucleotide sequence selected from the group consisting of nucleotides 3, 5, 7, 8, 16, 18, 25, 26, 29, 30, 35, 37, 40, 41, 43, 46, 50, 57, 60, 68, 71, 73, 74, 78, 81, 84, 86, 87, 90, 91, 93, 97, 98, 103, 105, 110, 111, 113, 114, 116, 117, 119, 120, 121, 125, 126, 128, 129, 131, 132, 133, 135, 137, 138, 139, 140, 141, 142, 143, 144, 145, 146, 147, 148, 149, 150, 151, 152, 153, 154, 155, 156, 157, 158, 159, 160, 161, 162, 163, 164, 165, 166, 167, 168, 169, 170, 171, 172, 173, 174, 175, 176, 177, 178, 179, 180, 181, 182, 183, 184, 18 , 141, 142, 143, 145, 146, 147, 149, 159, 161, 162, 165, 167, 169, 171, 176, 177, 179, 180, 181, 185, 186, 187, 195, 196, 197, 204, 206, 208, 211, 216, 218, 219, 225, 228, 230, 231, 235, 242, 243, 247, 250, 251, 252, 253, 254, 255, 257, 259, 260, 261, 262, 263, 265, 272, 273, 274, 279, 280, 281, 283, 284, 287, 2 92, 294, 296, 298, 299, 300, 303, 305, 307, 310, 311, 313, 314, 315, 319, 320, 321, 322, 323, 324, 334, 337, 339, 340, 342, 344, 345, 346, 347, 350, 359, 3 60, 361, 362, 363, 364, 368, 372, 375, 380, 382, ​​383, 384, 385, 387, 388, 391, 393, 395, 396, 397, 398, 401, 402, 403, 405, 406, 407, 410, 415, 416, 418, 42 1, 422, 423, 433, 434, 435, 438, 439, 446, 447, 449, 450, 452, 455, 457, 458, 460, 461, 462, 465, 466, 467, 469, 471, 473, 475, 476, 477, and 478, wherein the variant has a sequence identity of at least 60%, e.g., at least 65%, at least 70%, at least 75%, at least 80%, relative to the polypeptide of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, or 17.an α-amylase variant having at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% but less than 100% sequence identity, wherein the variant has α-amylase activity, and wherein the α-amylase variant has improved properties relative to the parent polypeptide; (b) a cleaning aid, preferably in an amount of 0.001 to 99.999, preferably 0.01 to 99.9 wt. %; The present invention provides a detergent composition comprising:

[0009] The present invention also provides a cleaning composition comprising: (a) Variants of a parent α-amylase polypeptide, comprising: i) a nucleotide sequence selected from the group consisting of: nucleotides 3, 5, 7, 8, 16, 18, 25, 26, 29, 30, 35, 37, 40, 41, 43, 46, 50, 57, 60, 68, 71, 73, 74, 78, 81, 84, 86, 87, 90, 91, 93, 97, 98, 103, 105, 110, 111, 113, 114, 116, 117, 119, 120, 121, 125, 126, 128, 129, 131, 132, 133, 135, 137, 138, 139, 140, 141, 142, 143, 144, 145, 146, 147, 148, 149, 150, 151, 152, 153, 154, 155, 156, 157, 158, 159, 160, 161, 162, 163, 164, 165, 166, 167, 168, 169, 170, 171, 172, 173, 174, 175, 176, 177, 178, 179, 180, 181, 182, 183, 184, 185, 186, 187 145, 146, 147, 149, 159, 161, 162, 165, 167, 169, 171, 176, 177, 179, 180, 181, 185, 186, 187, 195, 196, 197, 204, 206, 208, 211, 216, 218, 219, 225, 228, 2 30, 231, 235, 242, 243, 247, 250, 251, 252, 253, 254, 255, 257, 259, 260, 261, 262, 263, 265, 272, 273, 274, 279, 280, 281, 283, 284, 287, 292, 294, 296, 29 8, 299, 300, 303, 305, 307, 310, 311, 313, 314, 315, 319, 320, 321, 322, 323, 324, 334, 337, 339, 340, 342, 344, 345, 346, 347, 350, 359, 360, 361, 362, 363 , 364, 368, 372, 375, 380, 382, ​​383, 384, 385, 387, 388, 391, 393, 395, 396, 397, 398, 401, 402, 403, 405, 406, 407, 410, 415, 416, 418, 421, 422, 423, 433, (ii) has a substitution at one or more positions corresponding to positions R181, G182, D183, and G184 of the parent α-amylase, using SEQ ID NO: 1 for numbering; (iii) the α-amylase variant is selected from the group consisting of SEQ ID NOs: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 43, 43, 44, 44, 44, 44, 44, 45, 45, 45, 45, 45, 45, 46 ...7, 46, 47, 46, 47, 4712, 13, 14, 15, 16 or 17. A variant having at least 60%, such as at least 65%, for example at least 70%, such as at least 75%, for example at least 80%, such as at least 85%, for example at least 90%, such as at least 95%, for example at least 97%, such as at least 99%, but less than 100%, sequence identity to the amino acid sequence of the polypeptide of claim 12, 13, 14, 15, 16 or 17, (iv) the variant has α-amylase activity, and (v) the α-amylase variant has improved properties relative to the parent polypeptide. and a cleaning aid in an amount of preferably 0.001 to 99.999, preferably 0.01 to 99.9 wt %.

[0010] The present invention also provides a method of producing a cleaning composition, the method comprising: a) obtaining an amylase variant of a parent α-amylase polypeptide, the amylase variant comprising: a) a nucleotide sequence selected from the group consisting of nucleotides 3, 5, 7, 8, 16, 18, 25, 26, 29, 30, 35, 37, 40, 41, 43, 46, 50, 57, 60, 68, 71, 73, 74, 78, 81, 84, 86, 87, 90, 91, 93, 97, 98, 103, 105, 110, 111, 113, 114, 116, 117, 119, 120, 121, 125, 126, 128, 129, 130, 131, 132, 133, 134, 135, 136, 137, 138, 139, 140, 141, 142, 143, 144, 145, 146, 147, 148, 149, 150, 151, 152, 153, 154, 155, 156, 157, 158, 159, 200, 201, 202, 203, 204, 205, 206, 207, 208, 209, 210, 211, 212, 213, 214, 215, 216, 217, 218, 219, 9, 131, 132, 133, 135, 137, 138, 139, 140, 141, 142, 143, 145, 146, 147, 149, 159, 161, 162, 165, 167, 169, 171, 176, 177, 179, 180, 181, 185, 186, 187, 195, 196, 197, 204, 206, 208, 211, 216, 218, 219, 225, 228, 230, 231, 235, 242, 243, 247, 250, 251, 252, 253, 254, 255, 257, 259, 260, 261, 262, 263, 265, 272, 273, 274, 279, 280, 281, 283, 284, 287, 292, 294, 296, 298, 299, 300, 303, 305, 307, 310, 311, 313, 314, 315, 319, 320, 321, 322, 323, 324, 334, 337, 339, 340, 342, 344, 345, 346, 347, 350, 359, 360, 361, 362, 363, 364, 368, 372, 375, 380, 382, ​​383, 384, 385, 387, 388, 391, 393, 395, 396, 397, 398, 399, 400, 401, 402, 403, 404, 405, 406, 407, 408, 409, 410, 411, 412, 413, 414, 415, 416, 417, 418, 419, 420, 421, 422, 423, 424, 425, 426, 427, 428, 429, 430, 431, 432, 433, 434, 435, 436, 437, 438, 460, 461, 462, 465, 466, 467, 469, 471, 473, 475, 476, 477, and 478 of the parent α-amylase; and b) introducing substitutions at one or more positions corresponding to positions R181, G182, D183, R184, R185, R186, R187, R188, R189, R190, R191, R192, R193, R194, R195, R196, R197, R198, R199, R200, R201, R202, R203, R204, R205, R206, R207, R208, R210, R211, R212, R213, R214, R215, R216, R217, R218, R220, R221, R222, R223, R230, R231, R232, R233, R234, R235, R238, R239, R240, R246, R247, R249, R250, R252, R255, R257, R258, R260, R261, R262, R265, R266, R267, R269, R271, R273, R275, R276, R277, R278, R280, R281, R282, R283, R284, R285, R286, R287, R288, R289, R290, R291, R292, R2and G184, whereby an α-amylase variant of the parent α-amylase is obtained, the variant having at least 60%, such as at least 65%, for example at least 70%, such as at least 75%, for example at least 80%, such as at least 85%, for example at least 90%, such as at least 95%, for example at least 97%, such as at least 99%, but less than 100%, sequence identity to the amino acid sequence of a polypeptide of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16 or 17, wherein the variant has α-amylase activity, and wherein the α-amylase variant has improved properties relative to the parent polypeptide; and combining the α-amylase variant with a cleaning aid.

[0011] The present invention also provides a method for treating a surface, preferably a textile, comprising the steps of: (i) forming an aqueous cleaning solution comprising water, an amylase described herein, and a cleaning aid; (ii) treating the surface with an aqueous cleaning solution, preferably at a temperature of 5 or 60°C, preferably 10-40°C, or preferably at a maximum of 35°C, more preferably at a temperature of 30°C or less, or at a temperature of 20°C or less; (iii) rinsing the surface.

[0012] The compositions and methods herein are particularly useful for treating surfaces comprising fibers or fabrics, such as cotton and / or synthetic fibers (e.g., polyester, nylon, etc.), which may be in the form of fibers alone or in blends (e.g., polycotton). The present invention further relates to the use of the compositions or methods described above for cleaning or removing stains, particularly starch or other carbohydrate-containing stains. DETAILED DESCRIPTION OF THE INVENTION

[0013] definition Allelic variant: The term "allelic variant" refers to any of two or more alternative forms of a gene occupying the same chromosomal locus. Allelic variation arises naturally through mutation and can result in polymorphism within a population. A mutation in a gene can be silent (the encoded polypeptide is unchanged) or it can encode a polypeptide with an altered amino acid sequence. An allelic variant of a polypeptide is a polypeptide encoded by an allelic variant of a gene.

[0014] Alpha-amylase: The term "alpha-amylase" (alpha-1,4-glucan-4-glucanohydrolase, EC 3.2.1.1) constitutes a group of enzymes that catalyze the hydrolysis of starch and other linear and branched 1,4-glucoside oligosaccharides and polysaccharides. In one aspect, an alpha-amylase variant useful in the present invention has at least 60%, at least 70%, at least 80%, at least 90%, at least 95%, at least 99%, but less than 100%, of the alpha-amylase activity of the polypeptide of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, and / or 17.

[0015] Alpha-amylase activity: For purposes of the present invention, alpha-amylase activity is determined according to the procedure described in the Examples section. Alpha-amylase activity can be determined according to the method using the microswatch assay described in Example 2.

[0016] Amino Acid: As used herein, the term "amino acid" includes the 20 standard genetically encoded amino acids and their corresponding stereoisomers in the "d" form (as compared to the natural "l" form), omega amino acids, other naturally occurring amino acids, unconventional amino acids (e.g., α,α-disubstituted amino acids, N-alkyl amino acids, etc.), and chemically derivatized amino acids. Chemical derivatives of one or more amino acids can be obtained by reaction with a functional side group. Such derivatized molecules include, for example, molecules in which a free amino group is derivatized to form an amine hydrochloride, p-toluenesulfonyl group, carboxybenzoxy group, t-butyloxycarbonyl group, chloroacetyl group, or formyl group. Free carboxyl groups may be derivatized to form salts, methyl and ethyl esters, or other types of esters and hydrazides. Free hydroxyl groups may be derivatized to form O-acyl or O-alkyl derivatives. Chemical derivatives also include peptides containing naturally occurring amino acid derivatives of the 20 standard amino acids. For example, 4-hydroxyproline may be used in place of proline, 5-hydroxylysine may be used in place of lysine, 3-methylhistidine may be used in place of histidine, homoserine may be used in place of serine, and ornithine may be used in place of lysine. Derivatives also include peptides containing one or more additions or deletions, so long as the required activity is maintained. Other included modifications are terminal modifications such as amidation, amino-terminal acylation (e.g., acetylation or thioglycolic acid amidation), and terminal carboxylamidation (e.g., with ammonia or methylamine).

[0017] When an amino acid is specifically recited, such as "alanine" or "Ala" or "A," the term refers to both l-alanine and d-alanine unless otherwise specified. Other non-conventional amino acids may also be suitable components of the polypeptides of the invention, so long as the desired functional properties are retained by the polypeptide. For the peptides shown, each encoded amino acid residue is represented, where appropriate, by a single letter designation corresponding to the trivial name of the conventional amino acid. In one embodiment, the polypeptides of the invention comprise or consist of l-amino acids.

[0018] cDNA: The term "cDNA" refers to a DNA molecule that can be prepared by reverse transcription from a mature, spliced ​​mRNA molecule obtained from a eukaryotic cell. cDNA lacks intron sequences that may be present in the corresponding genomic DNA. The initial primary RNA transcript is a precursor to mRNA that is processed through a series of processes, including splicing, before appearing as a spliced ​​mature mRNA.

[0019] Coding sequence: The term "coding sequence" refers to a polynucleotide that directly specifies the amino acid sequence of a variant thereof. The boundaries of the coding sequence are generally determined by an open reading frame, and usually begin with a start codon such as ATG, GTG, and TTG and end with a stop codon such as TAA, TAG, and TGA. A coding sequence can be a DNA, cDNA, synthetic, or recombinant polynucleotide.

[0020] Control sequence: The term "control sequence" as used herein refers to a nucleic acid sequence necessary for expression of a polynucleotide encoding a variant. Each control sequence may be native (i.e., derived from the same gene) or foreign (i.e., derived from different genes) to the polynucleotide encoding the variant, or may be native or foreign to each other. Such control sequences include, but are not limited to, a leader, polyadenylation sequence, propeptide sequence, promoter, signal peptide sequence, and transcription terminator. At a minimum, control sequences include a promoter and transcriptional and translational stop signals. Control sequences may also be provided with linkers for the purpose of introducing specific restriction enzyme sites facilitating ligation of the control sequence with the coding region of the polynucleotide encoding the variant.

[0021] Corresponding: As used herein, the term "corresponding" refers to a method of determining a specific amino acid of a reference sequence, and a reference is made to a specific amino acid sequence. For example, for purposes of the present invention, when a reference is made to a specific amino acid position, one of skill in the art can align another amino acid sequence to the referenced amino acid sequence, thereby determining which specific amino acids may be of interest in the other amino acid sequence. Alignment of another amino acid sequence with a sequence set forth in SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, or 17, or with any other sequence listed herein, is described elsewhere herein. Alternative alignment methods may be used and are well known to those of skill in the art.

[0022] Enzyme detergent benefit: As used herein, the term "enzyme detergent benefit" refers to the beneficial effect that an enzyme can add to a detergent compared to the same detergent without the enzyme. Important detergent benefits that can be provided by enzymes are stain removal with little or no visible soiling after laundering and / or washing, prevention or reduction of redeposition of soils released during the laundering process (also known as anti-redeposition), and full or partial restoration of whiteness of textiles that were originally white but have acquired a grayish or yellowish appearance as a result of repeated use and washing (also known as whitening). Textile care benefits not directly related to catalytic stain removal or prevention of soil redeposition can also be important to enzyme detergent benefit. Examples of such fabric care benefits include preventing or reducing dye transfer from one fabric to another or to another part of the same fabric (also known as dye transfer prevention or backsoiling prevention), removing protruding or broken fibers from the fabric surface to reduce pilling tendency or remove existing pilling or fuzz (anti-pilling), improving fabric softness, clarifying fabric color, and removing particulate soils trapped in fabric or garment fibers. Enzyme bleaching is a further enzyme detergent benefit, and this catalytic activity is commonly used to catalyze the formation of bleaching ingredients such as hydrogen peroxide or other peroxides.

[0023] Expression: The term "expression" includes any step involved in the production of a variant, including but not limited to transcription, post-transcriptional modification, translation, post-translational modification, and secretion.

[0024] Expression vector: The term "expression vector" refers to a linear or circular DNA molecule that contains a polynucleotide encoding a variant, operably linked to additional nucleotides that effect its expression.

[0025] Fragment: The term "fragment" refers to a polypeptide having one or more (e.g., several) amino acids deleted from the amino and / or carboxyl terminus of the polypeptide of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, or 17, wherein the fragment has α-amylase activity. In one aspect, the fragment contains at least 200 contiguous amino acid residues of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, or 17, e.g., at least 300 contiguous amino acid residues, or at least 350 contiguous amino acid residues, or at least 400 contiguous amino acid residues, or at least 450 contiguous amino acid residues of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, or 17.

[0026] Hard surface cleaning: As used herein, the term "hard surface cleaning" refers to cleaning hard surfaces, which may include floors, tables, walls, roofs, etc., as well as the surfaces of hard objects such as automobiles (car washing) and dishes (dishwashing). Dishwashing includes, but is not limited to, cleaning dishes, cups, glasses, bowls, spoons, knives, forks, and other cutlery, serving utensils, ceramics, plastics, metals, china, glass, and acrylic.

[0027] High stringency: The term "high stringency conditions" refers to prehybridization and hybridization for 12-24 hours in 5X SSPE, 0.3% SDS, 200 micrograms / mL sheared and denatured salmon sperm DNA, and 50% formamide at 42°C according to standard Southern blotting procedures, using a probe at least 100 nucleotides in length. Finally, the carrier material is washed three times for 15 minutes each using 2X SSC, 0.2% SDS at 65°C. Host cell: The term "host cell" refers to any cell type that is amenable to transformation, transfection, transduction, etc., with a nucleic acid construct or expression vector containing a polynucleotide described herein. The term "host cell" encompasses any progeny of a parent cell that is not identical to the parent cell due to mutations that occur during replication.

[0028] Improved Properties: The term "improved properties" is defined herein as properties associated with a variant that are improved compared to the parent α-amylase. Such improved properties include increased amylolytic activity, improved catalytic efficiency, increased catalytic rate, increased chemical stability, increased oxidative stability, increased pH activity, increased pH stability, increased specific activity, increased substrate binding, increased substrate cleavage, increased substrate specificity, increased substrate stability, improved surface properties, increased thermal activity, improved thermal stability, improved cleaning performance such as soil performance, e.g., performance on starch-containing stains, stain removal, graying prevention, stability (e.g., thermal stability, pH stability, or stability in the presence of builders (including chelating agents)), and improved performance in powder, liquid, or gel detergent formulations. The improved properties include, but are not limited to, stability in a liquid or dishwashing composition, modified temperature-dependent performance and activity profile, pH activity, substrate specificity, product specificity, and chemical stability. The improved properties can be any of those defined and described herein. One preferred improved property herein is increased specific activity. Specifically, improved specific activity can refer to improved specific activity measured in a model detergent A composition. The improvement factor (IF) is greater than 1, preferably at least 1.1, or at least 1.2, or at least 1.3.

[0029] Improved cleaning performance: The term "improved cleaning performance" is defined herein to refer to an alteration of the cleaning performance of the parent α-amylase and / or amylase of the invention relative to SEQ ID NO: 1 or SEQ ID NO: 2. The alteration may be manifested, for example, as improved stain removal. Improved cleaning performance occurs when the amylase variant has improved properties relative to the parent polypeptide. This may be determined in a model A detergent composition. The improvement factor (IF) is preferably at least 1.1, at least 1.2, or at least 1.3.

[0030] Isolated: The term "isolated" refers to a substance in a form or environment that does not occur in nature. Non-limiting examples of isolated substances include: (1) any substance that does not occur in nature; (2) any substance, including, but not limited to, any enzyme, variant, nucleic acid, protein, peptide, or cofactor, that has been at least partially removed from one or all of the naturally occurring components with which it is associated; (3) any substance that has been artificially modified relative to the substance found in nature; or (4) any substance that has been modified by increasing the amount of the substance relative to other components with which it is naturally associated (e.g., multiple copies of the gene encoding the substance; use of a stronger promoter than that naturally associated with the gene encoding the substance). An isolated substance may be present in a fermentation broth sample. In one aspect, the present invention relates to an isolated α-amylase variant.

[0031] Isolated polynucleotide: The term "isolated polynucleotide" refers to a polynucleotide that has not been artificially modified. In one embodiment, the isolated polynucleotide is at least 1% pure, e.g., at least 5% pure, at least 10% pure, at least 20% pure, at least 40% pure, at least 60% pure, at least 80% pure, at least 90% pure, and at least 95% pure, as determined by agarose gel electrophoresis. The polynucleotide may be of genomic, cDNA, RNA, semisynthetic, synthetic origin, or any combination thereof.

[0032] Isolated variant: The term "isolated variant" refers to a variant that has been artificially modified. In one embodiment, the variant is at least 1% pure, e.g., at least 5% pure, at least 10% pure, at least 20% pure, at least 40% pure, at least 60% pure, at least 80% pure, and at least 90% pure, as determined by SDS-PAGE.

[0033] Low temperature: "Low temperature" is a temperature of 5 to 40°C, preferably 5 to 35°C, preferably 5 to 30°C, more preferably 5 to 25°C, more preferably 5 to 20°C, most preferably 5 to 15°C, especially 5 to 10°C. In a preferred embodiment, "low temperature" is a temperature of 10 to 35°C, preferably 10 to 30°C, or 10 to 25°C, or 10 to 20°C, or 10 to 15°C.

[0034] Mature polypeptide: The term "mature polypeptide" refers to a polypeptide in its final form after translation and any post-translational modifications, such as N-terminal processing, C-terminal truncation, glycosylation, phosphorylation, etc. It is known in the art that a host cell can produce a mixture of two or more different mature polypeptides (i.e., with different C-terminal and / or N-terminal amino acids) expressed by the same polynucleotide.

[0035] Mature polypeptide coding sequence: The term "mature polypeptide coding sequence" refers to a polynucleotide that encodes a mature polypeptide having alpha-amylase activity.

[0036] Modified: As used herein in the context of polypeptides, the term "modified" means that one or more amino acids in a reference amino acid sequence (i.e., SEQ ID NOs: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, or 17) are altered by substitution with another amino acid, by insertion of an amino acid, or by deletion, preferably by at least one deletion. The terms "modified," "altered," and "mutated" are used interchangeably and may constitute the same meaning and purpose.

[0037] Moderate stringency conditions: The term "moderate stringency conditions" refers to prehybridization and hybridization for 12-24 hours in 5X SSPE, 0.3% SDS, 200 micrograms / mL sheared and denatured salmon sperm DNA, and 35% formamide at 42°C according to standard Southern blotting procedures for probes at least 100 nucleotides in length. Finally, the carrier material is washed three times for 15 minutes each using 2X SSC, 0.2% SDS at 55°C.

[0038] Mutant: The term "mutant" refers to a polynucleotide that encodes a variant.

[0039] Nucleic Acid Construct: The term "nucleic acid construct" refers to a nucleic acid molecule, either single-stranded or double-stranded, that contains one or more regulatory sequences, that has been isolated from a naturally occurring gene or that has been modified to contain a segment of nucleic acid in a manner that is not otherwise found in nature, or that is synthetic. The term nucleic acid construct is synonymous with the term "expression cassette" when the nucleic acid construct contains the regulatory sequences required for expression of a coding sequence of the present invention.

[0040] Operably linked: The term "operably linked" refers to a configuration in which control sequences are positioned appropriately relative to a coding sequence of a polynucleotide so that the control sequences direct the expression of the coding sequence.

[0041] Parent or Parent α-Amylase: As used herein, the term "parent" or "parent α-amylase" refers to an α-amylase that is modified to produce a variant α-amylase of the invention. The term also refers to a polypeptide to which a variant of the invention is compared. A parent may be a naturally occurring (wild-type) polypeptide or a variant thereof prepared by any suitable means. For example, a parent protein may be a variant of a naturally occurring polypeptide that has been modified or altered in amino acid sequence. Thus, the parent α-amylase may have one or more (or one or more) amino acid substitutions, deletions, and / or insertions. Thus, the parent α-amylase may be a variant of the parent α-amylase. A parent may also be an allelic variant, which is a polypeptide encoded by any of two or more alternative forms of a gene occupying the same chromosomal locus. The term "parent" or "parent α-amylase" as used herein refers to an α-amylase of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16 or 17, or a hybrid thereof, or any α-amylase having at least 60% sequence identity to any of the polypeptides of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16 or 17. A parent amylase may also be a polypeptide comprising a fragment of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16 or 17.

[0042] Sequence identity: The relatedness between two amino acid sequences or between two nucleotide sequences is expressed by the parameter "sequence identity."

[0043] For purposes of the present invention, sequence identity between two amino acid sequences is determined using the Needleman-Wunsch algorithm (Needleman and Wunsch, 1970, J. Mol. Biol. 48:443-453), as implemented in the Needle program of the EMBOSS package (EMBOSS: The European Molecular Biology Open Software Suite, Rice et al., 2000, Trends Genet. 16:276-277), preferably version 5.0.0 or later. The parameters used may be a gap opening penalty of 10, a gap extension penalty of 0.5, and an EBLOSUM62 (EMBOSS version of BLOSUM62) substitution matrix. The Needle output labeled "longest identity" (obtained using the -nobrief option) is used as the percent identity, which is calculated as follows: (Identical residues × 100) / (length of alignment − total number of gaps in the alignment)

[0044] Alternatively, optional parameters used can be a gap opening penalty of 10, a gap extension penalty of 0.5, and an EDNAFULL (the EMBOSS version of NCBI NUC4.4) substitution matrix. The output of Needle labeled "longest identity" (obtained using the -nobrief option) is used as the percent identity, which is calculated as follows: (identical deoxyribonucleotides × 100) / (length of alignment − total number of gaps in alignment)

[0045] Starch removal process: The expression "starch removal process" relates to any kind of process in which starch is removed (or converted) in a cleaning process in which starch is removed from textiles, for example in textile cleaning, such as laundry. The starch removal process may also be hard surface cleaning, such as dishwashing, or a general cleaning process, such as industrial or institutional cleaning. The expression also generally includes other starch removal processes or starch conversions, ethanol production, starch liquefaction, textile desizing, paper and pulp production, beer production, and detergents.

[0046] Subsequence: The term "subsequence" refers to a polynucleotide having one or more (e.g., several) nucleotides deleted from the 5'- and / or 3'-end of a mature polypeptide coding sequence, wherein the subsequence encodes a fragment that has α-amylase activity.

[0047] Substantially pure polynucleotide: "Substantially pure polynucleotide" refers to a polynucleotide preparation that is free of other extraneous or unwanted nucleotides and is in a form suitable for use in a genetically engineered polypeptide production system. The term "substantially pure variant" contains less than 10%, less than 8%, less than 6%, less than 5%, less than 4%, less than 3%, less than 2%, less than 1%, and less than 0.5% by weight of other natively or recombinantly associated polynucleotide material. However, a substantially pure polynucleotide may contain naturally occurring 5'- and 3'-untranslated regions, such as promoters and terminators. In some embodiments, a substantially pure polynucleotide is preferably at least 90% pure, e.g., at least 92% pure, at least 94% pure, at least 95% pure, at least 96% pure, at least 97% pure, at least 98% pure, at least 99% pure, and at least 99.5% pure, as determined by weight. The polynucleotides of the present invention are preferably in a substantially pure form.

[0048] Substantially pure variant: The term "substantially pure variant" refers to a preparation containing no more than 10%, no more than 8%, no more than 6%, no more than 5%, no more than 4%, no more than 3%, no more than 2%, no more than 1%, and no more than 0.5% by weight of other natively or recombinantly associated polypeptide material. Preferably, the variant is at least 92% pure, e.g., at least 94% pure, at least 95% pure, at least 96% pure, at least 97% pure, at least 98% pure, at least 99% pure, at least 99.5% pure, and at least 100% pure, by weight of total polypeptide material present in the preparation. The variants of the present invention are preferably in substantially pure form. This can be achieved, for example, by preparing the variant by well-known recombinant methods or by classical purification techniques.

[0049] Textiles: Textile sample CS-27 is obtained from Center For Test materials BV (PO Box 120, 3133 KT Vlaardingen, the Netherlands).

[0050] Fabric care effect: The term "fabric care effect" as used herein is defined as not directly related to catalytic stain removal or preventing redeposition of soil, but is also important for enzyme detergent effect.Examples of such care effects include: preventing or reducing the transfer of dye from one fabric to another fabric or to another part of the same fabric (also known as dye transfer prevention or backsoiling prevention effect); removing protruding or broken fibers from the surface of a fabric to reduce pilling tendency or remove existing pilling or fuzz (also known as anti-pilling effect); improving fabric softness; clarifying the color of fabric; and removing particulate soil trapped in fabric fibers.Enzyme bleaching is another enzyme detergent effect, and this catalytic activity is generally used to catalyze the formation of bleaching components such as hydrogen peroxide or other peroxides or other bleaching substances.

[0051] Variant: The term "variant" or "polypeptide variant" or "α-amylase variant" refers to a polypeptide having α-amylase activity that contains modifications / mutations, i.e., substitutions, insertions, and / or deletions, at one or more (e.g., several) positions relative to a parent or wild-type amylase. Preferably, the variant contains fewer than 50 modifications, more preferably fewer than 30 modifications. Modified α-amylases are obtained by human intervention by modifying a parent α-amylase. A substitution refers to the replacement of an amino acid occupying a position with a different amino acid; a deletion refers to the removal of an amino acid occupying a position; and an insertion refers to the addition of 1 to 3 amino acids adjacent to and immediately following the amino acid occupying a position. The alpha-amylase variants herein have at least 20%, e.g., at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 95% but less than 100% of the alpha-amylase activity of the polypeptides set forth in SEQ ID NOs: 1-17.

[0052] "Very high stringency conditions" refers to prehybridization and hybridization for 12-24 hours in 5X SSPE, 0.3% SDS, 200 micrograms / mL sheared and denatured salmon sperm DNA, and 50% formamide at 42°C according to standard Southern blotting procedures, for probes at least 100 nucleotides in length. Finally, the carrier material is washed three times for 15 minutes each using 2X SSC, 0.2% SDS at 70°C.

[0053] "Very low stringency conditions" refers to prehybridization and hybridization for 12-24 hours in 5X SSPE, 0.3% SDS, 200 micrograms / mL sheared and denatured salmon sperm DNA, and 25% formamide at 42°C according to standard Southern blotting procedures, for probes at least 100 nucleotides in length. Finally, the carrier material is washed three times for 15 minutes each using 2X SSC, 0.2% SDS at 45°C.

[0054] Wild-type enzyme: The term "wild-type" α-amylase refers to an α-amylase expressed by a naturally occurring microorganism, such as a bacterium, yeast, or filamentous fungus, found in nature.

[0055] The term "washing performance" generally includes cleaning of hard surfaces, for example, in dishwashing, but also includes cleaning performance on textiles, such as laundry, and industrial and institutional cleaning. Improved washing performance can be measured by comparing the delta strengths as defined herein.

[0056] Variant designation conventions For purposes of the present invention, the polypeptide disclosed in SEQ ID NO: 1 is used to determine the corresponding amino acid residue in another α-amylase. Accordingly, all positions and specific substitutions mentioned refer to the numbering used in SEQ ID NO: 1. However, one skilled in the art will recognize that any other sequence disclosed herein may be used to determine the corresponding amino acid residue in another α-amylase polypeptide. The amino acid sequence of another α-amylase is aligned with the mature polypeptide disclosed in SEQ ID NO: 1, and based on the alignment, the amino acid position number corresponding to any amino acid residue in the mature polypeptide disclosed in SEQ ID NO: 1 is determined using the Needleman-Wunsch algorithm (Needleman and Wunsch, 1970, J. Mol. Biol. 48:443-453) as implemented in the Needle program in the EMBOSS package (EMBOSS: The European Molecular Biology Open Software Suite, Rice et al., 2000, Trends Genet. 16:276-277), preferably version 5.0.0 or higher. The parameters used are a gap opening penalty of 10, a gap extension penalty of 0.5, and the EBLOSUM62 (EMBOSS version of BLOSUM62) substitution matrix.

[0057] Identification of corresponding amino acid residues in other α-amylases can be performed using, but not limited to, MUSCLE (multiple sequence comparison by log prediction; version 3.5 or higher; Edgar, 2004, Nucleic Acids Research 32:1792-1797), MAFFT (version 6.857 or higher; Katoh and Kuma, 2002, Nucleic Acids Research 30:3059-3066; Katoh et al., 2005, Nucleic Acids Research 33:511-518; Katoh and Toh, 2007, Bioinformatics 23:372-374; Katoh et al., 2009, Methods in Molecular Biology 537:39-64; Katoh and Toh, 2010, Bioinformatics The degree of alignment can be determined by aligning multiple polypeptide sequences using several computer programs, including EMBOSS EMMA, which uses ClustalW (1.83 or higher; Thompson et al., 1994, Nucleic Acids Research 22:4673-4680), and EMBOSS EMMA, which uses ClustalW (1.83 or higher; Thompson et al., 1994, Nucleic Acids Research 22:4673-4680).

[0058] Other pairwise sequence comparison algorithms may be used when other α-amylases diverge from the polypeptide of SEQ ID NO: 1, such that traditional sequence-based comparisons fail to detect these relationships (Lindahl and Elofsson, 2000, J. Mol. Biol. 295:613-615). Greater sensitivity in sequence-based searches can be achieved using search programs that utilize probabilistic representations (profiles) of polypeptide families to search databases. For example, the PSI-BLAST program generates profiles through an iterative database search process and can detect distant homologs (Atschul et al., 1997, Nucleic Acids Res. 25:3389-3402). Even greater sensitivity can be achieved when a family or superfamily of polypeptides has more than one representation in a protein structure database. Programs such as GenTHREADER (Jones, 1999, J. Mol. Biol. 287:797-815; McGuffin and Jones, 2003, Bioinformatics 19:874-881) use information from a variety of sources (PSI-BLAST, secondary structure prediction, structural alignment profiles, and solvate potential) as input to neural networks that predict the structural fold of a query sequence. Similarly, the method of Gough et al., 2000, J. Mol. Biol. 313:903-919 can be used to align sequences of unknown structure with superfamily models present in the SCOP database. These alignments can then be used to generate homology models of the polypeptide, and such models can be assessed for accuracy using a variety of tools developed for that purpose.

[0059] For proteins of known structure, several tools and resources are available for searching and creating structural alignments. For example, the SCOP superfamily of proteins has been structurally aligned, and these alignments are accessible and downloadable. Various algorithms, such as distance alignment matrices (Holm and Sander, 1998, Proteins 33:88-96) or combinatorial extension (Shindyalov and Bourne, 1998, Protein Engineering 11:739-747), can be used to align two or more protein structures; implementations of these algorithms can then be used to query structural databases for the target structure to find potential structural homologs (e.g., Holm and Park, 2000, Bioinformatics 16:566-567).

[0060] For ease of reference when describing the α-amylase variants of the present invention, the following nomenclature is adopted: The generally accepted IUPAC one-letter or three-letter amino acid abbreviations are used.

[0061] Substitutions: For amino acid substitutions, the following nomenclature is used: original amino acid, position, substituted amino acid. Thus, for example, a substitution of threonine with alanine at position 226 is designated as "Thr226Ala" or "T226A." Multiple mutations are separated by adding a mark ("+"), e.g., "Gly205Arg+Ser411Phe" or "G205R+S411F" represent substitutions of glycine (G) with arginine (R) and serine (S) with phenylalanine (F) at positions 205 and 411, respectively.

[0062] Deletions: For amino acid deletions, use the following nomenclature: original amino acid, position, * Thus, the deletion of glycine at position 181 is expressed as "Ser181 *" or "S181 * Multiple deletions are separated by an addition sign ("+"), e.g., "Ser181 * +Thr182 * " or "S181 * +T182 * "

[0063] Insertions: For amino acid insertions, the following nomenclature is used: original amino acid, position, original amino acid, inserted amino acid. Thus, for example, an insertion of lysine after glycine at position 195 would be designated "Gly195GlyLys" or "G195GK." An insertion of multiple amino acids would be designated [original amino acid, position, original amino acid, inserted amino acid #1, inserted amino acid #2; etc.]. For example, an insertion of lysine and alanine after glycine at position 195 would be designated "Gly195GlyLysAla" or "G195GKA."

[0064] In such cases, the inserted amino acid residue is numbered by adding a lower case letter to the position number of the amino acid residue preceding the inserted amino acid residue. In the above example, the sequence could be:

[0065] [Table 1]

[0066] Multiple modifications: Variants containing multiple modifications are separated by adding an additional symbol ("+"), for example, "Arg170Tyr+Gly195Glu" or "R170Y+G195E" represent substitutions of arginine and glycine with tyrosine and glutamic acid at positions 170 and 195, respectively.

[0067] Differential modifications: When different modifications can be introduced at a position, the different modifications are separated by commas, for example, "Arg170Tyr,Glu" represents the substitution of arginine with tyrosine or glutamic acid at position 170. Thus, "Tyr167Gly,Ala+Arg170Gly,Ala" represents the following variant: It represents "Tyr167Gly+Arg170Gly", "Tyr167Gly+Arg170Ala", "Tyr167Ala+Arg170Gly", and "Tyr167Ala+Arg170Ala".

[0068] Detailed Description of the Invention variant The cleaning compositions of the present invention may be numbered as follows: 1, using SEQ ID NO: 1 for numbering: positions 3, 5, 7, 8, 16, 18, 25, 26, 29, 30, 35, 37, 40, 41, 43, 46, 50, 57, 60, 68, 71, 73, 74, 78, 81, 84, 86, 87, 90, 91, 93, 97, 98, 103, 105, 110, 111, 113, 114, 116, 117, 119, 120, 121, 125, 126, 128, 129, 131, 132, 133, 135, 137, 138, 139, 140, 141, 142, 143, 145, 146, 147, 149, 150, 151, 152, 153, 154, 155, 156, 157, 158, 159, 160, 161, 162, 163, 164, 165, 166, 167, 168, 169, 170, 171, 172, 173, 174, 175, 176, 177, 178, 179, 180, 181, 182, 183, 184, 185, 186, 187, 188, 189, 19 9, 161, 162, 165, 167, 169, 171, 176, 177, 179, 180, 181, 185, 186, 187, 195, 196, 197, 204, 206, 208, 211, 216, 218, 219, 225, 228, 230, 231, 235, 242, 243 3, 247, 250, 251, 252, 253, 254, 255, 257, 259, 260, 261, 262, 263, 265, 272, 273, 274, 279, 280, 281, 283, 284, 287, 292, 294, 296, 298, 299, 300, 303, 305 , 307, 310, 311, 313, 314, 315, 319, 320, 321, 322, 323, 324, 334, 337, 339, 340, 342, 344, 345, 346, 347, 350, 359, 360, 361, 362, 363, 364, 368, 372, 375 , 380, 382, ​​383, 384, 385, 387, 388, 391, 393, 395, 396, 397, 398, 401, 402, 403, 405, 406, 407, 410, 415, 416, 418, 421, 422, 423, 433, 434, 435, 438, 439, 446, 447, 449, 450, 452, 455, 457, 458, 460, 461, 462, 465, 466, 467, 469, 471, 473, 475, 476, 477, and 478, wherein the variants have a substitution at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, or a similar substitution at one or more positions corresponding to the polypeptide of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, or 17.at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% but less than 100% sequence identity, wherein the variant has α-amylase activity, and wherein the α-amylase variant has improved properties relative to the parent.

[0069] The variant has at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% but less than 100% sequence identity to the amino acid sequence of the parent α-amylase.

[0070] In another embodiment, the variant has at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, such as at least 96%, at least 97%, at least 98%, or at least 99% but less than 100% sequence identity to SEQ ID NO:1.

[0071] In another embodiment, the variant has at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, such as at least 96%, at least 97%, at least 98%, or at least 99% but less than 100% sequence identity to SEQ ID NO:2.

[0072] In another embodiment, the variant has at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, such as at least 96%, at least 97%, at least 98%, or at least 99% but less than 100% sequence identity to SEQ ID NO:3.

[0073] In another embodiment, the variant has at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, such as at least 96%, at least 97%, at least 98%, or at least 99% but less than 100% sequence identity to SEQ ID NO:4.

[0074] In another embodiment, the variant has at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, such as at least 96%, at least 97%, at least 98%, or at least 99% but less than 100% sequence identity to SEQ ID NO:5.

[0075] In another embodiment, the variant has at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, such as at least 96%, at least 97%, at least 98%, or at least 99% but less than 100% sequence identity to SEQ ID NO: 6.

[0076] In another embodiment, the variant has at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, such as at least 96%, at least 97%, at least 98%, or at least 99% but less than 100% sequence identity to SEQ ID NO: 7.

[0077] In another embodiment, the variant has at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, such as at least 96%, at least 97%, at least 98%, or at least 99% but less than 100% sequence identity to SEQ ID NO:8.

[0078] In another embodiment, the variant has at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, such as at least 96%, at least 97%, at least 98%, or at least 99% but less than 100% sequence identity to SEQ ID NO:9.

[0079] In another embodiment, the variant has at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, such as at least 96%, at least 97%, at least 98%, or at least 99% but less than 100% sequence identity to SEQ ID NO: 10.

[0080] In another embodiment, the variant has at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, such as at least 96%, at least 97%, at least 98%, or at least 99% but less than 100% sequence identity to SEQ ID NO: 12.

[0081] In another embodiment, the variant has at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, such as at least 96%, at least 97%, at least 98%, or at least 99% but less than 100% sequence identity to SEQ ID NO: 13.

[0082] In another embodiment, the variant has at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, such as at least 96%, at least 97%, at least 98%, or at least 99% but less than 100% sequence identity to SEQ ID NO: 14.

[0083] In another embodiment, the variant has at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, such as at least 96%, at least 97%, at least 98%, or at least 99% but less than 100% sequence identity to SEQ ID NO: 15.

[0084] In another embodiment, the variant has at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, such as at least 96%, at least 97%, at least 98%, or at least 99% but less than 100% sequence identity to SEQ ID NO: 16.

[0085] In another embodiment, the variant has at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, such as at least 96%, at least 97%, at least 98%, or at least 99% but less than 100% sequence identity to SEQ ID NO: 17.

[0086] In one aspect the number of modifications in a variant of the invention is 1 to 20, such as 1 to 10 or 1 to 5, such as 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10 modifications.

[0087] This variant is located at positions 3, 5, 7, 8, 16, 18, 25, 26, 29, 30, 35, 37, 40, 41, 43, 46, 50, 57, 60, 68, 71, 73, 74, 78, 81, 84, 86, 87, 90, 91, 93, 97, 98, 103, 105, 110, 111, 113, 114, 116, 117, 119, 120, 121, 125, 126, 128, 129, 131, 132, 133, 135, 137, 138, 139, 140, 141, 142, 143, 145, 146, 147, 149, 159, 160, 161, 162, 163, 164, 165, 166, 167, 168, 169, 170, 171, 172, 173, 174, 175, 176, 177, 178, 179, 180, 181, 182, 183, 184, 185, 186, 187, 188, 189, 190, 191, 192, 193, 194, 195, 196, 197, 198, 199, 200, 201, 202, 203, 2 61, 162, 165, 167, 169, 171, 176, 177, 179, 180, 181, 185, 186, 187, 195, 196, 197, 204, 206, 208, 211, 216, 218, 219, 225, 228, 230, 231, 235, 242, 243, 244 7, 250, 251, 252, 253, 254, 255, 257, 259, 260, 261, 262, 263, 265, 272, 273, 274, 279, 280, 281, 283, 284, 287, 292, 294, 296, 298, 299, 300, 303, 305, 307 , 310, 311, 313, 314, 315, 319, 320, 321, 322, 323, 324, 334, 337, 339, 340, 342, 344, 345, 346, 347, 350, 359, 360, 361, 362, 363, 364, 368, 372, 375, 380, 382, 383, 384, 385, 387, 388, 391, 393, 395, 396, 397, 398, 401, 402, 403, 405, 406, 407, 410, 415, 416, 418, 421, 422, 423, 433, 434, 435, 438, 439, 446, 447, 448, 450, 451, 452, 453, 454, 455, 456, 457, 458, 459, 460, 461, 462, 463, 464, 465, 466, 467, and 478, and further, the variant may comprise a substitution at two or more positions corresponding to any of SEQ ID NOs: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, or 17, wherein the substitution is at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%,having at least 96%, at least 97%, at least 98%, or at least 99% but less than 100% sequence identity.

[0088] This variant occurs at positions 3, 5, 7, 8, 16, 18, 25, 26, 29, 30, 35, 37, 40, 41, 43, 46, 50, 57, 60, 68, 71, 73, 74, 78, 81, 84, 86, 87, 90, 91, 93, 97, 98, 103, 105, 110, 111, 113, 114, 116, 117, 119, 120, 121, 125, 126, 128, 129, 131, 132, 133, 135, 137, 138, 139, 140, 141, 142, and 143. , 145, 146, 147, 149, 159, 161, 162, 165, 167, 169, 171, 176, 177, 179, 180, 181, 185, 186, 187, 195, 196, 197, 204, 206, 208, 211, 216, 218, 219, 225, 228, 230, 231, 235, 242, 243, 247, 250, 251, 252, 253, 254, 255, 257, 259, 260, 261, 262, 263, 265, 272, 273, 274, 279, 280, 281, 283, 284, 287, 292, 294, 296, 298, 299, 300, 303, 305, 307, 310, 311, 313, 314, 315, 319, 320, 321, 322, 323, 324, 334, 337, 339, 340, 342, 344, 345, 346, 347, 350, 359, 360, 361, 362, 363, 364, 368, 372, 375, 380, 382, ​​383, 384, 385, 387, 388, 389, 390, 391, 392, 393, 394, 395, 396, 397, 398, 399, 399, 400, 401, 402, 403, 404, 405, 406, 407, 408, 409, 410, 411, 412, 413, 414, 415, 416, 417, 418, 419, 420, 421, 422, 423, 424, 425, 426, 427, 428, 429, 430, 431, 432, 433, 434, 435, 436, 460, 461, 462, 465, 466, 467, 469, 471, 473, 475, 476, 477, and 478.

[0089] This variant occurs at positions 3, 5, 7, 8, 16, 18, 25, 26, 29, 30, 35, 37, 40, 41, 43, 46, 50, 57, 60, 68, 71, 73, 74, 78, 81, 84, 86, 87, 90, 91, 93, 97, 98, 103, 105, 110, 111, 113, 114, 116, 117, 119, 120, 121, 125, 126, 128, 129, 131, 132, 133, 135, 137, 138, 139, 140, 141, 142, and 143. , 145, 146, 147, 149, 159, 161, 162, 165, 167, 169, 171, 176, 177, 179, 180, 181, 185, 186, 187, 195, 196, 197, 204, 206, 208, 211, 216, 218, 219, 225, 228, 230, 231, 235, 242, 243, 247, 250, 251, 252, 253, 254, 255, 257, 259, 260, 261, 262, 263, 265, 272, 273, 274, 279, 280, 281, 283, 284, 287, 292, 294, 296, 298, 299, 300, 303, 305, 307, 310, 311, 313, 314, 315, 319, 320, 321, 322, 323, 324, 334, 337, 339, 340, 342, 344, 345, 346, 347, 350, 359, 360, 361, 362, 363, 364, 368, 372, 375, 380, 382, ​​383, 384, 385, 387, 388, 389, 390, 391, 392, 393, 394, 395, 396, 397, 398, 399, 399, 400, 401, 402, 403, 404, 405, 406, 407, 408, 409, 410, 411, 412, 413, 414, 415, 416, 417, 418, 419, 420, 421, 422, 423, 424, 425, 426, 427, 428, 429, 430, 431, 432, 433, 434, 435, 436, 460, 461, 462, 465, 466, 467, 469, 471, 473, 475, 476, 477, and 478.

[0090] This variant occurs at positions 3, 5, 7, 8, 16, 18, 25, 26, 29, 30, 35, 37, 40, 41, 43, 46, 50, 57, 60, 68, 71, 73, 74, 78, 81, 84, 86, 87, 90, 91, 93, 97, 98, 103, 105, 110, 111, 113, 114, 116, 117, 119, 120, 121, 125, 126, 128, 129, 131, 132, 133, 135, 137, 138, 139, 140, 141, 142, 143, and 145. , 146, 147, 149, 159, 161, 162, 165, 167, 169, 171, 176, 177, 179, 180, 181, 185, 186, 187, 195, 196, 197, 204, 206, 208, 211, 216, 218, 219, 225, 228, 230, 231, 235, 242, 243, 247, 250, 251, 252, 253, 254, 255, 257, 259, 260, 261, 262, 263, 265, 272, 273, 274, 279, 280, 281, 283, 284, 287, 292, 294, 296, 298, 299, 300, 303, 305, 307, 310, 311, 313, 314, 315, 319, 320, 321, 322, 323, 324, 334, 337, 339, 340, 342, 344, 345, 346, 347, 350, 359, 360, 361, 362, 363, 364, 368, 372, 375, 380, 382, ​​383, 384, 385, 387, 388, 391, 393, 394, 395, 396, 397, 398, 399, 399, 400, 401, 402, 403, 404, 405, 406, 407, 408, 409, 410, 411, 412, 413, 414, 415, 416, 417, 418, 419, 420, 421, 422, 423, 424, 425, 426, 427, 428, 429, 430, 431, 432, 433, 434, 435, 436, 437, 438, 439, 440, 441, 460, 461, 462, 465, 466, 467, 469, 471, 473, 475, 476, 477, and 478.

[0091] This variant occurs at positions 3, 5, 7, 8, 16, 18, 25, 26, 29, 30, 35, 37, 40, 41, 43, 46, 50, 57, 60, 68, 71, 73, 74, 78, 81, 84, 86, 87, 90, 91, 93, 97, 98, 103, 105, 110, 111, 113, 114, 116, 117, 119, 120, 121, 125, 126, 128, 129, 131, 132, 133, 135, 137, 138, 139, 140, 141, 142, and 143. , 145, 146, 147, 149, 159, 161, 162, 165, 167, 169, 171, 176, 177, 179, 180, 181, 185, 186, 187, 195, 196, 197, 204, 206, 208, 211, 216, 218, 219, 225, 228, 230, 231, 235, 242, 243, 247, 250, 251, 252, 253, 254, 255, 257, 259, 260, 261, 262, 263, 265, 272, 273, 274, 275, 276, 277, 278, 279, 280, 281, 282, 283, 284, 285, 286, 287, 288, 289, 290, 291, 292, 293, 300, 301, 302, 303, 304, 305, 306, 307, 308, 309, 310, 311, 312, 313, 314, 315, 316, 317, 318, 319, 320, 321, 322, 323, 324, 325, 326, 327, 328, 329 74, 279, 280, 281, 283, 284, 287, 292, 294, 296, 298, 299, 300, 303, 305, 307, 310, 311, 313, 314, 315, 319, 320, 321, 322, 323, 324, 334, 337, 339, 340, 342, 344, 345, 346, 347, 350, 359, 360, 361, 362, 363, 364, 368, 372, 375, 380, 382, ​​383, 384, 385, 387, 388 460, 461, 462, 465, 466, 467, 469, 471, 473, 475, 476, 477, and 478, up to 11 or more positions, corresponding to:

[0092] Preferred substitutions are, using SEQ ID NO: 1 for numbering, at positions N3A, N3C, N3D, N3E, N3F, N3G, N3H, N3L, N3P, N3Q, N3S, N3T, N3V, T5E, G7E, G7F, G7H, G7K, G7L, G7P, G7R, G7S, G7T, G7V, G7W, T8S, H16D, H16R, P18D, P18L, P18N, N25T, R26Y, D29N, D30F, R35P, R37A, R37N, T40A, T40C, T40D, T40E, T40G, T40H, T40I, T40K, T40N, T40V, T40W, A 41C, A41D, A41E, A41H, A41I, A41K, A41N, A41R, A41T, A41V, A41Y, W43E, P46 A, G50M, G50P, G50Q, G57A, G57S, A60E, A60S, A60V, E68F, E68Y, Q71F, G73D, G73H, G73I, G73L, G73Q, G73S, G73T, G73W, T74F, K78S, K78T, T81M, Q84I, Q8 4W, Q84Y, E86K, E86S, S87G, S87L, S87P, H90P, A91Q, A91V, A91W, K93V, V97C, V97H, Q98E, Q98F, Q98G, Q98I, Q98K, Q98L, Q98M, Q98R, Q98T, Q98Y, V103C, V 103G, V103S, M105F, G110K, G110R, A111E, A111Q, A113T, T114M, T114V, N11 6C, N116Y, V117F, A119H, A119L, A119Y, V120E, E121I, N125S, N126I, N128E , Q129T, I131Y, S132A, S132D, S132H, S132I, S132K, S132L, S132P, S132R, S1 32T, S132V, S132Y, G133E, G133L, G133T, G133V, Y135C, Y135D, Y135E, Y135 P, I137D, I137N, I137Q, I137W, I137Y, E138S, E138V, A139L, W140A, W140D, W 140M, W140N, W140P, W140S, W140T, W140V, T141G, K142E, K142N, F143H, F14 3I, F143M, F145G, F145H, F145I, F145L, F145S, P146A, P146H, P146N, G147D,G149F、G149K、W159A、W159E、H161W、F162T、V165P、W167D、W167G、W167P、Q169L、Q169P、Q169V、Q169Y、R171P、R176E、R176Q、R176V、R176Y、I177H、I177P、K179A、K179M、K179V、F180D、F180G、R181D、R181E、R181G、R181M、R181Y、K185T、K185V、K185W、A186D、W187T、N195Y、G196D、N197V、N197Y、A204F、A204H、A204L、V206N、M208D、P211C、P211M、E216L、R218Q、R219V、T225D、T225F、T225H、T225K、T225L、T225N、T225R、T225Y、L228V、L230C、L230F、D231G、D231T、D231V、I235A、K242L、Y243D、Y243M、Y243N、R247G、R247P、R247S、L250S、L250Y、T251D、H252P、V253F、V253S、R254D、R254F、R254H、R254P、R254T、R254V、R254W、N255C、N255F、N255P、T257A、T257C、T257D、T257E、T257G、T257P、T257Q、T257S、T257W、T257Y、K259P、K259V、K259W、K259Y、E260V、M261D、M261F、M261P、M261R、M261S、M261W、F262A、F262C、F262E、F262H、F262I、F262K、F262L、F262N、F262P、F262Q、F262R、F262S、F262T、F262V、F262W、A263E、A263H、A263W、A265Q、L272W、G273Q、A274D、L279I、N280P、K281H、K281I、K281S、N283P、W284A、W284D、W284E、W284G、W284I、W284K、W284M、W284N、W284P、W284Q、W284S、W284T、W284V、W284Y、S287I、S287K、S287L、S287P、S287R、S287T、S287V、S287Y、P292L、P292M、P292Y、H294N、H294S、N296T、Y298D、Y298E, N299T, A300G, S303A, S303C, S303D, S303E, S303F, S303G, S303H, S303L, S303M, S303N, S303Q, S303T, S303V, S303Y, G305E, Y307S, A310D, A310 N、K311A、K311C、L313Q、N314P、N314S、G315E、G315M、G315V、Q319C、Q319E、 K320S、K320W、H321F、H321K、H321L、H321R、H321T、H321V、H321W、P322C、P32 2E, P322H, P322T, P322Y, M323A, M323C, M323D, M323E, M323G, M323H, M323L, M323N, M323P, M323R, M323S, M323T, M323W, M323Y, H324A, H324I, H324P, H 324T, H324W, H324Y, S334V, G337V, L340M, S342A, S342G, S342T, V344A, V344C, V344I, Q345N, E346A, W347A, W347E, W347F, W347G, W347H, W347K, W347N W347P, W347R, P350A, P350C, P350N, P350T, R359C, R359F, R359G, R359M, R359S, R359W, E360D, E360H, E360P, E360T, E360W, Q361D, Q361H, Q361I, Q361 K、Q361L、Q361M、Q361N、Q361P、Q361R、Q361S、Q361T、Q361V、Q361W、Q361Y、 G362M、Y363A、Y363E、Y363F、Y363G、Y363I、Y363L、Y363Q、Y363R、Y363S、Y36 3T、Y363V、Y363W、P364C、Y368C、Y368Q、Y368R、Y368T、Y372C、Y372G、Y372N 、Y372T、P375Q、P375T、P380F、M382I、K383C、K383L、K383P、K383T、A384D、A 384N, K385N, K385V, D387C, D387P, D387R, D387W, P388I, E391M, R393K, R393M, N395C, N395D, N395K, N395V, N395W, F396E, F396G, F396T, A397V, Y398EQ401H, H402S, D403A, D403E, D403T, F405C, F405N, F405S, F405T, D406V , H407C, H407Q, H407T, I410H, I410M, I410R, I410W, R415C, R415L, R415 M, R415Y, E416D, E416F, E416L, E416N, N418A, N418P, N418V, H421A, H42 1G, H421L, H421P, H421Q, H421Y, P422D, P422G, P422I, P422N, P422Q, P4 22S, N423G, N423H, N423I, N423L, N423M, N423P, N423Q, N423V, N423W, N 423Y, G433A, G433N, G433S, P434I, P434L, G435C, G435L, G435M, G435T, G435V, K438A, K438I, K438Q, K438S, K438V, W439A, W439D, W439I, W439K , W439L, W439N, W439S, W439T, W439V, W439Y, K446A, K446F, A447D, A447 Y, Q449T, V450D, V450N, H452L, T455C, T455M, T455V, N457D, N457E, N45 7I, N457L, N457V, K458D, K458L, G460M, G460V, T461F, T461I, T461Q, T4 61V, V462C, N465A, N465C, N465E, N465G, N465H, N465L, N465P, N465T, A 466F, A466I, A466Y, D467I, W469A, W469E, W469H, W469L, W469N, W469Q, and wherein the variant is selected from substitutions at positions corresponding to W469R, W469S, W469Y, N471A, N471D, N471M, N471W, S473N, N475A, N475C, N475D, N475E, N475F, N475G, N475H, N475I, N475L, N475M, N475P, N475Q, N475S, N475T, N475V, N475W, G476F, G476L, G477A, ​​G477H, G477Q, G477S, G477W, S478A, S478F, and S478Q.or at least 60%, 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%, or at least 99% but less than 100% sequence identity to the polypeptide of SEQ ID NO:1 or SEQ ID NO:2 or SEQ ID NO:3 or SEQ ID NO:4 or SEQ ID NO:5 or SEQ ID NO:6 or SEQ ID NO:7 or SEQ ID NO:8 or SEQ ID NO:9 or SEQ ID NO:10 or SEQ ID NO:11 or SEQ ID NO:12 or SEQ ID NO:13 or SEQ ID NO:14 or SEQ ID NO:15 or SEQ ID NO:16 or SEQ ID NO:17.

[0093] In one aspect, the variants are located at positions N3A, N3C, N3D, N3E, N3F, N3G, N3H, N3L, N3P, N3Q, N3S, N3T, N3V, T5E, G7E, G7F, G7H, G7K, G7L, G7P, G7R, G7S, G7T, G7V, G7W, T8S, H16D, H16R, P18D, P18L, P18N, N25T, R26Y, D29N, D30F, R35P, R37A, R37N, T40A, T40C, T40D, T40E, T40G, T40H, T40I, T40K, T40N, T40V, T 40W, A41C, A41D, A41E, A41H, A41I, A41K, A41N, A41R, A41T, A41V, A41Y, W43 E, P46A, G50M, G50P, G50Q, G57A, G57S, A60E, A60S, A60V, E68F, E68Y, Q71F, G73D, G73H, G73I, G73L, G73Q, G73S, G73T, G73W, T74F, K78S, K78T, T81M, Q8 4I, Q84W, Q84Y, E86K, E86S, S87G, S87L, S87P, H90P, A91Q, A91V, A91W, K93V, V97C, V97H, Q98E, Q98F, Q98G, Q98I, Q98K, Q98L, Q98M, Q98R, Q98T, Q98Y, V1 03C, V103G, V103S, M105F, G110K, G110R, A111E, A111Q, A113T, T114M, T114 V, N116C, N116Y, V117F, A119H, A119L, A119Y, V120E, E121I, N125S, N126I, N128E, Q129T, I131Y, S132A, S132D, S132H, S132I, S132K, S132L, S132P, S13 2R, S132T, S132V, S132Y, G133E, G133L, G133T, G133V, Y135C, Y135D, Y135E , Y135P, I137D, I137N, I137Q, I137W, I137Y, E138S, E138V, A139L, W140A, W 140D, W140M, W140N, W140P, W140S, W140T, W140V, T141G, K142E, K142N, F14 3H, F143I, F143M, F145G, F145H, F145I, F145L, F145S, P146A, P146H, P146N,G147D、G149F、G149K、W159A、W159E、H161W、F162T、V165P、W167D、W167G、W167P、Q169L、Q169P、Q169V、Q169Y、R171P、R176E、R176Q、R176V、R176Y、I177H、I177P、K179A、K179M、K179V、F180D、F180G、R181D、R181E、R181G、R181M、R181Y、K185T、K185V、K185W、A186D、W187T、N195Y、G196D、N197V、N197Y、A204F、A204H、A204L、V206N、M208D、P211C、P211M、E216L、R218Q、R219V、T225D、T225F、T225H、T225K、T225L、T225N、T225R、T225Y、L228V、L230C、L230F、D231G、D231T、D231V、I235A、K242L、Y243D、Y243M、Y243N、R247G、R247P、R247S、L250S、L250Y、T251D、H252P、V253F、V253S、R254D、R254F、R254H、R254P、R254T、R254V、R254W、N255C、N255F、N255P、T257A、T257C、T257D、T257E、T257G、T257P、T257Q、T257S、T257W、T257Y、K259P、K259V、K259W、K259Y、E260V、M261D、M261F、M261P、M261R、M261S、M261W、F262A、F262C、F262E、F262H、F262I、F262K、F262L、F262N、F262P、F262Q、F262R、F262S、F262T、F262V、F262W、A263E、A263H、A263W、A265Q、L272W、G273Q、A274D、L279I、N280P、K281H、K281I、K281S、N283P、W284A、W284D、W284E、W284G、W284I、W284K、W284M、W284N、W284P、W284Q、W284S、W284T、W284V、W284Y、S287I、S287K、S287L、S287P、S287R、S287T、S287V、S287Y、P292L、P292M、P292Y、H294N、H294S、N296T、Y298D, Y298E, N299T, A300G, S303A, S303C, S303D, S303E, S303F, S303G, S303H, S303L, S303M, S303N, S303Q, S303T, S303V, S303Y, G305E, Y307S, A310 D, A310N, K311A, K311C, L313Q, N314P, N314S, G315E, G315M, G315V, Q319C, Q319E, K320S, K320W, H321F, H321K, H321L, H321R, H321T, H321V, H321W, P32 2C, P322E, P322H, P322T, P322Y, M323A, M323C, M323D, M323E, M323G, M323H, M323L, M323N, M323P, M323R, M323S, M323T, M323W, M323Y, H324A, H324I, H 324P, H324T, H324W, H324Y, S334V, G337V, L340M, S342A, S342G, S342T, V344A, V344C, V344I, Q345N, E346A, W347A, W347E, W347F, W347G, W347H, W347K W347N, W347P, W347R, P350A, P350C, P350N, P350T, R359C, R359F, R359G, R359M, R359S, R359W, E360D, E360H, E360P, E360T, E360W, Q361D, Q361H, Q361 I、Q361K、Q361L、Q361M、Q361N、Q361P、Q361R、Q361S、Q361T、Q361V、Q361W、 Q361Y、G362M、Y363A、Y363E、Y363F、Y363G、Y363I、Y363L、Y363Q、Y363R、Y36 3S, Y363T, Y363V, Y363W, P364C, Y368C, Y368Q, Y368R, Y368T, Y372C, Y372G, Y372N, Y372T, P375Q, P375T, P380F, M382I, K383C, K383L, K383P, K383T, A 384D, A384N, K385N, K385V, D387C, D387P, D387R, D387W, P388I, E391M, R393K, R393M, N395C, N395D, N395K, N395V, N395W, F396E, F396G, F396T, A397VY398E, Q401H, H402S, D403A, D403E, D403T, F405C, F405N, F405S, F405T, D4 06V, H407C, H407Q, H407T, I410H, I410M, I410R, I410W, R415C, R415L, R415M , R415Y, E416D, E416F, E416L, E416N, N418A, N418P, N418V, H421A, H421G, H 421L, H421P, H421Q, H421Y, P422D, P422G, P422I, P422N, P422Q, P422S, N423 G, N423H, N423I, N423L, N423M, N423P, N423Q, N423V, N423W, N423Y, G433A, G433N, G433S, P434I, P434L, G435C, G435L, G435M, G435T, G435V, K438A, K43 8I, K438Q, K438S, K438V, W439A, W439D, W439I, W439K, W439L, W439N, W439S , W439T, W439V, W439Y, K446A, K446F, A447D, A447Y, Q449T, V450D, V450N, H4 52L, T455C, T455M, T455V, N457D, N457E, N457I, N457L, N457V, K458D, K458 L, G460M, G460V, T461F, T461I, T461Q, T461V, V462C, N465A, N465C, N465E, N 465G, N465H, N465L, N465P, N465T, A466F, A466I, A466Y, D467I, W469A, W46 9E, W469H, W469L, W469N, W469Q, W469R, W469S, W469Y, N471A, N471D, N471M, and the variant comprises one or more of these substitutions at positions corresponding to N471W, S473N, N475A, N475C, N475D, N475E, N475F, N475G, N475H, N475I, N475L, N475M, N475P, N475Q, N475S, N475T, N475V, N475W, G476F, G476L, G477A, ​​G477H, G477Q, G477S, G477W, S478A, S478F, and S478Q, and further the variant has a nucleotide sequence at least 60%, 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%, or at least 99% but less than 100% sequence identity.

[0094] In one aspect, the variants are located at positions N3A, N3C, N3D, N3E, N3F, N3G, N3H, N3L, N3P, N3Q, N3S, N3T, N3V, T5E, G7E, G7F, G7H, G7K, G7L, G7P, G7R, G7S, G7T, G7V, G7W, T8S, H16D, H16R, P18D, P18L, P18N, N25T, R26Y, D29N, D30F, R35P, R37A, R37N, T40A, T40C, T40D, T40E, T40G, T40H, T40I, T40K, T40N, T40V, T 40W, A41C, A41D, A41E, A41H, A41I, A41K, A41N, A41R, A41T, A41V, A41Y, W43 E, P46A, G50M, G50P, G50Q, G57A, G57S, A60E, A60S, A60V, E68F, E68Y, Q71F, G73D, G73H, G73I, G73L, G73Q, G73S, G73T, G73W, T74F, K78S, K78T, T81M, Q8 4I, Q84W, Q84Y, E86K, E86S, S87G, S87L, S87P, H90P, A91Q, A91V, A91W, K93V, V97C, V97H, Q98E, Q98F, Q98G, Q98I, Q98K, Q98L, Q98M, Q98R, Q98T, Q98Y, V1 03C, V103G, V103S, M105F, G110K, G110R, A111E, A111Q, A113T, T114M, T114 V, N116C, N116Y, V117F, A119H, A119L, A119Y, V120E, E121I, N125S, N126I, N128E, Q129T, I131Y, S132A, S132D, S132H, S132I, S132K, S132L, S132P, S13 2R, S132T, S132V, S132Y, G133E, G133L, G133T, G133V, Y135C, Y135D, Y135E , Y135P, I137D, I137N, I137Q, I137W, I137Y, E138S, E138V, A139L, W140A, W 140D, W140M, W140N, W140P, W140S, W140T, W140V, T141G, K142E, K142N, F14 3H, F143I, F143M, F145G, F145H, F145I, F145L, F145S, P146A, P146H, P146N,G147D、G149F、G149K、W159A、W159E、H161W、F162T、V165P、W167D、W167G、W167P、Q169L、Q169P、Q169V、Q169Y、R171P、R176E、R176Q、R176V、R176Y、I177H、I177P、K179A、K179M、K179V、F180D、F180G、R181D、R181E、R181G、R181M、R181Y、K185T、K185V、K185W、A186D、W187T、N195Y、G196D、N197V、N197Y、A204F、A204H、A204L、V206N、M208D、P211C、P211M、E216L、R218Q、R219V、T225D、T225F、T225H、T225K、T225L、T225N、T225R、T225Y、L228V、L230C、L230F、D231G、D231T、D231V、I235A、K242L、Y243D、Y243M、Y243N、R247G、R247P、R247S、L250S、L250Y、T251D、H252P、V253F、V253S、R254D、R254F、R254H、R254P、R254T、R254V、R254W、N255C、N255F、N255P、T257A、T257C、T257D、T257E、T257G、T257P、T257Q、T257S、T257W、T257Y、K259P、K259V、K259W、K259Y、E260V、M261D、M261F、M261P、M261R、M261S、M261W、F262A、F262C、F262E、F262H、F262I、F262K、F262L、F262N、F262P、F262Q、F262R、F262S、F262T、F262V、F262W、A263E、A263H、A263W、A265Q、L272W、G273Q、A274D、L279I、N280P、K281H、K281I、K281S、N283P、W284A、W284D、W284E、W284G、W284I、W284K、W284M、W284N、W284P、W284Q、W284S、W284T、W284V、W284Y、S287I、S287K、S287L、S287P、S287R、S287T、S287V、S287Y、P292L、P292M、P292Y、H294N、H294S、N296T、Y298D, Y298E, N299T, A300G, S303A, S303C, S303D, S303E, S303F, S303G, S303H, S303L, S303M, S303N, S303Q, S303T, S303V, S303Y, G305E, Y307S, A310 D, A310N, K311A, K311C, L313Q, N314P, N314S, G315E, G315M, G315V, Q319C, Q319E, K320S, K320W, H321F, H321K, H321L, H321R, H321T, H321V, H321W, P32 2C, P322E, P322H, P322T, P322Y, M323A, M323C, M323D, M323E, M323G, M323H, M323L, M323N, M323P, M323R, M323S, M323T, M323W, M323Y, H324A, H324I, H 324P, H324T, H324W, H324Y, S334V, G337V, L340M, S342A, S342G, S342T, V344A, V344C, V344I, Q345N, E346A, W347A, W347E, W347F, W347G, W347H, W347K W347N, W347P, W347R, P350A, P350C, P350N, P350T, R359C, R359F, R359G, R359M, R359S, R359W, E360D, E360H, E360P, E360T, E360W, Q361D, Q361H, Q361 I、Q361K、Q361L、Q361M、Q361N、Q361P、Q361R、Q361S、Q361T、Q361V、Q361W、 Q361Y、G362M、Y363A、Y363E、Y363F、Y363G、Y363I、Y363L、Y363Q、Y363R、Y36 3S, Y363T, Y363V, Y363W, P364C, Y368C, Y368Q, Y368R, Y368T, Y372C, Y372G, Y372N, Y372T, P375Q, P375T, P380F, M382I, K383C, K383L, K383P, K383T, A 384D, A384N, K385N, K385V, D387C, D387P, D387R, D387W, P388I, E391M, R393K, R393M, N395C, N395D, N395K, N395V, N395W, F396E, F396G, F396T, A397VY398E, Q401H, H402S, D403A, D403E, D403T, F405C, F405N, F405S, F405T, D4 06V, H407C, H407Q, H407T, I410H, I410M, I410R, I410W, R415C, R415L, R415M , R415Y, E416D, E416F, E416L, E416N, N418A, N418P, N418V, H421A, H421G, H 421L, H421P, H421Q, H421Y, P422D, P422G, P422I, P422N, P422Q, P422S, N423 G, N423H, N423I, N423L, N423M, N423P, N423Q, N423V, N423W, N423Y, G433A, G433N, G433S, P434I, P434L, G435C, G435L, G435M, G435T, G435V, K438A, K43 8I, K438Q, K438S, K438V, W439A, W439D, W439I, W439K, W439L, W439N, W439S , W439T, W439V, W439Y, K446A, K446F, A447D, A447Y, Q449T, V450D, V450N, H4 52L, T455C, T455M, T455V, N457D, N457E, N457I, N457L, N457V, K458D, K458 L, G460M, G460V, T461F, T461I, T461Q, T461V, V462C, N465A, N465C, N465E, N 465G, N465H, N465L, N465P, N465T, A466F, A466I, A466Y, D467I, W469A, W46 9E, W469H, W469L, W469N, W469Q, W469R, W469S, W469Y, N471A, N471D, N471M, and the variant comprises one or more of these substitutions at positions corresponding to N471W, S473N, N475A, N475C, N475D, N475E, N475F, N475G, N475H, N475I, N475L, N475M, N475P, N475Q, N475S, N475T, N475V, N475W, G476F, G476L, G477A, ​​G477H, G477Q, G477S, G477W, S478A, S478F, and S478Q, and further the variant has a nucleotide sequence at least 60%, 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%, or at least 99% but less than 100% sequence identity.

[0095] In one aspect, the variants are located at positions N3A, N3C, N3D, N3E, N3F, N3G, N3H, N3L, N3P, N3Q, N3S, N3T, N3V, T5E, G7E, G7F, G7H, G7K, G7L, G7P, G7R, G7S, G7T, G7V, G7W, T8S, H16D, H16R, P18D, P18L, P18N, N25T, R26Y, D29N, D30F, R35P, R37A, R37N, T40A, T40C, T40D, T40E, T40G, T40H, T40I, T40K, T40N, T40V, T 40W, A41C, A41D, A41E, A41H, A41I, A41K, A41N, A41R, A41T, A41V, A41Y, W43 E, P46A, G50M, G50P, G50Q, G57A, G57S, A60E, A60S, A60V, E68F, E68Y, Q71F, G73D, G73H, G73I, G73L, G73Q, G73S, G73T, G73W, T74F, K78S, K78T, T81M, Q8 4I, Q84W, Q84Y, E86K, E86S, S87G, S87L, S87P, H90P, A91Q, A91V, A91W, K93V, V97C, V97H, Q98E, Q98F, Q98G, Q98I, Q98K, Q98L, Q98M, Q98R, Q98T, Q98Y, V1 03C, V103G, V103S, M105F, G110K, G110R, A111E, A111Q, A113T, T114M, T114 V, N116C, N116Y, V117F, A119H, A119L, A119Y, V120E, E121I, N125S, N126I, N128E, Q129T, I131Y, S132A, S132D, S132H, S132I, S132K, S132L, S132P, S13 2R, S132T, S132V, S132Y, G133E, G133L, G133T, G133V, Y135C, Y135D, Y135E , Y135P, I137D, I137N, I137Q, I137W, I137Y, E138S, E138V, A139L, W140A, W 140D, W140M, W140N, W140P, W140S, W140T, W140V, T141G, K142E, K142N, F14 3H, F143I, F143M, F145G, F145H, F145I, F145L, F145S, P146A, P146H, P146N,G147D、G149F、G149K、W159A、W159E、H161W、F162T、V165P、W167D、W167G、W167P、Q169L、Q169P、Q169V、Q169Y、R171P、R176E、R176Q、R176V、R176Y、I177H、I177P、K179A、K179M、K179V、F180D、F180G、R181D、R181E、R181G、R181M、R181Y、K185T、K185V、K185W、A186D、W187T、N195Y、G196D、N197V、N197Y、A204F、A204H、A204L、V206N、M208D、P211C、P211M、E216L、R218Q、R219V、T225D、T225F、T225H、T225K、T225L、T225N、T225R、T225Y、L228V、L230C、L230F、D231G、D231T、D231V、I235A、K242L、Y243D、Y243M、Y243N、R247G、R247P、R247S、L250S、L250Y、T251D、H252P、V253F、V253S、R254D、R254F、R254H、R254P、R254T、R254V、R254W、N255C、N255F、N255P、T257A、T257C、T257D、T257E、T257G、T257P、T257Q、T257S、T257W、T257Y、K259P、K259V、K259W、K259Y、E260V、M261D、M261F、M261P、M261R、M261S、M261W、F262A、F262C、F262E、F262H、F262I、F262K、F262L、F262N、F262P、F262Q、F262R、F262S、F262T、F262V、F262W、A263E、A263H、A263W、A265Q、L272W、G273Q、A274D、L279I、N280P、K281H、K281I、K281S、N283P、W284A、W284D、W284E、W284G、W284I、W284K、W284M、W284N、W284P、W284Q、W284S、W284T、W284V、W284Y、S287I、S287K、S287L、S287P、S287R、S287T、S287V、S287Y、P292L、P292M、P292Y、H294N、H294S、N296T、Y298D, Y298E, N299T, A300G, S303A, S303C, S303D, S303E, S303F, S303G, S303H, S303L, S303M, S303N, S303Q, S303T, S303V, S303Y, G305E, Y307S, A310 D, A310N, K311A, K311C, L313Q, N314P, N314S, G315E, G315M, G315V, Q319C, Q319E, K320S, K320W, H321F, H321K, H321L, H321R, H321T, H321V, H321W, P32 2C, P322E, P322H, P322T, P322Y, M323A, M323C, M323D, M323E, M323G, M323H, M323L, M323N, M323P, M323R, M323S, M323T, M323W, M323Y, H324A, H324I, H 324P, H324T, H324W, H324Y, S334V, G337V, L340M, S342A, S342G, S342T, V344A, V344C, V344I, Q345N, E346A, W347A, W347E, W347F, W347G, W347H, W347K W347N, W347P, W347R, P350A, P350C, P350N, P350T, R359C, R359F, R359G, R359M, R359S, R359W, E360D, E360H, E360P, E360T, E360W, Q361D, Q361H, Q361 I、Q361K、Q361L、Q361M、Q361N、Q361P、Q361R、Q361S、Q361T、Q361V、Q361W、 Q361Y、G362M、Y363A、Y363E、Y363F、Y363G、Y363I、Y363L、Y363Q、Y363R、Y36 3S, Y363T, Y363V, Y363W, P364C, Y368C, Y368Q, Y368R, Y368T, Y372C, Y372G, Y372N, Y372T, P375Q, P375T, P380F, M382I, K383C, K383L, K383P, K383T, A 384D, A384N, K385N, K385V, D387C, D387P, D387R, D387W, P388I, E391M, R393K, R393M, N395C, N395D, N395K, N395V, N395W, F396E, F396G, F396T, A397VY398E, Q401H, H402S, D403A, D403E, D403T, F405C, F405N, F405S, F405T, D4 06V, H407C, H407Q, H407T, I410H, I410M, I410R, I410W, R415C, R415L, R415 M, R415Y, E416D, E416F, E416L, E416N, N418A, N418P, N418V, H421A, H421G, H421L, H421P, H421Q, H421Y, P422D, P422G, P422I, P422N, P422Q, P422S, N42 3G, N423H, N423I, N423L, N423M, N423P, N423Q, N423V, N423W, N423Y, G433A , G433N, G433S, P434I, P434L, G435C, G435L, G435M, G435T, G435V, K438A, K4 38I, K438Q, K438S, K438V, W439A, W439D, W439I, W439K, W439L, W439N, W439 S, W439T, W439V, W439Y, K446A, K446F, A447D, A447Y, Q449T, V450D, V450N, H 452L, T455C, T455M, T455V, N457D, N457E, N457I, N457L, N457V, K458D, K45 8L, G460M, G460V, T461F, T461I, T461Q, T461V, V462C, N465A, N465C, N465E, N465G, N465H, N465L, N465P, N465T, A466F, A466I, A466Y, D467I, W469A, W4 69E, W469H, W469L, W469N, W469Q, W469R, W469S, W469Y, N471A, N471D, N471M , N471W, S473N, N475A, N475C, N475D, N475E, N475F, N475G, N475H, N475I, N475L, N475M, N475P, N475Q, N475S, N475T, N475V, N475W, G476F, G476L, G477A, ​​G477H, G477Q, G477S, G477W, S478A, S478F, and S478Q, and further the variant comprises one or more of these substitutions at positions corresponding to at least 60%, 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%, or at least 99% but less than 100% sequence identity.

[0096] In one aspect, the variants are located at positions N3A, N3C, N3D, N3E, N3F, N3G, N3H, N3L, N3P, N3Q, N3S, N3T, N3V, T5E, G7E, G7F, G7H, G7K, G7L, G7P, G7R, G7S, G7T, G7V, G7W, T8S, H16D, H16R, P18D, P18L, P18N, N25T, R26Y, D29N, D30F, R35P, R37A, R37N, T40A, T40C, T40D, T40E, T40G, T40H, T40I, T40K, T40N, T40V, T 40W, A41C, A41D, A41E, A41H, A41I, A41K, A41N, A41R, A41T, A41V, A41Y, W43 E, P46A, G50M, G50P, G50Q, G57A, G57S, A60E, A60S, A60V, E68F, E68Y, Q71F, G73D, G73H, G73I, G73L, G73Q, G73S, G73T, G73W, T74F, K78S, K78T, T81M, Q8 4I, Q84W, Q84Y, E86K, E86S, S87G, S87L, S87P, H90P, A91Q, A91V, A91W, K93V, V97C, V97H, Q98E, Q98F, Q98G, Q98I, Q98K, Q98L, Q98M, Q98R, Q98T, Q98Y, V1 03C, V103G, V103S, M105F, G110K, G110R, A111E, A111Q, A113T, T114M, T114 V, N116C, N116Y, V117F, A119H, A119L, A119Y, V120E, E121I, N125S, N126I, N128E, Q129T, I131Y, S132A, S132D, S132H, S132I, S132K, S132L, S132P, S13 2R, S132T, S132V, S132Y, G133E, G133L, G133T, G133V, Y135C, Y135D, Y135E , Y135P, I137D, I137N, I137Q, I137W, I137Y, E138S, E138V, A139L, W140A, W 140D, W140M, W140N, W140P, W140S, W140T, W140V, T141G, K142E, K142N, F14 3H, F143I, F143M, F145G, F145H, F145I, F145L, F145S, P146A, P146H, P146N,G147D、G149F、G149K、W159A、W159E、H161W、F162T、V165P、W167D、W167G、W167P、Q169L、Q169P、Q169V、Q169Y、R171P、R176E、R176Q、R176V、R176Y、I177H、I177P、K179A、K179M、K179V、F180D、F180G、R181D、R181E、R181G、R181M、R181Y、K185T、K185V、K185W、A186D、W187T、N195Y、G196D、N197V、N197Y、A204F、A204H、A204L、V206N、M208D、P211C、P211M、E216L、R218Q、R219V、T225D、T225F、T225H、T225K、T225L、T225N、T225R、T225Y、L228V、L230C、L230F、D231G、D231T、D231V、I235A、K242L、Y243D、Y243M、Y243N、R247G、R247P、R247S、L250S、L250Y、T251D、H252P、V253F、V253S、R254D、R254F、R254H、R254P、R254T、R254V、R254W、N255C、N255F、N255P、T257A、T257C、T257D、T257E、T257G、T257P、T257Q、T257S、T257W、T257Y、K259P、K259V、K259W、K259Y、E260V、M261D、M261F、M261P、M261R、M261S、M261W、F262A、F262C、F262E、F262H、F262I、F262K、F262L、F262N、F262P、F262Q、F262R、F262S、F262T、F262V、F262W、A263E、A263H、A263W、A265Q、L272W、G273Q、A274D、L279I、N280P、K281H、K281I、K281S、N283P、W284A、W284D、W284E、W284G、W284I、W284K、W284M、W284N、W284P、W284Q、W284S、W284T、W284V、W284Y、S287I、S287K、S287L、S287P、S287R、S287T、S287V、S287Y、P292L、P292M、P292Y、H294N、H294S、N296T、Y298D, Y298E, N299T, A300G, S303A, S303C, S303D, S303E, S303F, S303G, S303H, S303L, S303M, S303N, S303Q, S303T, S303V, S303Y, G305E, Y307S, A310 D, A310N, K311A, K311C, L313Q, N314P, N314S, G315E, G315M, G315V, Q319C, Q319E, K320S, K320W, H321F, H321K, H321L, H321R, H321T, H321V, H321W, P32 2C, P322E, P322H, P322T, P322Y, M323A, M323C, M323D, M323E, M323G, M323H, M323L, M323N, M323P, M323R, M323S, M323T, M323W, M323Y, H324A, H324I, H 324P, H324T, H324W, H324Y, S334V, G337V, L340M, S342A, S342G, S342T, V344A, V344C, V344I, Q345N, E346A, W347A, W347E, W347F, W347G, W347H, W347K W347N, W347P, W347R, P350A, P350C, P350N, P350T, R359C, R359F, R359G, R359M, R359S, R359W, E360D, E360H, E360P, E360T, E360W, Q361D, Q361H, Q361 I、Q361K、Q361L、Q361M、Q361N、Q361P、Q361R、Q361S、Q361T、Q361V、Q361W、 Q361Y、G362M、Y363A、Y363E、Y363F、Y363G、Y363I、Y363L、Y363Q、Y363R、Y36 3S, Y363T, Y363V, Y363W, P364C, Y368C, Y368Q, Y368R, Y368T, Y372C, Y372G, Y372N, Y372T, P375Q, P375T, P380F, M382I, K383C, K383L, K383P, K383T, A 384D, A384N, K385N, K385V, D387C, D387P, D387R, D387W, P388I, E391M, R393K, R393M, N395C, N395D, N395K, N395V, N395W, F396E, F396G, F396T, A397VY398E, Q401H, H402S, D403A, D403E, D403T, F405C, F405N, F405S, F405T, D4 06V, H407C, H407Q, H407T, I410H, I410M, I410R, I410W, R415C, R415L, R415 M, R415Y, E416D, E416F, E416L, E416N, N418A, N418P, N418V, H421A, H421G, H421L, H421P, H421Q, H421Y, P422D, P422G, P422I, P422N, P422Q, P422S, N42 3G, N423H, N423I, N423L, N423M, N423P, N423Q, N423V, N423W, N423Y, G433A , G433N, G433S, P434I, P434L, G435C, G435L, G435M, G435T, G435V, K438A, K4 38I, K438Q, K438S, K438V, W439A, W439D, W439I, W439K, W439L, W439N, W439 S, W439T, W439V, W439Y, K446A, K446F, A447D, A447Y, Q449T, V450D, V450N, H 452L, T455C, T455M, T455V, N457D, N457E, N457I, N457L, N457V, K458D, K45 8L, G460M, G460V, T461F, T461I, T461Q, T461V, V462C, N465A, N465C, N465E , N465G, N465H, N465L, N465P, N465T, A466F, A466I, A466Y, D467I, W469A, W 469E, W469H, W469L, W469N, W469Q, W469R, W469S, W469Y, N471A, N471D, N471 M, N471W, S473N, N475A, N475C, N475D, N475E, N475F, N475G, N475H, N475I, N475L, N475M, N475P, N475Q, N475S, N475T, N475V, N475W, G476F, G476L, G477A, ​​G477H, G477Q, G477S, G477W, S478A, S478F, and S478Q, and further the variant comprises one or more of these substitutions at positions corresponding to at least 60%, 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%, or at least 99% but less than 100% sequence identity.

[0097] In one aspect, the variants are located at positions N3A, N3C, N3D, N3E, N3F, N3G, N3H, N3L, N3P, N3Q, N3S, N3T, N3V, T5E, G7E, G7F, G7H, G7K, G7L, G7P, G7R, G7S, G7T, G7V, G7W, T8S, H16D, H16R, P18D, P18L, P18N, N25T, R26Y, D29N, D30F, R35P, R37A, R37N, T40A, T40C, T40D, T40E, T40G, T40H, T40I, T40K, T40N, T40V, T 40W, A41C, A41D, A41E, A41H, A41I, A41K, A41N, A41R, A41T, A41V, A41Y, W43 E, P46A, G50M, G50P, G50Q, G57A, G57S, A60E, A60S, A60V, E68F, E68Y, Q71F, G73D, G73H, G73I, G73L, G73Q, G73S, G73T, G73W, T74F, K78S, K78T, T81M, Q8 4I, Q84W, Q84Y, E86K, E86S, S87G, S87L, S87P, H90P, A91Q, A91V, A91W, K93V, V97C, V97H, Q98E, Q98F, Q98G, Q98I, Q98K, Q98L, Q98M, Q98R, Q98T, Q98Y, V1 03C, V103G, V103S, M105F, G110K, G110R, A111E, A111Q, A113T, T114M, T114 V, N116C, N116Y, V117F, A119H, A119L, A119Y, V120E, E121I, N125S, N126I, N128E, Q129T, I131Y, S132A, S132D, S132H, S132I, S132K, S132L, S132P, S13 2R, S132T, S132V, S132Y, G133E, G133L, G133T, G133V, Y135C, Y135D, Y135E , Y135P, I137D, I137N, I137Q, I137W, I137Y, E138S, E138V, A139L, W140A, W 140D, W140M, W140N, W140P, W140S, W140T, W140V, T141G, K142E, K142N, F14 3H, F143I, F143M, F145G, F145H, F145I, F145L, F145S, P146A, P146H, P146N,G147D、G149F、G149K、W159A、W159E、H161W、F162T、V165P、W167D、W167G、W167P、Q169L、Q169P、Q169V、Q169Y、R171P、R176E、R176Q、R176V、R176Y、I177H、I177P、K179A、K179M、K179V、F180D、F180G、R181D、R181E、R181G、R181M、R181Y、K185T、K185V、K185W、A186D、W187T、N195Y、G196D、N197V、N197Y、A204F、A204H、A204L、V206N、M208D、P211C、P211M、E216L、R218Q、R219V、T225D、T225F、T225H、T225K、T225L、T225N、T225R、T225Y、L228V、L230C、L230F、D231G、D231T、D231V、I235A、K242L、Y243D、Y243M、Y243N、R247G、R247P、R247S、L250S、L250Y、T251D、H252P、V253F、V253S、R254D、R254F、R254H、R254P、R254T、R254V、R254W、N255C、N255F、N255P、T257A、T257C、T257D、T257E、T257G、T257P、T257Q、T257S、T257W、T257Y、K259P、K259V、K259W、K259Y、E260V、M261D、M261F、M261P、M261R、M261S、M261W、F262A、F262C、F262E、F262H、F262I、F262K、F262L、F262N、F262P、F262Q、F262R、F262S、F262T、F262V、F262W、A263E、A263H、A263W、A265Q、L272W、G273Q、A274D、L279I、N280P、K281H、K281I、K281S、N283P、W284A、W284D、W284E、W284G、W284I、W284K、W284M、W284N、W284P、W284Q、W284S、W284T、W284V、W284Y、S287I、S287K、S287L、S287P、S287R、S287T、S287V、S287Y、P292L、P292M、P292Y、H294N、H294S、N296T、Y298D, Y298E, N299T, A300G, S303A, S303C, S303D, S303E, S303F, S303G, S303H, S303L, S303M, S303N, S303Q, S303T, S303V, S303Y, G305E, Y307S, A310 D, A310N, K311A, K311C, L313Q, N314P, N314S, G315E, G315M, G315V, Q319C, Q319E, K320S, K320W, H321F, H321K, H321L, H321R, H321T, H321V, H321W, P32 2C, P322E, P322H, P322T, P322Y, M323A, M323C, M323D, M323E, M323G, M323H, M323L, M323N, M323P, M323R, M323S, M323T, M323W, M323Y, H324A, H324I, H 324P, H324T, H324W, H324Y, S334V, G337V, L340M, S342A, S342G, S342T, V344A, V344C, V344I, Q345N, E346A, W347A, W347E, W347F, W347G, W347H, W347K W347N, W347P, W347R, P350A, P350C, P350N, P350T, R359C, R359F, R359G, R359M, R359S, R359W, E360D, E360H, E360P, E360T, E360W, Q361D, Q361H, Q361 I、Q361K、Q361L、Q361M、Q361N、Q361P、Q361R、Q361S、Q361T、Q361V、Q361W、 Q361Y、G362M、Y363A、Y363E、Y363F、Y363G、Y363I、Y363L、Y363Q、Y363R、Y36 3S, Y363T, Y363V, Y363W, P364C, Y368C, Y368Q, Y368R, Y368T, Y372C, Y372G, Y372N, Y372T, P375Q, P375T, P380F, M382I, K383C, K383L, K383P, K383T, A 384D, A384N, K385N, K385V, D387C, D387P, D387R, D387W, P388I, E391M, R393K, R393M, N395C, N395D, N395K, N395V, N395W, F396E, F396G, F396T, A397VY398E, Q401H, H402S, D403A, D403E, D403T, F405C, F405N, F405S, F405T, D4 06V, H407C, H407Q, H407T, I410H, I410M, I410R, I410W, R415C, R415L, R415M , R415Y, E416D, E416F, E416L, E416N, N418A, N418P, N418V, H421A, H421G, H 421L, H421P, H421Q, H421Y, P422D, P422G, P422I, P422N, P422Q, P422S, N423 G, N423H, N423I, N423L, N423M, N423P, N423Q, N423V, N423W, N423Y, G433A, G433N, G433S, P434I, P434L, G435C, G435L, G435M, G435T, G435V, K438A, K43 8I, K438Q, K438S, K438V, W439A, W439D, W439I, W439K, W439L, W439N, W439S , W439T, W439V, W439Y, K446A, K446F, A447D, A447Y, Q449T, V450D, V450N, H4 52L, T455C, T455M, T455V, N457D, N457E, N457I, N457L, N457V, K458D, K458 L, G460M, G460V, T461F, T461I, T461Q, T461V, V462C, N465A, N465C, N465E, N 465G, N465H, N465L, N465P, N465T, A466F, A466I, A466Y, D467I, W469A, W46 9E, W469H, W469L, W469N, W469Q, W469R, W469S, W469Y, N471A, N471D, N471M, and the variant comprises one or more of these substitutions at positions corresponding to N471W, S473N, N475A, N475C, N475D, N475E, N475F, N475G, N475H, N475I, N475L, N475M, N475P, N475Q, N475S, N475T, N475V, N475W, G476F, G476L, G477A, ​​G477H, G477Q, G477S, G477W, S478A, S478F, and S478Q, and further the variant has a nucleotide sequence at least 60%, 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%, or at least 99% but less than 100% sequence identity.

[0098] In one aspect, the variants are located at positions N3A, N3C, N3D, N3E, N3F, N3G, N3H, N3L, N3P, N3Q, N3S, N3T, N3V, T5E, G7E, G7F, G7H, G7K, G7L, G7P, G7R, G7S, G7T, G7V, G7W, T8S, H16D, H16R, P18D, P18L, P18N, N25T, R26Y, D29N, D30F, R35P, R37A, R37N, T40A, T40C, T40D, T40E, T40G, T40H, T40I, T40K, T40N, T40V, T 40W, A41C, A41D, A41E, A41H, A41I, A41K, A41N, A41R, A41T, A41V, A41Y, W43 E, P46A, G50M, G50P, G50Q, G57A, G57S, A60E, A60S, A60V, E68F, E68Y, Q71F, G73D, G73H, G73I, G73L, G73Q, G73S, G73T, G73W, T74F, K78S, K78T, T81M, Q8 4I, Q84W, Q84Y, E86K, E86S, S87G, S87L, S87P, H90P, A91Q, A91V, A91W, K93V, V97C, V97H, Q98E, Q98F, Q98G, Q98I, Q98K, Q98L, Q98M, Q98R, Q98T, Q98Y, V1 03C, V103G, V103S, M105F, G110K, G110R, A111E, A111Q, A113T, T114M, T114 V, N116C, N116Y, V117F, A119H, A119L, A119Y, V120E, E121I, N125S, N126I, N128E, Q129T, I131Y, S132A, S132D, S132H, S132I, S132K, S132L, S132P, S13 2R, S132T, S132V, S132Y, G133E, G133L, G133T, G133V, Y135C, Y135D, Y135E , Y135P, I137D, I137N, I137Q, I137W, I137Y, E138S, E138V, A139L, W140A, W 140D, W140M, W140N, W140P, W140S, W140T, W140V, T141G, K142E, K142N, F14 3H, F143I, F143M, F145G, F145H, F145I, F145L, F145S, P146A, P146H, P146N,G147D、G149F、G149K、W159A、W159E、H161W、F162T、V165P、W167D、W167G、W167P、Q169L、Q169P、Q169V、Q169Y、R171P、R176E、R176Q、R176V、R176Y、I177H、I177P、K179A、K179M、K179V、F180D、F180G、R181D、R181E、R181G、R181M、R181Y、K185T、K185V、K185W、A186D、W187T、N195Y、G196D、N197V、N197Y、A204F、A204H、A204L、V206N、M208D、P211C、P211M、E216L、R218Q、R219V、T225D、T225F、T225H、T225K、T225L、T225N、T225R、T225Y、L228V、L230C、L230F、D231G、D231T、D231V、I235A、K242L、Y243D、Y243M、Y243N、R247G、R247P、R247S、L250S、L250Y、T251D、H252P、V253F、V253S、R254D、R254F、R254H、R254P、R254T、R254V、R254W、N255C、N255F、N255P、T257A、T257C、T257D、T257E、T257G、T257P、T257Q、T257S、T257W、T257Y、K259P、K259V、K259W、K259Y、E260V、M261D、M261F、M261P、M261R、M261S、M261W、F262A、F262C、F262E、F262H、F262I、F262K、F262L、F262N、F262P、F262Q、F262R、F262S、F262T、F262V、F262W、A263E、A263H、A263W、A265Q、L272W、G273Q、A274D、L279I、N280P、K281H、K281I、K281S、N283P、W284A、W284D、W284E、W284G、W284I、W284K、W284M、W284N、W284P、W284Q、W284S、W284T、W284V、W284Y、S287I、S287K、S287L、S287P、S287R、S287T、S287V、S287Y、P292L、P292M、P292Y、H294N、H294S、N296T、Y298D, Y298E, N299T, A300G, S303A, S303C, S303D, S303E, S303F, S303G, S303H, S303L, S303M, S303N, S303Q, S303T, S303V, S303Y, G305E, Y307S, A310 D, A310N, K311A, K311C, L313Q, N314P, N314S, G315E, G315M, G315V, Q319C, Q319E, K320S, K320W, H321F, H321K, H321L, H321R, H321T, H321V, H321W, P32 2C, P322E, P322H, P322T, P322Y, M323A, M323C, M323D, M323E, M323G, M323H, M323L, M323N, M323P, M323R, M323S, M323T, M323W, M323Y, H324A, H324I, H 324P, H324T, H324W, H324Y, S334V, G337V, L340M, S342A, S342G, S342T, V344A, V344C, V344I, Q345N, E346A, W347A, W347E, W347F, W347G, W347H, W347K W347N, W347P, W347R, P350A, P350C, P350N, P350T, R359C, R359F, R359G, R359M, R359S, R359W, E360D, E360H, E360P, E360T, E360W, Q361D, Q361H, Q361 I、Q361K、Q361L、Q361M、Q361N、Q361P、Q361R、Q361S、Q361T、Q361V、Q361W、 Q361Y、G362M、Y363A、Y363E、Y363F、Y363G、Y363I、Y363L、Y363Q、Y363R、Y36 3S, Y363T, Y363V, Y363W, P364C, Y368C, Y368Q, Y368R, Y368T, Y372C, Y372G, Y372N, Y372T, P375Q, P375T, P380F, M382I, K383C, K383L, K383P, K383T, A 384D, A384N, K385N, K385V, D387C, D387P, D387R, D387W, P388I, E391M, R393K, R393M, N395C, N395D, N395K, N395V, N395W, F396E, F396G, F396T, A397VY398E, Q401H, H402S, D403A, D403E, D403T, F405C, F405N, F405S, F405T, D4 06V, H407C, H407Q, H407T, I410H, I410M, I410R, I410W, R415C, R415L, R415M , R415Y, E416D, E416F, E416L, E416N, N418A, N418P, N418V, H421A, H421G, H 421L, H421P, H421Q, H421Y, P422D, P422G, P422I, P422N, P422Q, P422S, N423 G, N423H, N423I, N423L, N423M, N423P, N423Q, N423V, N423W, N423Y, G433A, G433N, G433S, P434I, P434L, G435C, G435L, G435M, G435T, G435V, K438A, K43 8I, K438Q, K438S, K438V, W439A, W439D, W439I, W439K, W439L, W439N, W439S , W439T, W439V, W439Y, K446A, K446F, A447D, A447Y, Q449T, V450D, V450N, H4 52L, T455C, T455M, T455V, N457D, N457E, N457I, N457L, N457V, K458D, K458 L, G460M, G460V, T461F, T461I, T461Q, T461V, V462C, N465A, N465C, N465E, N 465G, N465H, N465L, N465P, N465T, A466F, A466I, A466Y, D467I, W469A, W46 9E, W469H, W469L, W469N, W469Q, W469R, W469S, W469Y, N471A, N471D, N471M, and the variant comprises one or more of these substitutions at positions corresponding to N471W, S473N, N475A, N475C, N475D, N475E, N475F, N475G, N475H, N475I, N475L, N475M, N475P, N475Q, N475S, N475T, N475V, N475W, G476F, G476L, G477A, ​​G477H, G477Q, G477S, G477W, S478A, S478F, and S478Q, and further the variant has a nucleotide sequence at least 60%, 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%, or at least 99% but less than 100% sequence identity.

[0099] In one aspect, the variants are located at positions N3A, N3C, N3D, N3E, N3F, N3G, N3H, N3L, N3P, N3Q, N3S, N3T, N3V, T5E, G7E, G7F, G7H, G7K, G7L, G7P, G7R, G7S, G7T, G7V, G7W, T8S, H16D, H16R, P18D, P18L, P18N, N25T, R26Y, D29N, D30F, R35P, R37A, R37N, T40A, T40C, T40D, T40E, T40G, T40H, T40I, T40K, T40N, T40V, T 40W, A41C, A41D, A41E, A41H, A41I, A41K, A41N, A41R, A41T, A41V, A41Y, W43 E, P46A, G50M, G50P, G50Q, G57A, G57S, A60E, A60S, A60V, E68F, E68Y, Q71F, G73D, G73H, G73I, G73L, G73Q, G73S, G73T, G73W, T74F, K78S, K78T, T81M, Q8 4I, Q84W, Q84Y, E86K, E86S, S87G, S87L, S87P, H90P, A91Q, A91V, A91W, K93V, V97C, V97H, Q98E, Q98F, Q98G, Q98I, Q98K, Q98L, Q98M, Q98R, Q98T, Q98Y, V1 03C, V103G, V103S, M105F, G110K, G110R, A111E, A111Q, A113T, T114M, T114 V, N116C, N116Y, V117F, A119H, A119L, A119Y, V120E, E121I, N125S, N126I, N128E, Q129T, I131Y, S132A, S132D, S132H, S132I, S132K, S132L, S132P, S13 2R, S132T, S132V, S132Y, G133E, G133L, G133T, G133V, Y135C, Y135D, Y135E , Y135P, I137D, I137N, I137Q, I137W, I137Y, E138S, E138V, A139L, W140A, W 140D, W140M, W140N, W140P, W140S, W140T, W140V, T141G, K142E, K142N, F14 3H, F143I, F143M, F145G, F145H, F145I, F145L, F145S, P146A, P146H, P146N,G147D、G149F、G149K、W159A、W159E、H161W、F162T、V165P、W167D、W167G、W167P、Q169L、Q169P、Q169V、Q169Y、R171P、R176E、R176Q、R176V、R176Y、I177H、I177P、K179A、K179M、K179V、F180D、F180G、R181D、R181E、R181G、R181M、R181Y、K185T、K185V、K185W、A186D、W187T、N195Y、G196D、N197V、N197Y、A204F、A204H、A204L、V206N、M208D、P211C、P211M、E216L、R218Q、R219V、T225D、T225F、T225H、T225K、T225L、T225N、T225R、T225Y、L228V、L230C、L230F、D231G、D231T、D231V、I235A、K242L、Y243D、Y243M、Y243N、R247G、R247P、R247S、L250S、L250Y、T251D、H252P、V253F、V253S、R254D、R254F、R254H、R254P、R254T、R254V、R254W、N255C、N255F、N255P、T257A、T257C、T257D、T257E、T257G、T257P、T257Q、T257S、T257W、T257Y、K259P、K259V、K259W、K259Y、E260V、M261D、M261F、M261P、M261R、M261S、M261W、F262A、F262C、F262E、F262H、F262I、F262K、F262L、F262N、F262P、F262Q、F262R、F262S、F262T、F262V、F262W、A263E、A263H、A263W、A265Q、L272W、G273Q、A274D、L279I、N280P、K281H、K281I、K281S、N283P、W284A、W284D、W284E、W284G、W284I、W284K、W284M、W284N、W284P、W284Q、W284S、W284T、W284V、W284Y、S287I、S287K、S287L、S287P、S287R、S287T、S287V、S287Y、P292L、P292M、P292Y、H294N、H294S、N296T、Y298D, Y298E, N299T, A300G, S303A, S303C, S303D, S303E, S303F, S303G, S303H, S303L, S303M, S303N, S303Q, S303T, S303V, S303Y, G305E, Y307S, A310 D, A310N, K311A, K311C, L313Q, N314P, N314S, G315E, G315M, G315V, Q319C, Q319E, K320S, K320W, H321F, H321K, H321L, H321R, H321T, H321V, H321W, P32 2C, P322E, P322H, P322T, P322Y, M323A, M323C, M323D, M323E, M323G, M323H, M323L, M323N, M323P, M323R, M323S, M323T, M323W, M323Y, H324A, H324I, H 324P, H324T, H324W, H324Y, S334V, G337V, L340M, S342A, S342G, S342T, V344A, V344C, V344I, Q345N, E346A, W347A, W347E, W347F, W347G, W347H, W347K W347N, W347P, W347R, P350A, P350C, P350N, P350T, R359C, R359F, R359G, R359M, R359S, R359W, E360D, E360H, E360P, E360T, E360W, Q361D, Q361H, Q361 I、Q361K、Q361L、Q361M、Q361N、Q361P、Q361R、Q361S、Q361T、Q361V、Q361W、 Q361Y、G362M、Y363A、Y363E、Y363F、Y363G、Y363I、Y363L、Y363Q、Y363R、Y36 3S, Y363T, Y363V, Y363W, P364C, Y368C, Y368Q, Y368R, Y368T, Y372C, Y372G, Y372N, Y372T, P375Q, P375T, P380F, M382I, K383C, K383L, K383P, K383T, A 384D, A384N, K385N, K385V, D387C, D387P, D387R, D387W, P388I, E391M, R393K, R393M, N395C, N395D, N395K, N395V, N395W, F396E, F396G, F396T, A397VY398E, Q401H, H402S, D403A, D403E, D403T, F405C, F405N, F405S, F405T, D4 06V, H407C, H407Q, H407T, I410H, I410M, I410R, I410W, R415C, R415L, R415M , R415Y, E416D, E416F, E416L, E416N, N418A, N418P, N418V, H421A, H421G, H 421L, H421P, H421Q, H421Y, P422D, P422G, P422I, P422N, P422Q, P422S, N423 G, N423H, N423I, N423L, N423M, N423P, N423Q, N423V, N423W, N423Y, G433A, G433N, G433S, P434I, P434L, G435C, G435L, G435M, G435T, G435V, K438A, K43 8I, K438Q, K438S, K438V, W439A, W439D, W439I, W439K, W439L, W439N, W439S , W439T, W439V, W439Y, K446A, K446F, A447D, A447Y, Q449T, V450D, V450N, H4 52L, T455C, T455M, T455V, N457D, N457E, N457I, N457L, N457V, K458D, K458 L, G460M, G460V, T461F, T461I, T461Q, T461V, V462C, N465A, N465C, N465E, N 465G, N465H, N465L, N465P, N465T, A466F, A466I, A466Y, D467I, W469A, W46 9E, W469H, W469L, W469N, W469Q, W469R, W469S, W469Y, N471A, N471D, N471M, and comprising one or more of these substitutions at positions corresponding to N471W, S473N, N475A, N475C, N475D, N475E, N475F, N475G, N475H, N475I, N475L, N475M, N475P, N475Q, N475S, N475T, N475V, N475W, G476F, G476L, G477A, ​​G477H, G477Q, G477S, G477W, S478A, S478F, and S478Q, and further wherein the variant has a nucleotide sequence at least 60%, 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%, or at least 99% but less than 100% sequence identity.

[0100] In one aspect, using SEQ ID NO: 1 for variant numbering, positions N3A, N3C, N3D, N3E, N3F, N3G, N3H, N3L, N3P, N3Q, N3S, N3T, N3V, T5E, G7E, G7F, G7H, G7K, G7L, G7P, G7R, G7S, G7T, G7V, G7W, T8S, H16D, H16R, P18D, P18L, P18N, N25T, R26Y, D29N, D30F, R35P, R37A, R37N, T40A, T40C, T40D, T40E, T40G, T40H, T40I, T40K, T40N, T40V, T40W , A41C, A41D, A41E, A41H, A41I, A41K, A41N, A41R, A41T, A41V, A41Y, W43E, P 46A, G50M, G50P, G50Q, G57A, G57S, A60E, A60S, A60V, E68F, E68Y, Q71F, G73D , G73H, G73I, G73L, G73Q, G73S, G73T, G73W, T74F, K78S, K78T, T81M, Q84I, Q 84W, Q84Y, E86K, E86S, S87G, S87L, S87P, H90P, A91Q, A91V, A91W, K93V, V97C , V97H, Q98E, Q98F, Q98G, Q98I, Q98K, Q98L, Q98M, Q98R, Q98T, Q98Y, V103C, V103G, V103S, M105F, G110K, G110R, A111E, A111Q, A113T, T114M, T114V, N11 6C, N116Y, V117F, A119H, A119L, A119Y, V120E, E121I, N125S, N126I, N128E , Q129T, I131Y, S132A, S132D, S132H, S132I, S132K, S132L, S132P, S132R, S1 32T, S132V, S132Y, G133E, G133L, G133T, G133V, Y135C, Y135D, Y135E, Y135 P, I137D, I137N, I137Q, I137W, I137Y, E138S, E138V, A139L, W140A, W140D, W 140M, W140N, W140P, W140S, W140T, W140V, T141G, K142E, K142N, F143H, F14 3I, F143M, F145G, F145H, F145I, F145L, F145S, P146A, P146H, P146N, G147D,G149F、G149K、W159A、W159E、H161W、F162T、V165P、W167D、W167G、W167P、Q169L、Q169P、Q169V、Q169Y、R171P、R176E、R176Q、R176V、R176Y、I177H、I177P、K179A、K179M、K179V、F180D、F180G、R181D、R181E、R181G、R181M、R181Y、K185T、K185V、K185W、A186D、W187T、N195Y、G196D、N197V、N197Y、A204F、A204H、A204L、V206N、M208D、P211C、P211M、E216L、R218Q、R219V、T225D、T225F、T225H、T225K、T225L、T225N、T225R、T225Y、L228V、L230C、L230F、D231G、D231T、D231V、I235A、K242L、Y243D、Y243M、Y243N、R247G、R247P、R247S、L250S、L250Y、T251D、H252P、V253F、V253S、R254D、R254F、R254H、R254P、R254T、R254V、R254W、N255C、N255F、N255P、T257A、T257C、T257D、T257E、T257G、T257P、T257Q、T257S、T257W、T257Y、K259P、K259V、K259W、K259Y、E260V、M261D、M261F、M261P、M261R、M261S、M261W、F262A、F262C、F262E、F262H、F262I、F262K、F262L、F262N、F262P、F262Q、F262R、F262S、F262T、F262V、F262W、A263E、A263H、A263W、A265Q、L272W、G273Q、A274D、L279I、N280P、K281H、K281I、K281S、N283P、W284A、W284D、W284E、W284G、W284I、W284K、W284M、W284N、W284P、W284Q、W284S、W284T、W284V、W284Y、S287I、S287K、S287L、S287P、S287R、S287T、S287V、S287Y、P292L、P292M、P292Y、H294N、H294S、N296T、Y298D、Y298E, N299T, A300G, S303A, S303C, S303D, S303E, S303F, S303G, S303H, S303L, S303M, S303N, S303Q, S303T, S303V, S303Y, G305E, Y307S, A310D, A310 N、K311A、K311C、L313Q、N314P、N314S、G315E、G315M、G315V、Q319C、Q319E、 K320S、K320W、H321F、H321K、H321L、H321R、H321T、H321V、H321W、P322C、P32 2E, P322H, P322T, P322Y, M323A, M323C, M323D, M323E, M323G, M323H, M323L, M323N, M323P, M323R, M323S, M323T, M323W, M323Y, H324A, H324I, H324P, H 324T, H324W, H324Y, S334V, G337V, L340M, S342A, S342G, S342T, V344A, V344C, V344I, Q345N, E346A, W347A, W347E, W347F, W347G, W347H, W347K, W347N W347P, W347R, P350A, P350C, P350N, P350T, R359C, R359F, R359G, R359M, R359S, R359W, E360D, E360H, E360P, E360T, E360W, Q361D, Q361H, Q361I, Q361 K、Q361L、Q361M、Q361N、Q361P、Q361R、Q361S、Q361T、Q361V、Q361W、Q361Y、 G362M、Y363A、Y363E、Y363F、Y363G、Y363I、Y363L、Y363Q、Y363R、Y363S、Y36 3T、Y363V、Y363W、P364C、Y368C、Y368Q、Y368R、Y368T、Y372C、Y372G、Y372N 、Y372T、P375Q、P375T、P380F、M382I、K383C、K383L、K383P、K383T、A384D、A 384N, K385N, K385V, D387C, D387P, D387R, D387W, P388I, E391M, R393K, R393M, N395C, N395D, N395K, N395V, N395W, F396E, F396G, F396T, A397V, Y398EQ401H, H402S, D403A, D403E, D403T, F405C, F405N, F405S, F405T, D406V, H4 07C, H407Q, H407T, I410H, I410M, I410R, I410W, R415C, R415L, R415M, R415 Y, E416D, E416F, E416L, E416N, N418A, N418P, N418V, H421A, H421G, H421L, H421P, H421Q, H421Y, P422D, P422G, P422I, P422N, P422Q, P422S, N423G, N4 23H, N423I, N423L, N423M, N423P, N423Q, N423V, N423W, N423Y, G433A, G433 N, G433S, P434I, P434L, G435C, G435L, G435M, G435T, G435V, K438A, K438I, K438Q, K438S, K438V, W439A, W439D, W439I, W439K, W439L, W439N, W439S, W4 39T, W439V, W439Y, K446A, K446F, A447D, A447Y, Q449T, V450D, V450N, H452L , T455C, T455M, T455V, N457D, N457E, N457I, N457L, N457V, K458D, K458L, G 460M, G460V, T461F, T461I, T461Q, T461V, V462C, N465A, N465C, N465E, N46 5G, N465H, N465L, N465P, N465T, A466F, A466I, A466Y, D467I, W469A, W469E , W469H, W469L, W469N, W469Q, W469R, W469S, W469Y, N471A, N471D, N471M, N and the variant comprises one or more of these substitutions at positions corresponding to 471W, S473N, N475A, N475C, N475D, N475E, N475F, N475G, N475H, N475I, N475L, N475M, N475P, N475Q, N475S, N475T, N475V, N475W, G476F, G476L, G477A, ​​G477H, G477Q, G477S, G477W, S478A, S478F, and S478Q, and further the variant has a sequence similar to that of the polypeptide of SEQ ID NO: 8 by at least 60%, 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%, or at least 99% but less than 100% sequence identity.

[0101] In one aspect, the variants are located at positions N3A, N3C, N3D, N3E, N3F, N3G, N3H, N3L, N3P, N3Q, N3S, N3T, N3V, T5E, G7E, G7F, G7H, G7K, G7L, G7P, G7R, G7S, G7T, G7V, G7W, T8S, H16D, H16R, P18D, P18L, P18N, N25T, R26Y, D29N, D30F, R35P, R37A, R37N, T40A, T40C, T40D, T40E, T40G, T40H, T40I, T40K, T40N, T40V, T 40W, A41C, A41D, A41E, A41H, A41I, A41K, A41N, A41R, A41T, A41V, A41Y, W43 E, P46A, G50M, G50P, G50Q, G57A, G57S, A60E, A60S, A60V, E68F, E68Y, Q71F, G73D, G73H, G73I, G73L, G73Q, G73S, G73T, G73W, T74F, K78S, K78T, T81M, Q8 4I, Q84W, Q84Y, E86K, E86S, S87G, S87L, S87P, H90P, A91Q, A91V, A91W, K93V, V97C, V97H, Q98E, Q98F, Q98G, Q98I, Q98K, Q98L, Q98M, Q98R, Q98T, Q98Y, V1 03C, V103G, V103S, M105F, G110K, G110R, A111E, A111Q, A113T, T114M, T114 V, N116C, N116Y, V117F, A119H, A119L, A119Y, V120E, E121I, N125S, N126I, N128E, Q129T, I131Y, S132A, S132D, S132H, S132I, S132K, S132L, S132P, S13 2R, S132T, S132V, S132Y, G133E, G133L, G133T, G133V, Y135C, Y135D, Y135E , Y135P, I137D, I137N, I137Q, I137W, I137Y, E138S, E138V, A139L, W140A, W 140D, W140M, W140N, W140P, W140S, W140T, W140V, T141G, K142E, K142N, F14 3H, F143I, F143M, F145G, F145H, F145I, F145L, F145S, P146A, P146H, P146N,G147D、G149F、G149K、W159A、W159E、H161W、F162T、V165P、W167D、W167G、W167P、Q169L、Q169P、Q169V、Q169Y、R171P、R176E、R176Q、R176V、R176Y、I177H、I177P、K179A、K179M、K179V、F180D、F180G、R181D、R181E、R181G、R181M、R181Y、K185T、K185V、K185W、A186D、W187T、N195Y、G196D、N197V、N197Y、A204F、A204H、A204L、V206N、M208D、P211C、P211M、E216L、R218Q、R219V、T225D、T225F、T225H、T225K、T225L、T225N、T225R、T225Y、L228V、L230C、L230F、D231G、D231T、D231V、I235A、K242L、Y243D、Y243M、Y243N、R247G、R247P、R247S、L250S、L250Y、T251D、H252P、V253F、V253S、R254D、R254F、R254H、R254P、R254T、R254V、R254W、N255C、N255F、N255P、T257A、T257C、T257D、T257E、T257G、T257P、T257Q、T257S、T257W、T257Y、K259P、K259V、K259W、K259Y、E260V、M261D、M261F、M261P、M261R、M261S、M261W、F262A、F262C、F262E、F262H、F262I、F262K、F262L、F262N、F262P、F262Q、F262R、F262S、F262T、F262V、F262W、A263E、A263H、A263W、A265Q、L272W、G273Q、A274D、L279I、N280P、K281H、K281I、K281S、N283P、W284A、W284D、W284E、W284G、W284I、W284K、W284M、W284N、W284P、W284Q、W284S、W284T、W284V、W284Y、S287I、S287K、S287L、S287P、S287R、S287T、S287V、S287Y、P292L、P292M、P292Y、H294N、H294S、N296T、Y298D, Y298E, N299T, A300G, S303A, S303C, S303D, S303E, S303F, S303G, S303H, S303L, S303M, S303N, S303Q, S303T, S303V, S303Y, G305E, Y307S, A310 D, A310N, K311A, K311C, L313Q, N314P, N314S, G315E, G315M, G315V, Q319C, Q319E, K320S, K320W, H321F, H321K, H321L, H321R, H321T, H321V, H321W, P32 2C, P322E, P322H, P322T, P322Y, M323A, M323C, M323D, M323E, M323G, M323H, M323L, M323N, M323P, M323R, M323S, M323T, M323W, M323Y, H324A, H324I, H 324P, H324T, H324W, H324Y, S334V, G337V, L340M, S342A, S342G, S342T, V344A, V344C, V344I, Q345N, E346A, W347A, W347E, W347F, W347G, W347H, W347K W347N, W347P, W347R, P350A, P350C, P350N, P350T, R359C, R359F, R359G, R359M, R359S, R359W, E360D, E360H, E360P, E360T, E360W, Q361D, Q361H, Q361 I、Q361K、Q361L、Q361M、Q361N、Q361P、Q361R、Q361S、Q361T、Q361V、Q361W、 Q361Y、G362M、Y363A、Y363E、Y363F、Y363G、Y363I、Y363L、Y363Q、Y363R、Y36 3S, Y363T, Y363V, Y363W, P364C, Y368C, Y368Q, Y368R, Y368T, Y372C, Y372G, Y372N, Y372T, P375Q, P375T, P380F, M382I, K383C, K383L, K383P, K383T, A 384D, A384N, K385N, K385V, D387C, D387P, D387R, D387W, P388I, E391M, R393K, R393M, N395C, N395D, N395K, N395V, N395W, F396E, F396G, F396T, A397VY398E, Q401H, H402S, D403A, D403E, D403T, F405C, F405N, F405S, F405T, D4 06V, H407C, H407Q, H407T, I410H, I410M, I410R, I410W, R415C, R415L, R415M , R415Y, E416D, E416F, E416L, E416N, N418A, N418P, N418V, H421A, H421G, H 421L, H421P, H421Q, H421Y, P422D, P422G, P422I, P422N, P422Q, P422S, N423 G, N423H, N423I, N423L, N423M, N423P, N423Q, N423V, N423W, N423Y, G433A, G433N, G433S, P434I, P434L, G435C, G435L, G435M, G435T, G435V, K438A, K43 8I, K438Q, K438S, K438V, W439A, W439D, W439I, W439K, W439L, W439N, W439S , W439T, W439V, W439Y, K446A, K446F, A447D, A447Y, Q449T, V450D, V450N, H4 52L, T455C, T455M, T455V, N457D, N457E, N457I, N457L, N457V, K458D, K458 L, G460M, G460V, T461F, T461I, T461Q, T461V, V462C, N465A, N465C, N465E, N 465G, N465H, N465L, N465P, N465T, A466F, A466I, A466Y, D467I, W469A, W46 9E, W469H, W469L, W469N, W469Q, W469R, W469S, W469Y, N471A, N471D, N471M, and the variant comprises one or more of these substitutions at positions corresponding to N471W, S473N, N475A, N475C, N475D, N475E, N475F, N475G, N475H, N475I, N475L, N475M, N475P, N475Q, N475S, N475T, N475V, N475W, G476F, G476L, G477A, ​​G477H, G477Q, G477S, G477W, S478A, S478F, and S478Q, and further the variant has a nucleotide sequence at least 60%, 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%, or at least 99% but less than 100% sequence identity.

[0102] In one aspect, the variants are located at positions N3A, N3C, N3D, N3E, N3F, N3G, N3H, N3L, N3P, N3Q, N3S, N3T, N3V, T5E, G7E, G7F, G7H, G7K, G7L, G7P, G7R, G7S, G7T, G7V, G7W, T8S, H16D, H16R, P18D, P18L, P18N, N25T, R26Y, D29N, D30F, R35P, R37A, R37N, T40A, T40C, T40D, T40E, T40G, T40H, T40I, T40K, T40N, T40V, T 40W, A41C, A41D, A41E, A41H, A41I, A41K, A41N, A41R, A41T, A41V, A41Y, W43 E, P46A, G50M, G50P, G50Q, G57A, G57S, A60E, A60S, A60V, E68F, E68Y, Q71F, G73D, G73H, G73I, G73L, G73Q, G73S, G73T, G73W, T74F, K78S, K78T, T81M, Q8 4I, Q84W, Q84Y, E86K, E86S, S87G, S87L, S87P, H90P, A91Q, A91V, A91W, K93V, V97C, V97H, Q98E, Q98F, Q98G, Q98I, Q98K, Q98L, Q98M, Q98R, Q98T, Q98Y, V1 03C, V103G, V103S, M105F, G110K, G110R, A111E, A111Q, A113T, T114M, T114 V, N116C, N116Y, V117F, A119H, A119L, A119Y, V120E, E121I, N125S, N126I, N128E, Q129T, I131Y, S132A, S132D, S132H, S132I, S132K, S132L, S132P, S13 2R, S132T, S132V, S132Y, G133E, G133L, G133T, G133V, Y135C, Y135D, Y135E , Y135P, I137D, I137N, I137Q, I137W, I137Y, E138S, E138V, A139L, W140A, W 140D, W140M, W140N, W140P, W140S, W140T, W140V, T141G, K142E, K142N, F14 3H, F143I, F143M, F145G, F145H, F145I, F145L, F145S, P146A, P146H, P146N,G147D、G149F、G149K、W159A、W159E、H161W、F162T、V165P、W167D、W167G、W167P、Q169L、Q169P、Q169V、Q169Y、R171P、R176E、R176Q、R176V、R176Y、I177H、I177P、K179A、K179M、K179V、F180D、F180G、R181D、R181E、R181G、R181M、R181Y、K185T、K185V、K185W、A186D、W187T、N195Y、G196D、N197V、N197Y、A204F、A204H、A204L、V206N、M208D、P211C、P211M、E216L、R218Q、R219V、T225D、T225F、T225H、T225K、T225L、T225N、T225R、T225Y、L228V、L230C、L230F、D231G、D231T、D231V、I235A、K242L、Y243D、Y243M、Y243N、R247G、R247P、R247S、L250S、L250Y、T251D、H252P、V253F、V253S、R254D、R254F、R254H、R254P、R254T、R254V、R254W、N255C、N255F、N255P、T257A、T257C、T257D、T257E、T257G、T257P、T257Q、T257S、T257W、T257Y、K259P、K259V、K259W、K259Y、E260V、M261D、M261F、M261P、M261R、M261S、M261W、F262A、F262C、F262E、F262H、F262I、F262K、F262L、F262N、F262P、F262Q、F262R、F262S、F262T、F262V、F262W、A263E、A263H、A263W、A265Q、L272W、G273Q、A274D、L279I、N280P、K281H、K281I、K281S、N283P、W284A、W284D、W284E、W284G、W284I、W284K、W284M、W284N、W284P、W284Q、W284S、W284T、W284V、W284Y、S287I、S287K、S287L、S287P、S287R、S287T、S287V、S287Y、P292L、P292M、P292Y、H294N、H294S、N296T、Y298D, Y298E, N299T, A300G, S303A, S303C, S303D, S303E, S303F, S303G, S303H, S303L, S303M, S303N, S303Q, S303T, S303V, S303Y, G305E, Y307S, A310 D, A310N, K311A, K311C, L313Q, N314P, N314S, G315E, G315M, G315V, Q319C, Q319E, K320S, K320W, H321F, H321K, H321L, H321R, H321T, H321V, H321W, P32 2C, P322E, P322H, P322T, P322Y, M323A, M323C, M323D, M323E, M323G, M323H, M323L, M323N, M323P, M323R, M323S, M323T, M323W, M323Y, H324A, H324I, H 324P, H324T, H324W, H324Y, S334V, G337V, L340M, S342A, S342G, S342T, V344A, V344C, V344I, Q345N, E346A, W347A, W347E, W347F, W347G, W347H, W347K W347N, W347P, W347R, P350A, P350C, P350N, P350T, R359C, R359F, R359G, R359M, R359S, R359W, E360D, E360H, E360P, E360T, E360W, Q361D, Q361H, Q361 I、Q361K、Q361L、Q361M、Q361N、Q361P、Q361R、Q361S、Q361T、Q361V、Q361W、 Q361Y、G362M、Y363A、Y363E、Y363F、Y363G、Y363I、Y363L、Y363Q、Y363R、Y36 3S, Y363T, Y363V, Y363W, P364C, Y368C, Y368Q, Y368R, Y368T, Y372C, Y372G, Y372N, Y372T, P375Q, P375T, P380F, M382I, K383C, K383L, K383P, K383T, A 384D, A384N, K385N, K385V, D387C, D387P, D387R, D387W, P388I, E391M, R393K, R393M, N395C, N395D, N395K, N395V, N395W, F396E, F396G, F396T, A397VY398E, Q401H, H402S, D403A, D403E, D403T, F405C, F405N, F405S, F405T, D4 06V, H407C, H407Q, H407T, I410H, I410M, I410R, I410W, R415C, R415L, R415M , R415Y, E416D, E416F, E416L, E416N, N418A, N418P, N418V, H421A, H421G, H 421L, H421P, H421Q, H421Y, P422D, P422G, P422I, P422N, P422Q, P422S, N423 G, N423H, N423I, N423L, N423M, N423P, N423Q, N423V, N423W, N423Y, G433A, G433N, G433S, P434I, P434L, G435C, G435L, G435M, G435T, G435V, K438A, K43 8I, K438Q, K438S, K438V, W439A, W439D, W439I, W439K, W439L, W439N, W439S , W439T, W439V, W439Y, K446A, K446F, A447D, A447Y, Q449T, V450D, V450N, H4 52L, T455C, T455M, T455V, N457D, N457E, N457I, N457L, N457V, K458D, K458 L, G460M, G460V, T461F, T461I, T461Q, T461V, V462C, N465A, N465C, N465E, N 465G, N465H, N465L, N465P, N465T, A466F, A466I, A466Y, D467I, W469A, W46 9E, W469H, W469L, W469N, W469Q, W469R, W469S, W469Y, N471A, N471D, N471M, and comprising one or more of these substitutions at positions corresponding to N471W, S473N, N475A, N475C, N475D, N475E, N475F, N475G, N475H, N475I, N475L, N475M, N475P, N475Q, N475S, N475T, N475V, N475W, G476F, G476L, G477A, ​​G477H, G477Q, G477S, G477W, S478A, S478F, and S478Q, and further wherein the variant has a sequence similar to that of the polypeptide of SEQ ID NO: 10, at least 60%, 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%, or at least 99% but less than 100% sequence identity.

[0103] In one aspect, the variants are located at positions N3A, N3C, N3D, N3E, N3F, N3G, N3H, N3L, N3P, N3Q, N3S, N3T, N3V, T5E, G7E, G7F, G7H, G7K, G7L, G7P, G7R, G7S, G7T, G7V, G7W, T8S, H16D, H16R, P18D, P18L, P18N, N25T, R26Y, D29N, D30F, R35P, R37A, R37N, T40A, T40C, T40D, T40E, T40G, T40H, T40I, T40K, T40N, T40V, T 40W, A41C, A41D, A41E, A41H, A41I, A41K, A41N, A41R, A41T, A41V, A41Y, W43 E, P46A, G50M, G50P, G50Q, G57A, G57S, A60E, A60S, A60V, E68F, E68Y, Q71F, G73D, G73H, G73I, G73L, G73Q, G73S, G73T, G73W, T74F, K78S, K78T, T81M, Q8 4I, Q84W, Q84Y, E86K, E86S, S87G, S87L, S87P, H90P, A91Q, A91V, A91W, K93V, V97C, V97H, Q98E, Q98F, Q98G, Q98I, Q98K, Q98L, Q98M, Q98R, Q98T, Q98Y, V1 03C, V103G, V103S, M105F, G110K, G110R, A111E, A111Q, A113T, T114M, T114 V, N116C, N116Y, V117F, A119H, A119L, A119Y, V120E, E121I, N125S, N126I, N128E, Q129T, I131Y, S132A, S132D, S132H, S132I, S132K, S132L, S132P, S13 2R, S132T, S132V, S132Y, G133E, G133L, G133T, G133V, Y135C, Y135D, Y135E , Y135P, I137D, I137N, I137Q, I137W, I137Y, E138S, E138V, A139L, W140A, W 140D, W140M, W140N, W140P, W140S, W140T, W140V, T141G, K142E, K142N, F14 3H, F143I, F143M, F145G, F145H, F145I, F145L, F145S, P146A, P146H, P146N,G147D、G149F、G149K、W159A、W159E、H161W、F162T、V165P、W167D、W167G、W167P、Q169L、Q169P、Q169V、Q169Y、R171P、R176E、R176Q、R176V、R176Y、I177H、I177P、K179A、K179M、K179V、F180D、F180G、R181D、R181E、R181G、R181M、R181Y、K185T、K185V、K185W、A186D、W187T、N195Y、G196D、N197V、N197Y、A204F、A204H、A204L、V206N、M208D、P211C、P211M、E216L、R218Q、R219V、T225D、T225F、T225H、T225K、T225L、T225N、T225R、T225Y、L228V、L230C、L230F、D231G、D231T、D231V、I235A、K242L、Y243D、Y243M、Y243N、R247G、R247P、R247S、L250S、L250Y、T251D、H252P、V253F、V253S、R254D、R254F、R254H、R254P、R254T、R254V、R254W、N255C、N255F、N255P、T257A、T257C、T257D、T257E、T257G、T257P、T257Q、T257S、T257W、T257Y、K259P、K259V、K259W、K259Y、E260V、M261D、M261F、M261P、M261R、M261S、M261W、F262A、F262C、F262E、F262H、F262I、F262K、F262L、F262N、F262P、F262Q、F262R、F262S、F262T、F262V、F262W、A263E、A263H、A263W、A265Q、L272W、G273Q、A274D、L279I、N280P、K281H、K281I、K281S、N283P、W284A、W284D、W284E、W284G、W284I、W284K、W284M、W284N、W284P、W284Q、W284S、W284T、W284V、W284Y、S287I、S287K、S287L、S287P、S287R、S287T、S287V、S287Y、P292L、P292M、P292Y、H294N、H294S、N296T、Y298D, Y298E, N299T, A300G, S303A, S303C, S303D, S303E, S303F, S303G, S303H, S303L, S303M, S303N, S303Q, S303T, S303V, S303Y, G305E, Y307S, A310 D, A310N, K311A, K311C, L313Q, N314P, N314S, G315E, G315M, G315V, Q319C, Q319E, K320S, K320W, H321F, H321K, H321L, H321R, H321T, H321V, H321W, P32 2C, P322E, P322H, P322T, P322Y, M323A, M323C, M323D, M323E, M323G, M323H, M323L, M323N, M323P, M323R, M323S, M323T, M323W, M323Y, H324A, H324I, H 324P, H324T, H324W, H324Y, S334V, G337V, L340M, S342A, S342G, S342T, V344A, V344C, V344I, Q345N, E346A, W347A, W347E, W347F, W347G, W347H, W347K W347N, W347P, W347R, P350A, P350C, P350N, P350T, R359C, R359F, R359G, R359M, R359S, R359W, E360D, E360H, E360P, E360T, E360W, Q361D, Q361H, Q361 I、Q361K、Q361L、Q361M、Q361N、Q361P、Q361R、Q361S、Q361T、Q361V、Q361W、 Q361Y、G362M、Y363A、Y363E、Y363F、Y363G、Y363I、Y363L、Y363Q、Y363R、Y36 3S, Y363T, Y363V, Y363W, P364C, Y368C, Y368Q, Y368R, Y368T, Y372C, Y372G, Y372N, Y372T, P375Q, P375T, P380F, M382I, K383C, K383L, K383P, K383T, A 384D, A384N, K385N, K385V, D387C, D387P, D387R, D387W, P388I, E391M, R393K, R393M, N395C, N395D, N395K, N395V, N395W, F396E, F396G, F396T, A397VY398E, Q401H, H402S, D403A, D403E, D403T, F405C, F405N, F405S, F405T, D4 06V, H407C, H407Q, H407T, I410H, I410M, I410R, I410W, R415C, R415L, R415M , R415Y, E416D, E416F, E416L, E416N, N418A, N418P, N418V, H421A, H421G, H 421L, H421P, H421Q, H421Y, P422D, P422G, P422I, P422N, P422Q, P422S, N423 G, N423H, N423I, N423L, N423M, N423P, N423Q, N423V, N423W, N423Y, G433A, G433N, G433S, P434I, P434L, G435C, G435L, G435M, G435T, G435V, K438A, K43 8I, K438Q, K438S, K438V, W439A, W439D, W439I, W439K, W439L, W439N, W439S , W439T, W439V, W439Y, K446A, K446F, A447D, A447Y, Q449T, V450D, V450N, H4 52L, T455C, T455M, T455V, N457D, N457E, N457I, N457L, N457V, K458D, K458 L, G460M, G460V, T461F, T461I, T461Q, T461V, V462C, N465A, N465C, N465E, N 465G, N465H, N465L, N465P, N465T, A466F, A466I, A466Y, D467I, W469A, W46 9E, W469H, W469L, W469N, W469Q, W469R, W469S, W469Y, N471A, N471D, N471M, and comprising one or more of these substitutions at positions corresponding to N471W, S473N, N475A, N475C, N475D, N475E, N475F, N475G, N475H, N475I, N475L, N475M, N475P, N475Q, N475S, N475T, N475V, N475W, G476F, G476L, G477A, ​​G477H, G477Q, G477S, G477W, S478A, S478F, and S478Q, and further wherein the variant has a sequence identity of at least 60%, 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%, or at least 99% but less than 100% sequence identity.

[0104] In one aspect, the variants are located at positions N3A, N3C, N3D, N3E, N3F, N3G, N3H, N3L, N3P, N3Q, N3S, N3T, N3V, T5E, G7E, G7F, G7H, G7K, G7L, G7P, G7R, G7S, G7T, G7V, G7W, T8S, H16D, H16R, P18D, P18L, P18N, N25T, R26Y, D29N, D30F, R35P, R37A, R37N, T40A, T40C, T40D, T40E, T40G, T40H, T40I, T40K, T40N, T40V, T 40W, A41C, A41D, A41E, A41H, A41I, A41K, A41N, A41R, A41T, A41V, A41Y, W43 E, P46A, G50M, G50P, G50Q, G57A, G57S, A60E, A60S, A60V, E68F, E68Y, Q71F, G73D, G73H, G73I, G73L, G73Q, G73S, G73T, G73W, T74F, K78S, K78T, T81M, Q8 4I, Q84W, Q84Y, E86K, E86S, S87G, S87L, S87P, H90P, A91Q, A91V, A91W, K93V, V97C, V97H, Q98E, Q98F, Q98G, Q98I, Q98K, Q98L, Q98M, Q98R, Q98T, Q98Y, V1 03C, V103G, V103S, M105F, G110K, G110R, A111E, A111Q, A113T, T114M, T114 V, N116C, N116Y, V117F, A119H, A119L, A119Y, V120E, E121I, N125S, N126I, N128E, Q129T, I131Y, S132A, S132D, S132H, S132I, S132K, S132L, S132P, S13 2R, S132T, S132V, S132Y, G133E, G133L, G133T, G133V, Y135C, Y135D, Y135E , Y135P, I137D, I137N, I137Q, I137W, I137Y, E138S, E138V, A139L, W140A, W 140D, W140M, W140N, W140P, W140S, W140T, W140V, T141G, K142E, K142N, F14 3H, F143I, F143M, F145G, F145H, F145I, F145L, F145S, P146A, P146H, P146N,G147D、G149F、G149K、W159A、W159E、H161W、F162T、V165P、W167D、W167G、W167P、Q169L、Q169P、Q169V、Q169Y、R171P、R176E、R176Q、R176V、R176Y、I177H、I177P、K179A、K179M、K179V、F180D、F180G、R181D、R181E、R181G、R181M、R181Y、K185T、K185V、K185W、A186D、W187T、N195Y、G196D、N197V、N197Y、A204F、A204H、A204L、V206N、M208D、P211C、P211M、E216L、R218Q、R219V、T225D、T225F、T225H、T225K、T225L、T225N、T225R、T225Y、L228V、L230C、L230F、D231G、D231T、D231V、I235A、K242L、Y243D、Y243M、Y243N、R247G、R247P、R247S、L250S、L250Y、T251D、H252P、V253F、V253S、R254D、R254F、R254H、R254P、R254T、R254V、R254W、N255C、N255F、N255P、T257A、T257C、T257D、T257E、T257G、T257P、T257Q、T257S、T257W、T257Y、K259P、K259V、K259W、K259Y、E260V、M261D、M261F、M261P、M261R、M261S、M261W、F262A、F262C、F262E、F262H、F262I、F262K、F262L、F262N、F262P、F262Q、F262R、F262S、F262T、F262V、F262W、A263E、A263H、A263W、A265Q、L272W、G273Q、A274D、L279I、N280P、K281H、K281I、K281S、N283P、W284A、W284D、W284E、W284G、W284I、W284K、W284M、W284N、W284P、W284Q、W284S、W284T、W284V、W284Y、S287I、S287K、S287L、S287P、S287R、S287T、S287V、S287Y、P292L、P292M、P292Y、H294N、H294S、N296T、Y298D, Y298E, N299T, A300G, S303A, S303C, S303D, S303E, S303F, S303G, S303H, S303L, S303M, S303N, S303Q, S303T, S303V, S303Y, G305E, Y307S, A310 D, A310N, K311A, K311C, L313Q, N314P, N314S, G315E, G315M, G315V, Q319C, Q319E, K320S, K320W, H321F, H321K, H321L, H321R, H321T, H321V, H321W, P32 2C, P322E, P322H, P322T, P322Y, M323A, M323C, M323D, M323E, M323G, M323H, M323L, M323N, M323P, M323R, M323S, M323T, M323W, M323Y, H324A, H324I, H 324P, H324T, H324W, H324Y, S334V, G337V, L340M, S342A, S342G, S342T, V344A, V344C, V344I, Q345N, E346A, W347A, W347E, W347F, W347G, W347H, W347K W347N, W347P, W347R, P350A, P350C, P350N, P350T, R359C, R359F, R359G, R359M, R359S, R359W, E360D, E360H, E360P, E360T, E360W, Q361D, Q361H, Q361 I、Q361K、Q361L、Q361M、Q361N、Q361P、Q361R、Q361S、Q361T、Q361V、Q361W、 Q361Y、G362M、Y363A、Y363E、Y363F、Y363G、Y363I、Y363L、Y363Q、Y363R、Y36 3S, Y363T, Y363V, Y363W, P364C, Y368C, Y368Q, Y368R, Y368T, Y372C, Y372G, Y372N, Y372T, P375Q, P375T, P380F, M382I, K383C, K383L, K383P, K383T, A 384D, A384N, K385N, K385V, D387C, D387P, D387R, D387W, P388I, E391M, R393K, R393M, N395C, N395D, N395K, N395V, N395W, F396E, F396G, F396T, A397VY398E, Q401H, H402S, D403A, D403E, D403T, F405C, F405N, F405S, F405T, D4 06V, H407C, H407Q, H407T, I410H, I410M, I410R, I410W, R415C, R415L, R415M , R415Y, E416D, E416F, E416L, E416N, N418A, N418P, N418V, H421A, H421G, H 421L, H421P, H421Q, H421Y, P422D, P422G, P422I, P422N, P422Q, P422S, N423 G, N423H, N423I, N423L, N423M, N423P, N423Q, N423V, N423W, N423Y, G433A, G433N, G433S, P434I, P434L, G435C, G435L, G435M, G435T, G435V, K438A, K43 8I, K438Q, K438S, K438V, W439A, W439D, W439I, W439K, W439L, W439N, W439S , W439T, W439V, W439Y, K446A, K446F, A447D, A447Y, Q449T, V450D, V450N, H4 52L, T455C, T455M, T455V, N457D, N457E, N457I, N457L, N457V, K458D, K458 L, G460M, G460V, T461F, T461I, T461Q, T461V, V462C, N465A, N465C, N465E, N 465G, N465H, N465L, N465P, N465T, A466F, A466I, A466Y, D467I, W469A, W46 9E, W469H, W469L, W469N, W469Q, W469R, W469S, W469Y, N471A, N471D, N471M, and comprising one or more of these substitutions at positions corresponding to N471W, S473N, N475A, N475C, N475D, N475E, N475F, N475G, N475H, N475I, N475L, N475M, N475P, N475Q, N475S, N475T, N475V, N475W, G476F, G476L, G477A, ​​G477H, G477Q, G477S, G477W, S478A, S478F, and S478Q, and further wherein the variant has a sequence similar to that of the polypeptide of SEQ ID NO: 12, at least 60%, 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%, or at least 99% but less than 100% sequence identity.

[0105] In one aspect, the variants are located at positions N3A, N3C, N3D, N3E, N3F, N3G, N3H, N3L, N3P, N3Q, N3S, N3T, N3V, T5E, G7E, G7F, G7H, G7K, G7L, G7P, G7R, G7S, G7T, G7V, G7W, T8S, H16D, H16R, P18D, P18L, P18N, N25T, R26Y, D29N, D30F, R35P, R37A, R37N, T40A, T40C, T40D, T40E, T40G, T40H, T40I, T40K, T40N, T40V, T 40W, A41C, A41D, A41E, A41H, A41I, A41K, A41N, A41R, A41T, A41V, A41Y, W43 E, P46A, G50M, G50P, G50Q, G57A, G57S, A60E, A60S, A60V, E68F, E68Y, Q71F, G73D, G73H, G73I, G73L, G73Q, G73S, G73T, G73W, T74F, K78S, K78T, T81M, Q8 4I, Q84W, Q84Y, E86K, E86S, S87G, S87L, S87P, H90P, A91Q, A91V, A91W, K93V, V97C, V97H, Q98E, Q98F, Q98G, Q98I, Q98K, Q98L, Q98M, Q98R, Q98T, Q98Y, V1 03C, V103G, V103S, M105F, G110K, G110R, A111E, A111Q, A113T, T114M, T114 V, N116C, N116Y, V117F, A119H, A119L, A119Y, V120E, E121I, N125S, N126I, N128E, Q129T, I131Y, S132A, S132D, S132H, S132I, S132K, S132L, S132P, S13 2R, S132T, S132V, S132Y, G133E, G133L, G133T, G133V, Y135C, Y135D, Y135E , Y135P, I137D, I137N, I137Q, I137W, I137Y, E138S, E138V, A139L, W140A, W 140D, W140M, W140N, W140P, W140S, W140T, W140V, T141G, K142E, K142N, F14 3H, F143I, F143M, F145G, F145H, F145I, F145L, F145S, P146A, P146H, P146N,G147D、G149F、G149K、W159A、W159E、H161W、F162T、V165P、W167D、W167G、W167P、Q169L、Q169P、Q169V、Q169Y、R171P、R176E、R176Q、R176V、R176Y、I177H、I177P、K179A、K179M、K179V、F180D、F180G、R181D、R181E、R181G、R181M、R181Y、K185T、K185V、K185W、A186D、W187T、N195Y、G196D、N197V、N197Y、A204F、A204H、A204L、V206N、M208D、P211C、P211M、E216L、R218Q、R219V、T225D、T225F、T225H、T225K、T225L、T225N、T225R、T225Y、L228V、L230C、L230F、D231G、D231T、D231V、I235A、K242L、Y243D、Y243M、Y243N、R247G、R247P、R247S、L250S、L250Y、T251D、H252P、V253F、V253S、R254D、R254F、R254H、R254P、R254T、R254V、R254W、N255C、N255F、N255P、T257A、T257C、T257D、T257E、T257G、T257P、T257Q、T257S、T257W、T257Y、K259P、K259V、K259W、K259Y、E260V、M261D、M261F、M261P、M261R、M261S、M261W、F262A、F262C、F262E、F262H、F262I、F262K、F262L、F262N、F262P、F262Q、F262R、F262S、F262T、F262V、F262W、A263E、A263H、A263W、A265Q、L272W、G273Q、A274D、L279I、N280P、K281H、K281I、K281S、N283P、W284A、W284D、W284E、W284G、W284I、W284K、W284M、W284N、W284P、W284Q、W284S、W284T、W284V、W284Y、S287I、S287K、S287L、S287P、S287R、S287T、S287V、S287Y、P292L、P292M、P292Y、H294N、H294S、N296T、Y298D, Y298E, N299T, A300G, S303A, S303C, S303D, S303E, S303F, S303G, S303H, S303L, S303M, S303N, S303Q, S303T, S303V, S303Y, G305E, Y307S, A310 D, A310N, K311A, K311C, L313Q, N314P, N314S, G315E, G315M, G315V, Q319C, Q319E, K320S, K320W, H321F, H321K, H321L, H321R, H321T, H321V, H321W, P32 2C, P322E, P322H, P322T, P322Y, M323A, M323C, M323D, M323E, M323G, M323H, M323L, M323N, M323P, M323R, M323S, M323T, M323W, M323Y, H324A, H324I, H 324P, H324T, H324W, H324Y, S334V, G337V, L340M, S342A, S342G, S342T, V344A, V344C, V344I, Q345N, E346A, W347A, W347E, W347F, W347G, W347H, W347K W347N, W347P, W347R, P350A, P350C, P350N, P350T, R359C, R359F, R359G, R359M, R359S, R359W, E360D, E360H, E360P, E360T, E360W, Q361D, Q361H, Q361 I、Q361K、Q361L、Q361M、Q361N、Q361P、Q361R、Q361S、Q361T、Q361V、Q361W、 Q361Y、G362M、Y363A、Y363E、Y363F、Y363G、Y363I、Y363L、Y363Q、Y363R、Y36 3S, Y363T, Y363V, Y363W, P364C, Y368C, Y368Q, Y368R, Y368T, Y372C, Y372G, Y372N, Y372T, P375Q, P375T, P380F, M382I, K383C, K383L, K383P, K383T, A 384D, A384N, K385N, K385V, D387C, D387P, D387R, D387W, P388I, E391M, R393K, R393M, N395C, N395D, N395K, N395V, N395W, F396E, F396G, F396T, A397VY398E, Q401H, H402S, D403A, D403E, D403T, F405C, F405N, F405S, F405T, D4 06V, H407C, H407Q, H407T, I410H, I410M, I410R, I410W, R415C, R415L, R415M , R415Y, E416D, E416F, E416L, E416N, N418A, N418P, N418V, H421A, H421G, H 421L, H421P, H421Q, H421Y, P422D, P422G, P422I, P422N, P422Q, P422S, N423 G, N423H, N423I, N423L, N423M, N423P, N423Q, N423V, N423W, N423Y, G433A, G433N, G433S, P434I, P434L, G435C, G435L, G435M, G435T, G435V, K438A, K43 8I, K438Q, K438S, K438V, W439A, W439D, W439I, W439K, W439L, W439N, W439S , W439T, W439V, W439Y, K446A, K446F, A447D, A447Y, Q449T, V450D, V450N, H4 52L, T455C, T455M, T455V, N457D, N457E, N457I, N457L, N457V, K458D, K458 L, G460M, G460V, T461F, T461I, T461Q, T461V, V462C, N465A, N465C, N465E, N 465G, N465H, N465L, N465P, N465T, A466F, A466I, A466Y, D467I, W469A, W46 9E, W469H, W469L, W469N, W469Q, W469R, W469S, W469Y, N471A, N471D, N471M, and comprising one or more of these substitutions at positions corresponding to N471W, S473N, N475A, N475C, N475D, N475E, N475F, N475G, N475H, N475I, N475L, N475M, N475P, N475Q, N475S, N475T, N475V, N475W, G476F, G476L, G477A, ​​G477H, G477Q, G477S, G477W, S478A, S478F, and S478Q, and further wherein the variant has a nucleotide sequence at least 60%, 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%, or at least 99% but less than 100% sequence identity.

[0106] In one aspect, the variants are located at positions N3A, N3C, N3D, N3E, N3F, N3G, N3H, N3L, N3P, N3Q, N3S, N3T, N3V, T5E, G7E, G7F, G7H, G7K, G7L, G7P, G7R, G7S, G7T, G7V, G7W, T8S, H16D, H16R, P18D, P18L, P18N, N25T, R26Y, D29N, D30F, R35P, R37A, R37N, T40A, T40C, T40D, T40E, T40G, T40H, T40I, T40K, T40N, T40V, T 40W, A41C, A41D, A41E, A41H, A41I, A41K, A41N, A41R, A41T, A41V, A41Y, W43 E, P46A, G50M, G50P, G50Q, G57A, G57S, A60E, A60S, A60V, E68F, E68Y, Q71F, G73D, G73H, G73I, G73L, G73Q, G73S, G73T, G73W, T74F, K78S, K78T, T81M, Q8 4I, Q84W, Q84Y, E86K, E86S, S87G, S87L, S87P, H90P, A91Q, A91V, A91W, K93V, V97C, V97H, Q98E, Q98F, Q98G, Q98I, Q98K, Q98L, Q98M, Q98R, Q98T, Q98Y, V1 03C, V103G, V103S, M105F, G110K, G110R, A111E, A111Q, A113T, T114M, T114 V, N116C, N116Y, V117F, A119H, A119L, A119Y, V120E, E121I, N125S, N126I, N128E, Q129T, I131Y, S132A, S132D, S132H, S132I, S132K, S132L, S132P, S13 2R, S132T, S132V, S132Y, G133E, G133L, G133T, G133V, Y135C, Y135D, Y135E , Y135P, I137D, I137N, I137Q, I137W, I137Y, E138S, E138V, A139L, W140A, W 140D, W140M, W140N, W140P, W140S, W140T, W140V, T141G, K142E, K142N, F14 3H, F143I, F143M, F145G, F145H, F145I, F145L, F145S, P146A, P146H, P146N,G147D、G149F、G149K、W159A、W159E、H161W、F162T、V165P、W167D、W167G、W167P、Q169L、Q169P、Q169V、Q169Y、R171P、R176E、R176Q、R176V、R176Y、I177H、I177P、K179A、K179M、K179V、F180D、F180G、R181D、R181E、R181G、R181M、R181Y、K185T、K185V、K185W、A186D、W187T、N195Y、G196D、N197V、N197Y、A204F、A204H、A204L、V206N、M208D、P211C、P211M、E216L、R218Q、R219V、T225D、T225F、T225H、T225K、T225L、T225N、T225R、T225Y、L228V、L230C、L230F、D231G、D231T、D231V、I235A、K242L、Y243D、Y243M、Y243N、R247G、R247P、R247S、L250S、L250Y、T251D、H252P、V253F、V253S、R254D、R254F、R254H、R254P、R254T、R254V、R254W、N255C、N255F、N255P、T257A、T257C、T257D、T257E、T257G、T257P、T257Q、T257S、T257W、T257Y、K259P、K259V、K259W、K259Y、E260V、M261D、M261F、M261P、M261R、M261S、M261W、F262A、F262C、F262E、F262H、F262I、F262K、F262L、F262N、F262P、F262Q、F262R、F262S、F262T、F262V、F262W、A263E、A263H、A263W、A265Q、L272W、G273Q、A274D、L279I、N280P、K281H、K281I、K281S、N283P、W284A、W284D、W284E、W284G、W284I、W284K、W284M、W284N、W284P、W284Q、W284S、W284T、W284V、W284Y、S287I、S287K、S287L、S287P、S287R、S287T、S287V、S287Y、P292L、P292M、P292Y、H294N、H294S、N296T、Y298D, Y298E, N299T, A300G, S303A, S303C, S303D, S303E, S303F, S303G, S303H, S303L, S303M, S303N, S303Q, S303T, S303V, S303Y, G305E, Y307S, A310 D, A310N, K311A, K311C, L313Q, N314P, N314S, G315E, G315M, G315V, Q319C, Q319E, K320S, K320W, H321F, H321K, H321L, H321R, H321T, H321V, H321W, P32 2C, P322E, P322H, P322T, P322Y, M323A, M323C, M323D, M323E, M323G, M323H, M323L, M323N, M323P, M323R, M323S, M323T, M323W, M323Y, H324A, H324I, H 324P, H324T, H324W, H324Y, S334V, G337V, L340M, S342A, S342G, S342T, V344A, V344C, V344I, Q345N, E346A, W347A, W347E, W347F, W347G, W347H, W347K W347N, W347P, W347R, P350A, P350C, P350N, P350T, R359C, R359F, R359G, R359M, R359S, R359W, E360D, E360H, E360P, E360T, E360W, Q361D, Q361H, Q361 I、Q361K、Q361L、Q361M、Q361N、Q361P、Q361R、Q361S、Q361T、Q361V、Q361W、 Q361Y、G362M、Y363A、Y363E、Y363F、Y363G、Y363I、Y363L、Y363Q、Y363R、Y36 3S, Y363T, Y363V, Y363W, P364C, Y368C, Y368Q, Y368R, Y368T, Y372C, Y372G, Y372N, Y372T, P375Q, P375T, P380F, M382I, K383C, K383L, K383P, K383T, A 384D, A384N, K385N, K385V, D387C, D387P, D387R, D387W, P388I, E391M, R393K, R393M, N395C, N395D, N395K, N395V, N395W, F396E, F396G, F396T, A397VY398E, Q401H, H402S, D403A, D403E, D403T, F405C, F405N, F405S, F405T, D4 06V, H407C, H407Q, H407T, I410H, I410M, I410R, I410W, R415C, R415L, R415M , R415Y, E416D, E416F, E416L, E416N, N418A, N418P, N418V, H421A, H421G, H 421L, H421P, H421Q, H421Y, P422D, P422G, P422I, P422N, P422Q, P422S, N423 G, N423H, N423I, N423L, N423M, N423P, N423Q, N423V, N423W, N423Y, G433A, G433N, G433S, P434I, P434L, G435C, G435L, G435M, G435T, G435V, K438A, K43 8I, K438Q, K438S, K438V, W439A, W439D, W439I, W439K, W439L, W439N, W439S , W439T, W439V, W439Y, K446A, K446F, A447D, A447Y, Q449T, V450D, V450N, H4 52L, T455C, T455M, T455V, N457D, N457E, N457I, N457L, N457V, K458D, K458 L, G460M, G460V, T461F, T461I, T461Q, T461V, V462C, N465A, N465C, N465E, N 465G, N465H, N465L, N465P, N465T, A466F, A466I, A466Y, D467I, W469A, W46 9E, W469H, W469L, W469N, W469Q, W469R, W469S, W469Y, N471A, N471D, N471M, and comprising one or more of these substitutions at positions corresponding to N471W, S473N, N475A, N475C, N475D, N475E, N475F, N475G, N475H, N475I, N475L, N475M, N475P, N475Q, N475S, N475T, N475V, N475W, G476F, G476L, G477A, ​​G477H, G477Q, G477S, G477W, S478A, S478F, and S478Q, and further wherein the variant has a sequence identity of at least 60%, 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%, or at least 99% but less than 100% sequence identity.

[0107] In one aspect, the variants are located at positions N3A, N3C, N3D, N3E, N3F, N3G, N3H, N3L, N3P, N3Q, N3S, N3T, N3V, T5E, G7E, G7F, G7H, G7K, G7L, G7P, G7R, G7S, G7T, G7V, G7W, T8S, H16D, H16R, P18D, P18L, P18N, N25T, R26Y, D29N, D30F, R35P, R37A, R37N, T40A, T40C, T40D, T40E, T40G, T40H, T40I, T40K, T40N, T40V, T 40W, A41C, A41D, A41E, A41H, A41I, A41K, A41N, A41R, A41T, A41V, A41Y, W43 E, P46A, G50M, G50P, G50Q, G57A, G57S, A60E, A60S, A60V, E68F, E68Y, Q71F, G73D, G73H, G73I, G73L, G73Q, G73S, G73T, G73W, T74F, K78S, K78T, T81M, Q8 4I, Q84W, Q84Y, E86K, E86S, S87G, S87L, S87P, H90P, A91Q, A91V, A91W, K93V, V97C, V97H, Q98E, Q98F, Q98G, Q98I, Q98K, Q98L, Q98M, Q98R, Q98T, Q98Y, V1 03C, V103G, V103S, M105F, G110K, G110R, A111E, A111Q, A113T, T114M, T114 V, N116C, N116Y, V117F, A119H, A119L, A119Y, V120E, E121I, N125S, N126I, N128E, Q129T, I131Y, S132A, S132D, S132H, S132I, S132K, S132L, S132P, S13 2R, S132T, S132V, S132Y, G133E, G133L, G133T, G133V, Y135C, Y135D, Y135E , Y135P, I137D, I137N, I137Q, I137W, I137Y, E138S, E138V, A139L, W140A, W 140D, W140M, W140N, W140P, W140S, W140T, W140V, T141G, K142E, K142N, F14 3H, F143I, F143M, F145G, F145H, F145I, F145L, F145S, P146A, P146H, P146N,G147D、G149F、G149K、W159A、W159E、H161W、F162T、V165P、W167D、W167G、W167P、Q169L、Q169P、Q169V、Q169Y、R171P、R176E、R176Q、R176V、R176Y、I177H、I177P、K179A、K179M、K179V、F180D、F180G、R181D、R181E、R181G、R181M、R181Y、K185T、K185V、K185W、A186D、W187T、N195Y、G196D、N197V、N197Y、A204F、A204H、A204L、V206N、M208D、P211C、P211M、E216L、R218Q、R219V、T225D、T225F、T225H、T225K、T225L、T225N、T225R、T225Y、L228V、L230C、L230F、D231G、D231T、D231V、I235A、K242L、Y243D、Y243M、Y243N、R247G、R247P、R247S、L250S、L250Y、T251D、H252P、V253F、V253S、R254D、R254F、R254H、R254P、R254T、R254V、R254W、N255C、N255F、N255P、T257A、T257C、T257D、T257E、T257G、T257P、T257Q、T257S、T257W、T257Y、K259P、K259V、K259W、K259Y、E260V、M261D、M261F、M261P、M261R、M261S、M261W、F262A、F262C、F262E、F262H、F262I、F262K、F262L、F262N、F262P、F262Q、F262R、F262S、F262T、F262V、F262W、A263E、A263H、A263W、A265Q、L272W、G273Q、A274D、L279I、N280P、K281H、K281I、K281S、N283P、W284A、W284D、W284E、W284G、W284I、W284K、W284M、W284N、W284P、W284Q、W284S、W284T、W284V、W284Y、S287I、S287K、S287L、S287P、S287R、S287T、S287V、S287Y、P292L、P292M、P292Y、H294N、H294S、N296T、Y298D, Y298E, N299T, A300G, S303A, S303C, S303D, S303E, S303F, S303G, S303H, S303L, S303M, S303N, S303Q, S303T, S303V, S303Y, G305E, Y307S, A310 D, A310N, K311A, K311C, L313Q, N314P, N314S, G315E, G315M, G315V, Q319C, Q319E, K320S, K320W, H321F, H321K, H321L, H321R, H321T, H321V, H321W, P32 2C, P322E, P322H, P322T, P322Y, M323A, M323C, M323D, M323E, M323G, M323H, M323L, M323N, M323P, M323R, M323S, M323T, M323W, M323Y, H324A, H324I, H 324P, H324T, H324W, H324Y, S334V, G337V, L340M, S342A, S342G, S342T, V344A, V344C, V344I, Q345N, E346A, W347A, W347E, W347F, W347G, W347H, W347K W347N, W347P, W347R, P350A, P350C, P350N, P350T, R359C, R359F, R359G, R359M, R359S, R359W, E360D, E360H, E360P, E360T, E360W, Q361D, Q361H, Q361 I、Q361K、Q361L、Q361M、Q361N、Q361P、Q361R、Q361S、Q361T、Q361V、Q361W、 Q361Y、G362M、Y363A、Y363E、Y363F、Y363G、Y363I、Y363L、Y363Q、Y363R、Y36 3S, Y363T, Y363V, Y363W, P364C, Y368C, Y368Q, Y368R, Y368T, Y372C, Y372G, Y372N, Y372T, P375Q, P375T, P380F, M382I, K383C, K383L, K383P, K383T, A 384D, A384N, K385N, K385V, D387C, D387P, D387R, D387W, P388I, E391M, R393K, R393M, N395C, N395D, N395K, N395V, N395W, F396E, F396G, F396T, A397VY398E, Q401H, H402S, D403A, D403E, D403T, F405C, F405N, F405S, F405T, D4 06V, H407C, H407Q, H407T, I410H, I410M, I410R, I410W, R415C, R415L, R415M , R415Y, E416D, E416F, E416L, E416N, N418A, N418P, N418V, H421A, H421G, H 421L, H421P, H421Q, H421Y, P422D, P422G, P422I, P422N, P422Q, P422S, N423 G, N423H, N423I, N423L, N423M, N423P, N423Q, N423V, N423W, N423Y, G433A, G433N, G433S, P434I, P434L, G435C, G435L, G435M, G435T, G435V, K438A, K43 8I, K438Q, K438S, K438V, W439A, W439D, W439I, W439K, W439L, W439N, W439S , W439T, W439V, W439Y, K446A, K446F, A447D, A447Y, Q449T, V450D, V450N, H4 52L, T455C, T455M, T455V, N457D, N457E, N457I, N457L, N457V, K458D, K458 L, G460M, G460V, T461F, T461I, T461Q, T461V, V462C, N465A, N465C, N465E, N 465G, N465H, N465L, N465P, N465T, A466F, A466I, A466Y, D467I, W469A, W46 9E, W469H, W469L, W469N, W469Q, W469R, W469S, W469Y, N471A, N471D, N471M, and comprising one or more of these substitutions at positions corresponding to N471W, S473N, N475A, N475C, N475D, N475E, N475F, N475G, N475H, N475I, N475L, N475M, N475P, N475Q, N475S, N475T, N475V, N475W, G476F, G476L, G477A, ​​G477H, G477Q, G477S, G477W, S478A, S478F, and S478Q, and further wherein the variant has a sequence identity of at least 60%, 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%, or at least 99% but less than 100% sequence identity.

[0108] In one aspect, the variants are located at positions N3A, N3C, N3D, N3E, N3F, N3G, N3H, N3L, N3P, N3Q, N3S, N3T, N3V, T5E, G7E, G7F, G7H, G7K, G7L, G7P, G7R, G7S, G7T, G7V, G7W, T8S, H16D, H16R, P18D, P18L, P18N, N25T, R26Y, D29N, D30F, R35P, R37A, R37N, T40A, T40C, T40D, T40E, T40G, T40H, T40I, T40K, T40N, T40V, T 40W, A41C, A41D, A41E, A41H, A41I, A41K, A41N, A41R, A41T, A41V, A41Y, W43 E, P46A, G50M, G50P, G50Q, G57A, G57S, A60E, A60S, A60V, E68F, E68Y, Q71F, G73D, G73H, G73I, G73L, G73Q, G73S, G73T, G73W, T74F, K78S, K78T, T81M, Q8 4I, Q84W, Q84Y, E86K, E86S, S87G, S87L, S87P, H90P, A91Q, A91V, A91W, K93V, V97C, V97H, Q98E, Q98F, Q98G, Q98I, Q98K, Q98L, Q98M, Q98R, Q98T, Q98Y, V1 03C, V103G, V103S, M105F, G110K, G110R, A111E, A111Q, A113T, T114M, T114 V, N116C, N116Y, V117F, A119H, A119L, A119Y, V120E, E121I, N125S, N126I, N128E, Q129T, I131Y, S132A, S132D, S132H, S132I, S132K, S132L, S132P, S13 2R, S132T, S132V, S132Y, G133E, G133L, G133T, G133V, Y135C, Y135D, Y135E , Y135P, I137D, I137N, I137Q, I137W, I137Y, E138S, E138V, A139L, W140A, W 140D, W140M, W140N, W140P, W140S, W140T, W140V, T141G, K142E, K142N, F14 3H, F143I, F143M, F145G, F145H, F145I, F145L, F145S, P146A, P146H, P146N,G147D、G149F、G149K、W159A、W159E、H161W、F162T、V165P、W167D、W167G、W167P、Q169L、Q169P、Q169V、Q169Y、R171P、R176E、R176Q、R176V、R176Y、I177H、I177P、K179A、K179M、K179V、F180D、F180G、R181D、R181E、R181G、R181M、R181Y、K185T、K185V、K185W、A186D、W187T、N195Y、G196D、N197V、N197Y、A204F、A204H、A204L、V206N、M208D、P211C、P211M、E216L、R218Q、R219V、T225D、T225F、T225H、T225K、T225L、T225N、T225R、T225Y、L228V、L230C、L230F、D231G、D231T、D231V、I235A、K242L、Y243D、Y243M、Y243N、R247G、R247P、R247S、L250S、L250Y、T251D、H252P、V253F、V253S、R254D、R254F、R254H、R254P、R254T、R254V、R254W、N255C、N255F、N255P、T257A、T257C、T257D、T257E、T257G、T257P、T257Q、T257S、T257W、T257Y、K259P、K259V、K259W、K259Y、E260V、M261D、M261F、M261P、M261R、M261S、M261W、F262A、F262C、F262E、F262H、F262I、F262K、F262L、F262N、F262P、F262Q、F262R、F262S、F262T、F262V、F262W、A263E、A263H、A263W、A265Q、L272W、G273Q、A274D、L279I、N280P、K281H、K281I、K281S、N283P、W284A、W284D、W284E、W284G、W284I、W284K、W284M、W284N、W284P、W284Q、W284S、W284T、W284V、W284Y、S287I、S287K、S287L、S287P、S287R、S287T、S287V、S287Y、P292L、P292M、P292Y、H294N、H294S、N296T、Y298D, Y298E, N299T, A300G, S303A, S303C, S303D, S303E, S303F, S303G, S303H, S303L, S303M, S303N, S303Q, S303T, S303V, S303Y, G305E, Y307S, A310 D, A310N, K311A, K311C, L313Q, N314P, N314S, G315E, G315M, G315V, Q319C, Q319E, K320S, K320W, H321F, H321K, H321L, H321R, H321T, H321V, H321W, P32 2C, P322E, P322H, P322T, P322Y, M323A, M323C, M323D, M323E, M323G, M323H, M323L, M323N, M323P, M323R, M323S, M323T, M323W, M323Y, H324A, H324I, H 324P, H324T, H324W, H324Y, S334V, G337V, L340M, S342A, S342G, S342T, V344A, V344C, V344I, Q345N, E346A, W347A, W347E, W347F, W347G, W347H, W347K W347N, W347P, W347R, P350A, P350C, P350N, P350T, R359C, R359F, R359G, R359M, R359S, R359W, E360D, E360H, E360P, E360T, E360W, Q361D, Q361H, Q361 I、Q361K、Q361L、Q361M、Q361N、Q361P、Q361R、Q361S、Q361T、Q361V、Q361W、 Q361Y、G362M、Y363A、Y363E、Y363F、Y363G、Y363I、Y363L、Y363Q、Y363R、Y36 3S, Y363T, Y363V, Y363W, P364C, Y368C, Y368Q, Y368R, Y368T, Y372C, Y372G, Y372N, Y372T, P375Q, P375T, P380F, M382I, K383C, K383L, K383P, K383T, A 384D, A384N, K385N, K385V, D387C, D387P, D387R, D387W, P388I, E391M, R393K, R393M, N395C, N395D, N395K, N395V, N395W, F396E, F396G, F396T, A397VY398E, Q401H, H402S, D403A, D403E, D403T, F405C, F405N, F405S, F405T, D4 06V, H407C, H407Q, H407T, I410H, I410M, I410R, I410W, R415C, R415L, R415M , R415Y, E416D, E416F, E416L, E416N, N418A, N418P, N418V, H421A, H421G, H 421L, H421P, H421Q, H421Y, P422D, P422G, P422I, P422N, P422Q, P422S, N423 G, N423H, N423I, N423L, N423M, N423P, N423Q, N423V, N423W, N423Y, G433A, G433N, G433S, P434I, P434L, G435C, G435L, G435M, G435T, G435V, K438A, K43 8I, K438Q, K438S, K438V, W439A, W439D, W439I, W439K, W439L, W439N, W439S , W439T, W439V, W439Y, K446A, K446F, A447D, A447Y, Q449T, V450D, V450N, H4 52L, T455C, T455M, T455V, N457D, N457E, N457I, N457L, N457V, K458D, K458 L, G460M, G460V, T461F, T461I, T461Q, T461V, V462C, N465A, N465C, N465E, N 465G, N465H, N465L, N465P, N465T, A466F, A466I, A466Y, D467I, W469A, W46 9E, W469H, W469L, W469N, W469Q, W469R, W469S, W469Y, N471A, N471D, N471M, and comprising one or more of these substitutions at positions corresponding to N471W, S473N, N475A, N475C, N475D, N475E, N475F, N475G, N475H, N475I, N475L, N475M, N475P, N475Q, N475S, N475T, N475V, N475W, G476F, G476L, G477A, ​​G477H, G477Q, G477S, G477W, S478A, S478F, and S478Q, and further wherein the variant has a sequence similar to that of the polypeptide of SEQ ID NO: 16, at least 60%, 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%, or at least 99% but less than 100% sequence identity.

[0109] In one aspect, the variants are located at positions N3A, N3C, N3D, N3E, N3F, N3G, N3H, N3L, N3P, N3Q, N3S, N3T, N3V, T5E, G7E, G7F, G7H, G7K, G7L, G7P, G7R, G7S, G7T, G7V, G7W, T8S, H16D, H16R, P18D, P18L, P18N, N25T, R26Y, D29N, D30F, R35P, R37A, R37N, T40A, T40C, T40D, T40E, T40G, T40H, T40I, T40K, T40N, T40V, T 40W, A41C, A41D, A41E, A41H, A41I, A41K, A41N, A41R, A41T, A41V, A41Y, W43 E, P46A, G50M, G50P, G50Q, G57A, G57S, A60E, A60S, A60V, E68F, E68Y, Q71F, G73D, G73H, G73I, G73L, G73Q, G73S, G73T, G73W, T74F, K78S, K78T, T81M, Q8 4I, Q84W, Q84Y, E86K, E86S, S87G, S87L, S87P, H90P, A91Q, A91V, A91W, K93V, V97C, V97H, Q98E, Q98F, Q98G, Q98I, Q98K, Q98L, Q98M, Q98R, Q98T, Q98Y, V1 03C, V103G, V103S, M105F, G110K, G110R, A111E, A111Q, A113T, T114M, T114 V, N116C, N116Y, V117F, A119H, A119L, A119Y, V120E, E121I, N125S, N126I, N128E, Q129T, I131Y, S132A, S132D, S132H, S132I, S132K, S132L, S132P, S13 2R, S132T, S132V, S132Y, G133E, G133L, G133T, G133V, Y135C, Y135D, Y135E , Y135P, I137D, I137N, I137Q, I137W, I137Y, E138S, E138V, A139L, W140A, W 140D, W140M, W140N, W140P, W140S, W140T, W140V, T141G, K142E, K142N, F14 3H, F143I, F143M, F145G, F145H, F145I, F145L, F145S, P146A, P146H, P146N,G147D、G149F、G149K、W159A、W159E、H161W、F162T、V165P、W167D、W167G、W167P、Q169L、Q169P、Q169V、Q169Y、R171P、R176E、R176Q、R176V、R176Y、I177H、I177P、K179A、K179M、K179V、F180D、F180G、R181D、R181E、R181G、R181M、R181Y、K185T、K185V、K185W、A186D、W187T、N195Y、G196D、N197V、N197Y、A204F、A204H、A204L、V206N、M208D、P211C、P211M、E216L、R218Q、R219V、T225D、T225F、T225H、T225K、T225L、T225N、T225R、T225Y、L228V、L230C、L230F、D231G、D231T、D231V、I235A、K242L、Y243D、Y243M、Y243N、R247G、R247P、R247S、L250S、L250Y、T251D、H252P、V253F、V253S、R254D、R254F、R254H、R254P、R254T、R254V、R254W、N255C、N255F、N255P、T257A、T257C、T257D、T257E、T257G、T257P、T257Q、T257S、T257W、T257Y、K259P、K259V、K259W、K259Y、E260V、M261D、M261F、M261P、M261R、M261S、M261W、F262A、F262C、F262E、F262H、F262I、F262K、F262L、F262N、F262P、F262Q、F262R、F262S、F262T、F262V、F262W、A263E、A263H、A263W、A265Q、L272W、G273Q、A274D、L279I、N280P、K281H、K281I、K281S、N283P、W284A、W284D、W284E、W284G、W284I、W284K、W284M、W284N、W284P、W284Q、W284S、W284T、W284V、W284Y、S287I、S287K、S287L、S287P、S287R、S287T、S287V、S287Y、P292L、P292M、P292Y、H294N、H294S、N296T、Y298D, Y298E, N299T, A300G, S303A, S303C, S303D, S303E, S303F, S303G, S303H, S303L, S303M, S303N, S303Q, S303T, S303V, S303Y, G305E, Y307S, A310 D, A310N, K311A, K311C, L313Q, N314P, N314S, G315E, G315M, G315V, Q319C, Q319E, K320S, K320W, H321F, H321K, H321L, H321R, H321T, H321V, H321W, P32 2C, P322E, P322H, P322T, P322Y, M323A, M323C, M323D, M323E, M323G, M323H, M323L, M323N, M323P, M323R, M323S, M323T, M323W, M323Y, H324A, H324I, H 324P, H324T, H324W, H324Y, S334V, G337V, L340M, S342A, S342G, S342T, V344A, V344C, V344I, Q345N, E346A, W347A, W347E, W347F, W347G, W347H, W347K W347N, W347P, W347R, P350A, P350C, P350N, P350T, R359C, R359F, R359G, R359M, R359S, R359W, E360D, E360H, E360P, E360T, E360W, Q361D, Q361H, Q361 I、Q361K、Q361L、Q361M、Q361N、Q361P、Q361R、Q361S、Q361T、Q361V、Q361W、 Q361Y、G362M、Y363A、Y363E、Y363F、Y363G、Y363I、Y363L、Y363Q、Y363R、Y36 3S, Y363T, Y363V, Y363W, P364C, Y368C, Y368Q, Y368R, Y368T, Y372C, Y372G, Y372N, Y372T, P375Q, P375T, P380F, M382I, K383C, K383L, K383P, K383T, A 384D, A384N, K385N, K385V, D387C, D387P, D387R, D387W, P388I, E391M, R393K, R393M, N395C, N395D, N395K, N395V, N395W, F396E, F396G, F396T, A397VY398E, Q401H, H402S, D403A, D403E, D403T, F405C, F405N, F405S, F405T, D4 06V, H407C, H407Q, H407T, I410H, I410M, I410R, I410W, R415C, R415L, R415M , R415Y, E416D, E416F, E416L, E416N, N418A, N418P, N418V, H421A, H421G, H 421L, H421P, H421Q, H421Y, P422D, P422G, P422I, P422N, P422Q, P422S, N423 G, N423H, N423I, N423L, N423M, N423P, N423Q, N423V, N423W, N423Y, G433A, G433N, G433S, P434I, P434L, G435C, G435L, G435M, G435T, G435V, K438A, K43 8I, K438Q, K438S, K438V, W439A, W439D, W439I, W439K, W439L, W439N, W439S , W439T, W439V, W439Y, K446A, K446F, A447D, A447Y, Q449T, V450D, V450N, H4 52L, T455C, T455M, T455V, N457D, N457E, N457I, N457L, N457V, K458D, K458 L, G460M, G460V, T461F, T461I, T461Q, T461V, V462C, N465A, N465C, N465E, N 465G, N465H, N465L, N465P, N465T, A466F, A466I, A466Y, D467I, W469A, W46 9E, W469H, W469L, W469N, W469Q, W469R, W469S, W469Y, N471A, N471D, N471M, and comprising one or more of these substitutions at positions corresponding to N471W, S473N, N475A, N475C, N475D, N475E, N475F, N475G, N475H, N475I, N475L, N475M, N475P, N475Q, N475S, N475T, N475V, N475W, G476F, G476L, G477A, ​​G477H, G477Q, G477S, G477W, S478A, S478F, and S478Q, and further wherein the variant has a sequence similar to that of the polypeptide of SEQ ID NO: 17, at least 60%, 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%, or at least 99% but less than 100% sequence identity.

[0110] The α-amylase variant has improved properties relative to the parent polypeptide, the improved properties being selected from the group consisting of improved catalytic efficiency, improved catalytic rate, increased chemical stability, increased oxidative stability, increased pH activity, increased pH stability, increased specific activity, increased stability under storage conditions, increased substrate binding, increased substrate cleavage, increased substrate specificity, increased substrate stability, improved surface properties, increased thermal activity, and increased thermostability. Preferably, the variant has improved properties relative to SEQ ID NO: 1 or SEQ ID NO: 2.

[0111] Preferably, the improved property is an increase in specific activity.

[0112] In one aspect, the improved property relative to the parent polypeptide is an increased specific activity in a model A detergent composition.

[0113] In one aspect, the improved property is an increase in specific activity in the Model A detergent composition, measured as an improvement factor (IF) > 1.0 relative to the parent polypeptide.

[0114] In one embodiment, the increased specific activity in the model A detergent composition, measured as an improvement factor (IF), is 1.1 or greater relative to the parent polypeptide.

[0115] In one embodiment, the increased specific activity in the model A detergent composition, measured as an improvement factor (IF), is 1.2 or greater relative to the parent polypeptide.

[0116] In one embodiment, the increased specific activity in the Model A detergent composition, measured as an improvement factor (IF), is 1.3 or greater relative to the parent polypeptide.

[0117] In one embodiment, the increased specific activity in the model A detergent composition, measured as an improvement factor (IF), is 1.4 or greater relative to the parent polypeptide.

[0118] In one embodiment, the increased specific activity in the Model A detergent composition, measured as an improvement factor (IF), is 1.5 or greater relative to the parent polypeptide.

[0119] In one embodiment, the increased specific activity in the model A detergent composition, measured as an improvement factor (IF), is 2.0 or greater relative to the parent polypeptide.

[0120] In one embodiment, the increased specific activity in the Model A detergent composition, measured as an improvement factor (IF), is 2.5 or greater relative to the parent polypeptide.

[0121] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 3. In another aspect, the amino acid at the position corresponding to position 3 is substituted with Ala, Arg, Asp, Cys, Gln, Glu, Gly, His, He, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variant comprises or consists of the substitutions N3A or N3C or N3D or N3E or N3F or N3G or N3H or N3L or N3P or N3Q or N3S or N3T or N3V of the polypeptide of SEQ ID NO: 1.

[0122] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 5. In another aspect, the amino acid at the position corresponding to position 5 is substituted with Ala, Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, He, Leu, Lys, Met, Phe, Pro, Ser, Trp, Tyr, or Val. In another aspect, the variant comprises or consists of the substitution T5E in the polypeptide of SEQ ID NO: 1.

[0123] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 7. In another aspect, the amino acid at the position corresponding to position 7 is substituted with Ala, Arg, Asn, Asp, Cys, Gln, Glu, His, He, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variant comprises or consists of the substitution G7E or G7F or G7H or G7K or G7L or G7P or G7R or G7S or G7T or G7V or G7W of the polypeptide of SEQ ID NO: 1.

[0124] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 8. In another aspect, the amino acid at the position corresponding to position 8 is substituted with Ala, Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe, Pro, Ser, Trp, Tyr, or Val. In another aspect, the variant comprises or consists of a substitution T8S of the polypeptide of SEQ ID NO: 1.

[0125] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 16. In another aspect, the amino acid at the position corresponding to position 16 is substituted with Ala, Arg, Asn, Asp, Cys, Gln, Glu, Gly, Ile, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variant comprises or consists of the substitution H16D or H16R in the polypeptide of SEQ ID NO: 1.

[0126] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 18. In another aspect, the amino acid at the position corresponding to position 18 is substituted with Ala, Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, He, Leu, Lys, Met, Phe, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variant comprises or consists of the substitution P18D or P18L or P18N in the polypeptide of SEQ ID NO: 1.

[0127] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 25. In another aspect, the amino acid at the position corresponding to position 25 is substituted with Ala, Arg, Asp, Cys, Gln, Glu, Gly, His, He, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variant comprises or consists of the substitution N25T in the polypeptide of SEQ ID NO: 1.

[0128] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 26. In another aspect, the amino acid at the position corresponding to position 26 is substituted with Ala, Asn, Asp, Cys, Gln, Glu, Gly, His, He, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variant comprises or consists of the substitution R26Y in the polypeptide of SEQ ID NO: 1.

[0129] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 29. In another aspect, the amino acid at the position corresponding to position 29 is substituted with Ala, Arg, Asn, Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variant comprises or consists of the substitution D29N in the polypeptide of SEQ ID NO: 1.

[0130] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 30. In another aspect, the amino acid at the position corresponding to position 30 is substituted with Ala, Arg, Asn, Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variant comprises or consists of the substitution D30F in the polypeptide of SEQ ID NO: 1.

[0131] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 35. In another aspect, the amino acid at the position corresponding to position 35 is substituted with Ala, Asn, Asp, Cys, Gln, Glu, Gly, His, He, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variant comprises or consists of the substitution R35P in the polypeptide of SEQ ID NO: 1.

[0132] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 37. In another aspect, the amino acid at the position corresponding to position 37 is substituted with Ala, Asn, Asp, Cys, Gln, Glu, Gly, His, He, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variant comprises or consists of the substitution R37A or R37N in the polypeptide of SEQ ID NO: 1.

[0133] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 40. In another aspect, the amino acid at the position corresponding to position 40 is substituted with Ala, Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, He, Leu, Lys, Met, Phe, Pro, Ser, Trp, Tyr, or Val. In another aspect, the variant comprises or consists of the substitution T40A or T40C or T40D or T40E or T40G or T40H or T40I or T40K or T40N or T40V or T40W in the polypeptide of SEQ ID NO: 1.

[0134] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 41. In another aspect, the amino acid at the position corresponding to position 41 is substituted with Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, He, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variant comprises or consists of the substitution A41C or A41D or A41E or A41H or A41I or A41K or A41N or A41R or A41T or A41V or A41Y in the polypeptide of SEQ ID NO: 1.

[0135] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 43. In another aspect, the amino acid at the position corresponding to position 43 is substituted with Ala, Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe, Pro, Ser, Thr, Tyr, or Val. In another aspect, the variant comprises or consists of the substitution W43E in the polypeptide of SEQ ID NO: 1.

[0136] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 46. In another aspect, the amino acid at the position corresponding to position 46 is substituted with Ala, Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, He, Leu, Lys, Met, Phe, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variant comprises or consists of the substitution P46A in the polypeptide of SEQ ID NO: 1.

[0137] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 50. In another aspect, the amino acid at the position corresponding to position 50 is substituted with Ala, Arg, Asn, Asp, Cys, Gln, Glu, His, Ile, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variant comprises or consists of the substitution G50M or G50P or G50Q in the polypeptide of SEQ ID NO: 1.

[0138] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 57. In another aspect, the amino acid at the position corresponding to position 57 is substituted with Ala, Arg, Asn, Asp, Cys, Gln, Glu, His, Ile, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variant comprises or consists of the substitution G57A or G57S in the polypeptide of SEQ ID NO: 1.

[0139] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 60. In another aspect, the amino acid at the position corresponding to position 60 is substituted with Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, He, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variant comprises or consists of the substitution A60E or A60S or A60V in the polypeptide of SEQ ID NO: 1.

[0140] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 68. In another aspect, the amino acid at the position corresponding to position 68 is substituted with Ala, Arg, Asn, Asp, Cys, Gln, Gly, His, He, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variant comprises or consists of the substitution E68F or E68Y in the polypeptide of SEQ ID NO: 1.

[0141] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 71. In another aspect, the amino acid at the position corresponding to position 71 is substituted with Ala, Arg, Asn, Asp, Cys, Glu, Gly, His, Ile, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variant comprises or consists of the substitution Q71F in the polypeptide of SEQ ID NO: 1.

[0142] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 73. In another aspect, the amino acid at the position corresponding to position 73 is substituted with Ala, Arg, Asn, Asp, Cys, Gln, Glu, His, He, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variant comprises or consists of the substitution G73D or G73H or G73I or G73L or G73Q or G73S or G73T or G73W of the polypeptide of SEQ ID NO: 1.

[0143] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 74. In another aspect, the amino acid at the position corresponding to position 74 is substituted with Ala, Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe, Pro, Ser, Trp, Tyr, or Val. In another aspect, the variant comprises or consists of the substitution T74F in the polypeptide of SEQ ID NO: 1.

[0144] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 78. In another aspect, the amino acid at the position corresponding to position 78 is substituted with Ala, Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, He, Leu, Met, Phe, Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variant comprises or consists of the substitution K78S or K78T in the polypeptide of SEQ ID NO: 1.

[0145] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 81. In another aspect, the amino acid at the position corresponding to position 81 is substituted with Ala, Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe, Pro, Ser, Trp, Tyr, or Val. In another aspect, the variant comprises or consists of the substitution T81M in the polypeptide of SEQ ID NO: 1.

[0146] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 84. In another aspect, the amino acid at the position corresponding to position 84 is substituted with Ala, Arg, Asn, Asp, Cys, Glu, Gly, His, He, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variant comprises or consists of the substitution Q84I or Q84W or Q84Y in the polypeptide of SEQ ID NO: 1.

[0147] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 86. In another aspect, the amino acid at the position corresponding to position 86 is substituted with Ala, Arg, Asn, Asp, Cys, Gln, Gly, His, He, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variant comprises or consists of the substitution E86K or E86S in the polypeptide of SEQ ID NO: 1.

[0148] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 87. In another aspect, the amino acid at the position corresponding to position 87 is substituted with Ala, Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, He, Leu, Lys, Met, Phe, Pro, Thr, Trp, Tyr, or Val. In another aspect, the variant comprises or consists of the substitution S87G or S87L or S87P in the polypeptide of SEQ ID NO: 1.

[0149] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 90. In another aspect, the amino acid at the position corresponding to position 90 is substituted with Ala, Arg, Asn, Asp, Cys, Gln, Glu, Gly, Ile, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variant comprises or consists of the substitution H90P in the polypeptide of SEQ ID NO: 1.

[0150] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 91. In another aspect, the amino acid at the position corresponding to position 91 is substituted with Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, He, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variant comprises or consists of the substitution A91Q or A91V or A91W in the polypeptide of SEQ ID NO: 1.

[0151] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 93. In another aspect, the amino acid at the position corresponding to position 93 is substituted with Ala, Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, He, Leu, Met, Phe, Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variant comprises or consists of the substitution K93V in the polypeptide of SEQ ID NO: 1.

[0152] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 97. In another aspect, the amino acid at the position corresponding to position 97 is substituted with Ala, Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, He, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp, or Tyr. In another aspect, the variant comprises or consists of the substitution V97C or V97H in the polypeptide of SEQ ID NO: 1.

[0153] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 98. In another aspect, the amino acid at the position corresponding to position 98 is substituted with Ala, Arg, Asn, Asp, Cys, Glu, Gly, His, He, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variant comprises or consists of the substitution Q98E or Q98F or Q98G or Q98I or Q98K or Q98L or Q98M or Q98R or Q98T or Q98Y in the polypeptide of SEQ ID NO: 1.

[0154] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 103. In another aspect, the amino acid at the position corresponding to position 103 is substituted with Ala, Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, He, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp, or Tyr. In another aspect, the variant comprises or consists of the substitution V103C or V10G or V103S in the polypeptide of SEQ ID NO: 1.

[0155] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 105. In another aspect, the amino acid at the position corresponding to position 105 is substituted with Ala, Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Phe, Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variant comprises or consists of the substitution M105F in the polypeptide of SEQ ID NO: 1.

[0156] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 110. In another aspect, the amino acid at the position corresponding to position 110 is substituted with Ala, Arg, Asn, Asp, Cys, Gln, Glu, His, Ile, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variant comprises or consists of the substitution G110K or G110R in the polypeptide of SEQ ID NO: 1.

[0157] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 111. In another aspect, the amino acid at the position corresponding to position 111 is substituted with Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variant comprises or consists of the substitution A111E or A111Q in the polypeptide of SEQ ID NO: 1.

[0158] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 113. In another aspect, the amino acid at the position corresponding to position 113 is substituted with Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, He, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variant comprises or consists of the substitution A113T in the polypeptide of SEQ ID NO: 1.

[0159] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 114. In another aspect, the amino acid at the position corresponding to position 114 is substituted with Ala, Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, He, Leu, Lys, Met, Phe, Pro, Ser, Trp, Tyr, or Val. In another aspect, the variant comprises or consists of the substitution T114M or T114V in the polypeptide of SEQ ID NO: 1.

[0160] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 116. In another aspect, the amino acid at the position corresponding to position 116 is substituted with Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, He, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variant comprises or consists of the substitution N116C or N116Y in the polypeptide of SEQ ID NO: 1.

[0161] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 117. In another aspect, the amino acid at the position corresponding to position 117 is substituted with Ala, Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp, or Tyr. In another aspect, the variant comprises or consists of the substitution V117F in the polypeptide of SEQ ID NO: 1.

[0162] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 119. In another aspect, the amino acid at the position corresponding to position 119 is substituted with Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, He, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variant comprises or consists of the substitution A119H or A119L or A119Y in the polypeptide of SEQ ID NO: 1.

[0163] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 120. In another aspect, the amino acid at the position corresponding to position 120 is substituted with Ala, Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp, or Tyr. In another aspect, the variant comprises or consists of the substitution V120E in the polypeptide of SEQ ID NO: 1.

[0164] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 121. In another aspect, the amino acid at the position corresponding to position 121 is substituted with Ala, Arg, Asn, Asp, Cys, Gln, Gly, His, Ile, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variant comprises or consists of the substitution E121I in the polypeptide of SEQ ID NO: 1.

[0165] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 125. In another aspect, the amino acid at the position corresponding to position 125 is substituted with Ala, Arg, Asp, Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variant comprises or consists of the substitution N125S in the polypeptide of SEQ ID NO: 1.

[0166] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 126. In another aspect, the amino acid at the position corresponding to position 126 is substituted with Ala, Arg, Asp, Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variant comprises or consists of the substitution N126I in the polypeptide of SEQ ID NO: 1.

[0167] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 128. In another aspect, the amino acid at the position corresponding to position 128 is substituted with Ala, Arg, Asp, Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variant comprises or consists of the substitution N128E in the polypeptide of SEQ ID NO: 1.

[0168] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 129. In another aspect, the amino acid at the position corresponding to position 129 is substituted with Ala, Arg, Asn, Asp, Cys, Glu, Gly, His, He, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variant comprises or consists of the substitution Q129T in the polypeptide of SEQ ID NO: 1.

[0169] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 131. In another aspect, the amino acid at the position corresponding to position 131 is substituted with Ala, Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variant comprises or consists of the substitution I131Y in the polypeptide of SEQ ID NO: 1.

[0170] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 132. In another aspect, the amino acid at the position corresponding to position 132 is substituted with Ala, Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, He, Leu, Lys, Met, Phe, Pro, Thr, Trp, Tyr, or Val. In another aspect, the variant comprises or consists of the substitution S132A or S132D or S132H or S132I or S132K or S132L or S132P or S132R or S132T or S132V or S132Y of the polypeptide of SEQ ID NO: 1.

[0171] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 133. In another aspect, the amino acid at the position corresponding to position 133 is substituted with Ala, Arg, Asn, Asp, Cys, Gln, Glu, His, He, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variant comprises or consists of the substitution G133E or G133L or G133T or G133V in the polypeptide of SEQ ID NO: 1.

[0172] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 135. In another aspect, the amino acid at the position corresponding to position 135 is substituted with Ala, Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, He, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp, or Val. In another aspect, the variant comprises or consists of the substitution Y135C or Y135D or Y135E or Y135P in the polypeptide of SEQ ID NO: 1.

[0173] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 137. In another aspect, the amino acid at the position corresponding to position 137 is substituted with Ala, Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variant comprises or consists of the substitution I137D or I137N or I137Q or I137W or I137Y in the polypeptide of SEQ ID NO: 1.

[0174] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 138. In another aspect, the amino acid at the position corresponding to position 138 is substituted with Ala, Arg, Asn, Asp, Cys, Gln, GIy, His, He, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variant comprises or consists of the substitution E138S or E138V in the polypeptide of SEQ ID NO: 1.

[0175] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 139. In another aspect, the amino acid at the position corresponding to position 139 is substituted with Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, He, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variant comprises or consists of the substitution A139L in the polypeptide of SEQ ID NO: 1.

[0176] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 140. In another aspect, the amino acid at the position corresponding to position 140 is substituted with Ala, Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, He, Leu, Lys, Met, Phe, Pro, Ser, Thr, Tyr, or Val. In another aspect, the variant comprises or consists of the substitution W140A or W140D or W140M or W140N or W140P or W140S or W140T or W140V in the polypeptide of SEQ ID NO: 1.

[0177] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 141. In another aspect, the amino acid at the position corresponding to position 141 is substituted with Ala, Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, He, Leu, Lys, Met, Phe, Pro, Ser, Trp, Tyr, or Val. In another aspect, the variant comprises or consists of the substitution T141G in the polypeptide of SEQ ID NO: 1.

[0178] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 142. In another aspect, the amino acid at the position corresponding to position 142 is substituted with Ala, Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, He, Leu, Met, Phe, Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variant comprises or consists of the substitution K142E or K142N in the polypeptide of SEQ ID NO: 1.

[0179] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 143. In another aspect, the amino acid at the position corresponding to position 143 is substituted with Ala, Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, He, Leu, Lys, Met, Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variant comprises or consists of the substitution F143H or F143I or F143M in the polypeptide of SEQ ID NO: 1.

[0180] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 145. In another aspect, the amino acid at the position corresponding to position 145 is substituted with Ala, Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, He, Leu, Lys, Met, Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variant comprises or consists of the substitution F145G or F145H or F145I or F145L or F145S of the polypeptide of SEQ ID NO: 1.

[0181] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 146. In another aspect, the amino acid at the position corresponding to position 146 is substituted with Ala, Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, He, Leu, Lys, Met, Phe, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variant comprises or consists of the substitution P146A or P146H or P146N in the polypeptide of SEQ ID NO: 1.

[0182] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 147. In another aspect, the amino acid at the position corresponding to position 147 is substituted with Ala, Arg, Asn, Asp, Cys, Gln, Glu, His, Ile, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variant comprises or consists of the substitution G147D in the polypeptide of SEQ ID NO: 1.

[0183] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 149. In another aspect, the amino acid at the position corresponding to position 149 is substituted with Ala, Arg, Asn, Asp, Cys, Gln, Glu, His, Ile, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variant comprises or consists of the substitution G149F or G149K in the polypeptide of SEQ ID NO: 1.

[0184] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 159. In another aspect, the amino acid at the position corresponding to position 159 is substituted with Ala, Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, He, Leu, Lys, Met, Phe, Pro, Ser, Thr, Tyr, or Val. In another aspect, the variant comprises or consists of the substitution W159A or W159E in the polypeptide of SEQ ID NO: 1.

[0185] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 161. In another aspect, the amino acid at the position corresponding to position 161 is substituted with Ala, Arg, Asn, Asp, Cys, Gln, Glu, Gly, Ile, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variant comprises or consists of the substitution H161W in the polypeptide of SEQ ID NO: 1.

[0186] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 162. In another aspect, the amino acid at the position corresponding to position 162 is substituted with Ala, Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, He, Leu, Lys, Met, Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variant comprises or consists of the substitution F162T in the polypeptide of SEQ ID NO: 1.

[0187] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 165. In another aspect, the amino acid at the position corresponding to position 165 is substituted with Ala, Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp, or Tyr. In another aspect, the variant comprises or consists of the substitution V165P in the polypeptide of SEQ ID NO: 1.

[0188] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 167. In another aspect, the amino acid at the position corresponding to position 167 is substituted with Ala, Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, He, Leu, Lys, Met, Phe, Pro, Ser, Thr, Tyr, or Val. In another aspect, the variant comprises or consists of the substitution W167D or W167G or W167P in the polypeptide of SEQ ID NO: 1.

[0189] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 169. In another aspect, the amino acid at the position corresponding to position 169 is substituted with Ala, Arg, Asn, Asp, Cys, Glu, Gly, His, He, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the parent comprises or consists of the substitution Q169L or Q169P or Q169V or Q169Y of the polypeptide of SEQ ID NO: 1.

[0190] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 171. In another aspect, the amino acid at the position corresponding to position 171 is substituted with Ala, Asn, Asp, Cys, Gln, Glu, Gly, His, He, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variant comprises or consists of the substitution R171P in the polypeptide of SEQ ID NO: 1.

[0191] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 176. In another aspect, the amino acid at the position corresponding to position 176 is substituted with Ala, Asn, Asp, Cys, Gln, Glu, Gly, His, He, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variant comprises or consists of the substitution R176E or R176Q or R176V or R176Y of the polypeptide of SEQ ID NO: 1.

[0192] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 177. In another aspect, the amino acid at the position corresponding to position 177 is substituted with Ala, Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variant comprises or consists of the substitution I177H or I177P in the polypeptide of SEQ ID NO: 1.

[0193] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 179. In another aspect, the amino acid at the position corresponding to position 179 is substituted with Ala, Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, He, Leu, Met, Phe, Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variant comprises or consists of the substitution K179A or K179M or K179V in the polypeptide of SEQ ID NO: 1.

[0194] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 180. In another aspect, the amino acid at the position corresponding to position 180 is substituted with Ala, Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, He, Leu, Lys, Met, Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variant comprises or consists of the substitution F180D or F180G in the polypeptide of SEQ ID NO: 1.

[0195] In another aspect, the variant comprises or consists of a deletion or substitution at a position corresponding to position 181. In another aspect, the amino acid at the position corresponding to position 181 is deleted from the polypeptide of SEQ ID NO: 1. In another aspect, the variant comprises or consists of a deletion or substitution at a position corresponding to position 181. In another aspect, the amino acid at the position corresponding to position 181 is deleted from the polypeptide of SEQ ID NO: 1. In another aspect, the variant comprises a deletion or substitution at a position corresponding to position 181 * In another aspect, the amino acid at the position corresponding to position 181 is substituted with Ala, Asn, Asp, Cys, Gln, Glu, Gly, His, He, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variant comprises or consists of the substitution R181D or R181E or R181G or R181M or R181Y of the polypeptide of SEQ ID NO: 1.

[0196] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 185. In another aspect, the amino acid at the position corresponding to position 185 is substituted with Ala, Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, He, Leu, Met, Phe, Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variant comprises or consists of the substitution K185T or K185V or K185W in the polypeptide of SEQ ID NO: 1.

[0197] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 186. In another aspect, the amino acid at the position corresponding to position 186 is substituted with Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, He, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variant comprises or consists of the substitution A186D in the polypeptide of SEQ ID NO: 1.

[0198] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 187. In another aspect, the amino acid at the position corresponding to position 187 is substituted with Ala, Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, He, Leu, Lys, Met, Phe, Pro, Ser, Thr, Tyr, or Val. In another aspect, the variant comprises or consists of the substitution W187T in the polypeptide of SEQ ID NO: 1.

[0199] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 195. In another aspect, the amino acid at the position corresponding to position 195 is substituted with Ala, Arg, Asp, Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variant comprises or consists of the substitution N195Y in the polypeptide of SEQ ID NO: 1.

[0200] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 196. In another aspect, the amino acid at the position corresponding to position 196 is substituted with Ala, Arg, Asn, Asp, Cys, Gln, Glu, His, He, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variant comprises or consists of the substitution G196D in the polypeptide of SEQ ID NO: 1.

[0201] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 197. In another aspect, the amino acid at the position corresponding to position 197 is substituted with Ala, Arg, Asp, Cys, Gln, Glu, Gly, His, He, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variant comprises or consists of the substitution N197V or N197Y in the polypeptide of SEQ ID NO: 1.

[0202] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 204. In another aspect, the amino acid at the position corresponding to position 204 is substituted with Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, He, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variant comprises or consists of the substitution A204F or A204H or A204L in the polypeptide of SEQ ID NO: 1.

[0203] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 206. In another aspect, the amino acid at the position corresponding to position 206 is substituted with Ala, Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, He, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp, or Tyr. In another aspect, the variant comprises or consists of the substitution V206N in the polypeptide of SEQ ID NO: 1.

[0204] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 208. In another aspect, the amino acid at the position corresponding to position 208 is substituted with Ala, Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Phe, Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variant comprises or consists of the substitution M208D in the polypeptide of SEQ ID NO: 1.

[0205] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 211. In another aspect, the amino acid at the position corresponding to position 211 is substituted with Ala, Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, He, Leu, Lys, Met, Phe, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variant comprises or consists of the substitution P211C or P211M in the polypeptide of SEQ ID NO: 1.

[0206] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 216. In another aspect, the amino acid at the position corresponding to position 216 is substituted with Ala, Arg, Asn, Asp, Cys, Gln, Gly, His, He, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variant comprises or consists of the substitution E216L in the polypeptide of SEQ ID NO: 1.

[0207] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 218. In another aspect, the amino acid at the position corresponding to position 218 is substituted with Ala, Asn, Asp, Cys, Gln, Glu, Gly, His, He, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variant comprises or consists of the substitution R218Q in the polypeptide of SEQ ID NO: 1.

[0208] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 219. In another aspect, the amino acid at the position corresponding to position 219 is substituted with Ala, Asn, Asp, Cys, Gln, Glu, Gly, His, He, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variant comprises or consists of the substitution R219V in the polypeptide of SEQ ID NO: 1.

[0209] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 225. In another aspect, the amino acid at the position corresponding to position 225 is substituted with Ala, Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, He, Leu, Lys, Met, Phe, Pro, Ser, Trp, Tyr, or Val. In another aspect, the variant comprises or consists of the substitution T225D or T225F or T225H or T225K or T225L or T225N or T225R or T225Y of the polypeptide of SEQ ID NO: 1.

[0210] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 228. In another aspect, the amino acid at the position corresponding to position 228 is substituted with Ala, Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, He, Lys, Met, Phe, Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variant comprises or consists of the substitution L228V in the polypeptide of SEQ ID NO: 1.

[0211] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 230. In another aspect, the amino acid at the position corresponding to position 230 is substituted with Ala, Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, He, Lys, Met, Phe, Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variant comprises or consists of the substitution L230C or L230F in the polypeptide of SEQ ID NO: 1.

[0212] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 231. In another aspect, the amino acid at the position corresponding to position 231 is substituted with Ala, Arg, Asn, Cys, Gln, Glu, Gly, His, He, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variant comprises or consists of the substitution D213G or D231T or D231V in the polypeptide of SEQ ID NO: 1.

[0213] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 235. In another aspect, the amino acid at the position corresponding to position 235 is substituted with Ala, Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variant comprises or consists of the substitution I235A in the polypeptide of SEQ ID NO: 1.

[0214] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 242. In another aspect, the amino acid at the position corresponding to position 242 is substituted with Ala, Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, Ile, Leu, Met, Phe, Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variant comprises or consists of the substitution K242L in the polypeptide of SEQ ID NO: 1.

[0215] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 243. In another aspect, the amino acid at the position corresponding to position 243 is substituted with Ala, Asn, Asp, Cys, Gln, Glu, Gly, His, He, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variant comprises or consists of the substitution Y243D or Y243M or Y243N in the polypeptide of SEQ ID NO: 1.

[0216] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 247. In another aspect, the amino acid at the position corresponding to position 247 is substituted with Ala, Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, He, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variant comprises or consists of the substitution R247G or R247P or R247S in the polypeptide of SEQ ID NO: 1.

[0217] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 250. In another aspect, the amino acid at the position corresponding to position 250 is substituted with Ala, Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, He, Lys, Met, Phe, Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variant comprises or consists of the substitution L250S or L250Y in the polypeptide of SEQ ID NO: 1.

[0218] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 251. In another aspect, the amino acid at the position corresponding to position 251 is substituted with Ala, Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, He, Leu, Lys, Met, Phe, Pro, Ser, Trp, Tyr, or Val. In another aspect, the variant comprises or consists of the substitution T251D in the polypeptide of SEQ ID NO: 1.

[0219] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 252. In another aspect, the amino acid at the position corresponding to position 252 is substituted with Ala, Arg, Asn, Asp, Cys, Gln, Glu, Gly, Ile, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variant comprises or consists of the substitution H252P in the polypeptide of SEQ ID NO: 1.

[0220] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 253. In another aspect, the amino acid at the position corresponding to position 253 is substituted with Ala, Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, He, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp, or Tyr. In another aspect, the variant comprises or consists of the substitution V253S or V253F in the polypeptide of SEQ ID NO: 1.

[0221] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 254. In another aspect, the amino acid at the position corresponding to position 254 is substituted with Ala, Asn, Asp, Cys, Gln, Glu, Gly, His, He, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variant comprises or consists of the substitution R254D or R254F or R254H or R254P or R254T or R254V or R254W of the polypeptide of SEQ ID NO: 1.

[0222] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 255. In another aspect, the amino acid at the position corresponding to position 255 is substituted with Ala, Arg, Asp, Cys, Gln, Glu, Gly, His, He, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variant comprises or consists of the substitution N255C or N255F or N255P in the polypeptide of SEQ ID NO: 1.

[0223] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 257. In another aspect, the amino acid at the position corresponding to position 257 is substituted with Ala, Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, He, Leu, Lys, Met, Phe, Pro, Ser, Trp, Tyr, or Val. In another aspect, the variant comprises or consists of the substitution T257A or T257C or T257D or T257E or T257G or T257P or T257Q or T257S or T257W or T257Y in the polypeptide of SEQ ID NO: 1.

[0224] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 259. In another aspect, the amino acid at the position corresponding to position 259 is substituted with Ala, Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, He, Leu, Met, Phe, Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variant comprises or consists of the substitution K259P or K259V or K259W or K259Y in the polypeptide of SEQ ID NO: 1.

[0225] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 260. In another aspect, the amino acid at the position corresponding to position 260 is substituted with Ala, Arg, Asn, Asp, Cys, Gln, Gly, His, He, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variant comprises or consists of the substitution E260V in the polypeptide of SEQ ID NO: 1.

[0226] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 261. In another aspect, the amino acid at the position corresponding to position 261 is substituted with Ala, Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, He, Leu, Lys, Phe, Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variant comprises or consists of the substitution M261D or M261F or M261P or M261R or M261S or M261W of the polypeptide of SEQ ID NO: 1.

[0227] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 262. In another aspect, the amino acid at the position corresponding to position 262 is substituted with Ala, Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, He, Leu, Lys, Met, Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variant comprises or consists of the substitution F262A or F262C or F262E or F262H or F262I or F262K or F262L or F262N or F262P or F262Q or F262R or F262S or F262T or F262V or F262W of the polypeptide of SEQ ID NO: 1.

[0228] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 263. In another aspect, the amino acid at the position corresponding to position 263 is substituted with Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, He, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variant comprises or consists of the substitution A263E or A263H or A263W in the polypeptide of SEQ ID NO: 1.

[0229] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 265. In another aspect, the amino acid at the position corresponding to position 265 is substituted with Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, He, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variant comprises or consists of the substitution A265Q in the polypeptide of SEQ ID NO: 1.

[0230] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 272. In another aspect, the amino acid at the position corresponding to position 272 is substituted with Ala, Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, He, Lys, Met, Phe, Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variant comprises or consists of the substitution L272W in the polypeptide of SEQ ID NO: 1.

[0231] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 273. In another aspect, the amino acid at the position corresponding to position 273 is substituted with Ala, Arg, Asn, Asp, Cys, Gln, Glu, His, He, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variant comprises or consists of the substitution G273Q in the polypeptide of SEQ ID NO: 1.

[0232] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 274. In another aspect, the amino acid at the position corresponding to position 274 is substituted with Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, He, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variant comprises or consists of the substitution A274D in the polypeptide of SEQ ID NO: 1.

[0233] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 279. In another aspect, the amino acid at the position corresponding to position 279 is substituted with Ala, Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, He, Lys, Met, Phe, Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variant comprises or consists of the substitution L279I in the polypeptide of SEQ ID NO: 1.

[0234] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 280. In another aspect, the amino acid at the position corresponding to position 280 is substituted with Ala, Arg, Asp, Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variant comprises or consists of the substitution N280P in the polypeptide of SEQ ID NO: 1.

[0235] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 281. In another aspect, the amino acid at the position corresponding to position 281 is substituted with Ala, Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, He, Leu, Met, Phe, Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variant comprises or consists of the substitution K281H or K281I or K281S in the polypeptide of SEQ ID NO: 1.

[0236] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 283. In another aspect, the amino acid at the position corresponding to position 283 is substituted with Ala, Arg, Asp, Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variant comprises or consists of the substitution N283P in the polypeptide of SEQ ID NO: 1.

[0237] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 284. In another aspect, the amino acid at the position corresponding to position 284 is substituted with Ala, Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, He, Leu, Lys, Met, Phe, Pro, Ser, Thr, Tyr, or Val. In another aspect, the variant comprises or consists of the substitution W284A or W284D or W284E or W284G or W284I or W284K or W284M or W284N or W284P or W284Q or W284S or W284T or W284V or W284Y in the polypeptide of SEQ ID NO: 1.

[0238] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 287. In another aspect, the amino acid at the position corresponding to position 287 is substituted with Ala, Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, He, Leu, Lys, Met, Phe, Pro, Thr, Trp, Tyr, or Val. In another aspect, the variant comprises or consists of the substitution S287I or S287K or S287L or S287P or S287R or S287T or S287V or S287Y of the polypeptide of SEQ ID NO: 1.

[0239] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 292. In another aspect, the amino acid at the position corresponding to position 292 is substituted with Ala, Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, He, Leu, Lys, Met, Phe, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variant comprises or consists of the substitution P292L or P292M or P292Y in the polypeptide of SEQ ID NO: 1.

[0240] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 294. In another aspect, the amino acid at the position corresponding to position 294 is substituted with Ala, Arg, Asn, Asp, Cys, Gln, Glu, Gly, Ile, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variant comprises or consists of the substitution H294N or H249S in the polypeptide of SEQ ID NO: 1.

[0241] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 296. In another aspect, the amino acid at the position corresponding to position 296 is substituted with Ala, Arg, Asp, Cys, Gln, Glu, Gly, His, He, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variant comprises or consists of the substitution N296T in the polypeptide of SEQ ID NO: 1.

[0242] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 298. In another aspect, the amino acid at the position corresponding to position 298 is substituted with Ala, Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, He, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp, or Val. In another aspect, the variant comprises or consists of the substitution Y298D or Y298E in the polypeptide of SEQ ID NO: 1.

[0243] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 299. In another aspect, the amino acid at the position corresponding to position 299 is substituted with Ala, Arg, Asp, Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variant comprises or consists of the substitution N299T in the polypeptide of SEQ ID NO: 1.

[0244] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 300. In another aspect, the amino acid at the position corresponding to position 300 is substituted with Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, He, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variant comprises or consists of the substitution A300G in the polypeptide of SEQ ID NO: 1.

[0245] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 303. In another aspect, the amino acid at the position corresponding to position 303 is substituted with Ala, Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, He, Leu, Lys, Met, Phe, Pro, Thr, Trp, Tyr, or Val. In another aspect, the variant comprises or consists of the substitution S303A or S303C or S303D or S303E or S303F or S303G or S303H or S303L or S303M or S303N or S303Q or S303T or S303V or S303Y of the polypeptide of SEQ ID NO: 1.

[0246] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 305. In another aspect, the amino acid at the position corresponding to position 305 is substituted with Ala, Arg, Asn, Asp, Cys, Gln, Glu, His, Ile, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variant comprises or consists of the substitution G305E in the polypeptide of SEQ ID NO: 1.

[0247] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 307. In another aspect, the amino acid at the position corresponding to position 307 is substituted with Ala, Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, He, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp, or Val. In another aspect, the variant comprises or consists of the substitution Y307S in the polypeptide of SEQ ID NO: 1.

[0248] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 310. In another aspect, the amino acid at the position corresponding to position 310 is substituted with Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, He, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variant comprises or consists of the substitution A310D or A310N in the polypeptide of SEQ ID NO: 1.

[0249] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 311. In another aspect, the amino acid at the position corresponding to position 311 is substituted with Ala, Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, He, Leu, Met, Phe, Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variant comprises or consists of the substitution K311C or K311C in the polypeptide of SEQ ID NO: 1.

[0250] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 313. In another aspect, the amino acid at the position corresponding to position 313 is substituted with Ala, Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, He, Lys, Met, Phe, Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variant comprises or consists of the substitution L313Q in the polypeptide of SEQ ID NO: 1.

[0251] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 314. In another aspect, the amino acid at the position corresponding to position 314 is substituted with Ala, Arg, Asp, Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variant comprises or consists of the substitution N314S or N314P in the polypeptide of SEQ ID NO: 1.

[0252] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 315. In another aspect, the amino acid at the position corresponding to position 315 is substituted with Ala, Arg, Asn, Asp, Cys, Gln, Glu, His, Ile, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variant comprises or consists of the substitution G315E or G315M or G315V in the polypeptide of SEQ ID NO: 1.

[0253] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 319. In another aspect, the amino acid at the position corresponding to position 319 is substituted with Ala, Arg, Asn, Asp, Cys, Glu, Gly, His, He, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variant comprises or consists of the substitution Q319C or Q319E in the polypeptide of SEQ ID NO: 1.

[0254] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 320. In another aspect, the amino acid at the position corresponding to position 320 is substituted with Ala, Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, He, Leu, Met, Phe, Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variant comprises or consists of the substitution K320S or K320W in the polypeptide of SEQ ID NO: 1.

[0255] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 321. In another aspect, the amino acid at the position corresponding to position 321 is substituted with Ala, Arg, Asn, Asp, Cys, Gln, Glu, Gly, Ile, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variant comprises or consists of the substitution H321F or H321K or H321L or H321R or H321T or H321V or H321W in the polypeptide of SEQ ID NO: 1.

[0256] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 322. In another aspect, the amino acid at the position corresponding to position 322 is substituted with Ala, Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, He, Leu, Lys, Met, Phe, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variant comprises or consists of the substitution P322C or P322E or P322H or P322T or P322Y of the polypeptide of SEQ ID NO: 1.

[0257] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 323. In another aspect, the amino acid at the position corresponding to position 323 is substituted with Ala, Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, He, Leu, Lys, Phe, Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variant comprises or consists of the substitution M323A or M323C or M323D or M323E or M323G or M323H or M323L or M323N or M323P or M323R or M323S or M323T or M323W or M323Y in the polypeptide of SEQ ID NO: 1.

[0258] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 324. In another aspect, the amino acid at the position corresponding to position 324 is substituted with Ala, Arg, Asn, Asp, Cys, Gln, Glu, Gly, Ile, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variant comprises or consists of the substitution H324A or H324I or H324P or H324T or H324W or H324Y in the polypeptide of SEQ ID NO: 1.

[0259] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 334. In another aspect, the amino acid at the position corresponding to position 334 is substituted with Ala, Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, He, Leu, Lys, Met, Phe, Pro, Thr, Trp, Tyr, or Val. In another aspect, the variant comprises or consists of the substitution S334V in the polypeptide of SEQ ID NO: 1.

[0260] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 337. In another aspect, the amino acid at the position corresponding to position 337 is substituted with Ala, Arg, Asn, Asp, Cys, Gln, Glu, His, Ile, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variant comprises or consists of the substitution G337V in the polypeptide of SEQ ID NO: 1.

[0261] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 340. In another aspect, the amino acid at the position corresponding to position 340 is substituted with Ala, Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, He, Lys, Met, Phe, Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variant comprises or consists of the substitution L340M in the polypeptide of SEQ ID NO: 1.

[0262] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 342. In another aspect, the amino acid at the position corresponding to position 342 is substituted with Ala, Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, He, Leu, Lys, Met, Phe, Pro, Thr, Trp, Tyr, or Val. In another aspect, the variant comprises or consists of the substitution S342A or S342G or S342T of the polypeptide of SEQ ID NO: 1.

[0263] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 344. In another aspect, the amino acid at the position corresponding to position 344 is substituted with Ala, Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, He, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp, or Tyr. In another aspect, the variant comprises or consists of the substitution V344A or V344C or V344I of the polypeptide of SEQ ID NO: 1.

[0264] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 345. In another aspect, the amino acid at the position corresponding to position 345 is substituted with Ala, Arg, Asn, Asp, Cys, Glu, Gly, His, He, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variant comprises or consists of the substitution Q345N in the polypeptide of SEQ ID NO: 1.

[0265] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 346. In another aspect, the amino acid at the position corresponding to position 346 is substituted with Ala, Arg, Asn, Asp, Cys, Gln, GIy, His, He, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variant comprises or consists of the substitution E346A in the polypeptide of SEQ ID NO: 1.

[0266] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 347. In another aspect, the amino acid at the position corresponding to position 347 is substituted with Ala, Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, He, Leu, Lys, Met, Phe, Pro, Ser, Thr, Tyr, or Val. In another aspect, the variant comprises or consists of the substitution W347A or W347E or W347F or W347G or W347H or W347K or W347N or W347P or W347R in the polypeptide of SEQ ID NO: 1.

[0267] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 350. In another aspect, the amino acid at the position corresponding to position 350 is substituted with Ala, Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, He, Leu, Lys, Met, Phe, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variant comprises or consists of the substitution P350A or P350C or P350N or P350T of the polypeptide of SEQ ID NO: 1.

[0268] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 359. In another aspect, the amino acid at the position corresponding to position 359 is substituted with Ala, Asn, Asp, Cys, Gln, Glu, Gly, His, He, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variant comprises or consists of the substitution R359C or R359F or R359G or R359M or R359S or R359W of the polypeptide of SEQ ID NO: 1.

[0269] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 360. In another aspect, the amino acid at the position corresponding to position 360 is substituted with Ala, Arg, Asn, Asp, Cys, Gln, GIy, His, He, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variant comprises or consists of the substitution E360D or E360H or E360P or E360T or E360W in the polypeptide of SEQ ID NO: 1.

[0270] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 361. In another aspect, the amino acid at the position corresponding to position 361 is substituted with Ala, Arg, Asn, Asp, Cys, Glu, Gly, His, He, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variant comprises or consists of the substitution Q361D or Q361H or Q361I or Q361K or Q361L or Q361M or Q361N or Q361P or Q361R or Q361S or Q361T or Q361V or Q361W or Q361Y of the polypeptide of SEQ ID NO: 1.

[0271] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 362. In another aspect, the amino acid at the position corresponding to position 362 is substituted with Ala, Arg, Asn, Asp, Cys, Gln, Glu, His, Ile, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variant comprises or consists of the substitution G326M in the polypeptide of SEQ ID NO: 1.

[0272] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 362. In another aspect, the amino acid at the position corresponding to position 362 is substituted with Ala, Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, He, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp, or Val. In another aspect, the variant comprises or consists of the substitution Y363A or Y363E or Y363F or Y363G or Y363I or Y363L or Y363Q or Y363R or Y363S or Y363T or Y363V or Y363W of the polypeptide of SEQ ID NO: 1.

[0273] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 364. In another aspect, the amino acid at the position corresponding to position 364 is substituted with Ala, Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, He, Leu, Lys, Met, Phe, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variant comprises or consists of the substitution P364C in the polypeptide of SEQ ID NO: 1.

[0274] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 368. In another aspect, the amino acid at the position corresponding to position 368 is substituted with Ala, Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, He, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp, or Val. In another aspect, the variant comprises or consists of the substitution Y368C or Y368Q or Y368R or Y368T of the polypeptide of SEQ ID NO: 1.

[0275] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 372. In another aspect, the amino acid at the position corresponding to position 372 is substituted with Ala, Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, He, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp, or Val. In another aspect, the variant comprises or consists of the substitutions Y372C, Y327G, Y327N, Y327T in the polypeptide of SEQ ID NO: 1.

[0276] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 375. In another aspect, the amino acid at the position corresponding to position 375 is substituted with Ala, Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, He, Leu, Lys, Met, Phe, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variant comprises or consists of the substitution P375Q or P375T in the polypeptide of SEQ ID NO: 1.

[0277] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 380. In another aspect, the amino acid at the position corresponding to position 380 is substituted with Ala, Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, He, Leu, Lys, Met, Phe, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variant comprises or consists of the substitution P380F in the polypeptide of SEQ ID NO: 1.

[0278] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 382. In another aspect, the amino acid at the position corresponding to position 382 is substituted with Ala, Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Phe, Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variant comprises or consists of the substitution M328I in the polypeptide of SEQ ID NO: 1.

[0279] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 383. In another aspect, the amino acid at the position corresponding to position 383 is substituted with Ala, Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, He, Leu, Met, Phe, Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variant comprises or consists of the substitution K383C or K383L or K383P or K383T of the polypeptide of SEQ ID NO: 1.

[0280] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 384. In another aspect, the amino acid at the position corresponding to position 384 is substituted with Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, He, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variant comprises or consists of the substitution A384D or A384N in the polypeptide of SEQ ID NO: 1.

[0281] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 385. In another aspect, the amino acid at the position corresponding to position 385 is substituted with Ala, Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, He, Leu, Met, Phe, Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variant comprises or consists of the substitution K385N or K385V in the polypeptide of SEQ ID NO: 1.

[0282] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 387. In another aspect, the amino acid at the position corresponding to position 387 is substituted with Ala, Arg, Asn, Cys, Gln, Glu, Gly, His, He, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variant comprises or consists of the substitution D387C or D387P or D387R or D387W in the polypeptide of SEQ ID NO: 1.

[0283] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 388. In another aspect, the amino acid at the position corresponding to position 388 is substituted with Ala, Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, He, Leu, Lys, Met, Phe, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variant comprises or consists of the substitution P388I in the polypeptide of SEQ ID NO: 1.

[0284] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 391. In another aspect, the amino acid at the position corresponding to position 391 is substituted with Ala, Arg, Asn, Asp, Cys, Gln, GIy, His, He, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variant comprises or consists of the substitution E399M in the polypeptide of SEQ ID NO: 1.

[0285] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 393. In another aspect, the amino acid at the position corresponding to position 393 is substituted with Ala, Asn, Asp, Cys, Gln, Glu, Gly, His, He, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variant comprises or consists of the substitution R393K or R393M in the polypeptide of SEQ ID NO: 1.

[0286] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 395. In another aspect, the amino acid at the position corresponding to position 395 is substituted with Ala, Arg, Asp, Cys, Gln, Glu, Gly, His, He, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variant comprises or consists of the substitution N395C or N395D or N395K or N395V or N395W of the polypeptide of SEQ ID NO: 1.

[0287] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 396. In another aspect, the amino acid at the position corresponding to position 396 is substituted with Ala, Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, He, Leu, Lys, Met, Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variant comprises or consists of the substitution F396E or F396G or F396T in the polypeptide of SEQ ID NO: 1.

[0288] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 397. In another aspect, the amino acid at the position corresponding to position 397 is substituted with Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, He, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variant comprises or consists of the substitution A379V in the polypeptide of SEQ ID NO: 1.

[0289] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 398. In another aspect, the amino acid at the position corresponding to position 398 is substituted with Ala, Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, He, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp, or Val. In another aspect, the variant comprises or consists of the substitution Y398E in the polypeptide of SEQ ID NO: 1.

[0290] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 401. In another aspect, the amino acid at the position corresponding to position 401 is substituted with Ala, Arg, Asn, Asp, Cys, Glu, Gly, His, He, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variant comprises or consists of the substitution Q401H in the polypeptide of SEQ ID NO: 1.

[0291] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 402. In another aspect, the amino acid at the position corresponding to position 402 is substituted with Ala, Arg, Asn, Asp, Cys, Gln, Glu, Gly, Ile, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variant comprises or consists of the substitution H402S in the polypeptide of SEQ ID NO: 1.

[0292] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 403. In another aspect, the amino acid at the position corresponding to position 403 is substituted with Ala, Arg, Asn, Cys, Gln, Glu, Gly, His, He, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variant comprises or consists of the substitution D403A or D403E or D403T of the polypeptide of SEQ ID NO: 1.

[0293] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 405. In another aspect, the amino acid at the position corresponding to position 405 is substituted with Ala, Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, He, Leu, Lys, Met, Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variant comprises or consists of the substitution F405C or F405N or F405S or F405T of the polypeptide of SEQ ID NO: 1.

[0294] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 406. In another aspect, the amino acid at the position corresponding to position 406 is substituted with Ala, Arg, Asn, Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variant comprises or consists of the substitution D406V in the polypeptide of SEQ ID NO: 1.

[0295] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 407. In another aspect, the amino acid at the position corresponding to position 407 is substituted with Ala, Arg, Asn, Asp, Cys, Gln, Glu, Gly, Ile, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variant comprises or consists of the substitution H407C or H407Q or H407T of the polypeptide of SEQ ID NO: 1.

[0296] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 407. In another aspect, the amino acid at the position corresponding to position 407 is substituted with Ala, Arg, Asn, Asp, Cys, Gln, Glu, Gly, Ile, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variant comprises or consists of the substitution H407C or H407Q or H407T of the polypeptide of SEQ ID NO: 1.

[0297] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 410. In another aspect, the amino acid at the position corresponding to position 410 is substituted with Ala, Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variant comprises or consists of the substitution I410H or I410M or I410R or I410W in the polypeptide of SEQ ID NO: 1.

[0298] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 415. In another aspect, the amino acid at the position corresponding to position 415 is substituted with Ala, Asn, Asp, Cys, Gln, Glu, Gly, His, He, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variant comprises or consists of the substitution R415C or R415L or R415M or R415Y of the polypeptide of SEQ ID NO: 1.

[0299] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 416. In another aspect, the amino acid at the position corresponding to position 416 is substituted with Ala, Arg, Asn, Asp, Cys, Gln, GIy, His, He, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variant comprises or consists of the substitution E416D or E416F or E416L or E416N in the polypeptide of SEQ ID NO: 1.

[0300] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 418. In another aspect, the amino acid at the position corresponding to position 418 is substituted with Ala, Arg, Asp, Cys, Gln, Glu, Gly, His, He, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variant comprises or consists of the substitution N418A or N418P or N418V in the polypeptide of SEQ ID NO: 1.

[0301] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 421. In another aspect, the amino acid at the position corresponding to position 421 is substituted with Ala, Arg, Asn, Asp, Cys, Gln, Glu, Gly, Ile, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variant comprises or consists of the substitution H421A or H421G or H421L or H421P or H421Q or H421Y in the polypeptide of SEQ ID NO: 1.

[0302] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 422. In another aspect, the amino acid at the position corresponding to position 422 is substituted with Ala, Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, He, Leu, Lys, Met, Phe, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variant comprises or consists of the substitution P422D or P422G or P422I or P422N or P422Q or P422S of the polypeptide of SEQ ID NO: 1.

[0303] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 423. In another aspect, the amino acid at the position corresponding to position 423 is substituted with Ala, Arg, Asp, Cys, Gln, Glu, Gly, His, He, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variant comprises or consists of the substitution N423G or N423H or N423I or N423L or N423M or N423P or N423Q or N423V or N423W or N423Y of the polypeptide of SEQ ID NO: 1.

[0304] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 433. In another aspect, the amino acid at the position corresponding to position 433 is substituted with Ala, Arg, Asn, Asp, Cys, Gln, Glu, His, He, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variant comprises or consists of the substitution G433A or G433N or G433S of the polypeptide of SEQ ID NO: 1.

[0305] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 434. In another aspect, the amino acid at the position corresponding to position 434 is substituted with Ala, Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, He, Leu, Lys, Met, Phe, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variant comprises or consists of the substitution P434I or P434L in the polypeptide of SEQ ID NO: 1.

[0306] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 435. In another aspect, the amino acid at the position corresponding to position 435 is substituted with Ala, Arg, Asn, Asp, Cys, Gln, Glu, His, He, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variant comprises or consists of the substitution G435C or G435L or G435M or G435T or G435V of the polypeptide of SEQ ID NO: 1.

[0307] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 438. In another aspect, the amino acid at the position corresponding to position 438 is substituted with Ala, Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, He, Leu, Met, Phe, Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variant comprises or consists of the substitution K438A or K438I or K438Q or K438S or K438V of the polypeptide of SEQ ID NO: 1.

[0308] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 439. In another aspect, the amino acid at the position corresponding to position 439 is substituted with Ala, Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, He, Leu, Lys, Met, Phe, Pro, Ser, Thr, Tyr, or Val. In another aspect, the variant comprises or consists of the substitution W439A or W439D or W439I or W439K or W439L or W439N or W439S or W439T or W439V or W439Y in the polypeptide of SEQ ID NO: 1.

[0309] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 446. In another aspect, the amino acid at the position corresponding to position 446 is substituted with Ala, Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, He, Leu, Met, Phe, Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variant comprises or consists of the substitution K446A or K446F in the polypeptide of SEQ ID NO: 1.

[0310] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 447. In another aspect, the amino acid at the position corresponding to position 447 is substituted with Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, He, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variant comprises or consists of the substitution A447D or A447Y in the polypeptide of SEQ ID NO: 1.

[0311] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 449. In another aspect, the amino acid at the position corresponding to position 449 is substituted with Ala, Arg, Asn, Asp, Cys, Glu, Gly, His, He, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variant comprises or consists of the substitution Q449T in the polypeptide of SEQ ID NO: 1.

[0312] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 450. In another aspect, the amino acid at the position corresponding to position 450 is substituted with Ala, Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp, or Tyr. In another aspect, the variant comprises or consists of the substitution V450D or V450N in the polypeptide of SEQ ID NO: 1.

[0313] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 452. In another aspect, the amino acid at the position corresponding to position 452 is substituted with Ala, Arg, Asn, Asp, Cys, Gln, Glu, Gly, Ile, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variant comprises or consists of the substitution H452L in the polypeptide of SEQ ID NO: 1.

[0314] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 455. In another aspect, the amino acid at the position corresponding to position 455 is substituted with Ala, Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, He, Leu, Lys, Met, Phe, Pro, Ser, Trp, Tyr, or Val. In another aspect, the variant comprises or consists of the substitution T455C or T455M or T455V in the polypeptide of SEQ ID NO: 1.

[0315] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 457. In another aspect, the amino acid at the position corresponding to position 457 is substituted with Ala, Arg, Asp, Cys, Gln, Glu, Gly, His, He, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variant comprises or consists of the substitution N457D or N457E or N457I or N457L or N457V of the polypeptide of SEQ ID NO: 1.

[0316] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 458. In another aspect, the amino acid at the position corresponding to position 458 is substituted with Ala, Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, He, Leu, Met, Phe, Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variant comprises or consists of the substitution K458D or K458L in the polypeptide of SEQ ID NO: 1.

[0317] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 460. In another aspect, the amino acid at the position corresponding to position 460 is substituted with Ala, Arg, Asn, Asp, Cys, Gln, Glu, His, Ile, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variant comprises or consists of the substitution G460M or G460V in the polypeptide of SEQ ID NO: 1.

[0318] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 461. In another aspect, the amino acid at the position corresponding to position 461 is substituted with Ala, Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, He, Leu, Lys, Met, Phe, Pro, Ser, Trp, Tyr, or Val. In another aspect, the variant comprises or consists of the substitution T461F or T461I or T461Q or T461V in the polypeptide of SEQ ID NO: 1.

[0319] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 462. In another aspect, the amino acid at the position corresponding to position 462 is substituted with Ala, Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, He, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp, or Tyr. In another aspect, the variant comprises or consists of the substitution V462C in the polypeptide of SEQ ID NO: 1.

[0320] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 465. In another aspect, the amino acid at the position corresponding to position 465 is substituted with Ala, Arg, Asp, Cys, Gln, Glu, Gly, His, He, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variant comprises or consists of the substitution N465A or N465C or N465E or N465G or N465H or N465L or N465P or N465T of the polypeptide of SEQ ID NO: 1.

[0321] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 466. In another aspect, the amino acid at the position corresponding to position 466 is substituted with Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, He, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variant comprises or consists of the substitution A466F or A466I or A466Y in the polypeptide of SEQ ID NO: 1.

[0322] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 467. In another aspect, the amino acid at the position corresponding to position 467 is substituted with Ala, Arg, Asn, Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variant comprises or consists of the substitution D467I in the polypeptide of SEQ ID NO: 1.

[0323] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 469. In another aspect, the amino acid at the position corresponding to position 469 is substituted with Ala, Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, He, Leu, Lys, Met, Phe, Pro, Ser, Thr, Tyr, or Val. In another aspect, the variant comprises or consists of the substitution W469A or W469E or W469H or W469L or W469N or W469Q or W469R or W469S or W469Y in the polypeptide of SEQ ID NO: 1.

[0324] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 471. In another aspect, the amino acid at the position corresponding to position 471 is substituted with Ala, Arg, Asp, Cys, Gln, Glu, Gly, His, He, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variant comprises or consists of the substitution N471A or N471D or N471M or N471W in the polypeptide of SEQ ID NO: 1.

[0325] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 473. In another aspect, the amino acid at the position corresponding to position 473 is substituted with Ala, Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe, Pro, Thr, Trp, Tyr, or Val. In another aspect, the variant comprises or consists of the substitution S473N in the polypeptide of SEQ ID NO: 1.

[0326] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 475. In another aspect, the amino acid at the position corresponding to position 475 is substituted with Ala, Arg, Asp, Cys, Gln, Glu, Gly, His, He, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variant comprises or consists of the substitution N475A or N475C or N475D or N475E or N475F or N475G or N475H or N475I or N475L or N475M or N475P or N475Q or N475S or N475T or N475V or N475W of the polypeptide of SEQ ID NO: 1.

[0327] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 476. In another aspect, the amino acid at the position corresponding to position 476 is substituted with Ala, Arg, Asn, Asp, Cys, Gln, Glu, His, He, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variant comprises or consists of the substitution G476F or G476L in the polypeptide of SEQ ID NO: 1.

[0328] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 477. In another aspect, the amino acid at the position corresponding to position 477 is substituted with Ala, Arg, Asn, Asp, Cys, Gln, Glu, His, He, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp, Tyr, or Val. In another aspect, the variant comprises or consists of the substitution G477A or G477H or G477Q or G477S or G477W of the polypeptide of SEQ ID NO: 1.

[0329] In another aspect, the variant comprises or consists of a substitution at a position corresponding to position 478. In another aspect, the amino acid at the position corresponding to position 478 is substituted with Ala, Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, He, Leu, Lys, Met, Phe, Pro, Thr, Trp, Tyr, or Val. In another aspect, the variant comprises or consists of the substitution S478A or S478F or S478Q of the polypeptide of SEQ ID NO: 1.

[0330] In one aspect, the α-amylase variants are selected from the group consisting of α-amylases at positions 3, 5, 7, 8, 16, 18, 25, 26, 29, 30, 35, 37, 40, 41, 43, 46, 50, 57, 60, 68, 71, 73, 74, 78, 81, 84, 86, 87, 90, 91, 93, 97, 98, 103, 105, 110, 111, 113, 114, 116, 117, 119, 120, 121, 125, 126, 128, 129, 131, 132, 133, 135, 137, 138, 139, 140, 141, 142, 143, 144, 145, 146, 147, 148, 149, 150, 151, 152, 153, 154, 155, 156, 157, 158, 159, 200, 201, 202, 203, 204, 205, 206, 207, 208, 209, 210, 211, 212, 213, 214, 215, 216, 217, 218, 219, 220, 221, 222, 223, 224, 225, 226, 228, 229 9, 140, 141, 142, 143, 145, 146, 147, 149, 159, 161, 162, 165, 167, 169, 171, 176, 177, 179, 180, 181, 185, 186, 187, 195, 196, 197, 204, 206, 208, 211, 216, 218, 219, 225, 228, 230, 231, 235, 242, 243, 247, 250, 251, 252, 253, 254, 255, 257, 259, 260, 261, 262, 263, 265, 272, 273, 274, 279, 280, 281, 283, 284, 287, 292, 294, 296, 298, 299, 300, 303, 305, 307, 310, 311, 313, 314, 315, 319, 320, 321, 322, 323, 324, 334, 337, 339, 340, 342, 344, 345, 346, 347, 350, 359, 360, 361, 362, 363, 364, 368, 372, 375, 380, 382, ​​383, 384, 385, 387, 388, 391, 39 3, 395, 396, 397, 398, 401, 402, 403, 405, 406, 407, 410, 415, 416, 418, 421, 422, 423, 433, 434, 435, 438, 439, 446, 447, 449, 450, 452, 455, 457, 458, 460, 461, 462, 465, 466, 467, 469, 471, 473, 475, 476, 477, and 478, and further wherein the α-amylase variant has an improvement factor (IF) of >1.and the variant has an increased specific activity measured as 1, and further has at least 60%, 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%, or at least 99% but less than 100% sequence identity to the polypeptide of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, or 17.

[0331] Preferably, the one or more substitutions are at positions N3A, N3C, N3D, N3E, N3F, N3G, N3H, N3L, N3P, N3Q, N3S, N3T, N3V, T5E, G7E, G7F, G7H, G7K, G7L, G7P, G7R, G7S, G7T, G7V, G7W, T8S, H16D, H16R, P18D, P18L, P18N, N25T, R26Y, D29N, D30F, R35P, R37A, R37N, T40A, T40C, T40D, T40E, T40G, T40H, T40I, T40K, T40N, T40 V, T40W, A41C, A41D, A41E, A41H, A41I, A41K, A41N, A41R, A41T, A41V, A41Y, W43E, P46A, G50M, G50P, G50Q, G57A, G57S, A60E, A60S, A60V, E68F, E68Y, Q71 F, G73D, G73H, G73I, G73L, G73Q, G73S, G73T, G73W, T74F, K78S, K78T, T81M, Q84I, Q84W, Q84Y, E86K, E86S, S87G, S87L, S87P, H90P, A91Q, A91V, A91W, K93 V, V97C, V97H, Q98E, Q98F, Q98G, Q98I, Q98K, Q98L, Q98M, Q98R, Q98T, Q98Y, V103C, V103G, V103S, M105F, G110K, G110R, A111E, A111Q, A113T, T114M, T11 4V, N116C, N116Y, V117F, A119H, A119L, A119Y, V120E, E121I, N125S, N126I , N128E, Q129T, I131Y, S132A, S132D, S132H, S132I, S132K, S132L, S132P, S1 32R, S132T, S132V, S132Y, G133E, G133L, G133T, G133V, Y135C, Y135D, Y135 E, Y135P, I137D, I137N, I137Q, I137W, I137Y, E138S, E138V, A139L, W140A, W 140D, W140M, W140N, W140P, W140S, W140T, W140V, T141G, K142E, K142N, F14 3H, F143I, F143M, F145G, F145H, F145I, F145L, F145S, P146A, P146H, P146N,G147D、G149F、G149K、W159A、W159E、H161W、F162T、V165P、W167D、W167G、W167P、Q169L、Q169P、Q169V、Q169Y、R171P、R176E、R176Q、R176V、R176Y、I177H、I177P、K179A、K179M、K179V、F180D、F180G、R181D、R181E、R181G、R181M、R181Y、K185T、K185V、K185W、A186D、W187T、N195Y、G196D、N197V、N197Y、A204F、A204H、A204L、V206N、M208D、P211C、P211M、E216L、R218Q、R219V、T225D、T225F、T225H、T225K、T225L、T225N、T225R、T225Y、L228V、L230C、L230F、D231G、D231T、D231V、I235A、K242L、Y243D、Y243M、Y243N、R247G、R247P、R247S、L250S、L250Y、T251D、H252P、V253F、V253S、R254D、R254F、R254H、R254P、R254T、R254V、R254W、N255C、N255F、N255P、T257A、T257C、T257D、T257E、T257G、T257P、T257Q、T257S、T257W、T257Y、K259P、K259V、K259W、K259Y、E260V、M261D、M261F、M261P、M261R、M261S、M261W、F262A、F262C、F262E、F262H、F262I、F262K、F262L、F262N、F262P、F262Q、F262R、F262S、F262T、F262V、F262W、A263E、A263H、A263W、A265Q、L272W、G273Q、A274D、L279I、N280P、K281H、K281I、K281S、N283P、W284A、W284D、W284E、W284G、W284I、W284K、W284M、W284N、W284P、W284Q、W284S、W284T、W284V、W284Y、S287I、S287K、S287L、S287P、S287R、S287T、S287V、S287Y、P292L、P292M、P292Y、H294N、H294S、N296T、Y298D, Y298E, N299T, A300G, S303A, S303C, S303D, S303E, S303F, S303G, S303H, S303L, S303M, S303N, S303Q, S303T, S303V, S303Y, G305E, Y307S, A310 D, A310N, K311A, K311C, L313Q, N314P, N314S, G315E, G315M, G315V, Q319C, Q319E, K320S, K320W, H321F, H321K, H321L, H321R, H321T, H321V, H321W, P32 2C, P322E, P322H, P322T, P322Y, M323A, M323C, M323D, M323E, M323G, M323H, M323L, M323N, M323P, M323R, M323S, M323T, M323W, M323Y, H324A, H324I, H 324P, H324T, H324W, H324Y, S334V, G337V, L340M, S342A, S342G, S342T, V344A, V344C, V344I, Q345N, E346A, W347A, W347E, W347F, W347G, W347H, W347K W347N, W347P, W347R, P350A, P350C, P350N, P350T, R359C, R359F, R359G, R359M, R359S, R359W, E360D, E360H, E360P, E360T, E360W, Q361D, Q361H, Q361 I、Q361K、Q361L、Q361M、Q361N、Q361P、Q361R、Q361S、Q361T、Q361V、Q361W、 Q361Y、G362M、Y363A、Y363E、Y363F、Y363G、Y363I、Y363L、Y363Q、Y363R、Y36 3S, Y363T, Y363V, Y363W, P364C, Y368C, Y368Q, Y368R, Y368T, Y372C, Y372G, Y372N, Y372T, P375Q, P375T, P380F, M382I, K383C, K383L, K383P, K383T, A 384D, A384N, K385N, K385V, D387C, D387P, D387R, D387W, P388I, E391M, R393K, R393M, N395C, N395D, N395K, N395V, N395W, F396E, F396G, F396T, A397VY398E, Q401H, H402S, D403A, D403E, D403T, F405C, F405N, F405S, F405T , D406V, H407C, H407Q, H407T, I410H, I410M, I410R, I410W, R415C, R415 L, R415M, R415Y, E416D, E416F, E416L, E416N, N418A, N418P, N418V, H42 1A, H421G, H421L, H421P, H421Q, H421Y, P422D, P422G, P422I, P422N, P42 2Q, P422S, N423G, N423H, N423I, N423L, N423M, N423P, N423Q, N423V, N4 23W, N423Y, G433A, G433N, G433S, P434I, P434L, G435C, G435L, G435M, G 435T, G435V, K438A, K438I, K438Q, K438S, K438V, W439A, W439D, W439I, W439K, W439L, W439N, W439S, W439T, W439V, W439Y, K446A, K446F, A447D, A447Y, Q449T, V450D, V450N, H452L, T455C, T455M, T455V, N457D, N457E , N457I, N457L, N457V, K458D, K458L, G460M, G460V, T461F, T461I, T461 Q, T461V, V462C, N465A, N465C, N465E, N465G, N465H, N465L, N465P, N46 5T, A466F, A466I, A466Y, D467I, W469A, W469E, W469H, W469L, W469N, W46 and the amino acid sequence of ...

[0332] Preferably, the α-amylase variant comprises the amino acids N3A, N3C, N3D, N3E, N3F, N3G, N3H, N3L, N3P, N3Q, N3S, N3T, N3V, T5E, G7E, G7F, G7H, G7K, G7L, G7P, G7R, G7S, G7T, G7V, G7W, T8S, H16D, H16R, P18D, P18L, P18N, N25T, R26Y, D29N, D30F, R35P, R37A, R37N, T40A, T40C, T40D, T40E, T40G, T40H, T40I, T40K, T40L, T40R ... 0N, T40V, T40W, A41C, A41D, A41E, A41H, A41I, A41K, A41N, A41R, A41T, A41V , A41Y, W43E, P46A, G50M, G50P, G50Q, G57A, G57S, A60E, A60S, A60V, E68F, E6 8Y, Q71F, G73D, G73H, G73I, G73L, G73Q, G73S, G73T, G73W, T74F, K78S, K78T , T81M, Q84I, Q84W, Q84Y, E86K, E86S, S87G, S87L, S87P, H90P, A91Q, A91V, A9 1W, K93V, V97C, V97H, Q98E, Q98F, Q98G, Q98I, Q98K, Q98L, Q98M, Q98R, Q98T , Q98Y, V103C, V103G, V103S, M105F, G110K, G110R, A111E, A111Q, A113T, T11 4M, T114V, N116C, N116Y, V117F, A119H, A119L, A119Y, V120E, E121I, N125S , N126I, N128E, Q129T, I131Y, S132A, S132D, S132H, S132I, S132K, S132L, S1 32P, S132R, S132T, S132V, S132Y, G133E, G133L, G133T, G133V, Y135C, Y135 D, Y135E, Y135P, I137D, I137N, I137Q, I137W, I137Y, E138S, E138V, A139L, W 140A, W140D, W140M, W140N, W140P, W140S, W140T, W140V, T141G, K142E, K14 2N, F143H, F143I, F143M, F145G, F145H, F145I, F145L, F145S, P146A, P146H,P146N、G147D、G149F、G149K、W159A、W159E、H161W、F162T、V165P、W167D、W167G、W167P、Q169L、Q169P、Q169V、Q169Y、R171P、R176E、R176Q、R176V、R176Y、I177H、I177P、K179A、K179M、K179V、F180D、F180G、R181D、R181E、R181G、R181M、R181Y、K185T、K185V、K185W、A186D、W187T、N195Y、G196D、N197V、N197Y、A204F、A204H、A204L、V206N、M208D、P211C、P211M、E216L、R218Q、R219V、T225D、T225F、T225H、T225K、T225L、T225N、T225R、T225Y、L228V、L230C、L230F、D231G、D231T、D231V、I235A、K242L、Y243D、Y243M、Y243N、R247G、R247P、R247S、L250S、L250Y、T251D、H252P、V253F、V253S、R254D、R254F、R254H、R254P、R254T、R254V、R254W、N255C、N255F、N255P、T257A、T257C、T257D、T257E、T257G、T257P、T257Q、T257S、T257W、T257Y、K259P、K259V、K259W、K259Y、E260V、M261D、M261F、M261P、M261R、M261S、M261W、F262A、F262C、F262E、F262H、F262I、F262K、F262L、F262N、F262P、F262Q、F262R、F262S、F262T、F262V、F262W、A263E、A263H、A263W、A265Q、L272W、G273Q、A274D、L279I、N280P、K281H、K281I、K281S、N283P、W284A、W284D、W284E、W284G、W284I、W284K、W284M、W284N、W284P、W284Q、W284S、W284T、W284V、W284Y、S287I、S287K、S287L、S287P、S287R、S287T、S287V、S287Y、P292L、P292M、P292Y、H294N、H294S、N296T, Y298D, Y298E, N299T, A300G, S303A, S303C, S303D, S303E, S303F, S303G, S303H, S303L, S303M, S303N, S303Q, S303T, S303V, S303Y, G305E, Y307 S、A310D、A310N、K311A、K311C、L313Q、N314P、N314S、G315E、G315M、G315V、 Q319C、Q319E、K320S、K320W、H321F、H321K、H321L、H321R、H321T、H321V、H32 1W, P322C, P322E, P322H, P322T, P322Y, M323A, M323C, M323D, M323E, M323G, M323H, M323L, M323N, M323P, M323R, M323S, M323T, M323W, M323Y, H324A, H 324I, H324P, H324T, H324W, H324Y, S334V, G337V, L340M, S342A, S342G, S342T, V344A, V344C, V344I, Q345N, E346A, W347A, W347E, W347F, W347G, W347H W347K, W347N, W347P, W347R, P350A, P350C, P350N, P350T, R359C, R359F, R359G, R359M, R359S, R359W, E360D, E360H, E360P, E360T, E360W, Q361D, Q361 H、Q361I、Q361K、Q361L、Q361M、Q361N、Q361P、Q361R、Q361S、Q361T、Q361V、 Q361W、Q361Y、G362M、Y363A、Y363E、Y363F、Y363G、Y363I、Y363L、Y363Q、Y36 3R, Y363S, Y363T, Y363V, Y363W, P364C, Y368C, Y368Q, Y368R, Y368T, Y372C, Y372G, Y372N, Y372T, P375Q, P375T, P380F, M382I, K383C, K383L, K383P, K 383T, A384D, A384N, K385N, K385V, D387C, D387P, D387R, D387W, P388I, E391M, R393K, R393M, N395C, N395D, N395K, N395V, N395W, F396E, F396G, F396TA397V, Y398E, Q401H, H402S, D403A, D403E, D403T, F405C, F405N, F405S, F405T, D406V, H407C, H407Q, H407T, I410H, I410M, I410R, I410W, R415C, R 415L, R415M, R415Y, E416D, E416F, E416L, E416N, N418A, N418P, N418V, H4 21A, H421G, H421L, H421P, H421Q, H421Y, P422D, P422G, P422I, P422N, P42 2Q, P422S, N423G, N423H, N423I, N423L, N423M, N423P, N423Q, N423V, N42 3W, N423Y, G433A, G433N, G433S, P434I, P434L, G435C, G435L, G435M, G435 T, G435V, K438A, K438I, K438Q, K438S, K438V, W439A, W439D, W439I, W439K , W439L, W439N, W439S, W439T, W439V, W439Y, K446A, K446F, A447D, A447Y, Q449T, V450D, V450N, H452L, T455C, T455M, T455V, N457D, N457E, N457I, N457L, N457V, K458D, K458L, G460M, G460V, T461F, T461I, T461Q, T461V, V 462C, N465A, N465C, N465E, N465G, N465H, N465L, N465P, N465T, A466F, A4 66I, A466Y, D467I, W469A, W469E, W469H, W469L, W469N, W469Q, W469R, W46 and preferably the α-amylase variant comprises one or more of these substitutions at positions corresponding to: 9S, W469Y, N471A, N471D, N471M, N471W, S473N, N475A, N475C, N475D, N475E, N475F, N475G, N475H, N475I, N475L, N475M, N475P, N475Q, N475S, N475T, N475V, N475W, G476F, G476L, G477A, ​​G477H, G477Q, G477S, G477W, S478A, S478F and S478Q.It has an increase in specific activity in the Model A detergent composition measured as an improvement factor (IF) > 1.1.

[0333] Preferably, the α-amylase variant comprises the amino acids N3A, N3C, N3D, N3E, N3F, N3G, N3H, N3L, N3P, N3Q, N3S, N3T, N3V, T5E, G7E, G7F, G7H, G7K, G7L, G7P, G7R, G7S, G7T, G7V, G7W, T8S, H16D, H16R, P18D, P18N, N25T, R26Y, D29N, D30F, R37A, R37N, T40A, T40C, T40D, T40E, T40G, T40H, T40I, T40K, T40N, T40V ... 0W, A41C, A41D, A41E, A41H, A41I, A41K, A41N, A41R, A41T, A41V, A41Y, W43E , P46A, G50M, G50P, G50Q, G57A, G57S, A60E, A60S, A60V, E68F, Q71F, G73D, G 73H, G73I, G73L, G73Q, G73S, G73T, G73W, K78S, K78T, T81M, Q84I, Q84W, Q84 Y, E86K, E86S, S87G, S87L, S87P, H90P, A91Q, A91V, A91W, K93V, V97C, V97H, Q 98E, Q98F, Q98G, Q98I, Q98K, Q98L, Q98M, Q98R, Q98T, Q98Y, V103C, V103G, V 103S, M105F, G110K, G110R, A111E, A111Q, A113T, T114M, T114V, N116Y, V11 7F, A119H, A119L, A119Y, V120E, E121I, N125S, N126I, N128E, Q129T, I131Y , S132A, S132D, S132H, S132I, S132K, S132L, S132P, S132R, S132T, S132V, S1 32Y, G133E, G133L, G133T, G133V, Y135C, Y135D, Y135E, Y135P, I137D, I137 N, I137Q, I137W, I137Y, E138S, E138V, A139L, W140A, W140D, W140M, W140N, W 140P, W140S, W140T, W140V, T141G, K142E, K142N, F143H, F143I, F143M, F14 5G, F145H, F145I, F145L, F145S, P146A, P146H, P146N, G147D, G149F, G149K,W159A、H161W、F162T、V165P、W167D、W167G、W167P、Q169V、Q169Y、R171P、R176E、R176Q、R176V、R176Y、I177H、I177P、K179A、K179M、K179V、F180D、F180G、R181D、R181E、R181G、R181M、R181Y、K185V、K185W、A186D、W187T、N195Y、G196D、N197V、N197Y、A204F、A204H、A204L、M208D、P211C、P211M、E216L、R219V、T225D、T225F、T225H、T225K、T225L、T225N、T225R、T225Y、L228V、L230C、L230F、D231G、D231T、D231V、I235A、K242L、Y243D、Y243M、Y243N、R247G、R247P、L250S、L250Y、T251D、H252P、V253F、V253S、R254D、R254F、R254H、R254P、R254T、R254V、R254W、N255C、N255F、N255P、T257A、T257C、T257D、T257E、T257G、T257P、T257Q、T257S、T257W、T257Y、K259P、K259V、K259W、K259Y、E260V、M261D、M261F、M261P、M261R、M261S、M261W、F262A、F262C、F262E、F262H、F262I、F262K、F262L、F262N、F262P、F262Q、F262R、F262S、F262T、F262V、F262W、A263E、A263H、A263W、A265Q、L272W、G273Q、L279I、N280P、K281H、K281I、K281S、N283P、W284A、W284D、W284E、W284G、W284I、W284K、W284M、W284N、W284P、W284Q、W284S、W284T、W284V、W284Y、S287I、S287K、S287L、S287P、S287R、S287T、S287V、S287Y、P292L、P292M、P292Y、H294N、H294S、N296T、Y298D、Y298E、N299T、A300G、S303A、S303C、S303D、S303E、S303F、S303G、S303H、S303L, S303M, S303N, S303Q, S303T, S303V, S303Y, G305E, Y307S, A310D, A310N, K311A, K311C, L313Q, N314P, N314S, G315E, G315M, G315V, Q319C, Q319 E, K320S, K320W, H321F, H321K, H321L, H321R, H321T, H321V, H321W, P322C, P322E, P322H, P322T, P322Y, M323A, M323C, M323D, M323E, M323G, M323H, M32 3L, M323N, M323P, M323R, M323S, M323Y, H324A, H324I, H324P, H324T, H324W, H324Y, S334V, G337V, L340M, S342A, S342G, S342T, V344A, V344C, V344I, Q 345N, E346A, W347A, W347E, W347F, W347G, W347H, W347K, W347N, W347P, W347R, P350A, P350C, P350N, P350T, R359C, R359F, R359G, R359M, R359W, E360D E360H、E360P、E360T、E360W、Q361D、Q361H、Q361I、Q361K、Q361L、Q361M、Q3 61N、Q361P、Q361R、Q361S、Q361T、Q361V、Q361W、Q361Y、G362M、Y363A、Y363 E, Y363F, Y363G, Y363I, Y363L, Y363Q, Y363S, Y363T, Y363V, Y363W, P364C, Y368C, Y368Q, Y368R, Y368T, Y372C, Y372G, Y372N, Y372T, P375Q, P375T, P38 0F, M382I, K383C, K383L, K383P, K383T, A384D, A384N, K385N, K385V, D387C, D387P, D387R, D387W, P388I, E391M, R393K, R393M, N395C, N395D, N395K, N 395V, N395W, F396E, F396G, F396T, A397V, Y398E, Q401H, H402S, D403A, D403E, D403T, F405C, F405N, F405S, F405T, D406V, H407C, H407Q, H407T, I410HI410M, I410R, I410W, R415C, R415L, R415M, R415Y, E416D, E416F, E416L, E416N, N418A, N418P, N418V, H421A, H421G, H421L, H421P, H421 Q, H421Y, P422D, P422G, P422I, P422N, P422Q, P422S, N423G, N423H, N423I, N423L, N423M, N423Q, N423V, N423W, N423Y, G433A, G433S, P4 34I, P434L, G435C, G435L, G435M, G435T, G435V, K438A, K438I, K438Q, K438S, K438V, W439A, W439D, W439I, W439K, W439L, W439N, W439S, W439T, W439V, W439Y, K446A, K446F, A447D, A447Y, Q449T, V450D, V450N, H452L, T455C, T455M, T455V, N457D, N457E, N457I, N457L, N457 V, K458D, K458L, G460M, T461F, T461I, T461Q, T461V, V462C, N465A, N465C, N465E, N465G, N465H, N465L, N465P, N465T, A466F, A466I, A4 66Y, D467I, W469A, W469E, W469H, W469L, W469N, W469Q, W469R, W469Y, N471A, N471D, N471M, N471W, S473N, N475A, N475C, N475D, N475E, The α-amylase variants comprise one or more of these substitutions at positions corresponding to N475F, N475G, N475H, N475I, N475L, N475M, N475P, N475Q, N475S, N475T, N475V, N475W, G476F, G476L, G477A, ​​G477H, G477Q, G477S, G477W, S478A, S478F, and S478Q, and preferably have an increased specific activity in a Model A detergent composition, as measured by an improvement factor (IF) of >1.2.

[0334] Preferably, the α-amylase variant comprises the amino acids N3A, N3C, N3D, N3E, N3F, N3G, N3H, N3L, N3P, N3Q, N3V, T5E, G7E, G7F, G7H, G7L, G7R, G7S, G7T, G7V, G7W, H16D, R26Y, D29N, D30F, R37A, R37N, T40A, T40C, T40D, T40E, T40G, T40H, T40I, T40N, T40V, T40W, A41C, A41D, A41E, A41H, A41I, A41K, A41N, A41R ... 1Y, P46A, G50Q, G57A, G57S, A60S, A60V, E68F, G73D, G73H, G73I, G73L, G73Q , G73S, G73T, G73W, K78T, T81M, Q84I, Q84W, E86S, S87G, H90P, Q98F, Q98I, Q9 8K, Q98L, Q98M, Q98Y, V103C, V103G, G110K, G110R, A111E, A111Q, A119L, A1 19Y, V120E, N125S, N128E, I131Y, S132A, S132D, S132H, S132I, S132K, S132L , S132R, S132V, S132Y, G133E, Y135C, Y135D, Y135E, Y135P, I137N, I137W, E 138S, E138V, A139L, W140A, W140D, W140M, W140N, W140P, W140S, W140T, K14 2E, F143H, F143I, F145I, P146A, P146H, P146N, G147D, G149K, H161W, V165P , W167D, W167P, Q169V, R171P, R176E, R176Q, R176Y, I177H, K179A, K179V, R1 81E, R181G, R181M, R181Y, A186D, W187T, G196D, N197V, N197Y, A204H, A204 L, P211C, P211M, T225D, T225F, T225H, T225K, T225L, T225N, T225R, T225Y, L 230C, L230F, D231V, Y243D, Y243N, R247G, R247P, L250S, H252P, V253F, R25 4T, R254V, R254W, N255P, T257A, T257C, T257D, T257E, T257G, T257W, K259P,K259V、K259W、E260V、M261D、M261F、M261P、M261R、M261S、M261W、F262A、F2 62C、F262E、F262H、F262I、F262K、F262L、F262N、F262P、F262Q、F262R、F262 S、F262T、F262V、F262W、A263E、A263H、A263W、A265Q、L279I、K281I、N283P、 W284A、W284D、W284E、W284G、W284I、W284K、W284M、W284N、W284P、W284Q、W28 4S、W284T、W284V、W284Y、S287K、S287L、S287R、S287T、S287V、S287Y、H294S 、N296T、Y298D、N299T、S303A、S303C、S303D、S303E、S303F、S303G、S303H、S 303L, S303M, S303N, S303Q, S303T, S303V, S303Y, G305E, Y307S, A310N, K311A, K311C, L313Q, N314P, N314S, G315E, G315V, Q319E, K320W, H321F, H321K H321L, H321V, P322C, P322E, P322H, P322T, P322Y, M323A, M323C, M323D, M323E, M323G, M323H, M323L, M323N, M323P, M323R, M323S, M323Y, H324A, H324 I、H324P、H324T、H324W、H324Y、G337V、L340M、S342G、V344C、E346A、W347E、 W347F、W347G、W347H、W347K、W347N、W347P、W347R、P350C、P350N、P350T、R35 9G、R359W、E360D、E360T、E360W、Q361D、Q361H、Q361I、Q361K、Q361L、Q361M 、Q361N、Q361P、Q361R、Q361T、Q361V、Q361W、Q361Y、G362M、Y363A、Y363F、Y 363G, Y363I, Y363Q, Y363S, Y363T, Y363V, Y363W, P364C, Y368C, Y368Q, Y368R, Y372C, Y372G, P375Q, P375T, P380F, K383C, K383T, A384D, K385N, K385VD387C, D387P, D387R, E391M, R393K, R393M, N395C, N395D, N395W, F396E, F396G, F396T, Y398E, H402S, D403A, D403T, F405C, F405N, F405 S, F405T, D406V, H407C, H407Q, H407T, I410H, I410M, I410W, R415C, R415L, R415M, R415Y, E416F, E416L, E416N, N418A, N418P, H421A, H4 21G, H421L, H421P, H421Q, H421Y, P422D, P422G, P422N, P422S, N423G, N423I, N423L, N423M, N423Q, N423V, N423W, N423Y, G433S, P434I, P434L, G435M, G435T, G435V, K438I, K438Q, K438S, K438V, W439A, W439D, W439I, W439K, W439L, W439N, W439S, W439T, W439V, K446A, K446 F, A447D, A447Y, H452L, T455V, N457D, N457E, N457I, N457L, N457V, K458D, K458L, G460M, T461F, T461I, T461Q, T461V, V462C, N465A, N4 65E, N465G, N465L, A466F, A466I, A466Y, D467I, W469A, W469E, W469H, W469L, W469Q, W469R, W469Y, N471A, N471D, N471M, N475A, N475C, comprising one or more of these substitutions at positions corresponding to N475D, N475E, N475F, N475G, N475H, N475I, N475L, N475M, N475P, N475Q, N475S, N475T, N475V, N475W, G476L, G477A, ​​G477H, G477Q, G477S, G477W, S478A, and S478F, preferably the α-amylase variant has an increased specific activity in the Model A detergent composition as measured by an improvement factor (IF) of >1.3. Preferably, the α-amylase variant is selected from the group consisting of the α-amylase variants at positions N3A, N3C, N3D, N3E, N3F, N3G, N3H, N3L, N3V, G7F, G7H, G7R, G7V, G7W, R26Y ...D29N、R37A、R37N、T40C、T40D、T40E、T40G、T40I、T40N、T40V、A41C、A41D、A41E、A41K、A41N、A41R、A41Y、G73D、G73H、G73I、G73L、G73Q、G73S、G73T、G73W、K78T、Q84I、Q84W、S87G、Q98I、Q98L、V103C、G110K、A111Q、A119Y、N125S、N128E、S132D、S132I、S132L、S132R、Y135C、Y135E、I137W、E138S、E138V、A139L、W140A、W140D、W140M、W140N、W140P、W140S、W140T、K142E、F143I、F145I、P146H、G149K、H161W、V165P、W167D、W167P、Q169V、R176Q、I177H、P211C、T225F、T225H、T225K、T225R、T225Y、L230C、Y243D、R247G、V253F、N255P、T257A、T257D、T257E、K259P、K259V、M261D、M261R、M261W、F262A、F262C、F262E、F262H、F262I、F262K、F262L、F262N、F262P、F262Q、F262R、F262S、F262W、A263H、L279I、N283P、W284D、W284E、W284I、W284K、W284M、W284N、W284P、W284Q、W284S、W284V、S287K、S287L、S287R、S287Y、N296T、S303A、S303C、S303D、S303E、S303F、S303G、S303H、S303M、S303N、S303Q、S303T、S303V、S303Y、G305E、K311C、L313Q、N314P、G315V、H321K、H321L、H321V、P322C、P322E、P322H、P322T、P322Y、M323A、M323D、M323E、M323G、M323L、M323P、M323S、M323Y、H324A、H324I、H324P、H324W、L340M、S342G、E346A、W347F、W347K、W347N、W347P、W347R、R359W、E360W、Q361D、Q361H、Q361I、Q361K、Q361L、Q361M、Q361P、Q361R、Q361V, Q361W, Q361Y, G362M, Y363A, Y363G, Y363T, Y363V, Y363W, P364C, Y368C, Y368Q, Y372C, Y372G, K38 3T, A384D, D387C, D387P, E391M, R393K, R393M, N395D, N395W, F396E, F396T, D403A, F405C, F405N, F405S, D406V, H407C, H407T, I410H, I410M, I410W, R415C, R415L, R415M, R415Y, N418P, H421A, H421L, H421P, H42 1Q, P422D, P422G, N423L, N423V, N423W, N423Y, G433S, G435M, K438V, W439A, W439I, W439K, W439S, W439T, W439V, K446A, A447Y, H452L, T455V, N457E, N457I, N457V, K458D, T461F, T461Q, N465A, N465E, N465G, A46 6I, D467I, W469A, W469E, W469Y, N471A, N471D, N471M, N475A, N475C, N475D, N475E, N475F, N475G, N475H, The α-amylase variants comprising one or more of these substitutions at positions corresponding to N475L, N475M, N475P, N475Q, N475S, N475T, N475V, N475W, G477A, ​​G477H, G477Q, G477S, G477W, S478A preferably have an increased specific activity in the Model A detergent composition as measured by an improvement factor (IF) of >1.4. Preferably, the α-amylase variant comprises the amino acids at positions N3A, N3C, N3D, N3E, N3G, N3L, G7F, R26Y, R37A, R37N, T40C, T40D, T40G, T40I, T40N, T40V, A41C, A41D, A41K, A41N, A41R, G73D, G73I, using SEQ ID NO: 1 for numbering. , G73L, G73Q, G73S, G73T, G73W, K78T, Q84I, G110K, A111Q, N128E, Y135E, A139L, W140A, W14 0M, K142E, F145I, G149K, V165P, W167P, R176Q, I177H, T225F, T225H, T225K, T225Y, L230C,R247G, T257D, T257E, M261W, F262A, F262E, F262K, F262N, F262P, F262Q, F26 2R, F262S, F262W, N283P, W284D, W284P, W284S, W284V, S287Y, S303A, S303C, S303D, S303E, S303F, S303G, S303H, S303M, S303N, S303Q, S303T, S303V, S303Y, L313Q, N314P, H321 K, H321L, H321V, P322C, P322H, P322Y, M323A, M323D, M323Y, H324P, H324W, L340M, S342G, E346A, W34 7N, W347P, W347R, R359W, E360W, Q361K, Q361L, Q361M, Q361R, Q361W, Q361Y, Y363G, Y363V, Y363W, Y3 68C, Y372C, K383T, D387P, E391M, R393K, R393M, N395D, N395W, D403A, F405C, F405S, D406V, H407C, H 407T, I410H, R415C, R415L, R415M, R415Y, N418P, H421A, H421L, H421P, P422D, N423L, N423V, G435M, W439A, K446A, H452L, T455V, N457I, K458D, N465E, N471A, N471D, N471M, N475A, N475C, N475D, N475E , N475F, N475G, N475H, N475L, N475P, N475Q, N475V, N475W, G477A, ​​G477H, G477Q, and G477S, and preferably the α-amylase variant has an increased specific activity in a Model A detergent composition as measured by an improvement factor (IF) of >1.5.

[0335] Preferably, the α-amylase variant comprises one or more of these substitutions at positions corresponding to positions R37A, R176Q, T225Y, F262P, N283P, S303C, S303D, S303G, S303M, S303N, S303T, S303V, S303Y Y368C, N395D, H407T, I410H, R415C, H421A, G435M, N475C, G477A, ​​and G477H, using SEQ ID NO: 1 for numbering, and preferably the α-amylase variant has an increased specific activity in a Model A detergent composition, measured as an improvement factor (IF) of > 2.0.

[0336] Preferably, the α-amylase variants of the invention comprise one or more of these substitutions at positions corresponding to positions S303C, S303T, Y368C, and H421A, using SEQ ID NO: 1 for numbering, and preferably the α-amylase variants have an increased specific activity in a Model A detergent composition, measured as an improvement factor (IF) of >2.5.

[0337] The increase in specific activity, measured as an improvement factor (IF) > 1.1 as described herein, can be compared to any parent, specifically a parent polypeptide selected from SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 5, SEQ ID NO: 6, SEQ ID NO: 7, SEQ ID NO: 8, SEQ ID NO: 9, SEQ ID NO: 10, SEQ ID NO: 11, SEQ ID NO: 12, SEQ ID NO: 13, SEQ ID NO: 14, SEQ ID NO: 15, SEQ ID NO: 16 or SEQ ID NO: 17.

[0338] The α-amylase variant may further comprise deletions at one or more positions corresponding to positions R181, G182, D183, and G184, using SEQ ID NO: 1 for numbering.

[0339] Preferably, the α-amylase variant is R181 * +G182 * , R181 * +D183 * , R181 * +G184* , G182 * +D183 * , G182 * +G184 * , or D183 * +G184 * and preferably D183. * +G184 * Includes.

[0340] The variant may further comprise one or more additional substitutions and / or deletions at one or more (eg, several) other positions.

[0341] The amino acid changes can be minor, i.e., conservative amino acid substitutions or insertions that do not significantly affect protein folding and / or activity, small deletions, typically 1-30 amino acids, small amino- or carboxy-terminal extensions such as an amino-terminal methionine residue, small linker peptides of up to 20-25 residues, or small extensions that facilitate purification by altering net charge or another function, such as poly-histidine tracts, antigenic epitopes, or binding domains.

[0342] Thus, while the α-amylase variant may contain only one particular substitution that provides an improved property according to the present invention, it may still have additional modifications that have at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% but less than 100% sequence identity to the polypeptide of amino acid sequence SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, or 17. These additional modifications preferably do not significantly alter the improved properties of the α-amylase.

[0343] Examples of conservative substitutions are those within the group of basic amino acids (arginine, lysine, and histidine), acidic amino acids (glutamic acid and aspartic acid), polar amino acids (glutamine and asparagine), hydrophobic amino acids (leucine, isoleucine, and valine), aromatic amino acids (phenylalanine, tryptophan, and tyrosine), and low molecular weight amino acids (glycine, alanine, serine, threonine, and methionine). Amino acid substitutions that generally do not change the specific activity are known in the art and are described, for example, in H. Neurath and R.L. Hill, 1979, The Proteins, Academic Press, New York. Common substitutions 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.

[0344] Essential amino acids in a polypeptide can be identified by procedures known in the art, such as site-directed mutagenesis or alanine-scanning mutagenesis (Cunningham and Wells, 1989, Science 244:1081-1085). In the latter technique, single alanine mutations are introduced at every residue in the molecule, and the resulting mutant molecules are tested for protease activity to identify amino acid residues important for the molecule's activity. See 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, as determined by techniques such as nuclear magnetic resonance, crystallography, electron diffraction, or photoaffinity labeling, in conjunction with mutations of amino acids at putative contact sites. See, e.g., 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.

[0345] In one embodiment, the variant has an improved (increased) specific activity compared to the parent polypeptide.

[0346] In one embodiment, the variant has improved (increased) stability under storage conditions compared to the parent polypeptide, preferably SEQ ID NO:1 and / or SEQ ID NO:2.

[0347] In one embodiment, the variant has improved (increased) thermostability compared to the parent polypeptide, preferably SEQ ID NO:1 and / or SEQ ID NO:2.

[0348] In one embodiment, the variant has improved (increased) cleaning performance compared to the parent polypeptide, preferably SEQ ID NO:1 and / or SEQ ID NO:2.

[0349] Parent α-amylase The parent α-amylase may be a polypeptide having at least 60% sequence identity to any one of the polypeptides of SEQ ID NOs: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16 or 17.

[0350] In one embodiment, the parent α-amylase has at least 60%, e.g., at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 87%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to a polypeptide having α-amylase activity as set forth in SEQ ID NO: 1. In one embodiment, the amino acid sequence of the parent differs from the polypeptide of SEQ ID NO: 1 by 10 or fewer amino acids, e.g., by 5 amino acids, 4 amino acids, 3 amino acids, 2 amino acids, or 1 amino acid. The parent preferably comprises or consists of the amino acid sequence of SEQ ID NO: 1. In one embodiment, the parent comprises or consists of the polypeptide of SEQ ID NO: 1. In another embodiment, the parent is an allelic variant of the polypeptide of SEQ ID NO: 1.

[0351] In one embodiment, the parent α-amylase has at least 60%, e.g., at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 87%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to a polypeptide having α-amylase activity as set forth in SEQ ID NO: 2. In one embodiment, the amino acid sequence of the parent differs from the polypeptide of SEQ ID NO: 2 by 10 or fewer amino acids, e.g., by 5 amino acids, 4 amino acids, 3 amino acids, 2 amino acids, or 1 amino acid. The parent preferably comprises or consists of the amino acid sequence of SEQ ID NO: 2. In one embodiment, the parent comprises or consists of the polypeptide of SEQ ID NO: 2. In another embodiment, the parent is an allelic variant of the polypeptide of SEQ ID NO: 2.

[0352] In one embodiment, the parent α-amylase has at least 60%, e.g., at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 87%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to a polypeptide having α-amylase activity as set forth in SEQ ID NO: 3. In one embodiment, the amino acid sequence of the parent differs from the polypeptide of SEQ ID NO: 3 by 10 or fewer amino acids, e.g., by 5 amino acids, 4 amino acids, 3 amino acids, 2 amino acids, or 1 amino acid. The parent preferably comprises or consists of the amino acid sequence of SEQ ID NO: 3. In one embodiment, the parent comprises or consists of the polypeptide of SEQ ID NO: 3. In another embodiment, the parent is an allelic variant of the polypeptide of SEQ ID NO: 3.

[0353] In one embodiment, the parent α-amylase has at least 60%, e.g., at least 65%, at least 70%, at least 80%, at least 85%, at least 87%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to a polypeptide having α-amylase activity as set forth in SEQ ID NO: 4. In one embodiment, the amino acid sequence of the parent differs from the polypeptide of SEQ ID NO: 4 by 10 or fewer amino acids, e.g., by 5 amino acids, 4 amino acids, 3 amino acids, 2 amino acids, or 1 amino acid. The parent preferably comprises or consists of the amino acid sequence of SEQ ID NO: 4. In one embodiment, the parent comprises or consists of the polypeptide of SEQ ID NO: 4. In another embodiment, the parent is an allelic variant of the polypeptide of SEQ ID NO: 4.

[0354] In one embodiment, the parent α-amylase has at least 60%, e.g., at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 87%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to a polypeptide having α-amylase activity as set forth in SEQ ID NO: 5. In one embodiment, the amino acid sequence of the parent differs from the polypeptide of SEQ ID NO: 5 by 10 or fewer amino acids, e.g., by 5 amino acids, 4 amino acids, 3 amino acids, 2 amino acids, or 1 amino acid. The parent preferably comprises or consists of the amino acid sequence of SEQ ID NO: 5. In one embodiment, the parent comprises or consists of the polypeptide of SEQ ID NO: 5. In another embodiment, the parent is an allelic variant of the polypeptide of SEQ ID NO: 5.

[0355] In one embodiment, the parent α-amylase has at least 60%, e.g., at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 87%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to a polypeptide having α-amylase activity as set forth in SEQ ID NO: 6. In one embodiment, the amino acid sequence of the parent differs from the polypeptide of SEQ ID NO: 6 by 10 or fewer amino acids, e.g., by 5 amino acids, 4 amino acids, 3 amino acids, 2 amino acids, or 1 amino acid. The parent preferably comprises or consists of the amino acid sequence of SEQ ID NO: 6. In one embodiment, the parent comprises or consists of the polypeptide of SEQ ID NO: 6. In another embodiment, the parent is an allelic variant of the polypeptide of SEQ ID NO: 6.

[0356] In one embodiment, the parent α-amylase has at least 60%, e.g., at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 87%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to a polypeptide having α-amylase activity as set forth in SEQ ID NO: 7. In one embodiment, the amino acid sequence of the parent differs from the polypeptide of SEQ ID NO: 7 by 10 or fewer amino acids, e.g., by 5 amino acids, 4 amino acids, 3 amino acids, 2 amino acids, or 1 amino acid. The parent preferably comprises or consists of the amino acid sequence of SEQ ID NO: 7. In one embodiment, the parent comprises or consists of the polypeptide of SEQ ID NO: 7. In another embodiment, the parent is an allelic variant of the polypeptide of SEQ ID NO: 7.

[0357] In one embodiment, the parent α-amylase has at least 60%, e.g., at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 87%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to a polypeptide having α-amylase activity as set forth in SEQ ID NO: 8. In one embodiment, the amino acid sequence of the parent differs from the polypeptide of SEQ ID NO: 8 by 10 or fewer amino acids, e.g., by 5 amino acids, 4 amino acids, 3 amino acids, 2 amino acids, or 1 amino acid. The parent preferably comprises or consists of the amino acid sequence of SEQ ID NO: 8. In one embodiment, the parent comprises or consists of the polypeptide of SEQ ID NO: 8. In another embodiment, the parent is an allelic variant of the polypeptide of SEQ ID NO: 8.

[0358] In one embodiment, the parent α-amylase has at least 60%, e.g., at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 87%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to a polypeptide having α-amylase activity as set forth in SEQ ID NO: 9. In one embodiment, the amino acid sequence of the parent differs from the polypeptide of SEQ ID NO: 9 by 10 or fewer amino acids, e.g., by 5 amino acids, 4 amino acids, 3 amino acids, 2 amino acids, or 1 amino acid. The parent preferably comprises or consists of the amino acid sequence of SEQ ID NO: 9. In one embodiment, the parent comprises or consists of the polypeptide of SEQ ID NO: 9. In another embodiment, the parent is an allelic variant of the polypeptide of SEQ ID NO: 9.

[0359] In one embodiment, the parent α-amylase has at least 60%, such as at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 87%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to a polypeptide having α-amylase activity as set forth in SEQ ID NO: 10. In one embodiment, the amino acid sequence of the parent differs from the polypeptide of SEQ ID NO: 10 by 10 or fewer amino acids, such as by 5 amino acids, 4 amino acids, 3 amino acids, 2 amino acids, or 1 amino acid. The parent preferably comprises or consists of the amino acid sequence of SEQ ID NO: 10. In one embodiment, the parent comprises or consists of the polypeptide of SEQ ID NO: 10. In another embodiment, the parent is an allelic variant of the polypeptide of SEQ ID NO: 10.

[0360] In one embodiment, the parent α-amylase has at least 60%, such as at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 87%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to a polypeptide having α-amylase activity as set forth in SEQ ID NO: 11. In one embodiment, the amino acid sequence of the parent differs from the polypeptide of SEQ ID NO: 11 by 10 or fewer amino acids, such as by 5 amino acids, 4 amino acids, 3 amino acids, 2 amino acids, or 1 amino acid. The parent preferably comprises or consists of the amino acid sequence of SEQ ID NO: 11. In one embodiment, the parent comprises or consists of the polypeptide of SEQ ID NO: 11. In another embodiment, the parent is an allelic variant of the polypeptide of SEQ ID NO: 11.

[0361] In one embodiment, the parent α-amylase has at least 60%, e.g., at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 87%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to a polypeptide having α-amylase activity as set forth in SEQ ID NO: 12. In one embodiment, the amino acid sequence of the parent differs from the polypeptide of SEQ ID NO: 12 by 10 or fewer amino acids, e.g., by 5 amino acids, 4 amino acids, 3 amino acids, 2 amino acids, or 1 amino acid. The parent preferably comprises or consists of the amino acid sequence of SEQ ID NO: 12. In one embodiment, the parent comprises or consists of the polypeptide of SEQ ID NO: 12. In another embodiment, the parent is an allelic variant of the polypeptide of SEQ ID NO: 12.

[0362] In one embodiment, the parent α-amylase has at least 60%, e.g., at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 87%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to a polypeptide having α-amylase activity as set forth in SEQ ID NO: 13. In one embodiment, the amino acid sequence of the parent differs from the polypeptide of SEQ ID NO: 13 by 10 or fewer amino acids, e.g., by 5 amino acids, 4 amino acids, 3 amino acids, 2 amino acids, or 1 amino acid. The parent preferably comprises or consists of the amino acid sequence of SEQ ID NO: 13. In one embodiment, the parent comprises or consists of the polypeptide of SEQ ID NO: 13. In another embodiment, the parent is an allelic variant of the polypeptide of SEQ ID NO: 13.

[0363] In one embodiment, the parent α-amylase has at least 60%, e.g., at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 87%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to a polypeptide having α-amylase activity as set forth in SEQ ID NO: 14. In one embodiment, the amino acid sequence of the parent differs from the polypeptide of SEQ ID NO: 14 by 10 or fewer amino acids, e.g., by 5 amino acids, 4 amino acids, 3 amino acids, 2 amino acids, or 1 amino acid. The parent preferably comprises or consists of the amino acid sequence of SEQ ID NO: 14. In one embodiment, the parent comprises or consists of the polypeptide of SEQ ID NO: 14. In another embodiment, the parent is an allelic variant of the polypeptide of SEQ ID NO: 14.

[0364] In one embodiment, the parent α-amylase has at least 60%, such as at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 87%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to a polypeptide having α-amylase activity as set forth in SEQ ID NO: 15. In one embodiment, the amino acid sequence of the parent differs from the polypeptide of SEQ ID NO: 15 by 10 or fewer amino acids, such as by 5 amino acids, 4 amino acids, 3 amino acids, 2 amino acids, or 1 amino acid. The parent preferably comprises or consists of the amino acid sequence of SEQ ID NO: 15. In one embodiment, the parent comprises or consists of the polypeptide of SEQ ID NO: 15. In another embodiment, the parent is an allelic variant of the polypeptide of SEQ ID NO: 15.

[0365] In one embodiment, the parent α-amylase has at least 60%, e.g., at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 87%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to a polypeptide having α-amylase activity as set forth in SEQ ID NO: 16. In one embodiment, the amino acid sequence of the parent differs from the polypeptide of SEQ ID NO: 16 by 10 or fewer amino acids, e.g., by 5 amino acids, 4 amino acids, 3 amino acids, 2 amino acids, or 1 amino acid. The parent preferably comprises or consists of the amino acid sequence of SEQ ID NO: 16. In one embodiment, the parent comprises or consists of the polypeptide of SEQ ID NO: 16. In another embodiment, the parent is an allelic variant of the polypeptide of SEQ ID NO: 16.

[0366] In one embodiment, the parent α-amylase has at least 60%, such as at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 87%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to a polypeptide having α-amylase activity as set forth in SEQ ID NO: 17. In one embodiment, the amino acid sequence of the parent differs from the polypeptide of SEQ ID NO: 17 by 10 or fewer amino acids, such as by 5 amino acids, 4 amino acids, 3 amino acids, 2 amino acids, or 1 amino acid. The parent preferably comprises or consists of the amino acid sequence of SEQ ID NO: 17. In one embodiment, the parent comprises or consists of the polypeptide of SEQ ID NO: 17. In another embodiment, the parent is an allelic variant of the polypeptide of SEQ ID NO: 17.

[0367] The amino acid sequences of SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:9, SEQ ID NO:10, SEQ ID NO:11, SEQ ID NO:12, SEQ ID NO:13, SEQ ID NO:14, SEQ ID NO:15, SEQ ID NO:16, or SEQ ID NO:17, or active fragments thereof, can be used to design nucleic acid probes to identify and clone parent-encoding DNA from strains of different genera or species, according to methods well known in the art. In particular, such probes can be used to hybridize the genome or cDNA of the desired genus or species, followed by conventional Southern blotting procedures to identify and isolate the corresponding gene therein. Such probes can be significantly shorter than the entire sequence, but should be at least 14, e.g., at least 25, at least 35, or at least 70 nucleotides in length. Preferably, the nucleic acid probe is at least 100 nucleotides in length, or at least 200 nucleotides, at least 300 nucleotides, at least 400 nucleotides, at least 500 nucleotides, at least 600 nucleotides, at least 700 nucleotides, at least 800 nucleotides, or at least 900 nucleotides in length. Both DNA and RNA can be used. The probe can be a nucleic acid probe for a corresponding gene (e.g., 32 P, 3 H, 35 The probes are typically labeled for detection (with S, biotin, or avidin). Such probes are encompassed by the present invention.

[0368] Genomic DNA or cDNA libraries prepared from such other organisms can be hybridized with the above-described probes and screened for DNA encoding the parent. Genomic or other DNA from such other organisms can be separated by agarose or polyacrylamide electrophoresis, or other separation techniques. DNA from the library or separated DNA can be transferred and immobilized on nitrocellulose or other suitable carrier material for use in Southern blotting.

[0369] For purposes of the present invention, hybridization refers to the hybridization of a polynucleotide under low to very high stringency conditions to a labeled nucleotide probe corresponding to a polynucleotide encoding SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:9, SEQ ID NO:10, SEQ ID NO:11, SEQ ID NO:12, SEQ ID NO:13, SEQ ID NO:14, SEQ ID NO:15, SEQ ID NO:16, SEQ ID NO:17, or a subsequence thereof. Molecules to which the probe hybridizes can be detected using, for example, X-ray film or other detection means known in the art. In one aspect, the nucleic acid probe is a polynucleotide encoding the polypeptide of SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:9, SEQ ID NO:10, SEQ ID NO:11, SEQ ID NO:12, SEQ ID NO:13, SEQ ID NO:14, SEQ ID NO:15, SEQ ID NO:16, or SEQ ID NO:17, or an active fragment thereof. For long probes, at least 100 nucleotides in length, very low to very high stringency conditions are optimally defined as prehybridization and hybridization at 42°C in 5X SSPE, 0.3% SDS, 200 micrograms / mL of sheared denatured salmon sperm DNA, and either 25% formamide for very low stringency, 35% formamide for medium to upper-medium stringency, or 50% formamide for high stringency, followed by standard Southern blotting procedures for 12 to 24 hours. Finally, the carrier material is washed three times for 15 minutes each with 2x SSC, 0.2% SDS at 45°C (very low stringency), 50°C (low stringency), 55°C (medium stringency), 60°C (medium-high stringency), 65°C (high stringency), or 70°C (very high stringency).For short probes, approximately 15 to 70 nucleotides in length, stringency conditions are optimally defined as 12 to 24 hours of prehybridization and hybridization following standard Southern blotting procedures, in 0.9 M NaCl, 0.09 M Tris-HCl pH 7.6, 6 mM EDTA, 0.5% NP-40, 1X Denhardt's solution, 1 mM sodium pyrophosphate, 1 mM monobasic sodium phosphate, 0.1 mM ATP, and 0.2 mg of yeast RNA per mL, at approximately 5 to 10°C below the calculated Tm, using calculations according to Bolton and McCarthy (1962, Proc. Natl. Acad. Sci. USA 48:1390). Finally, the carrier material is washed once for 15 minutes in 6X SCC and 0.1% SDS, and the calculated Tm is reached. m Wash twice for 15 minutes each with 6X SSC at a temperature approximately 5°C to approximately 10°C lower than the standard.

[0370] The parent may be derived from any genus of microorganism. For purposes of the present invention, the term "derived from," when used herein in reference to a given source, shall mean that the parent encoded by the polynucleotide is produced by that source or by a cell into which a polynucleotide from that source has been inserted. In one embodiment, the parent is secreted extracellularly. The parent may be a bacterial α-amylase. For example, the parent may be a Gram-positive bacterial polypeptide, such as Bacillus, Clostridium, Enterococcus, Geobacillus, Lactobacillus, Lactococcus, Oceanobacillus, Staphylococcus, Streptococcus, or Streptomyces. or a Gram-negative bacterial polypeptide, such as an alpha-amylase from Campylobacter, E. coli, Flavobacterium, Fusobacterium, Helicobacter, Ilyobacter, Neissera, Pseudomonas, Salmonella, or Ureaplasma.In one aspect, the parent is Bacillus alkalophilus, Bacillus amyloliquefaciens, Bacillus brevis, Bacillus circulans, Bacillus clausii, Bacillus coagulans, Bacillus firmus, Bacillus lautus, Bacillus lentus, Bacillus licheniformis, Bacillus megaterium, Bacillus pumilus, Bacillus stearothermophilus, Bacillus In another embodiment, the parent is an α-amylase from Streptococcus equisimilis, Streptococcus pyogenes, Streptococcus uberis, or Streptococcus equi subsp. zooepidemicus. In another aspect, the parent is an α-amylase from Streptomyces achromogenes, Streptomyces avermitilis, Streptomyces coelicolor, Streptomyces griseus, or Streptomyces lividans.

[0371] The parent may also be a fungal α-amylase, for example, a yeast α-amylase, such as an α-amylase from the genera Candida, Kluyveromyces, Pichia, Saccharomyces, Schizosaccharomyces, or Yarrowia. For example, the parent can be a fungal α-amylase, such as Acremonium, Agaricus, Alternaria, Aspergillus, Aureobasidium, Botryosphaeria, Ceripoliopsis, Chaetomidium, Chrysosporium, Claviceps, Cochiobolos, Coprinopsis, Coptotermes, Corynascus, Cryphonectria, Cryptococcus, Diplodia, Exidia, Filibasidium, Fusarium, Gibberella, Holomastigotoides, Humicola, Irpex, Lentinula, Leptospaeria. , Magnaporthe, Melanocarpus, Meripilus, Mucor, Myceliophysora, Neurospora, Paecilomyces, Penicillium, Phanerochaete, Piromyces, Poitrasia, Pseudoplectania, Pseudotrichonympha, Rhizomucor, Schizophyllium, Scythallidium, Talaromyces, Thermoascus, Thielavia, Tolypocladium, Trichoderma, Trichophaea, Verticillium, Volvariella, or Xylaria. In another aspect, the parent may be an α-amylase from Saccharomyces carlsbergensis, Saccharomyces cerevisiae, Saccharomyces diastaticus, Saccharomyces douglasii, Saccharomyces kluyveri, Saccharomyces norbemsis, or Saccharomyces obiformis.Parents include Acremonium cellulolyticus, Aspergillus aculitus, Aspergillus awamori, Aspergillus foetidus, Aspergillus fumigatus, Aspergillus japonicus, Aspergillus nidulans, Aspergillus niger, Aspergillus oryzae, Chrysosporium inops, Chrysosporium keratinophilum, Chrysosporium lucknowense, Chrysosporium merdarium, Chrysosporium pannicola, and Chrysosporium queenstrandicum. queenslandicum, Chrysosporium tropicum, Chrysosporium zonatum, Fusarium bactridioides, Fusarium cerealis, Fusarium crookwellense, Fusarium culmorum, Fusarium graminearum, Fusarium graminum, Fusarium heterosporum, Fusarium negundi, Fusarium oxysporum, Fusarium reticulantum reticulatum, Fusarium roseum, Fusarium sambucinum, Fusarium sarcochroum,Fusarium sporotrichioides, Fusarium sulphureum, Fusarium torulosum, Fusarium trichothecioides, Fusarium venenatum, Humicola grisea, Humicola insolens, Humicola lanuginosa, Irpex lacteus, Mucor miehei, Myceliophthora thermophila, Neurospora crassa crassa, Penicillium funiculosum, Penicillium purpurogenum, Phanerochaete chrysosporium, Thielavia achromatica, Thielavia albomyces, Thielavia albopilosa, Thielavia australeinsis, Thielavia fimeti, Thielavia microspora, Thielavia ovispora, Thielavia peruviana, Thielavia setosa setosa), Thielavia spededonium, Thielavia subthermophila, Thielavia terrestris, Trichoderma harzianum, Trichoderma koningii, Trichoderma longibrachiatum, Trichoderma reesei,or Trichoderma viride α-amylase.

[0372] In another aspect, the parent is a Bacillus sp. α-amylase, for example, the α-amylase of SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:9, SEQ ID NO:10, SEQ ID NO:11, SEQ ID NO:12, SEQ ID NO:13, SEQ ID NO:14, SEQ ID NO:15, SEQ ID NO:16, or SEQ ID NO:17.

[0373] For the aforementioned species, it will be understood that the present invention encompasses perfect and imperfect forms, as well as other taxonomic equivalents, such as anamorphs, regardless of the species name by which they are known. Those skilled in the art will readily recognize the identity of appropriate equivalents. Strains of these species are readily available to the public in numerous culture collections, such as the American Type Culture Collection (ATCC), Deutsche Sammlung von Mikroorganismen und Zellkulturen GmbH (DSM), Centraalbureau Voor Schimmelcultures (CBS), and the Agricultural Research Service, Culture Collection, Northern Regional Research Center (NRRL).

[0374] Parents can be identified and obtained from other sources, including microorganisms isolated from natural sources (e.g., soil, compost, water, etc.), or DNA samples obtained directly from natural materials (e.g., soil, compost, water, etc.) using the probes described above. Techniques for isolating microorganisms and DNA directly from natural habitats are well known in the art. Polynucleotides encoding the parents can then be obtained by similarly screening genomic or cDNA libraries of other microorganisms, or mixed DNA samples. Once the polynucleotides encoding the parents are detected with the probes, the polynucleotides can be isolated or cloned using techniques known to those skilled in the art (see, for example, Sambrook et al., 1989, supra).

[0375] The parent may be a hybrid polypeptide in which a portion of one polypeptide is fused at the N-terminus or C-terminus of a portion of another polypeptide. The parent may also be a fusion polypeptide or a cleavable fusion polypeptide in which one polypeptide is fused at the N-terminus or C-terminus of another polypeptide. A fusion polypeptide is produced by fusing a polynucleotide encoding one polypeptide to a polynucleotide encoding another polypeptide. Techniques for producing fusion polypeptides are known in the art and involve ligating coding sequences encoding the polypeptides so that they are in frame, and expression of the fused polypeptides is under the control of the same promoter and terminator. Fusion proteins may also be constructed using intein technology, in which the fusion is generated post-translationally (Cooper et al., 1993, EMBO J. 12:2575-2583; Dawson et al., 1994, Science 266:776-779). The fusion polypeptide may further include a cleavage site between the two polypeptides. Upon secretion of the fusion protein, the site is cleaved, releasing the two polypeptides.Examples of cleavage sites include Martin et al., 2003, J. Ind. Microbiol. Biotechnol. 3:568-576, Svetina et al., 2000, J. Biotechnol. 76: 245-251, Rasmussen-Wilson et al. al., 1997, Appl.Environ.Microbiol.63:3488-3493, Ward et al., 1995, Biotechnology 13:498-503 and Contreras et al., 1991, Biotechnology 9:378-381, Eaton et al., 1986, Biochemistry 25:505-512, Collins-Racie et al., 1995, Biotechnology 13:982-987, Carter et al. al., 1989, Proteins: Structure, Function, and Genetics 6:240-248 and Stevens, 2003, Drug Discovery World 4:35-48.

[0376] Preparation of variants Variants with α-amylase activity include (a) the amino acid sequence of positions 3, 5, 7, 8, 16, 18, 25, 26, 29, 30, 35, 37, 40, 41, 43, 46, 50, 57, 60, 68, 71, 73, 74, 78, 81, 84, 86, 87, 90, 91, 93, 97, 98, 103, 105, 110, 111, 113, 114, 116, 117, 119, 120, 121, 125, 126, 128, 129, 131, 132, 133, 135, 137, 138, 139, 140, 141, 142, 143, 144, 145, 146, 147, 148, 149, 150, 151, 152, 153, 154, 155, 156, 157, 158, 159, 160, 161, 162, 163, 164, 165, 166, 167, 168, 169, 170, 171, 172, 173, 174, 175, 176, 177, 178, 179, 180, 181, 182, 183, 184, 185, 186, 187, 188, 189, 19 42, 143, 145, 146, 147, 149, 159, 161, 162, 165, 167, 169, 171, 176, 177, 179, 180, 181, 185, 186, 187, 195, 196, 197, 204, 206, 208, 211, 216, 218, 219, 225, 228, 230, 231, ...

Claims

1. 1. A cleaning composition comprising: a) an α-amylase variant of a parent α-amylase having α-amylase activity, comprising one or more of the following substitutions, using SEQ ID NO: 1 for numbering: G7E, G7F, G7H, G7K, G7L, G7P, G7R, G7S, G7T, G7V, G7W, said variant having at least 90% but less than 100% sequence identity to the parent polypeptide of SEQ ID NO: 1, said variant having α-amylase activity, and said α-amylase variant having improved properties relative to the parent polypeptide of SEQ ID NO: 1; 1. A cleaning composition comprising: an α-amylase variant, said variant further comprising one or more of the following substitutions, using SEQ ID NO:1 for numbering: R37A, R176Q, T225Y, F262P, N283P, S303C, S303D, S303G, S303M, S303N, S303T, S303V, S303Y, Y368C, N395D, H407T, I410H, R415C, H421A, G435M, N475C, G477A, ​​and G477H; and b) a cleaning aid.

2. The composition of claim 1, wherein the composition comprises from 0.1% to 60% by weight of one or more surfactants.

3. The composition described in claim 2, wherein the surfactant comprises an anionic surfactant.

4. The composition of any one of claims 1 to 3, wherein the improved property relative to the parent polypeptide is an increased specific activity.

5. 5. The composition of any one of claims 1 to 4, wherein the improved property is an increase in specific activity measured as an improvement factor (IF) > 1.0 when compared to the parent polypeptide of SEQ ID NO: 1 having α-amylase activity.

6. 6. The composition of any one of claims 1 to 5, wherein the α-amylase variant further comprises a deletion at one position corresponding to positions R181, G182, D183, and G184, using SEQ ID NO: 1 for numbering.

7. The composition of any one of claims 1 to 6, wherein the α-amylase variant according to any one of claims 1 to 6 has at least 95% but less than 100% sequence identity to the amino acid sequence of the parent α-amylase polypeptide of SEQ ID NO:

1.

8. The composition according to any one of claims 1 to 7, comprising at least one additional enzyme.

9. A composition according to any one of claims 1 to 8, wherein the composition is a liquid or powder laundry detergent composition.

10. A composition according to any one of claims 1 to 8, wherein the composition is a liquid or powder dishwashing composition.

11. 11. A method of treating a surface, comprising: (i) forming an aqueous cleaning solution comprising water and the cleaning composition of any one of claims 1 to 10; (ii) treating the surface with the aqueous cleaning solution; and (iii) rinsing the surface.

12. Use of a composition according to any one of claims 1 to 10 in a cleaning process which is laundry or hard surface cleaning.

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