Polypeptides having protease activity for use in detergent compositions

Abstract

In the present invention new protease enzymes are provided. More specifically, genetically engineered protease enzymes, compositions comprising the enzymes, and methods of making and using the enzymes or compositions comprising the enzymes are provided.

Description

[0001] BASF SE

[0002] 1

[0003] Polypeptides having protease activity for use in detergent compositions

[0004] Field of the invention

[0005] In the present invention new protease enzymes are provided. More specifically, genetically engineered protease enzymes, compositions comprising the enzymes, and methods of making and using the enzymes or compositions comprising the enzymes are provided.

[0006] Background of the invention

[0007] Enzymes are increasingly used in various applications as sustainable alternatives to petrochemistry. Enzymes are biodegradable and can be catalytically active already at lower temperatures, which results in reduction of energy consumption. In particular, in the detergent industry enzymes are implemented in washing formulations to improve cleaning efficiency and / or reduce energy consumption in a washing step.

[0008] Proteases are enzymes capable of hydrolyzing proteins. Thus, proteases have been employed in the removal of protein stains and have been added to detergent compositions for this purpose. In detergent applications the proteases shall be stable at elevated temperatures and / or within the denaturing conditions of the detergents and the wash liquor.

[0009] WO 2016 / 09671 1 and WO 2016 / 096714 describe a subtilase variant having improved stability and / or improved wash performance in liquid detergents compared to the parent subtilase and detergents containing the variant. WO 2016 / 001450 and WO 2020 / 002255 discloses subtilase variants with increased stability. WO 2010 / 056640 also describes subtilisin variants. US 6,376,450 discloses multiple-substituted protease variants which provide improved and enhanced cleaning ability. US 2020 / 172890 A1 discloses performance-enhanced and storage-stable protease variants. WO 99 / 20770 A2, WO 03 / 062381 A2, WO 2011 / 140364 A1 , WO 201 1 / 0721 17 A1 , WO 201 1 / 072099 A2, WO 2012 / 151534 A1 , WO 2015 / 144932 A1 , WO 2017 / 207762 A1 , WO 2017 / 192692A1 , WO 2018 / 069158 A 1 and WO 2022 / 225696 A2 also disclose protease variants.

[0010] Nevertheless, there is still a need for new protease enzymes, which are stable in harsh detergent compositions and show a good washing performance. Brief summary of the invention

[0011] The present invention is directed to a variant polypeptide having protease activity or a fragment of said polypeptide having protease activity, wherein:

[0012] (i) the polypeptide or fragment thereof has an amino acid sequence which is at least 82%, but less than 100%, identical to the amino acid sequence as set forth in SEQ ID NO: 1 and

[0013] (ii) the polypeptide or fragment thereof comprises an amino acid substitution at two or more amino acid residues selected from the group consisting of 38, 59 and 212 compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2, wherein the amino acid substitutions 38F, 38H, 38N, 59A, 59H, 59N, 59S, 212H and 212M are excluded and wherein the fragment of the variant polypeptide com5 to prises 100 to 259 consecutive amino acids of the full-length variant polypeptide.

[0014] The present invention is directed to a variant polypeptide having protease activity or a fragment of said polypeptide having protease activity, wherein:

[0015] (I) the polypeptide or fragment thereof has an amino acid sequence which is at least 82%, but less than 100%, identical to the amino acid sequence as set forth in SEQ ID NO: 1 and

[0016] (ii) the polypeptide or fragment thereof comprises (A) an amino acid substitution at amino acid residue 212 and (B) an amino acid substitution at amino acid residue 38 and / or 59 compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2, wherein the amino acid substitutions 38F, 38H, 38N, 59A, 59H, 59N, 59S, 212H and 212M are excluded and wherein the fragment of the variant polypeptide comprises 100 to 259 consecutive amino acids of the full-length variant polypeptide.

[0017] The present invention further relates to a polynucleotide encoding said variant polypeptide and a composition comprising said variant polypeptide.

[0018] Detailed description of the invention

[0019] The present invention may be understood more readily by reference to the following detailed description of the embodiments of the invention and the examples included herein.

[0020] Although the present invention will be described with respect to particular embodiments, this description is not to be construed in a limiting sense.

[0021] Definitions

[0022] Unless otherwise noted, the terms used herein are to be understood according to conventional usage by those of ordinary skill in the relevant art. Before describing in detail exemplary embodiments of the present invention, definitions important for understanding the present invention are given. Unless stated otherwise or apparent from the nature of the definition, the definitions apply to all compounds, methods and uses described herein.

[0023] As used in this specification and in the appended claims, the singular forms of "a" and "an" also include the respective plurals unless the context clearly dictates otherwise.

[0024] In the context of the present invention, the terms "about" and "approximately" denote an interval of accuracy that a person skilled in the art will understand to still ensure the technical effect of the feature in question. The term typically indicates a deviation from the indicated numerical value of ±20 %, preferably ±15 %, more preferably ±10 %, and even more preferably ±5 %.

[0025] Furthermore, the terms "first", "second", "third" or "(a)", "(b)", "(c)", "(d)" etc. and the like in the description and in the claims, are used for distinguishing between similar elements and not necessarily for describing a sequential or chronological order. It is to be understood that the terms so used are interchangeable under appropriate circumstances and that the embodiments of the invention described herein are capable of operation in other sequences than described or illustrated herein. In case the terms "first", "second", "third" or "(a)", "(b)", "(c)", "(d)", "I", "II" etc. relate to steps of a method or use or assay there is no time or time interval coherence between the steps, i.e. the steps may be carried out simultaneously or there may be time intervals of seconds, minutes, hours, days, weeks, months or even years between such steps, unless otherwise indicated in the application as set forth herein above or below.

[0026] Throughout this application, various publications are referenced. The disclosure of all of these publications and those references cited within those publications in their entireties are hereby incorporated by reference into this application in order to more fully describe the state of the art to which this invention pertains.

[0027] It is to be understood that the term "comprising" is not limiting. For the purposes of the present invention the term "consisting of" is considered to be a preferred embodiment of the term "comprising". If hereinafter a group is defined to comprise at least a certain number of members, this is meant to also encompass a group which consists of these members only.

[0028] “Amino acid substitutions” are described by providing the original amino acid followed by the number of the position within the amino acid sequence, followed by the substituted amino acid. For example, the substitution of histidine at position 120 with alanine is designated as “His120Ala” or “H120A”. Substitutions can also be described by merely naming the resulting amino acid in the variant without specifying the amino acid of the parent at this position, e.g., by using “X120A” or “120A” or “Xaa120Ala” or “120Ala”.

[0029] Variants comprising multiple substitutions are separated by “+”, e.g., “Arg170Tyr- 3ly195Glu”, “R170Y+G195E” or “X170Y+X195E” representing a substitution of arginine and glycine at positions 170 and 195 with tyrosine and gl utamic acid, respectively. Alternatively, multiple substitutions may be separated by space or a comma, e.g., “R170Y G195E” or “R170Y, G195E” respectively. Where different alternative substitutions can be introduced at a position, the different substitutions are separated by a comma, e.g., “Arg170Tyr,Glu” and “R170T,E”, respectively, represents a substitution of arginine at position 170 with tyrosine or glutamic acid. Alternative substitutions at a particular position can also be indicated as “X120A,G,H”, “120A,G,H”, “X120A / G / H”, or“120A / G / H”. Alternatively, different substitutions may be indicated in brackets, e.g., “Arg170[Tyr, Gly]” or “Arg170{Tyr,Gly}” or in short “R170 [Y,G]” or“R170 {Y,G}”.

[0030] The numbering of the amino acid residues of the proteases described herein is as commonly used for proteases in the field (cf. P.N. Bryan, Biochimica et Biophysica Acta 1543 (2000), 203-222, cf. p. 204, left col., 3rdpara.) according to the numbering of the BPN’ subtilisin protease from Bacillus amyloliquefaciens the sequence of which is shown in SEQ ID NO: 2 (i.e., according to the numbering of SEQ ID NO: 2 or according to “BPN’ numbering”).

[0031] In an alternative way, one can describe the amino acid positions with reference to the numbering of SEQ ID NO: 1 or SEQ ID NO: 3, i.e., according to the numbering of SEQ ID NO: 1 or according to the numbering of SEQ ID NO: 3. Table 1 below shows the amino acid numbering according to SEQ ID NO: 2 and the numbering of the corresponding amino acids in the sequences according to SEQ ID NO: 1 or 3:

[0032] Table 1

[0033] The term “native” (or naturally or wild-type or endogenous) cell or organism or polynucleotide or polypeptide refers to the cell or organism or polynucleotide or polypeptide as found in nature (i.e., without there being any human intervention).

[0034] The term "heterologous” (or exogenous or foreign or recombinant or non-native or non-natural) polypeptide is defined herein as a polypeptide that is not native to the host cell, a polypeptide native to the host cell in which structural modifications, e.g., deletions, substitutions, and / or insertions, have been made by recombinant DNA techniques to alter the native polypeptide, or a polypeptide native to the host cell whose expression is quantitatively altered or whose expression is directed from a genomic location different from the native host cell as a result of manipulation of the DNA of the host cell by recombinant DNA techniques, e.g., a stronger promoter. Similarly, the term “heterologous” (or exogenous orforeign or recombinant or non-native or non-natural) polynucleotide refers to a polynucleotide that is not native to the host cell, a polynucleotide native to the host cell in which structural modifications, e.g., deletions, substitutions, and / or insertions, have been made by recombinant DNA techniques to alter the native polynucleotide, or a polynucleotide native to the host cell whose expression is quantitatively altered as a result of manipulation of the regulatory elements of the polynucleotide by recombinant DNA techniques, e.g., a stronger promoter, or a polynucleotide native to the host cell, but integrated not within its natural genetic environment as a result of genetic manipulation by recombinant DNA techniques. With respect to the relation between two or more polynucleotide sequences or the relation between two or more amino acid sequences, the term "heterologous” is used to characterize that the two or more polynucleotide sequences or two or more amino acid sequences are naturally not occurring in the specific combination with each other.

[0035] For the purpose of the invention, "recombinant" (or transgenic) with regard to a cell or an organism means that the cell or organism contains a heterologous polynucleotide, which is introduced by man using gene technology. With regard to a polynucleotide “recombinant” includes all constructs produced by using gene technology / recombinant DNA techniques in which either

[0036] (a) the sequence of the polynucleotide or a part thereof, or

[0037] (b) one or more genetic control sequences, which are operably linked to the polynucleotide, including but not limited to a promoter, or

[0038] (c) both a) and b) are not located in their wild-type genetic environment or have been modified by man.

[0039] A "synthetic" compound is obtained by in vitro chemical and / or enzymatic synthesis.

[0040] Variant polynucleotide and variant polypeptide sequences may be defined by their sequence identity when compared to a parent sequence. Sequence identity usually is provided as “% sequence identity” or“% identity”. For calculation of sequence identities, in a first step a sequence alignment is produced. According to this invention, a pairwise global alignment is produced, meaning that two sequences are aligned over their complete length, which is usually produced by using a mathematical approach, called alignment algorithm.

[0041] According to the invention, the alignment is generated by using the algorithm of Needleman and Wunsch (J. Mol. Biol. (1970) 48, p. 443-453). Preferably, the program “NEEDLE” (The European Molecular Biology Open Software Suite (EMBOSS)) is used for the purposes of the current invention, with using the programs default parameter (polynucleotides: gap open=10.0, gap extend=0.5 and matrix=EDNAFULL; polypeptides: gap open=10.0, gap extend=0.5 and matrix=EBLOSUM62). After aligning two sequences, in a second step, an identity value is determined from the alignment produced. For this purpose, the %-identity is calculated by dividing the number of identical residues by the length of the alignment region which is showing the respective sequence of the present invention over its complete length multiplied with 100: %-identity = (identical residues / length of the alignment region which is showing the respective sequence of the present invention over its complete length) *100.

[0042] A special aspect concerning amino acid substitutions are conservative mutations which often appear to have a minimal effect on protein folding resulting in substantially maintained enzyme properties of the respective enzyme variant compared to the enzyme properties of the parent enzyme. Conservative mutations are those where one amino acid is exchanged with a similar amino acid. Such an exchange most probably does not change enzyme properties.

[0043] Herein the following conservative exchanges are considered:

[0044] Amino acid A is similar to amino acids S

[0045] Amino acid D is similar to amino acids E; N

[0046] Amino acid E is similar to amino acids D; K; Q

[0047] Amino acid F is similar to amino acids W; Y

[0048] Amino acid H is similar to amino acids N; Y

[0049] Amino acid I is similar to amino acids L; M; V

[0050] Amino acid K is similar to amino acids E; Q; R

[0051] Amino acid L is similar to amino acids I; M; V Amino acid M is similar to amino acids I; L; V Amino acid N is similar to amino acids D; H; S Amino acid Q is similar to amino acids E; K; R Amino acid R is similar to amino acids K; Q Amino acid S is similar to amino acids A; N; T Amino acid T is similar to amino acids S Amino acid V is similar to amino acids I; L; M Amino acid W is similar to amino acids F; Y Amino acid Y is similar to amino acids F; H; W Conservative amino acid substitutions may occur over the full length of the sequence of a polypeptide sequence of a functional protein such as an enzyme. Preferably, such mutations are not pertaining the functional domains of an en- zyme, more preferably conservative mutations are not pertaining the catalytic centers of an enzyme.

[0052] A "fragment" or “subsequence” as used herein refers to a portion of an amino acid sequence. Polypeptides comprising a deletion of one or more amino acids at the N terminus and / or the C terminus of the polypeptide and essentially retain protease activity are herein designated as "functional fragments". Preferably, the functional fragment has at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 76%, at least 77%, at least 78%, at least 79%, at least 80% identical, at least 81 %, at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least

[0053] 87%, at least 88%, at least 89%, at least 90%, at least 91 %, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 98.5 %, at least 99%, or at least 99.5% of the length of the original full length amino acid sequence. Preferably, the functional fragment comprises 100 to 259 consecutive amino acids of the full-length variant polypeptide. Also preferably, the functional fragment retains at least 75%, at least 76%, at least 77%, at least 78%, at least 79%, at least 80% identical, at least 81 %, at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91 %, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 98.5 %, at least 99%, at least 99.5% or at least 100% of the enzyme activity of the original full length amino acid sequence. The functional fragment comprises consecutive amino acids compared to the original full length amino acid sequence, respectively. In the context of the present invention, the "original full length amino acid sequence" is an amino acid sequence which is at least 82% identical to the amino acid sequence according to SEQ ID NO: 1 and which has the same length as the amino acid sequence according to SEQ ID NO: 1 .

[0054] In one embodiment, the polypeptide of the present invention does not comprise any internal deletions compared to the amino acid sequence according to SEQ ID NO: 1 . However, as discussed above, the polypeptide of the present invention may comprise a deletion of one or more amino acids at the N terminus and / or the C terminus of the polypeptide. “Genetic construct” or “expression cassette” as used herein, is a nucleic acid molecule composed of at least one sequence of interest to be expressed, operably linked to one or more control sequences (at least to a promoter) as described herein.

[0055] The term “vector” as used herein comprises any kind of construct suitable to carry foreign polynucleotide sequences for transfer to another cell, or for stable or transient expression within a given cell. The term “vector” as used herein encompasses any kind of cloning vehicles, such as but not limited to plasmids, phagemids, viral vectors (e.g., phages), bacteriophage, baculoviruses, cosmids, fosmids, artificial chromosomes, and any other vectors specific for specific hosts of interest. Foreign polynucleotide sequences usually comprise a coding sequence which may be referred to herein as “gene of interest”. The gene of interest may comprise introns and exons, depending on the kind of origin or destination of host cell.

[0056] The term “introduction of a polynucleotide” or “transformation of a polynucleotide” as referred to herein encompasses the transfer of an exogenous polynucleotide into a host cell, irrespective of the method used for transfer. That is, the term “transformation of a polynucleotide” as used herein is independent from vector, shuttle system, or host cell, and it not only relates to the polynucleotide transfer method of transformation as known in the art (cf. , for example, Sam- brook, J. et al. (1989) Molecular Cloning: A Laboratory Manual, 2nd Ed., Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY), but it encompasses any further kind of polynucleotide transfer methods such as, but not limited to, transduction or transfection.

[0057] A polynucleotide encoding a polypeptide may be “expressed”. The term “expression” or “gene expression” means the transcription of a specific gene or specific genes or specific nucleic acid construct. The term “expression” or “gene expression” means the transcription of a gene or genes or genetic construct into structural RNA (e.g., rRNA, tRNA) or mRNA with or without subsequent translation of the latter into a protein. The process includes transcription of DNA and processing of the resulting mRNA product.

[0058] The term “purification” or “purifying” refers to a process in which at least one component, e.g., a protein of interest, is separated from at least another component, e.g., a particulate matter of a fermentation broth, and transferred into a different compartment or phase, wherein the different compartments or phases do not necessarily need to be separated by a physical barrier. Examples of such different compartments are two compartments separated by a filtration membrane or cloth, i.e., filtrate and retentate; examples of such different phases are pellet and supernatant or cake and filtrate, respectively. The resulting solution after purifying the enzyme of interest from the fermentation broth is called herein “purified enzyme solution”.

[0059] “Protein formulation” (or “enzyme preparation” or “polypeptide formulation”), e.g., “protease formulation”, means any non-complex formulation comprising a small number of ingredients, preferably, 2-8 components, wherein the ingredients serve the purpose of stabilizing the proteins / polypeptides comprised in the protein / polypeptide formulation and / or the stabilization of the protein / polypeptide formulation itself. Preferably, the non-complex protein / polypeptide formulation comprises the protein / polypeptide in higher concentrations than the complex formulation, e.g., than a detergent formulation. Thus, preferably the non-complex protein formulation is a concentrated protein / polypeptide formulation. Preferably, non-complex protein / polypeptide formulations comprise 20 to 120 mg / g active enzyme, whereas complex formulations, like detergent compositions, comprise 0.002 to 10 mg / g active enzyme. In contrast to a non-complex formulation, a complex formulation means herein a formulation comprising a higher number of ingredients, preferably, 9-30 components, wherein the ingredients serve the purpose of stabilizing the proteins comprised in the protein formulation and / or the stabilization of the protein formulation itself, but additionally the complex formulation comprises components that serve the purpose of the complex formulation, e.g., a detergent composition. An example for a non- complex protein formulation is a concentrated enzyme composition that is used as a stock-solution to prepare a complex formulation, e.g., a detergent composition, wherein in the detergent compositions other compounds are present that serve the cleaning purpose of the detergent composition, e.g., surfactants and / or chelating agents.

[0060] “Enzyme properties” include, but are not limited to, catalytic activity, substrate / cofactor specificity, product specificity, stability in the course of time, thermostability, pH stability, and chemical stability. “Enzymatic activity” or “catalytic activity” means the catalytic effect exerted by an enzyme, expressed as units per milligram of enzyme (specific activity) or molecules of substrate transformed per minute per molecule of enzyme (molecular activity). Enzymatic activity can be specified by the enzyme’s actual function, e.g., proteases exerting proteolytic activity by catalyzing hydrolytic cleavage of peptide bonds, lipases exerting lipolytic activity by hydrolytic cleavage of ester bonds, amylases activity involves hydrolysis of glycosidic linkages in polysaccharides, etc. According to the invention, the enzymatic activity is proteolytic activity which can be determined by using Succinyl-Ala-Ala-Pro-Phe-p-nitroanilide (Suc-AAPF-pNA; see e.g. DelMar et al. (1979), Analytical Biochem 99, 316-320) or Suc-AAPF-AMC (7-amido-4-methylcoumarin) as substrate. pNA or AMC is cleaved from the substrate molecule by proteolytic cleavage, resulting in release of yellow color of free pNA or change of fluorescence properties which can be quantified either by measuring emission at 460 nm after excitation at 380 nm. Other methods using for example casein as a proteinaceous substrate are known to those skilled in the art. The term “enzyme stability” according to the current invention relates to the retention of enzymatic activity as a function of time during storage or operation. Retention of enzymatic activity as a function of time during storage is called “storage stability” and is preferred within the context of the invention. To determine and quantify changes in catalytic activity of enzymes stored or used under certain conditions over time, the enzymatic activity is measured under defined conditions at time zero (100%) and at a certain point in time later (x%). By comparison of the values measured, the residual activity of the enzyme can be determined in its extent. The extent of the residual activity of the enzyme determines an enzyme’s stability or non -stability.

[0061] “Enzyme inhibitors” as used herein are compounds that slow down or halt enzymatic activity. Enzyme inhibitors frequently also stabilize the enzyme in its three-dimensional structure. Hence, enzyme inhibitors usually also act as “enzyme stabilizers”.

[0062] “pH stability” refers to the ability of an enzyme to exert enzymatic activity after exposure to a certain pH value.

[0063] The terms “detergent stability” (also called herein “residual activity in a detergent” or “storage stability in a detergent composition”) refer to the ability of an enzyme to exert catalytic activity or wash performance after storage in a detergent composition, preferably, at a temperature of 37 °C, 45 °C, or 50 °C for up to 14 days in a detergent composition (preferably, in Model B detergent as described herein). Most preferably, “detergent stability” is determined by measuring residual activity of the protease after storage at a temperature of 45 °C for 18 hours and / or 3 days in a detergent composition (preferably, in Model B detergent as described herein).

[0064] As used herein, "wash performance" (also called herein “cleaning performance”) of an enzyme refers to the contribution of the enzyme to the cleaning performance of a detergent composition, i.e. the cleaning performance added to the detergent composition by the performance of the enzyme. The term “wash performance” is used herein similarly for laundry and hard surface cleaning. Wash performance is compared under relevant washing conditions. The term "relevant washing conditions" is used herein to indicate the conditions, particularly washing temperature, time, washing mechanics, sud concentration, type ofdetergent and water hardness, actually used in households in a detergent market segment. The term "improved wash performance" is used to indicate that a better end result is obtained in stain removal under relevant washing conditions, or that less enzyme, on weight basis, is needed to obtain the same end result relative to the corresponding control conditions.

[0065] As used herein, the term "specific performance" refers to the cleaning and removal of specific stains or soils per unit of active enzyme. In some embodiments, the specific performance is determined using stains or soils such as egg, egg yolk, milk, grass, minced meat blood, chocolate sauce, baby food, sebum, etc.

[0066] “Detergent composition” or “detergent” means compositions designated for cleaning soiled material. Detergent compositions are complex formulations as furtherdefined herein. Detergent compositions according to the invention include detergent compositions for different applications such as laundry and hard surface cleaning. Detergent compositions according to the invention also include biodegradable detergent compositions. The term “detergent component” is defined herein to mean a type of chemical, which can be used in detergent compositions. A typical detergent component is a surfactant. "Surfactant" (synonymously used herein with “surface active agent”) means an organic chemical that, when added to a liquid, changes the properties of that liquid at an interface. According to its ionic charge, a surfactant is called non-ionic, anionic, cationic, or amphoteric. The term “effective amount of a detergent component” includes amounts of certain components to provide effective stain removal and / or effective cleaning conditions (e.g. pH, temperature, water hardness, quantity of foaming), amounts of certain components to effectively provide optical benefits (e.g. optical brightening, dye transfer inhibition, color care), and amounts of certain components to effectively aid the processing (maintain physical characteristics during processing, storage and use; e.g. rheology modifiers, hydrotropes, desiccants). Detergent compositions typically have a protease concentration of 0.002 to 10 mg / g active enzyme.

[0067] The term “laundry” or “laundering” relates to both household laundering and industrial laundering and means the process of treating textiles and / or fabrics with a solution containing a detergent composition of the present invention. The laundering process may be carried out by using technical devices such as a household or an industrial washing machine. Alternatively, the laundering process may be done by hand.

[0068] The term “textile” means any textile material including yarns (thread made of natural or synthetic fibers used for knitting or weaving), yarn intermediates, fibers, non-woven materials, natural materials, synthetic materials, as well as fabrics made of these materials such as garments, cloths and other articles. The terms “fabric” (a textile made by weaving, knitting orfelting fibers) or “garment” (any article of clothing made of textile) as used herein, are intended to include the broader term textile as well.

[0069] The term “fibers” includes natural fibers, synthetic fibers, and mixtures thereof. Examples of natural fibers are of plant (such as flax, jute and cotton) or animal origin, comprising proteins like collagen, keratin and fibroin (e.g. silk, sheep’s wool, angora, mohair, cashmere). Examples for fibers of synthetic origin are polyurethane fibers such as Spandex® or Lycra®, polyester fibers, polyolefins such as elastofin, or polyamide fibers such as nylon. Fibers may be single fibers or parts of textiles such as knitwear, woven or non-woven fabrics.

[0070] The term “hard surface cleaning” relates to both household hard surface cleaning and industrial hard surface cleaning and means the process of treating hard surfaces with a solution containing a detergent composition of the present invention. Hard surfaces may include any hard surfaces in the household or industry, such as floors, furnishing, walls, sanitary ceramics, glass, metallic surfaces including cutlery or dishes and medical devices such as diagnostic instruments, trays, pans, holders, racks, forceps, scissors, shears, saws (e.g. bone saws and their blades), hemostats, knives, chisels, rongeurs, files, nippers, drills, drill bits, rasps, burrs, spreaders, breakers, elevators, clamps, needle holders, carriers, clips, hooks, gouges, curettes, retractors, straightener, punches, extractors, scoops, keratomes, spatulas, expressors, trocars, dilators, cages, glassware, tubing, catheters, cannulas, plugs, stents, endoscopes, artho- scopes and related equipment. A particular form of hard surface cleaning is dishwashing, particularly automatic dishwashing (ADW).

[0071] The term “dish wash” refers to all forms of washing dishes, e.g. by hand or automatic dish wash. Washing dishes includes, but is not limited to, the cleaning of all forms of crockery such as plates, cups, glasses, bowls, all forms of cutlery such as spoons, knives, forks and serving utensils as well as ceramics, plastics such as melamine, metals, china, glass and acrylics. Cleaning performance is evaluated under relevant cleaning conditions. The term "relevant cleaning conditions" herein refers to the conditions, particularly cleaning temperature, time, cleaning mechanics, suds concentration, type of detergent and water hardness, actually used in laundry machines, automatic dish washers or in manual cleaning processes.

[0072] The term “medical device cleaning” refers to the cleaning step in reprocessing reusable medical devices. Medical device cleaning methods can be divided into two categories, manual and mechanical / automated cleaning methods. Manual cleaning is used when mechanical units are not available or medical devices to be cleaned are too fragile or difficult to clean with a mechanical unit. Mechanical / automated cleaning methods remove soiling and microorganisms through an automated cleaning and rinsing process, this includes ultrasonic cleaning and washing.

[0073] In the field of detergency, usually the term “stains” is used with reference to laundry, e.g., cleaning of textiles, fabric, or fibers, whereas the term “soils” is usually used with reference to hard surface cleaning, e.g., cleaning of dishes and cutlery. However, herein the terms “stain” and “soil” shall be used interchangeably.

[0074] A “sequestering builder” as used herein is different from a precipitating builder in that no significant amount of precipitate is formed when the builder is used in an amount sufficient to combine with all of the calcium ions in an aqueous solution with 7 °dH hardness (German hardness) initially at neutral pH. A “strong builder” is classified as high efficiency chelators that can bind the divalent cations such as Ca2+ strongly with a logarithmic stability constant (Log Kca) of the cation / chelator complex of above 4, particular above 5, above 6 or above 7. The stability constants are determined at an ionic strength of 0.1 M and at a temperature of 25°C. A ..strong sequestering builder” combines both of the above- mentioned properties. Strong sequestering builders include, but are not limited to, Ethylenediaminetetraacetic acid (EDTA), Ethylene diamine tetra(methylene phosphonic acid (EDTMP), Nitrilo trimethylene phosphonic acid (NTMP), Diethylenetriamine Penta(Methylene Phosphonic acid) (DTPMP), methylglycinediacetic acid (MGDA), Nitrilotriacetic acid (NTA), 1 -Hydroxy Ethylidene-1 ,1 -Diphosphonic acid (HEDP), sodium tripolyphosphate (STPP), iminodisuccinic acid (IDS), N,N-diacetic acid tetra sodium salt (GLDA), pyrophosphate and ethylenediaminedisuccinic acid (EDDS), preferably MGDA and / or EDDS.

[0075] An “antimicrobial agent” is a chemical compound that kills microorganisms or inhibits their growth or reproduction. Microorganisms can be bacteria, yeasts or molds. A “preservative” is an antimicrobial agent which may be added to aqueous products and compositions to maintain the original performance, characteristics and integrity of the products and compositions by killing contaminating microorganisms or inhibiting their growth.

[0076] A composition “essentially devoid” of a compound shall mean herein that the respective compound is not added to the composition on purpose, meaning that at most non-effective amounts are present, most preferably 0% of the compound are contained in the composition. Detailed description

[0077] In the present invention new protease enzymes are provided. More specifically, variants of a parent protease, methods of making the variant proteases, compositions comprising the protease variants, and methods of using the variant proteases or compositions comprising the variant proteases are provided.

[0078] Protease Variant

[0079] The present invention is directed to a polypeptide having protease activity or a functional fragment of said polypeptide having protease activity, wherein:

[0080] (i) the polypeptide has an amino acid sequence which is at least 82%, but less than 100%, identical to the amino acid sequence as set forth in SEQ ID NO: 1 and

[0081] (ii) the polypeptide or fragment thereof comprises an amino acid substitution at two or more amino acid residues selected from the group consisting of 38, 59 and 212 compared to the amino acid sequence as set forth in SEQ ID NO:

[0082] 3 and referring to the numbering of SEQ ID NO: 2, wherein the amino acid substitutions 38F, 38H, 38N, 59A, 59H, 59N, 59S, 212H and 212M are excluded and wherein the fragment of the variant polypeptide comprises 100 to 259 consecutive amino acids of the full-length variant polypeptide.

[0083] The present invention is also directed to a polypeptide having protease activity, wherein:

[0084] (I) the polypeptide has an amino acid sequence which is at least 82%, but less than 100%, identical to the amino acid sequence as set forth in SEQ ID NO: 1 and

[0085] (ii) the polypeptide comprises an amino acid substitution at two or more amino acid residues selected from the group consisting of 38, 59 and 212 compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2, wherein the amino acid substitutions 38F, 38H, 38N, 59A, 59H, 59N, 59S, 212H and 212M are excluded.

[0086] The present invention is directed to a polypeptide having protease activity or a functional fragment of said polypeptide having protease activity, wherein:

[0087] (I) the polypeptide has an amino acid sequence which is at least 82%, but less than 100%, identical to the amino acid sequence as set forth in SEQ ID NO: 1 and

[0088] (ii) the polypeptide or fragment thereof comprises (A) an amino acid substitution at amino acid residue 212 and (B) an amino acid substitution at amino acid residue 38 and / or 59 compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2, wherein the amino acid substitutions 38F, 38H, 38N, 59A, 59H, 59N, 59S, 212H and 212M are excluded and wherein the fragment of the variant polypeptide comprises 100 to 259 consecutive amino acids of the full-length variant polypeptide.

[0089] The present invention is also directed to a polypeptide having protease activity, wherein: (i) the polypeptide has an amino acid sequence which is at least 82%, but less than 100%, identical to the amino acid sequence as set forth in SEQ ID NO: 1 and

[0090] (ii) the polypeptide or fragment thereof comprises (A) an amino acid substitution at amino acid residue 212 and (B) an amino acid substitution at amino acid residue 38 and / or 59 compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2, wherein the amino acid substitutions 38F, 38H, 38N, 59A, 59H, 59N, 59S, 212H and 212M are excluded.

[0091] The present invention is directed to a polypeptide having protease activity or a functional fragment of said polypeptide having protease activity, wherein:

[0092] (I) the polypeptide has an amino acid sequence which is at least 82%, but less than 100%, identical to the amino acid sequence as set forth in SEQ ID NO: 1 and

[0093] (ii) the polypeptide or fragment thereof comprises two or more amino acid substitutions selected from the group consisting of X38R / K / W, X59K / E and X212G / R / K / P / A / F / W / N / Y / D compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2, wherein the fragment of the variant polypeptide comprises 100 to 259 consecutive amino acids of the full-length variant polypeptide.

[0094] The present invention is directed to a polypeptide having protease activity, wherein:

[0095] (I) the polypeptide has an amino acid sequence which is at least 82%, but less than 100%, identical to the amino acid sequence as set forth in SEQ ID NO: 1 and

[0096] (ii) the polypeptide comprises two or more amino acid substitutions selected from the group consisting of X38R / K / W, X59K / E and X212G / R / K / P / A / F / W / N / Y / D compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2.

[0097] The present invention is directed to a polypeptide having protease activity or a functional fragment of said polypeptide having protease activity, wherein:

[0098] (I) the polypeptide has an amino acid sequence which is at least 82%, but less than 100%, identical to the amino acid sequence as set forth in SEQ ID NO: 1 and

[0099] (ii) the polypeptide or fragment thereof comprises (A) the amino acid substitution X212G / M / R / K / P / A / F / H / W / N / Y / D and (B) the amino acid substitution X38R / K / W and / or X59K / E compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2, wherein the fragment of the variant polypeptide comprises 100 to 259 consecutive amino acids of the full-length variant polypeptide.

[0100] The present invention is directed to a polypeptide having protease activity, wherein:

[0101] (I) the polypeptide has an amino acid sequence which is at least 82%, but less than 100%, identical to the amino acid sequence as set forth in SEQ ID NO: 1 and (ii) the polypeptide comprises (A) the amino acid substitution X212G / M / R / K / P / A / F / H / W / N / Y / D and (B) the amino acid substitution X38R / K / W and / or X59K / E compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2.

[0102] The present invention is directed to a polypeptide having protease activity or a functional fragment of said polypeptide having protease activity, wherein:

[0103] (i) the polypeptide has an amino acid sequence which is at least 82%, but less than 975%, identical to the amino acid sequence as set forth in SEQ ID NO: 1 and

[0104] (ii) the polypeptide or fragment thereof comprises an amino acid substitution at two or more amino acid residues selected from the group consisting of 38, 59 and 212 compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2, wherein the amino acid substitutions 38F, 38H, 38N, 59A, 59H, 59N, 59S, 212H and 212M are excluded and wherein the fragment of the variant polypeptide comprises 100 to 259 consecutive amino acids of the full-length variant polypeptide.

[0105] The present invention is also directed to a polypeptide having protease activity, wherein:

[0106] (I) the polypeptide has an amino acid sequence which is at least 82%, but less than 97.5%, identical to the amino acid sequence as set forth in SEQ ID NO: 1 and

[0107] (ii) the polypeptide comprises an amino acid substitution at two or more amino acid residues selected from the group consisting of 38, 59 and 212 compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2, wherein the amino acid substitutions 38F, 38H, 38N, 59A, 59H, 59N, 59S, 212H and 212M are excluded.

[0108] The present invention is directed to a polypeptide having protease activity or a functional fragment of said polypeptide having protease activity, wherein:

[0109] (I) the polypeptide has an amino acid sequence which is at least 82%, but less than 97.5%, identical to the amino acid sequence as set forth in SEQ ID NO: 1 and

[0110] (ii) the polypeptide or fragment thereof comprises (A) an amino acid substitution at amino acid residue 212 and (B) an amino acid substitution at amino acid residue 38 and / or 59 compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2, wherein the amino acid substitutions 38F, 38H, 38N, 59A, 59H, 59N, 59S, 212H and 212M are excluded and wherein the fragment of the variant polypeptide comprises 100 to 259 consecutive amino acids of the full-length variant polypeptide.

[0111] The present invention is also directed to a polypeptide having protease activity, wherein:

[0112] (I) the polypeptide has an amino acid sequence which is at least 82%, but less than 97.5%, identical to the amino acid sequence as set forth in SEQ ID NO: 1 and (ii) the polypeptide comprises (A) an amino acid substitution at amino acid residue 212 and (B) an amino acid substitution at amino acid residue 38 and / or 59 compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2, wherein the amino acid substitutions 38F, 38H, 38N, 59A, 59H, 59N, 59S, 212H and 212M are excluded.

[0113] The present invention is directed to a polypeptide having protease activity or a functional fragment of said polypeptide having protease activity, wherein:

[0114] (i) the polypeptide has an amino acid sequence which is at least 82%, but less than 97.5%, identical to the amino acid sequence as set forth in SEQ ID NO: 1 and

[0115] (ii) the polypeptide or fragment thereof comprises two or more amino acid substitutions selected from the group consisting of X38R / K / W, X59K / E and X212G / R / K / P / A / F / W / N / Y / D compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2, wherein the fragment of the variant polypeptide comprises 100 to 259 consecutive amino acids of the full-length variant polypeptide.

[0116] The present invention is directed to a polypeptide having protease activity, wherein:

[0117] (I) the polypeptide has an amino acid sequence which is at least 82%, but less than 97.5%, identical to the amino acid sequence as set forth in SEQ ID NO: 1 and

[0118] (ii) the polypeptide comprises two or more amino acid substitutions selected from the group consisting of X38R / K / W, X59K / E and X212G / R / K / P / A / F / W / N / Y / D compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2.

[0119] The present invention is directed to a polypeptide having protease activity or a functional fragment of said polypeptide having protease activity, wherein:

[0120] (I) the polypeptide has an amino acid sequence which is at least 82%, but less than 97.5%, identical to the amino acid sequence as set forth in SEQ ID NO: 1 and

[0121] (ii) the polypeptide or fragment thereof comprises (A) the amino acid substitution X212G / M / R / K / P / A / F / H / W / N / Y / D and (B) the amino acid substitution X38R / K / W and / or X59K / E compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2, wherein the fragment of the variant polypeptide comprises 100 to 259 consecutive amino acids of the full-length variant polypeptide.

[0122] The present invention is directed to a polypeptide having protease activity, wherein:

[0123] (I) the polypeptide has an amino acid sequence which is at least 82%, but less than 97.5%, identical to the amino acid sequence as set forth in SEQ ID NO: 1 and

[0124] (ii) the polypeptide comprises (A) the amino acid substitution X212G / M / R / K / P / A / F / H / W / N / Y / D and (B) the amino acid substitution X38R / K / W and / or X59K / E compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2. The present invention is directed to a polypeptide having protease activity or a functional fragment of said polypeptide having protease activity, wherein:

[0125] (I) the polypeptide has an amino acid sequence which is at least 82%, but less than 100%, identical to the amino acid sequence as set forth in SEQ ID NO: 1 and

[0126] (ii) the polypeptide or fragment thereof comprises an amino acid substitution at two or more amino acid residues selected from the group consisting of 38, 59 and 212 compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2, wherein the amino acid substitutions 38F, 38H, 38N, 59A, 59H, 59N, 59S, 212H and 212M are excluded, wherein the fragment of the variant polypeptide comprises 100 to 259 consecutive amino acids of the full-length variant polypeptide and wherein the variant polypeptide comprises amino acid residue D or E at position 101 , preferably E at position 101 , referring to the numbering of SEQ ID NO: 2.

[0127] The present invention is also directed to a polypeptide having protease activity, wherein:

[0128] (I) the polypeptide has an amino acid sequence which is at least 82%, but less than 100%, identical to the amino acid sequence as set forth in SEQ ID NO: 1 and

[0129] (ii) the polypeptide comprises an amino acid substitution at two or more amino acid residues selected from the group consisting of 38, 59 and 212 compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2, wherein the amino acid substitutions 38F, 38H, 38N, 59A, 59H, 59N, 59S, 212H and 212M are excluded and wherein the variant polypeptide comprises amino acid residue D or E at position 101 , preferably E at position 101 , referring to the numbering of SEQ ID NO: 2.

[0130] The present invention is directed to a polypeptide having protease activity or a functional fragment of said polypeptide having protease activity, wherein:

[0131] (I) the polypeptide has an amino acid sequence which is at least 82%, but less than 100%, identical to the amino acid sequence as set forth in SEQ ID NO: 1 and

[0132] (ii) the polypeptide or fragment thereof comprises two or more amino acid substitutions selected from the group consisting of X38R / K / W, X59K / E and X212G / R / K / P / A / F / W / N / Y / D compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2, wherein the fragment of the variant polypeptide comprises 100 to 259 consecutive amino acids of the full-length variant polypeptide and wherein the variant polypeptide comprises amino acid residue D or E at position 101 , preferably E at position 101 , referring to the numbering of SEQ ID NO: 2.

[0133] The present invention is directed to a polypeptide having protease activity, wherein:

[0134] (I) the polypeptide has an amino acid sequence which is at least 82%, but less than 100%, identical to the amino acid sequence as set forth in SEQ ID NO: 1 and (ii) the polypeptide comprises two or more amino acid substitutions selected from the group consisting of X38R / K / W, X59K / E and X212G / R / K / P / A / F / W / N / Y / D compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2, wherein the variant polypeptide comprises amino acid residue D or E at position 101 , preferably E at position 101 , referring to the numbering of SEQ ID NO: 2.

[0135] The present invention is directed to a polypeptide having protease activity or a functional fragment of said polypeptide having protease activity, wherein:

[0136] (I) the polypeptide has an amino acid sequence which is at least 82%, but less than 975%, identical to the amino acid sequence as set forth in SEQ ID NO: 1 and

[0137] (ii) the polypeptide or fragment thereof comprises an amino acid substitution at two or more amino acid residues selected from the group consisting of 38, 59 and 212 compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2, wherein the amino acid substitutions 38F, 38H, 38N, 59A, 59H, 59N, 59S, 212H and 212M are excluded, wherein the fragment of the variant polypeptide comprises 100 to 259 consecutive amino acids of the full-length variant polypeptide and wherein the variant polypeptide comprises amino acid residue D or E at position 101 , preferably E at position 101 , referring to the numbering of SEQ ID NO: 2.

[0138] The present invention is also directed to a polypeptide having protease activity, wherein:

[0139] (I) the polypeptide has an amino acid sequence which is at least 82%, but less than 975%, identical to the amino acid sequence as set forth in SEQ ID NO: 1 and

[0140] (ii) the polypeptide comprises an amino acid substitution at two or more amino acid residues selected from the group consisting of 38, 59 and 212 compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2, wherein the amino acid substitutions 38F, 38H, 38N, 59A, 59H, 59N, 59S, 212H and 212M are excluded and wherein the variant polypeptide comprises amino acid residue D or E at position 101 , preferably E at position 101 , referring to the numbering of SEQ ID NO: 2.

[0141] The present invention is directed to a polypeptide having protease activity or a functional fragment of said polypeptide having protease activity, wherein:

[0142] (I) the polypeptide has an amino acid sequence which is at least 82%, but less than 975%, identical to the amino acid sequence as set forth in SEQ ID NO: 1 and

[0143] (ii) the polypeptide or fragment thereof comprises two or more amino acid substitutions selected from the group consisting of X38R / K / W, X59K / E and X212G / R / K / P / A / F / W / N / Y / D compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2, wherein the fragment of the variant polypeptide comprises 100 to 259 consecutive amino acids of the full-length variant polypeptide and wherein the variant polypeptide comprises amino acid residue D or E at position 101 , preferably E at position 101 , referring to the numbering of SEQ ID NO: 2. The present invention is directed to a polypeptide having protease activity, wherein:

[0144] (i) the polypeptide has an amino acid sequence which is at least 82%, but less than 975%, identical to the amino acid sequence as set forth in SEQ ID NO: 1 and

[0145] (ii) the polypeptide comprises two or more amino acid substitutions selected from the group consisting of X38R / K / W, X59K / E and X212G / R / K / P / A / F / W / N / Y / D compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2, wherein the variant polypeptide comprises amino acid residue D or E at position 101 , preferably E at position 101 , referring to the numbering of SEQ ID NO: 2.

[0146] The variant polypeptide having protease activity of the present invention is a non-naturally occurring protease. Preferably, the variant polypeptide having protease activity of the present invention is a purified, isolated, synthetic, and / or recombinant protease. Preferably, the variant polypeptide having protease activity of the present invention is a purified and recombinant protease.

[0147] Proteases according to the invention have “proteolytic activity” or “protease activity”. “Proteolytic activity” or “protease activity” describes the capability for the hydrolysis of peptide bonds in polypeptides. Protease activity may be determined by assays for measurement of protease activity which are known to those skilled in the art. The methods for analyzing proteolytic activity are well-known in the literature (see e.g. Gupta et al. (2002), Appl. Microbiol. Biotechnol. 60: 381 -395). For instance, proteolytic activity can be determined by using Succinyl-Ala-Ala-Pro-Phe-p-nitroanilide (Suc-AAPF-pNA; see e.g. DelMar et al. (1979), Analytical Biochem 99, 316-320) or Suc-AAPF-AMC (7-amido-4- methylcoumarin) as substrate. pNA or AMC is cleaved from the substrate molecule by proteolytic cleavage, resulting in release of yellow color of free pNA or change of fluorescence properties which can be quantified either by measuring emission at 460 nm after excitation at 380 nm.

[0148] The variant polypeptide of the present invention preferably has a sequence identity of at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 90.5%, at least 91 %, at least 91 .5%, at least 92%, at least 92.5%, at least 93%, at least 93.5%, at least 94%, at least 94.5%, at least 95%, at least 95.1 %, at least 95.2%, at least 95.3%, at least 95.4%, at least 95.5%, at least 95.6%, at least 95.7%, at least 95.8%, at least 95.9% at least 96%, at least 96.1 %, at least 96.2%, at least 96.3%, at least 96.4%, at least 96.5%, at least 96.6%, at least 96.7%, at least 96.8%, at least 96.9%, at least 97%, at least 97.1 %, at least 97.2%, at least 97.3%, at least 97.4%, at least 97.5%, at least 97.6%, at least 97.7%, at least 97.8%, at least 97.9%, but less than 100% to the amino acid sequence according to SEQ ID NO: 1 and comprises an amino acid substitution at two or more amino acid residues selected from the group consisting of 38, 59 and 212 compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2, wherein the amino acid substitutions 38F, 38H, 38N, 59A, 59H, 59N, 59S, 212H and 212M are excluded and wherein the fragment of the variant polypeptide comprises 100 to 259 consecutive amino acids of the full-length variant polypeptide.

[0149] The variant polypeptide of the present invention preferably has a sequence identity of at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 90.5%, at least 91 %, at least 91 .5%, at least 92%, at least 92.5%, at least 93%, at least 93.5%, at least 94%, at least 94.5%, at least 95%, at least 95.1 %, at least 95.2%, at least 95.3%, at least 95.4%, at least 95.5%, at least 95.6%, at least 95.7%, at least 95.8%, at least 95.9% at least 96%, at least 96.1 %, at least 96.2%, at least 96.3%, at least 96.4%, at least 96.5%, at least 96.6%, at least 96.7%, at least 96.8%, at least 96.9%, at least 97%, at least 97.1 %, at least 97.2%, at least 97.3%, at least 97.4%, at least 97.5%, at least 97.6%, at least 97.7%, at least 97.8%, at least 97.9%, but less than 100% to the amino acid sequence according to SEQ ID NO: 1 and comprises (A) an amino acid substitution at amino acid residue 212 and (B) an amino acid substitution at amino acid residue 38 and / or 59 compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2, wherein the amino acid substitutions 38F, 38H, 38N, 59A, 59H, 59N, 59S, 212H and 212M are excluded and wherein the fragment of the variant polypeptide comprises 100 to 259 consecutive amino acids of the full-length variant polypeptide. In one embodiment the variant polypeptide of the present invention preferably has a sequence identity of at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 90.5%, at least 91 %, at least 91 .5%, at least 92%, at least 92.5%, at least 93%, at least 93.5%, at least 94%, at least 94.5%, at least 95%, at least 95.1 %, at least 95.2%, at least 95.3%, at least 95.4%, at least 95.5%, at least 95.6%, at least 95.7%, at least 95.8%, at least 95.9% at least 96%, at least 96.1 %, at least 96.2%, at least 96.3%, at least 96.4%, at least 96.5%, at least 96.6%, at least 96.7%, at least 96.8%, at least 96.9%, at least 97%, at least 97.1 %, at least 97.2%, at least 97.3%, at least 97.4%, at least 97.5%, at least 97.6%, at least 97.7%, at least 97.8%, at least 97.9%, but less than 100% to the amino acid sequence according to SEQ ID NO: 1 and comprises two or more amino acid substitutions selected from the group consisting of X38R / K / W, X59K / E and X212G / R / K / P / A / F / W / N / Y / D compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2.

[0150] In one embodiment the variant polypeptide of the present invention preferably has a sequence identity of at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 90.5%, at least 91 %, at least 91 .5%, at least 92%, at least 92.5%, at least 93%, at least 93.5%, at least 94%, at least 94.5%, at least 95%, at least 95.1 %, at least 95.2%, at least 95.3%, at least 95.4%, at least 95.5%, at least 95.6%, at least 95.7%, at least 95.8%, at least 95.9% at least 96%, at least 96.1 %, at least 96.2%, at least 96.3%, at least 96.4%, at least 96.5%, at least 96.6%, at least 96.7%, at least 96.8%, at least 96.9%, at least 97%, at least 97.1 %, at least 97.2%, at least 97.3%, at least 97.4%, at least 97.5%, at least 97.6%, at least 97.7%, at least 97.8%, at least 97.9%, but less than 100% to the amino acid sequence according to SEQ ID NO: 1 and comprises (A) the amino acid substitution X212G / R / K / P / A / F / W / N / Y / D and (B) the amino acid substitution X38R / K / W and / or X59K / E compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2.

[0151] In one embodiment the variant polypeptide of the present invention preferably has a sequence identity of at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 90.5%, at least 91 %, at least 91 .5%, at least 92%, at least 92.5%, at least 93%, at least 93.5%, at least 94%, at least 94.5%, at least 95%, at least 95.1 %, at least 95.2%, at least 95.3%, at least 95.4%, at least 95.5%, at least 95.6%, at least 95.7%, at least 95.8%, at least 95.9% at least 96%, at least 96.1 %, at least 96.2%, at least 96.3%, at least 96.4%, at least 96.5%, at least 96.6%, at least 96.7%, at least 96.8%, at least 96.9%, at least 97%, at least 97.1 %, at least 97.2%, at least 97.3%, at least 97.4%, at least 97.5%, at least 97.6%, at least 97.7%, at least 97.8%, at least 97.9%, but less than 100% to the amino acid sequence according to SEQ ID NO: 1 and comprises two or more amino acid substitutions selected from the group consisting of X38R / K / W, X59K / E and X212P / A / G / R / K / F / Y compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2.

[0152] In one embodiment the variant polypeptide of the present invention preferably has a sequence identity of at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 90.5%, at least 91 %, at least 91 .5%, at least 92%, at least 92.5%, at least 93%, at least 93.5%, at least 94%, at least 94.5%, at least 95%, at least 95.1 %, at least 95.2%, at least 95.3%, at least 95.4%, at least 95.5%, at least 95.6%, at least 95.7%, at least 95.8%, at least 95.9% at least 96%, at least 96.1 %, at least 96.2%, at least 96.3%, at least 96.4%, at least 96.5%, at least 96.6%, at least 96.7%, at least 96.8%, at least 96.9%, at least 97%, at least 97.1 %, at least 97.2%, at least 97.3%, at least 97.4%, at least 97.5%, at least 97.6%, at least 97.7%, at least 97.8%, at least 97.9%, but less than 100% to the amino acid sequence according to SEQ ID NO: 1 and comprises two or more amino acid substitutions selected from the group consisting of T38R / K / W, Q59K / E and S212G / R / K / P / A / F / W / N / Y / D compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2.

[0153] In one embodiment the variant polypeptide of the present invention preferably has a sequence identity of at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 90.5%, at least 91 %, at least 91 .5%, at least 92%, at least 92.5%, at least 93%, at least 93.5%, at least 94%, at least 94.5%, at least 95%, at least 95.1 %, at least 95.2%, at least 95.3%, at least 95.4%, at least 95.5%, at least 95.6%, at least 95.7%, at least 95.8%, at least 95.9% at least 96%, at least 96.1 %, at least 96.2%, at least 96.3%, at least 96.4%, at least 96.5%, at least 96.6%, at least 96.7%, at least 96.8%, at least 96.9%, at least 97%, at least 97.1 %, at least 97.2%, at least 97.3%, at least 97.4%, at least 97.5%, at least 97.6%, at least 97.7%, at least 97.8%, at least 97.9%, but less than 100% to the amino acid sequence according to SEQ ID NO: 1 and comprises two or more amino acid substitutions selected from the group consisting of T38R / K / W, Q59K / E and S212P / A / G / R / K / F / Y compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2. The variant polypeptide of the present invention preferably has a sequence identity of at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 90.5%, at least 91 %, at least 91 .5%, at least 92%, at least 92.5%, at least 93%, at least 93.5%, at least 94%, at least 94.5%, at least 95%, at least 95.1 %, at least 95.2%, at least 95.3%, at least 95.4%, at least 95.5%, at least 95.6%, at least 95.7%, at least 95.8%, at least 95.9% at least 96%, at least 96.1 %, at least 96.2%, at least 96.3%, at least 96.4%, at least 96.5%, at least 96.6%, at least 96.7%, at least 96.8%, at least 96.9%, at least 97%, at least 97.1 %, at least 97.2%, at least 97.3%, at least 97.4%, at least 97.5%, at least 97.6%, at least 97.7%, at least 97.8%, at least 97.9%, but less than 100% to the amino acid sequence according to SEQ ID NO: 1 and comprises an amino acid substitution at two or more amino acid residues selected from the group consisting of 38, 59 and 212 compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2, wherein the amino acid substitutions 38F, 38H, 38N, 59A, 59H, 59N, 59S, 212H and 212M are excluded, wherein the fragment of the variant polypeptide comprises 100 to 259 consecutive amino acids of the full-length variant polypeptide and wherein said variant polypeptide comprises amino acid residue D or E at position 101 , preferably E at position 101 , referring to the numbering of SEQ ID NO: 2.

[0154] In one embodiment the variant polypeptide of the present invention preferably has a sequence identity of at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 90.5%, at least 91 %, at least 91 .5%, at least 92%, at least 92.5%, at least 93%, at least 93.5%, at least 94%, at least 94.5%, at least 95%, at least 95.1 %, at least 95.2%, at least 95.3%, at least 95.4%, at least 95.5%, at least 95.6%, at least 95.7%, at least 95.8%, at least 95.9% at least 96%, at least 96.1 %, at least 96.2%, at least 96.3%, at least 96.4%, at least 96.5%, at least 96.6%, at least 96.7%, at least 96.8%, at least 96.9%, at least 97%, at least 97.1 %, at least 97.2%, at least 97.3%, at least 97.4%, at least 97.5%, at least 97.6%, at least 97.7%, at least 97.8%, at least 97.9%, but less than 100% to the amino acid sequence according to SEQ ID NO: 1 and comprises two or more amino acid substitutions selected from the group consisting of X38R / K / W, X59K / E and X212G / R / K / P / A / F / W / N / Y / D compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2, wherein said variant polypeptide comprises amino acid residue D or E at position 101 , preferably E at position 101 , referring to the numbering of SEQ ID NO: 2.

[0155] In one embodiment the variant polypeptide of the present invention preferably has a sequence identity of at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 90.5%, at least 91 %, at least 91 .5%, at least 92%, at least 92.5%, at least 93%, at least 93.5%, at least 94%, at least 94.5%, at least 95%, at least 95.1 %, at least 95.2%, at least 95.3%, at least 95.4%, at least 95.5%, at least 95.6%, at least 95.7%, at least 95.8%, at least 95.9% at least 96%, at least 96.1 %, at least 96.2%, at least 96.3%, at least 96.4%, at least 96.5%, at least 96.6%, at least 96.7%, at least 96.8%, at least 96.9%, at least 97%, at least 97.1 %, at least 97.2%, at least 97.3%, at least 97.4%, at least 97.5%, at least 97.6%, at least 97.7%, at least 97.8%, at least 97.9%, but less than 100% to the amino acid sequence according to SEQ ID NO: 1 and comprises two or more amino acid substitutions selected from the group consisting of X38R / K / H / W, X59K / E and X212P / A / G / R / K / F / Y / M compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2, wherein said variant polypeptide comprises amino acid residue D or E at position 101 , preferably E at position 101 , referring to the numbering of SEQ ID NO: 2.

[0156] In one embodiment the variant polypeptide of the present invention preferably has a sequence identity of at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 90.5%, at least 91 %, at least 91 .5%, at least 92%, at least 92.5%, at least 93%, at least 93.5%, at least 94%, at least 94.5%, at least 95%, at least 95.1 %, at least 95.2%, at least 95.3%, at least 95.4%, at least 95.5%, at least 95.6%, at least 95.7%, at least 95.8%, at least 95.9% at least 96%, at least 96.1 %, at least 96.2%, at least 96.3%, at least 96.4%, at least 96.5%, at least 96.6%, at least 96.7%, at least 96.8%, at least 96.9%, at least 97%, at least 97.1 %, at least 97.2%, at least 97.3%, at least 97.4%, at least 97.5%, at least 97.6%, at least 97.7%, at least 97.8%, at least 97.9%, but less than 100% to the amino acid sequence according to SEQ ID NO: 1 and comprises two or more amino acid substitutions selected from the group consisting of T38R / K / W, Q59K / E and S212G / R / K / P / A / F / W / N / Y / D compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2, wherein said variant polypeptide comprises amino acid residue D or E at position 101 , preferably E at position 101 , referring to the numbering of SEQ ID NO: 2.

[0157] In one embodiment the variant polypeptide of the present invention preferably has a sequence identity of at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 90.5%, at least 91 %, at least 91 .5%, at least 92%, at least 92.5%, at least 93%, at least 93.5%, at least 94%, at least 94.5%, at least 95%, at least 95.1 %, at least 95.2%, at least 95.3%, at least 95.4%, at least 95.5%, at least 95.6%, at least 95.7%, at least 95.8%, at least 95.9% at least 96%, at least 96.1 %, at least 96.2%, at least 96.3%, at least 96.4%, at least 96.5%, at least 96.6%, at least 96.7%, at least 96.8%, at least 96.9%, at least 97%, at least 97.1 %, at least 97.2%, at least 97.3%, at least 97.4%, at least 97.5%, at least 97.6%, at least 97.7%, at least 97.8%, at least 97.9%, but less than 100% to the amino acid sequence according to SEQ ID NO: 1 and comprises two or more amino acid substitutions selected from the group consisting of T38R / K / H / W, Q59K / E and S212P / A / G / R / K / F / Y / M compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2, wherein said variant polypeptide comprises amino acid residue D or E at position 101 , preferably E at position 101 , referring to the numbering of SEQ ID NO: 2.

[0158] The variant polypeptide of the present invention preferably has a sequence identity of 82% to 98%, 82.5% to 98%, 83% to 98%, 83.5% to 98%, 84% to 98%, 84.5% to 98%, 85% to 98%, 85.5% to 98%, 86% to 98%, 86.5% to 98%, 87% to 98%, 87.5% to 98%, 88% to 98%, 88.5% to 98%, 89% to 98%, 89.5% to 98%, 90% to 98%, 90.3% to 98%, 90.5% to 98%, 90.7% to 98%, 91 % to 98%, 91 .3% to 98%, 91 .5% to 98%, 91 .7% to 98%, 92% to 98%, 92.3% to 98%, 92.5% to 98%, 92.7% to 98%, 93% to 98%, 93.3% to 98%, 93.5% to 98%, 93.7% to 98%, 94% to 98%, 94.3% to 98%, 94.5% to 98%, 94.7% to 98%, 95% to 98%, 95.1 % to 98%, 95.2% to 98%, 95.3% to 98%, 95.4% to 98%, 95.5% to 98% to the amino acid sequence according to SEQ ID NO: 1 and comprises an amino acid substitution at two or more amino acid residues selected from the group consisting of 38, 59 and 212 compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2, wherein the amino acid substitutions 38F, 38H, 38N, 59A, 59H, 59N, 59S, 212H and 212M are excluded and wherein the fragment of the variant polypeptide comprises 100 to 259 consecutive amino acids of the full-length variant polypeptide.

[0159] The variant polypeptide of the present invention preferably has a sequence identity of 82% to 98%, 82.5% to 98%, 83% to 98%, 83.5% to 98%, 84% to 98%, 84.5% to 98%, 85% to 98%, 85.5% to 98%, 86% to 98%, 86.5% to 98%, 87% to 98%, 87.5% to 98%, 88% to 98%, 88.5% to 98%, 89% to 98%, 89.5% to 98%, 90% to 98%, 90.3% to 98%, 90.5% to 98%, 90.7% to 98%, 91 % to 98%, 91 .3% to 98%, 91 .5% to 98%, 91 .7% to 98%, 92% to 98%, 92.3% to 98%, 92.5% to 98%, 92.7% to 98%, 93% to 98%, 93.3% to 98%, 93.5% to 98%, 93.7% to 98%, 94% to 98%, 94.3% to 98%, 94.5% to 98%, 94.7% to 98%, 95% to 98%, 95.1 % to 98%, 95.2% to 98%, 95.3% to 98%, 95.4% to 98%, 95.5% to 98% to the amino acid sequence according to SEQ ID NO: 1 and comprises (A) an amino acid substitution at amino acid residue 212 and (B) an amino acid substitution at amino acid residue 38 and / or 59 compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2, wherein the amino acid substitutions 38F, 38H, 38N, 59A, 59H, 59N, 59S, 212H and 212M are excluded and wherein the fragment of the variant polypeptide comprises 100 to 259 consecutive amino acids of the full-length variant polypeptide. In one embodiment the variant polypeptide of the present invention preferably has a sequence identity of 82% to 98%, 82.5% to 98%, 83% to 98%, 83.5% to 98%, 84% to 98%, 84.5% to 98%, 85% to 98%, 85.5% to 98%, 86% to 98%, 86.5% to 98%, 87% to 98%, 87.5% to 98%, 88% to 98%, 88.5% to 98%, 89% to 98%, 89.5% to 98%, 90% to 98%, 90.3% to 98%, 90.5% to 98%, 90.7% to 98%, 91 % to 98%, 91 .3% to 98%, 91 .5% to 98%, 91 .7% to 98%, 92% to 98%, 92.3% to 98%, 92.5% to 98%, 92.7% to 98%, 93% to 98%, 93.3% to 98%, 93.5% to 98%, 93.7% to 98%, 94% to 98%, 94.3% to 98%, 94.5% to 98%, 94.7% to 98%, 95% to 98%, 95.1 % to 98%, 95.2% to 98%, 95.3% to 98%, 95.4% to 98%, 95.5% to 98% to the amino acid sequence according to SEQ ID NO: 1 and comprises two or more amino acid substitutions selected from the group consisting of X38R / K / W, X59K / E and X212G / R / K / P / A / F / W / N / Y / D compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2.

[0160] In one embodiment the variant polypeptide of the present invention preferably has a sequence identity of 82% to 98%, 82.5% to 98%, 83% to 98%, 83.5% to 98%, 84% to 98%, 84.5% to 98%, 85% to 98%, 85.5% to 98%, 86% to 98%, 86.5% to 98%, 87% to 98%, 87.5% to 98%, 88% to 98%, 88.5% to 98%, 89% to 98%, 89.5% to 98%, 90% to 98%, 90.3% to 98%, 90.5% to 98%, 90.7% to 98%, 91 % to 98%, 91 .3% to 98%, 91 .5% to 98%, 91 .7% to 98%, 92% to 98%, 92.3% to 98%, 92.5% to 98%, 92.7% to 98%, 93% to 98%, 93.3% to 98%, 93.5% to 98%, 93.7% to 98%, 94% to 98%, 94.3% to 98%, 94.5% to 98%, 94.7% to 98%, 95% to 98%, 95.1 % to 98%, 95.2% to 98%, 95.3% to 98%, 95.4% to 98%, 95.5% to 98% to the amino acid sequence according to SEQ ID NO: 1 and comprises (A) the amino acid substitution X212G / R / K / P / A / F / W / N / Y / D and (B) the amino acid substitution X38R / K / W and / orX59K / E compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2.

[0161] In one embodiment the variant polypeptide of the present invention preferably has a sequence identity of 82% to 98%, 82.5% to 98%, 83% to 98%, 83.5% to 98%, 84% to 98%, 84.5% to 98%, 85% to 98%, 85.5% to 98%, 86% to 98%, 86.5% to 98%, 87% to 98%, 87.5% to 98%, 88% to 98%, 88.5% to 98%, 89% to 98%, 89.5% to 98%, 90% to 98%, 90.3% to 98%, 90.5% to 98%, 90.7% to 98%, 91 % to 98%, 91 .3% to 98%, 91 .5% to 98%, 91 .7% to 98%, 92% to 98%, 92.3% to 98%, 92.5% to 98%, 92.7% to 98%, 93% to 98%, 93.3% to 98%, 93.5% to 98%, 93.7% to 98%, 94% to 98%, 94.3% to 98%, 94.5% to 98%, 94.7% to 98%, 95% to 98%, 95.1 % to 98%, 95.2% to 98%, 95.3% to 98%, 95.4% to 98%, 95.5% to 98% to the amino acid sequence according to SEQ ID NO: 1 and comprises two or more amino acid substitutions selected from the group consisting of X38R / K / W, X59K / E and X212P / A / G / R / K / F / Y compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2. In one embodiment the variant polypeptide of the present invention preferably has a sequence identity of 82% to 98%, 82.5% to 98%, 83% to 98%, 83.5% to 98%, 84% to 98%, 84.5% to 98%, 85% to 98%, 85.5% to 98%, 86% to 98%, 86.5% to 98%, 87% to 98%, 87.5% to 98%, 88% to 98%, 88.5% to 98%, 89% to 98%, 89.5% to 98%, 90% to 98%, 90.3% to 98%, 90.5% to 98%, 90.7% to 98%, 91 % to 98%, 91 .3% to 98%, 91 .5% to 98%, 91 .7% to 98%, 92% to 98%, 92.3% to 98%, 92.5% to 98%, 92.7% to 98%, 93% to 98%, 93.3% to 98%, 93.5% to 98%, 93.7% to 98%, 94% to 98%, 94.3% to 98%, 94.5% to 98%, 94.7% to 98%, 95% to 98%, 95.1 % to 98%, 95.2% to 98%, 95.3% to 98%, 95.4% to 98%, 95.5% to 98% to the amino acid sequence according to SEQ ID NO: 1 and comprises two or more amino acid substitutions selected from the group consisting of T38R / K / W, Q59K / E and S212G / R / K / P / A / F / W / N / Y / D compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2.

[0162] In one embodiment the variant polypeptide of the present invention preferably has a sequence identity of 82% to 98%, 82.5% to 98%, 83% to 98%, 83.5% to 98%, 84% to 98%, 84.5% to 98%, 85% to 98%, 85.5% to 98%, 86% to 98%, 86.5% to 98%, 87% to 98%, 87.5% to 98%, 88% to 98%, 88.5% to 98%, 89% to 98%, 89.5% to 98%, 90% to 98%, 90.3% to 98%, 90.5% to 98%, 90.7% to 98%, 91 % to 98%, 91 .3% to 98%, 91 .5% to 98%, 91 .7% to 98%, 92% to 98%, 92.3% to 98%, 92.5% to 98%, 92.7% to 98%, 93% to 98%, 93.3% to 98%, 93.5% to 98%, 93.7% to 98%, 94% to 98%, 94.3% to 98%, 94.5% to 98%, 94.7% to 98%, 95% to 98%, 95.1 % to 98%, 95.2% to 98%, 95.3% to 98%, 95.4% to 98%, 95.5% to 98% to the amino acid sequence according to SEQ ID NO: 1 and comprises two or more amino acid substitutions selected from the group consisting of T38R / K / W, Q59K / E and S212P / A / G / R / K / F / Y compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2.

[0163] The variant polypeptide of the present invention preferably has a sequence identity of 82% to 98%, 82.5% to 98%, 83% to 98%, 83.5% to 98%, 84% to 98%, 84.5% to 98%, 85% to 98%, 85.5% to 98%, 86% to 98%, 86.5% to 98%, 87% to 98%, 87.5% to 98%, 88% to 98%, 88.5% to 98%, 89% to 98%, 89.5% to 98%, 90% to 98%, 90.3% to 98%, 90.5% to 98%, 90.7% to 98%, 91 % to 98%, 91 .3% to 98%, 91 .5% to 98%, 91 .7% to 98%, 92% to 98%, 92.3% to 98%, 92.5% to 98%, 92.7% to 98%, 93% to 98%, 93.3% to 98%, 93.5% to 98%, 93.7% to 98%, 94% to 98%, 94.3% to 98%, 94.5% to 98%, 94.7% to 98%, 95% to 98%, 95.1 % to 98%, 95.2% to 98%, 95.3% to 98%, 95.4% to 98%, 95.5% to 98% to the amino acid sequence according to SEQ ID NO: 1 and comprises an amino acid substitution at two or more amino acid residues selected from the group consisting of 38, 59 and 212 compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2, wherein the amino acid substitutions 38F, 38H, 38N, 59A, 59H, 59N, 59S, 212H and 212M are excluded, wherein the fragment of the variant polypeptide comprises 100 to 259 consecutive amino acids of the full-length variant polypeptide and wherein said variant polypeptide comprises amino acid residue D or E at position 101 , preferably E at position 101 , referring to the numbering of SEQ ID NO: 2.

[0164] In one embodiment the variant polypeptide of the present invention preferably has a sequence identity of 82% to 98%, 82.5% to 98%, 83% to 98%, 83.5% to 98%, 84% to 98%, 84.5% to 98%, 85% to 98%, 85.5% to 98%, 86% to 98%, 86.5% to 98%, 87% to 98%, 87.5% to 98%, 88% to 98%, 88.5% to 98%, 89% to 98%, 89.5% to 98%, 90% to 98%, 90.3% to 98%, 90.5% to 98%, 90.7% to 98%, 91 % to 98%, 91 .3% to 98%, 91 .5% to 98%, 91 .7% to 98%, 92% to 98%, 92.3% to 98%, 92.5% to 98%, 92.7% to 98%, 93% to 98%, 93.3% to 98%, 93.5% to 98%, 93.7% to 98%, 94% to 98%, 94.3% to 98%, 94.5% to 98%, 94.7% to 98%, 95% to 98%, 95.1 % to 98%, 95.2% to 98%, 95.3% to 98%, 95.4% to 98%, 95.5% to 98% to the amino acid sequence according to SEQ ID NO: 1 and comprises two or more amino acid substitutions selected from the group consisting of X38R / K / W, X59K / E and X212G / R / K / P / A / F / W / N / Y / D compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2, wherein said variant polypeptide comprises amino acid residue D or E at position 101 , preferably E at position 101 , referring to the numbering of SEQ ID NO: 2.

[0165] In one embodiment the variant polypeptide of the present invention preferably has a sequence identity of 82% to 98%, 82.5% to 98%, 83% to 98%, 83.5% to 98%, 84% to 98%, 84.5% to 98%, 85% to 98%, 85.5% to 98%, 86% to 98%, 86.5% to 98%, 87% to 98%, 87.5% to 98%, 88% to 98%, 88.5% to 98%, 89% to 98%, 89.5% to 98%, 90% to 98%, 90.3% to 98%, 90.5% to 98%, 90.7% to 98%, 91 % to 98%, 91 .3% to 98%, 91 .5% to 98%, 91 .7% to 98%, 92% to 98%, 92.3% to 98%, 92.5% to 98%, 92.7% to 98%, 93% to 98%, 93.3% to 98%, 93.5% to 98%, 93.7% to 98%, 94% to 98%, 94.3% to 98%, 94.5% to 98%, 94.7% to 98%, 95% to 98%, 95.1 % to 98%, 95.2% to 98%, 95.3% to 98%, 95.4% to 98%, 95.5% to 98% to the amino acid sequence according to SEQ ID NO: 1 and comprises two or more amino acid substitutions selected from the group consisting of X38R / K / W, X59K / E and X212P / A / G / R / K / F / Y compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2, wherein said variant polypeptide comprises amino acid residue D or E at position 101 , preferably E at position 101 , referring to the numbering of SEQ ID NO: 2.

[0166] In one embodiment the variant polypeptide of the present invention preferably has a sequence identity of 82% to 98%, 82.5% to 98%, 83% to 98%, 83.5% to 98%, 84% to 98%, 84.5% to 98%, 85% to 98%, 85.5% to 98%, 86% to 98%, 86.5% to 98%, 87% to 98%, 87.5% to 98%, 88% to 98%, 88.5% to 98%, 89% to 98%, 89.5% to 98%, 90% to 98%, 90.3% to 98%, 90.5% to 98%, 90.7% to 98%, 91 % to 98%, 91 .3% to 98%, 91 .5% to 98%, 91 .7% to 98%, 92% to 98%, 92.3% to 98%, 92.5% to 98%, 92.7% to 98%, 93% to 98%, 93.3% to 98%, 93.5% to 98%, 93.7% to 98%, 94% to 98%, 94.3% to 98%, 94.5% to 98%, 94.7% to 98%, 95% to 98%, 95.1 % to 98%, 95.2% to 98%, 95.3% to 98%, 95.4% to 98%, 95.5% to 98% to the amino acid sequence according to SEQ ID NO: 1 and comprises two or more amino acid substitutions selected from the group consisting of T38R / K / W, Q59K / E and S212G / R / K / P / A / F / W / N / Y / D compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2, wherein said variant polypeptide comprises amino acid residue D or E at position 101 , preferably E at position 101 , referring to the numbering of SEQ ID NO: 2.

[0167] In one embodiment the variant polypeptide of the present invention preferably has a sequence identity of 82% to 98%, 82.5% to 98%, 83% to 98%, 83.5% to 98%, 84% to 98%, 84.5% to 98%, 85% to 98%, 85.5% to 98%, 86% to 98%, 86.5% to 98%, 87% to 98%, 87.5% to 98%, 88% to 98%, 88.5% to 98%, 89% to 98%, 89.5% to 98%, 90% to 98%, 90.3% to 98%, 90.5% to 98%, 90.7% to 98%, 91 % to 98%, 91 .3% to 98%, 91 .5% to 98%, 91 .7% to 98%, 92% to 98%, 92.3% to 98%, 92.5% to 98%, 92.7% to 98%, 93% to 98%, 93.3% to 98%, 93.5% to 98%, 93.7% to 98%, 94% to 98%, 94.3% to 98%, 94.5% to 98%, 94.7% to 98%, 95% to 98%, 95.1 % to 98%, 95.2% to 98%, 95.3% to 98%, 95.4% to 98%, 95.5% to 98% to the amino acid sequence according to SEQ ID NO: 1 and comprises two or more amino acid substitutions selected from the group consisting of T38R / K / W, Q59K / E and S212P / A / G / R / K / F / Y compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2, wherein said variant polypeptide comprises amino acid residue D or E at position 101 , preferably E at position 101 , referring to the numbering of SEQ ID NO: 2.

[0168] In one embodiment the variant polypeptide of the present invention preferably has a sequence identity of at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 90.5%, at least 91 %, at least 91 .5%, at least 92%, at least 92.5%, at least 93%, at least 93.5%, at least 94%, at least 94.5%, at least 95%, at least 95.1 %, at least 95.2%, at least 95.3%, at least 95.4%, at least 95.5%, at least 95.6%, at least 95.7%, at least 95.8%, at least 95.9% at least 96%, at least 96.1 %, at least 96.2%, at least 96.3%, at least 96.4%, at least 96.5%, at least 96.6%, at least 96.7%, at least 96.8%, at least 96.9%, at least 97%, at least 97.1 %, at least 97.2%, at least 97.3%, at least 97.4%, at least 97.5%, at least 97.6%, at least 97.7%, at least 97.8%, at least 97.9%, but less than 100% to the amino acid sequence according to SEQ ID NO: 1 and one of the following combinations of amino acid substitutions:

[0169] (a) X38R / K / H / W and X59K / E;

[0170] (b) X38R / K / W and X212G / R / K / P / A / F / W / N / Y / D;

[0171] (c) X59K / E and X212G / R / K / P / A / F / W / N / Y / D; or

[0172] (d) X38R / K / H / W, X59K / E and X212G / M / R / K / P / A / F / H / W / N / Y / D compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2. In one embodiment the variant polypeptide of the present invention preferably has a sequence identity of at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 90.5%, at least 91 %, at least 91 .5%, at least 92%, at least 92.5%, at least 93%, at least 93.5%, at least 94%, at least 94.5%, at least 95%, at least 95.1 %, at least 95.2%, at least 95.3%, at least 95.4%, at least 95.5%, at least 95.6%, at least 95.7%, at least 95.8%, at least 95.9% at least 96%, at least 96.1 %, at least 96.2%, at least 96.3%, at least 96.4%, at least 96.5%, at least 96.6%, at least 96.7%, at least 96.8%, at least 96.9%, at least 97%, at least 97.1 %, at least 97.2%, at least 97.3%, at least 97.4%, at least 97.5%, at least 97.6%, at least 97.7%, at least 97.8%, at least 97.9%, but less than 100% to the amino acid sequence according to SEQ ID NO: 1 and one of the following combinations of amino acid substitutions:

[0173] (a) X38R / K / W and X212G / R / K / P / A / F / W / N / Y / D;

[0174] (b) X59K / E and X212G / R / K / P / A / F / W / N / Y / D; or

[0175] (c) X38R / K / H / W, X59K / E and X212G / M / R / K / P / A / F / H / W / N / Y / D compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2. In one embodiment the variant polypeptide of the present invention preferably has a sequence identity of at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 90.5%, at least 91 %, at least 91 .5%, at least 92%, at least 92.5%, at least 93%, at least 93.5%, at least 94%, at least 94.5%, at least 95%, at least 95.1 %, at least 95.2%, at least 95.3%, at least 95.4%, at least 95.5%, at least 95.6%, at least 95.7%, at least 95.8%, at least 95.9% at least 96%, at least 96.1 %, at least 96.2%, at least 96.3%, at least 96.4%, at least 96.5%, at least 96.6%, at least 96.7%, at least 96.8%, at least 96.9%, at least 97%, at least 97.1 %, at least 97.2%, at least 97.3%, at least 97.4%, at least 97.5%, at least 97.6%, at least 97.7%, at least 97.8%, at least 97.9%, but less than 100% to the amino acid sequence according to SEQ ID NO: 1 and one of the following combinations of amino acid substitutions:

[0176] (a) X38W and X212F;

[0177] (b) X38K and X212F;

[0178] (c) X38W and X212P;

[0179] (d) X38W and X212R;

[0180] (e) X38K and X212P;

[0181] (f) X59K and X212P;

[0182] (g) X38K, X59K and X212P; or

[0183] (h) X38W, X59K and X212P compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2. In one embodiment the variant polypeptide of the present invention preferably has a sequence identity of at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 90.5%, at least 91 %, at least 91 .5%, at least 92%, at least 92.5%, at least 93%, at least 93.5%, at least 94%, at least 94.5%, at least 95%, at least 95.1 %, at least 95.2%, at least 95.3%, at least 95.4%, at least 95.5%, at least 95.6%, at least 95.7%, at least 95.8%, at least 95.9% at least 96%, at least 96.1 %, at least 96.2%, at least 96.3%, at least 96.4%, at least 96.5%, at least 96.6%, at least 96.7%, at least 96.8%, at least 96.9%, at least 97%, at least 97.1 %, at least 97.2%, at least 97.3%, at least 97.4%, at least 97.5%, at least 97.6%, at least 97.7%, at least 97.8%, at least 97.9%, but less than 100% to the amino acid sequence according to SEQ ID NO: 1 and one of the following combinations of amino acid substitutions:

[0184] (a) T38R / K / H / W and Q59K / E;

[0185] (b) T38R / K / W and S212G / R / K / P / A / F / W / N / Y / D;

[0186] (c) Q59K / E and S212G / M / R / K / P / A / F / H / W / N / Y / D; or

[0187] (d) T38R / K / H / W, Q59K / E and S212G / M / R / K / P / A / F / H / W / N / Y / D compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2. In one embodiment the variant polypeptide of the present invention preferably has a sequence identity of at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 90.5%, at least 91 %, at least 91 .5%, at least 92%, at least 92.5%, at least 93%, at least 93.5%, at least 94%, at least 94.5%, at least 95%, at least 95.1 %, at least 95.2%, at least 95.3%, at least 95.4%, at least 95.5%, at least 95.6%, at least 95.7%, at least 95.8%, at least 95.9% at least 96%, at least 96.1 %, at least 96.2%, at least 96.3%, at least 96.4%, at least 96.5%, at least 96.6%, at least 96.7%, at least 96.8%, at least 96.9%, at least 97%, at least 97.1 %, at least 97.2%, at least 97.3%, at least 97.4%, at least 97.5%, at least 97.6%, at least 97.7%, at least 97.8%, at least 97.9%, but less than 100% to the amino acid sequence according to SEQ ID NO: 1 and one of the following combinations of amino acid substitutions:

[0188] (a) X38R / K / H / W and X59K / E;

[0189] (b) X38R / K / W and X212P / A / G / R / K / F / Y;

[0190] (c) X59K / E and X212P / A / G / R / K / F / Y / M; or

[0191] (d) X38R / K / H / W,38R / K / H / W, X59K / E and X212P / A / G / R / K / F / Y / M compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2. In one embodiment the variant polypeptide of the present invention preferably has a sequence identity of at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 90.5%, at least 91 %, at least 91 .5%, at least 92%, at least 92.5%, at least 93%, at least 93.5%, at least 94%, at least 94.5%, at least 95%, at least 95.1 %, at least 95.2%, at least 95.3%, at least 95.4%, at least 95.5%, at least 95.6%, at least 95.7%, at least 95.8%, at least 95.9% at least 96%, at least 96.1 %, at least 96.2%, at least 96.3%, at least 96.4%, at least 96.5%, at least 96.6%, at least 96.7%, at least 96.8%, at least 96.9%, at least 97%, at least 97.1 %, at least 97.2%, at least 97.3%, at least 97.4%, at least 97.5%, at least 97.6%, at least 97.7%, at least 97.8%, at least 97.9%, but less than 100% to the amino acid sequence according to SEQ ID NO: 1 and one of the following combinations of amino acid substitutions:

[0192] (a) T38R / K / H / W and Q59K / E;

[0193] (b) T38R / K / W and S212P / A / G / R / K / F / Y;

[0194] (c) Q59K / E and S212P / A / G / R / K / F / Y / M; or

[0195] (d) T38R / K / H / W, Q59K / E and S212P / A / G / R / K / F / Y / M compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2. In one embodiment the variant polypeptide of the present invention preferably has a sequence identity of at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 90.5%, at least 91 %, at least 91 .5%, at least 92%, at least 92.5%, at least 93%, at least 93.5%, at least 94%, at least 94.5%, at least 95%, at least 95.1 %, at least 95.2%, at least 95.3%, at least 95.4%, at least 95.5%, at least 95.6%, at least 95.7%, at least 95.8%, at least 95.9% at least 96%, at least 96.1 %, at least 96.2%, at least 96.3%, at least 96.4%, at least 96.5%, at least 96.6%, at least 96.7%, at least 96.8%, at least 96.9%, at least 97%, at least 97.1 %, at least 97.2%, at least 97.3%, at least 97.4%, at least 97.5%, at least 97.6%, at least 97.7%, at least 97.8%, at least 97.9%, but less than 100% to the amino acid sequence according to SEQ ID NO: 1 and one of the following combinations of amino acid substitutions:

[0196] (a) X38R / K / H / W and X59K / E;

[0197] (b) X38R / K / W and X212G / R / K / P / A / F / W / N / Y / D;

[0198] (c) X59K / E and X212G / M / R / K / P / A / F / H / W / N / Y / D; or

[0199] (d) X38R / K / H / W, X59K / E and X212G / M / R / K / P / A / F / H / W / N / Y / D compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2, wherein said variant polypeptide comprises amino acid residue D or E at position 101 , preferably E at position 101 , referring to the numbering of SEQ ID NO: 2.

[0200] In one embodiment the variant polypeptide of the present invention preferably has a sequence identity of at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 90.5%, at least 91 %, at least 91 .5%, at least 92%, at least 92.5%, at least 93%, at least 93.5%, at least 94%, at least 94.5%, at least 95%, at least 95.1 %, at least 95.2%, at least 95.3%, at least 95.4%, at least 95.5%, at least 95.6%, at least 95.7%, at least 95.8%, at least 95.9% at least 96%, at least 96.1 %, at least 96.2%, at least 96.3%, at least 96.4%, at least 96.5%, at least 96.6%, at least 96.7%, at least 96.8%, at least 96.9%, at least 97%, at least 97.1 %, at least 97.2%, at least 97.3%, at least 97.4%, at least 97.5%, at least 97.6%, at least 97.7%, at least 97.8%, at least 97.9%, but less than 100% to the amino acid sequence according to SEQ ID NO: 1 and one of the following combinations of amino acid substitutions:

[0201] (a) T38R / K / H / W and Q59K / E;

[0202] (b) T38R / K / W and S212G / R / K / P / A / F / W / N / Y / D;

[0203] (c) Q59K / E and S212G / M / R / K / P / A / F / H / W / N / Y / D; or

[0204] (d) T38R / K / H / W, Q59K / E and S212G / M / R / K / P / A / F / H / W / N / Y / D compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2, wherein said variant polypeptide comprises amino acid residue D or E at position 101 , preferably E at position 101 , referring to the numbering of SEQ ID NO: 2.

[0205] In one embodiment the variant polypeptide of the present invention preferably has a sequence identity of at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 90.5%, at least 91 %, at least 91 .5%, at least 92%, at least 92.5%, at least 93%, at least 93.5%, at least 94%, at least 94.5%, at least 95%, at least 95.1 %, at least 95.2%, at least 95.3%, at least 95.4%, at least 95.5%, at least 95.6%, at least 95.7%, at least 95.8%, at least 95.9% at least 96%, at least 96.1 %, at least 96.2%, at least 96.3%, at least 96.4%, at least 96.5%, at least 96.6%, at least 96.7%, at least 96.8%, at least 96.9%, at least 97%, at least 97.1 %, at least 97.2%, at least 97.3%, at least 97.4%, at least 97.5%, at least 97.6%, at least 97.7%, at least 97.8%, at least 97.9%, but less than 100% to the amino acid sequence according to SEQ ID NO: 1 and one of the following combinations of amino acid substitutions: (a) X38R / K / H / W and X59K / E;

[0206] (b) X38R / K / W and X212P / A / G / R / K / F / Y;

[0207] (c) X59K / E and X212P / A / G / R / K / F / Y / M; or

[0208] (d) X38R / K / H / W, X59K / E and X212P / A / G / R / K / F / Y / M compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2, wherein said variant polypeptide comprises amino acid residue D or E at position 101 , preferably E at position 101 , referring to the numbering of SEQ ID NO: 2.

[0209] In one embodiment the variant polypeptide of the present invention preferably has a sequence identity of at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 90.5%, at least 91 %, at least 91 .5%, at least 92%, at least 92.5%, at least 93%, at least 93.5%, at least 94%, at least 94.5%, at least 95%, at least 95.1 %, at least 95.2%, at least 95.3%, at least 95.4%, at least 95.5%, at least 95.6%, at least 95.7%, at least 95.8%, at least 95.9% at least 96%, at least 96.1 %, at least 96.2%, at least 96.3%, at least 96.4%, at least 96.5%, at least 96.6%, at least 96.7%, at least 96.8%, at least 96.9%, at least 97%, at least 97.1 %, at least 97.2%, at least 97.3%, at least 97.4%, at least 97.5%, at least 97.6%, at least 97.7%, at least 97.8%, at least 97.9%, but less than 100% to the amino acid sequence according to SEQ ID NO: 1 and one of the following combinations of amino acid substitutions:

[0210] (a) T38R / K / H / W and Q59K / E;

[0211] (b) T38R / K / W and S212P / A / G / R / K / F / Y;

[0212] (c) Q59K / E and S212P / A / G / R / K / F / Y / M; or

[0213] (d) T38R / K / H / W, Q59K / E and S212P / A / G / R / K / F / Y / M compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2, wherein said variant polypeptide comprises amino acid residue D or E at position 101 , preferably E at position 101 , referring to the numbering of SEQ ID NO: 2.

[0214] In one embodiment the variant polypeptide of the present invention preferably has a sequence identity of at least 82%, but less than 100% to the amino acid sequence according to SEQ ID NO: 1 and one of the following combinations of amino acid substitutions:

[0215] (a) X38R / K / H / W and X59K / E;

[0216] (b) X38R / K / W and X212G / R / K / P / A / F / W / N / Y / D;

[0217] (c) X59K / E and X212G / R / K / P / A / F / W / N / Y / D; or

[0218] (d) X38R / K / H / W, X59K / E and X212G / R / K / P / A / F / W / N / Y / D compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2. In one embodiment the variant polypeptide of the present invention preferably has a sequence identity of at least 82%, but less than 100% to the amino acid sequence according to SEQ ID NO: 1 and one of the following combinations of amino acid substitutions: (a) T38R / K / H / W and Q59K / E;

[0219] (b) T38R / K / W and S212G / R / K / P / A / F / W / N / Y / D;

[0220] (c) Q59K / E and S212G / M / R / K / P / A / F / H / W / N / Y / D; or

[0221] (d) T38R / K / H / W, Q59K / E and S212G / M / R / K / P / A / F / H / W / N / Y / D compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2. In one embodiment the variant polypeptide of the present invention preferably has a sequence identity of at least 82%, but less than 100% to the amino acid sequence according to SEQ ID NO: 1 and one of the following combinations of amino acid substitutions:

[0222] (a) X38R / K / H / W and X59K / E;

[0223] (b) X38R / K / W and X212P / A / G / R / K / F / Y;

[0224] (c) X59K / E and X212P / A / G / R / K / F / Y / M; or

[0225] (d) X38R / K / H / W, X59K / E and X212P / A / G / R / K / F / Y / M compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2. In one embodiment the variant polypeptide of the present invention preferably has a sequence identity of at least 82%, but less than 100% to the amino acid sequence according to SEQ ID NO: 1 and one of the following combinations of amino acid substitutions:

[0226] (a) T38R / K / H / W and Q59K / E;

[0227] (b) T38R / K / W and S212P / A / G / R / K / F / Y;

[0228] (c) Q59K / E and S212P / A / G / R / K / F / Y / M; or

[0229] (d) T38R / K / H / W, Q59K / E and S212P / A / G / R / K / F / Y / M compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2. In one embodiment the variant polypeptide of the present invention preferably has a sequence identity of at least 82%, but less than 100% to the amino acid sequence according to SEQ ID NO: 1 and one of the following combinations of amino acid substitutions:

[0230] (a) X38R / K / H / W and X59K / E;

[0231] (b) X38R / K / W and X212G / R / K / P / A / F / W / N / Y / D;

[0232] (c) X59K / E and X212G / R / K / P / A / F / W / N / Y / D; or

[0233] (d) X38R / K / H / W, X59K / E and X212G / R / K / P / A / F / W / N / Y / D compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2, wherein said variant polypeptide comprises amino acid residue D or E at position 101 , preferably E at position 101 , referring to the numbering of SEQ ID NO: 2.

[0234] In one embodiment the variant polypeptide of the present invention preferably has a sequence identity of at least 82%, but less than 100% to the amino acid sequence according to SEQ ID NO: 1 and one of the following combinations of amino acid substitutions: (a) T38R / K / H / W and Q59K / E;

[0235] (b) T38R / K / W and S212G / R / K / P / A / F / W / N / Y / D;

[0236] (c) Q59K / E and S212G / M / R / K / P / A / F / H / W / N / Y / D; or

[0237] (d) T38R / K / H / W, Q59K / E and S212G / M / R / K / P / A / F / H / W / N / Y / D compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2, wherein said variant polypeptide comprises amino acid residue D or E at position 101 , preferably E at position 101 , referring to the numbering of SEQ ID NO: 2.

[0238] In one embodiment the variant polypeptide of the present invention preferably has a sequence identity of at least 82%, but less than 100% to the amino acid sequence according to SEQ ID NO: 1 and one of the following combinations of amino acid substitutions:

[0239] (a) X38R / K / H / W and X59K / E;

[0240] (b) X38R / K / W and X212P / A / G / R / K / F / Y;

[0241] (c) X59K / E and X212P / A / G / R / K / F / Y / M; or

[0242] (d) X38R / K / H / W, X59K / E and X212P / A / G / R / K / F / Y / M compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2, wherein said variant polypeptide comprises amino acid residue D or E at position 101 , preferably E at position 101 , referring to the numbering of SEQ ID NO: 2.

[0243] In one embodiment the variant polypeptide of the present invention preferably has a sequence identity of at least 82%, but less than 100% to the amino acid sequence according to SEQ ID NO: 1 and one of the following combinations of amino acid substitutions:

[0244] (a) T38R / K / H / W and Q59K / E;

[0245] (b) T38R / K / W and S212P / A / G / R / K / F / Y;

[0246] (c) Q59K / E and S212P / A / G / R / K / F / Y / M; or

[0247] (d) T38R / K / H / W, Q59K / E and S212P / A / G / R / K / F / Y / M compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2, wherein said variant polypeptide comprises amino acid residue D or E at position 101 , preferably E at position 101 , referring to the numbering of SEQ ID NO: 2.

[0248] In one embodiment the variant polypeptide of the present invention preferably has a sequence identity of at least 95.5%, but less than 100% to the amino acid sequence according to SEQ ID NO: 1 and one of the following combinations of amino acid substitutions:

[0249] (a) X38R / K / H / W and X59K / E;

[0250] (b) X38R / K / W and X212G / R / K / P / A / F / W / N / Y / D;

[0251] (c) X59K / E and X212G / R / K / P / A / F / W / N / Y / D; or

[0252] (d) X38R / K / H / W, X59K / E and X212G / R / K / P / A / F / W / N / Y / D compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2. In one embodiment the variant polypeptide of the present invention preferably has a sequence identity of at least 95.5%, but less than 100% to the amino acid sequence according to SEQ ID NO: 1 and one of the following combinations of amino acid substitutions:

[0253] (a) X38W and X212F;

[0254] (b) X38K and X212F;

[0255] (c) X38W and X212P;

[0256] (d) X38W and X212R;

[0257] (e) X38K and X212P;

[0258] (f) X59K and X212P;

[0259] (g) X38K, X59K, X212P; or

[0260] (h) X38W, X59K, X212P compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2. In one embodiment the variant polypeptide of the present invention preferably has a sequence identity of at least 95.5%, but less than 100% to the amino acid sequence according to SEQ ID NO: 1 and one of the following combinations of amino acid substitutions:

[0261] (a) T38R / K / H / W and Q59K / E;

[0262] (b) T38R / K / W and S212G / R / K / P / A / F / W / N / Y / D;

[0263] (c) Q59K / E and S212G / M / R / K / P / A / F / H / W / N / Y / D; or

[0264] (d) T38R / K / H / W, Q59K / E and S212G / M / R / K / P / A / F / H / W / N / Y / D compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2. In one embodiment the variant polypeptide of the present invention preferably has a sequence identity of at least 95.5%, but less than 100% to the amino acid sequence according to SEQ ID NO: 1 and one of the following combinations of amino acid substitutions:

[0265] (a) X38R / K / H / W and X59K / E;

[0266] (b) X38R / K / W and X212P / A / G / R / K / F / Y;

[0267] (c) X59K / E and X212P / A / G / R / K / F / Y / M; or

[0268] (d) X38R / K / H / W, X59K / E and X212P / A / G / R / K / F / Y / M compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2. In one embodiment the variant polypeptide of the present invention preferably has a sequence identity of at least 95.5%, but less than 100% to the amino acid sequence according to SEQ ID NO: 1 and one of the following combinations of amino acid substitutions:

[0269] (a) T38R / K / H / W and Q59K / E;

[0270] (b) T38R / K / W and S212P / A / G / R / K / F / Y; (c) Q59K / E and S212P / A / G / R / K / F / Y / M; or

[0271] (d) T38R / K / H / W, Q59K / E and S212P / A / G / R / K / F / Y / M compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2. In one embodiment the variant polypeptide of the present invention preferably has a sequence identity of at least 95.5%, but less than 100% to the amino acid sequence according to SEQ ID NO: 1 and one of the following combinations of amino acid substitutions:

[0272] (a) X38R / K / H / W and X59K / E;

[0273] (b) X38R / K / W and X212G / R / K / P / A / F / W / N / Y / D;

[0274] (c) X59K / E and X212G / R / K / P / A / F / W / N / Y / D; or

[0275] (d) X38R / K / H / W, X59K / E and X212G / R / K / P / A / F / W / N / Y / D compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2, wherein said variant polypeptide comprises amino acid residue D or E at position 101 , preferably E at position 101 , referring to the numbering of SEQ ID NO: 2.

[0276] In one embodiment the variant polypeptide of the present invention preferably has a sequence identity of at least 95.5%, but less than 100% to the amino acid sequence according to SEQ ID NO: 1 and one of the following combinations of amino acid substitutions:

[0277] (a) T38R / K / H / W and Q59K / E;

[0278] (b) T38R / K / W and S212G / R / K / P / A / F / W / N / Y / D;

[0279] (c) Q59K / E and S212G / M / R / K / P / A / F / H / W / N / Y / D; or

[0280] (d) T38R / K / H / W, Q59K / E and S212G / M / R / K / P / A / F / H / W / N / Y / D compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2, wherein said variant polypeptide comprises amino acid residue D or E at position 101 , preferably E at position 101 , referring to the numbering of SEQ ID NO: 2.

[0281] In one embodiment the variant polypeptide of the present invention preferably has a sequence identity of at least 95.5%, but less than 100% to the amino acid sequence according to SEQ ID NO: 1 and one of the following combinations of amino acid substitutions:

[0282] (a) X38R / K / H / W and X59K / E;

[0283] (b) X38R / K / W and X212P / A / G / R / K / F / Y;

[0284] (c) X59K / E and X212P / A / G / R / K / F / Y / M; or

[0285] (d) X38R / K / H / W, X59K / E and X212P / A / G / R / K / F / Y / M compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2, wherein said variant polypeptide comprises amino acid residue D or E at position 101 , preferably E at position 101 , referring to the numbering of SEQ ID NO: 2. In one embodiment the variant polypeptide of the present invention preferably has a sequence identity of at least 95.5%, but less than 100% to the amino acid sequence according to SEQ ID NO: 1 and one of the following combinations of amino acid substitutions:

[0286] (a) T38R / K / H / W and Q59K / E;

[0287] (b) T38R / K / W and S212P / A / G / R / K / F / Y;

[0288] (c) Q59K / E and S212P / A / G / R / K / F / Y / M; or

[0289] (d) T38R / K / H / W, Q59K / E and S212P / A / G / R / K / F / Y / M compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2, wherein said variant polypeptide comprises amino acid residue D or E at position 101 , preferably E at position 101 , referring to the numbering of SEQ ID NO: 2.

[0290] In one embodiment the variant polypeptide of the present invention preferably has a sequence identity of 82% to 98%, 82.5% to 98%, 83% to 98%, 83.5% to 98%, 84% to 98%, 84.5% to 98%, 85% to 98%, 85.5% to 98%, 86% to 98%, 86.5% to 98%, 87% to 98%, 87.5% to 98%, 88% to 98%, 88.5% to 98%, 89% to 98%, 89.5% to 98%, 90% to 98%, 90.3% to 98%, 90.5% to 98%, 90.7% to 98%, 91 % to 98%, 91 .3% to 98%, 91 .5% to 98%, 91 .7% to 98%, 92% to 98%, 92.3% to 98%, 92.5% to 98%, 92.7% to 98%, 93% to 98%, 93.3% to 98%, 93.5% to 98%, 93.7% to 98%, 94% to 98%, 94.3% to 98%, 94.5% to 98%, 94.7% to 98%, 95% to 98%, 95.1 % to 98%, 95.2% to 98%, 95.3% to 98%, 95.4% to 98%, 95.5% to 98% to the amino acid sequence according to SEQ ID NO: 1 and one of the following combinations of amino acid substitutions:

[0291] (a) X38R / K / H / W and X59K / E;

[0292] (b) X38R / K / W and X212G / R / K / P / A / F / W / N / Y / D;

[0293] (c) X59K / E and X212G / R / K / P / A / F / W / N / Y / D; or

[0294] (d) X38R / K / H / W, X59K / E and X212G / R / K / P / A / F / W / N / Y / D compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2. In one embodiment the variant polypeptide of the present invention preferably has a sequence identity of 82% to 98%, 82.5% to 98%, 83% to 98%, 83.5% to 98%, 84% to 98%, 84.5% to 98%, 85% to 98%, 85.5% to 98%, 86% to 98%, 86.5% to 98%, 87% to 98%, 87.5% to 98%, 88% to 98%, 88.5% to 98%, 89% to 98%, 89.5% to 98%, 90% to 98%, 90.3% to 98%, 90.5% to 98%, 90.7% to 98%, 91 % to 98%, 91 .3% to 98%, 91 .5% to 98%, 91 .7% to 98%, 92% to 98%, 92.3% to 98%, 92.5% to 98%, 92.7% to 98%, 93% to 98%, 93.3% to 98%, 93.5% to 98%, 93.7% to 98%, 94% to 98%, 94.3% to 98%, 94.5% to 98%, 94.7% to 98%, 95% to 98%, 95.1 % to 98%, 95.2% to 98%, 95.3% to 98%, 95.4% to 98%, 95.5% to 98% to the amino acid sequence according to SEQ ID NO: 1 and one of the following combinations of amino acid substitutions:

[0295] (a) X38W and X212F;

[0296] (b) X38K and X212F;

[0297] (c) X38W and X212P; (d) X38W and X212R;

[0298] (e) X38K and X212P;

[0299] (f) X59K and X212P;

[0300] (g) X38K, X59K, X212P; or

[0301] (h) X38W, X59K, X212P compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2. In one embodiment the variant polypeptide of the present invention preferably has a sequence identity of 82% to 98%, 82.5% to 98%, 83% to 98%, 83.5% to 98%, 84% to 98%, 84.5% to 98%, 85% to 98%, 85.5% to 98%, 86% to 98%, 86.5% to 98%, 87% to 98%, 87.5% to 98%, 88% to 98%, 88.5% to 98%, 89% to 98%, 89.5% to 98%, 90% to 98%, 90.3% to 98%, 90.5% to 98%, 90.7% to 98%, 91 % to 98%, 91 .3% to 98%, 91 .5% to 98%, 91 .7% to 98%, 92% to 98%, 92.3% to 98%, 92.5% to 98%, 92.7% to 98%, 93% to 98%, 93.3% to 98%, 93.5% to 98%, 93.7% to 98%, 94% to 98%, 94.3% to 98%, 94.5% to 98%, 94.7% to 98%, 95% to 98%, 95.1 % to 98%, 95.2% to 98%, 95.3% to 98%, 95.4% to 98%, 95.5% to 98% to the amino acid sequence according to SEQ ID NO: 1 and one of the following combinations of amino acid substitutions:

[0302] (a) T38R / K / H / W and Q59K / E;

[0303] (b) T38R / K / W and S212G / R / K / P / A / F / W / N / Y / D;

[0304] (c) Q59K / E and S212G / M / R / K / P / A / F / H / W / N / Y / D; or

[0305] (d) T38R / K / H / W, Q59K / E and S212G / M / R / K / P / A / F / H / W / N / Y / D compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2. In one embodiment the variant polypeptide of the present invention preferably has a sequence identity of 82% to 98%, 82.5% to 98%, 83% to 98%, 83.5% to 98%, 84% to 98%, 84.5% to 98%, 85% to 98%, 85.5% to 98%, 86% to 98%, 86.5% to 98%, 87% to 98%, 87.5% to 98%, 88% to 98%, 88.5% to 98%, 89% to 98%, 89.5% to 98%, 90% to 98%, 90.3% to 98%, 90.5% to 98%, 90.7% to 98%, 91 % to 98%, 91 .3% to 98%, 91 .5% to 98%, 91 .7% to 98%, 92% to 98%, 92.3% to 98%, 92.5% to 98%, 92.7% to 98%, 93% to 98%, 93.3% to 98%, 93.5% to 98%, 93.7% to 98%, 94% to 98%, 94.3% to 98%, 94.5% to 98%, 94.7% to 98%, 95% to 98%, 95.1 % to 98%, 95.2% to 98%, 95.3% to 98%, 95.4% to 98%, 95.5% to 98% to the amino acid sequence according to SEQ ID NO: 1 and one of the following combinations of amino acid substitutions:

[0306] (a) X38R / K / H / W and X59K / E;

[0307] (b) X38R / K / W and X212P / A / G / R / K / F / Y;

[0308] (c) X59K / E and X212P / A / G / R / K / F / Y / M; or

[0309] (d) X38R / K / H / W, X59K / E and X212P / A / G / R / K / F / Y / M compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2. In one embodiment the variant polypeptide of the present invention preferably has a sequence identity of 82% to 98%, 82.5% to 98%, 83% to 98%, 83.5% to 98%, 84% to 98%, 84.5% to 98%, 85% to 98%, 85.5% to 98%, 86% to 98%, 86.5% to 98%, 87% to 98%, 87.5% to 98%, 88% to 98%, 88.5% to 98%, 89% to 98%, 89.5% to 98%, 90% to 98%, 90.3% to 98%, 90.5% to 98%, 90.7% to 98%, 91 % to 98%, 91 .3% to 98%, 91 .5% to 98%, 91 .7% to 98%, 92% to 98%, 92.3% to 98%, 92.5% to 98%, 92.7% to 98%, 93% to 98%, 93.3% to 98%, 93.5% to 98%, 93.7% to 98%, 94% to 98%, 94.3% to 98%, 94.5% to 98%, 94.7% to 98%, 95% to 98%, 95.1 % to 98%, 95.2% to 98%, 95.3% to 98%, 95.4% to 98%, 95.5% to 98% to the amino acid sequence according to SEQ ID NO: 1 and one of the following combinations of amino acid substitutions:

[0310] (a) T38R / K / H / W and Q59K / E;

[0311] (b) T38R / K / W and S212P / A / G / R / K / F / Y;

[0312] (c) Q59K / E and S212P / A / G / R / K / F / Y / M; or

[0313] (d) T38R / K / H / W, Q59K / E and S212P / A / G / R / K / F / Y / M compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2. In one embodiment the variant polypeptide of the present invention preferably has a sequence identity of 82% to 98%, 82.5% to 98%, 83% to 98%, 83.5% to 98%, 84% to 98%, 84.5% to 98%, 85% to 98%, 85.5% to 98%, 86% to 98%, 86.5% to 98%, 87% to 98%, 87.5% to 98%, 88% to 98%, 88.5% to 98%, 89% to 98%, 89.5% to 98%, 90% to 98%, 90.3% to 98%, 90.5% to 98%, 90.7% to 98%, 91 % to 98%, 91 .3% to 98%, 91 .5% to 98%, 91 .7% to 98%, 92% to 98%, 92.3% to 98%, 92.5% to 98%, 92.7% to 98%, 93% to 98%, 93.3% to 98%, 93.5% to 98%, 93.7% to 98%, 94% to 98%, 94.3% to 98%, 94.5% to 98%, 94.7% to 98%, 95% to 98%, 95.1 % to 98%, 95.2% to 98%, 95.3% to 98%, 95.4% to 98%, 95.5% to 98% to the amino acid sequence according to SEQ ID NO: 1 and one of the following combinations of amino acid substitutions:

[0314] (a) X38R / K / H / W and X59K / E;

[0315] (b) X38R / K / W and X212G / R / K / P / A / F / W / N / Y / D;

[0316] (c) X59K / E and X212G / R / K / P / A / F / W / N / Y / D; or

[0317] (d) X38R / K / H / W, X59K / E and X212G / R / K / P / A / F / W / N / Y / D compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2, wherein said variant polypeptide comprises amino acid residue D or E at position 101 , preferably E at position 101 , referring to the numbering of SEQ ID NO: 2.

[0318] In one embodiment the variant polypeptide of the present invention preferably has a sequence identity of 82% to 98%, 82.5% to 98%, 83% to 98%, 83.5% to 98%, 84% to 98%, 84.5% to 98%, 85% to 98%, 85.5% to 98%, 86% to 98%, 86.5% to 98%, 87% to 98%, 87.5% to 98%, 88% to 98%, 88.5% to 98%, 89% to 98%, 89.5% to 98%, 90% to 98%, 90.3% to 98%, 90.5% to 98%, 90.7% to 98%, 91 % to 98%, 91 .3% to 98%, 91 .5% to 98%, 91 .7% to 98%, 92% to 98%, 92.3% to 98%, 92.5% to 98%, 92.7% to 98%, 93% to 98%, 93.3% to 98%, 93.5% to 98%, 93.7% to 98%, 94% to 98%, 94.3% to 98%, 94.5% to 98%, 94.7% to 98%, 95% to 98%, 95.1 % to 98%, 95.2% to 98%, 95.3% to 98%, 95.4% to 98%, 95.5% to 98% to the amino acid sequence according to SEQ ID NO: 1 and one of the following combinations of amino acid substitutions: (a) T38R / K / H / W and Q59K / E;

[0319] (b) T38R / K / W and S212G / R / K / P / A / F / W / N / Y / D;

[0320] (c) Q59K / E and S212G / M / R / K / P / A / F / H / W / N / Y / D; or

[0321] (d) T38R / K / H / W, Q59K / E and S212G / M / R / K / P / A / F / H / W / N / Y / D compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2, wherein said variant polypeptide comprises amino acid residue D or E at position 101 , preferably E at position 101 , referring to the numbering of SEQ ID NO: 2.

[0322] In one embodiment the variant polypeptide of the present invention preferably has a sequence identity of 82% to 98%, 82.5% to 98%, 83% to 98%, 83.5% to 98%, 84% to 98%, 84.5% to 98%, 85% to 98%, 85.5% to 98%, 86% to 98%, 86.5% to 98%, 87% to 98%, 87.5% to 98%, 88% to 98%, 88.5% to 98%, 89% to 98%, 89.5% to 98%, 90% to 98%, 90.3% to 98%, 90.5% to 98%, 90.7% to 98%, 91 % to 98%, 91 .3% to 98%, 91 .5% to 98%, 91 .7% to 98%, 92% to 98%, 92.3% to 98%, 92.5% to 98%, 92.7% to 98%, 93% to 98%, 93.3% to 98%, 93.5% to 98%, 93.7% to 98%, 94% to 98%, 94.3% to 98%, 94.5% to 98%, 94.7% to 98%, 95% to 98%, 95.1 % to 98%, 95.2% to 98%, 95.3% to 98%, 95.4% to 98%, 95.5% to 98% to the amino acid sequence according to SEQ ID NO: 1 and one of the following combinations of amino acid substitutions:

[0323] (a) X38R / K / H / W and X59K / E;

[0324] (b) X38R / K / W and X212P / A / G / R / K / F / Y;

[0325] (c) X59K / E and X212P / A / G / R / K / F / Y / M; or

[0326] (d) X38R / K / H / W, X59K / E and X212P / A / G / R / K / F / Y / M compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2, wherein said variant polypeptide comprises amino acid residue D or E at position 101 , preferably E at position 101 , referring to the numbering of SEQ ID NO: 2.

[0327] In one embodiment the variant polypeptide of the present invention preferably has a sequence identity of 82% to 98%, 82.5% to 98%, 83% to 98%, 83.5% to 98%, 84% to 98%, 84.5% to 98%, 85% to 98%, 85.5% to 98%, 86% to 98%, 86.5% to 98%, 87% to 98%, 87.5% to 98%, 88% to 98%, 88.5% to 98%, 89% to 98%, 89.5% to 98%, 90% to 98%, 90.3% to 98%, 90.5% to 98%, 90.7% to 98%, 91 % to 98%, 91 .3% to 98%, 91 .5% to 98%, 91 .7% to 98%, 92% to 98%, 92.3% to 98%, 92.5% to 98%, 92.7% to 98%, 93% to 98%, 93.3% to 98%, 93.5% to 98%, 93.7% to 98%, 94% to 98%, 94.3% to 98%, 94.5% to 98%, 94.7% to 98%, 95% to 98%, 95.1 % to 98%, 95.2% to 98%, 95.3% to 98%, 95.4% to 98%, 95.5% to 98% to the amino acid sequence according to SEQ ID NO: 1 and one of the following combinations of amino acid substitutions:

[0328] (a) T38R / K / H / W and Q59K / E;

[0329] (b) T38R / K / W and S212P / A / G / R / K / F / Y;

[0330] (c) Q59K / E and S212P / A / G / R / K / F / Y / M; or

[0331] (d) T38R / K / H / W, Q59K / E and S212P / A / G / R / K / F / Y / M compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2, wherein said variant polypeptide comprises amino acid residue D or E at position 101 , preferably E at position 101 , referring to the numbering of SEQ ID NO: 2.

[0332] In one embodiment the variant polypeptide of the present invention preferably has a sequence identity of 82% to 98% to the amino acid sequence according to SEQ ID NO: 1 and one of the following combinations of amino acid substitutions:

[0333] (a) X38R / K / H / W and X59K / E;

[0334] (b) X38R / K / W and X212G / R / K / P / A / F / W / N / Y / D;

[0335] (c) X59K / E and X212G / R / K / P / A / F / W / N / Y / D; or

[0336] (d) X38R / K / H / W, X59K / E and X212G / R / K / P / A / F / W / N / Y / D, compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2. In one embodiment the variant polypeptide of the present invention preferably has a sequence identity of 82% to 98% to the amino acid sequence according to SEQ ID NO: 1 and one of the following combinations of amino acid substitutions:

[0337] (a) X38W and X212F;

[0338] (b) X38K and X212F;

[0339] (c) X38W and X212P;

[0340] (d) X38W and X212R;

[0341] (e) X38K and X212P;

[0342] (f) X59K and X212P;

[0343] (g) X38K, X59K, X212P; or

[0344] (h) X38W, X59K, X212P compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2. In one embodiment the variant polypeptide of the present invention preferably has a sequence identity of 82% to 98 to the amino acid sequence according to SEQ ID NO: 1 and one of the following combinations of amino acid substitutions:

[0345] (a) T38R / K / H / W and Q59K / E;

[0346] (b) T38R / K / W and S212G / R / K / P / A / F / W / N / Y / D;

[0347] (c) Q59K / E and S212G / M / R / K / P / A / F / H / W / N / Y / D; or

[0348] (d) T38R / K / H / W, Q59K / E and S212G / M / R / K / P / A / F / H / W / N / Y / D, compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2. In one embodiment the variant polypeptide of the present invention preferably has a sequence identity of 82% to 98% to the amino acid sequence according to SEQ ID NO: 1 and one of the following combinations of amino acid substitutions: (a) X38R / K / H / W and X59K / E;

[0349] (b) X38R / K / W and X212P / A / G / R / K / F / Y;

[0350] (c) X59K / E and X212P / A / G / R / K / F / Y / M; or

[0351] (d) X38R / K / H / W, X59K / E and X212P / A / G / R / K / F / Y / M, compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2. In one embodiment the variant polypeptide of the present invention preferably has a sequence identity of 82% to 98% to the amino acid sequence according to SEQ ID NO: 1 and one of the following combinations of amino acid substitutions:

[0352] (a) T38R / K / H / W and Q59K / E;

[0353] (b) T38R / K / W and S212P / A / G / R / K / F / Y;

[0354] (c) Q59K / E and S212P / A / G / R / K / F / Y / M; or

[0355] (d) T38R / K / H / W, Q59K / E and S212P / A / G / R / K / F / Y / M, compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2. In one embodiment the variant polypeptide of the present invention preferably has a sequence identity of 82% to 98% to the amino acid sequence according to SEQ ID NO: 1 and one of the following combinations of amino acid substitutions:

[0356] (a) X38R / K / H / W and X59K / E;

[0357] (b) X38R / K / W and X212G / R / K / P / A / F / W / N / Y / D;

[0358] (c) X59K / E and X212G / M / R / K / P / A / F / H / W / N / Y / D; or

[0359] (d) X38R / K / H / W, X59K / E and X212G / M / R / K / P / A / F / H / W / N / Y / D, compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2, wherein said variant polypeptide comprises amino acid residue D or E at position 101 , preferably E at position 101 , referring to the numbering of SEQ ID NO: 2.

[0360] In one embodiment the variant polypeptide of the present invention preferably has a sequence identity of 82% to 98 to the amino acid sequence according to SEQ ID NO: 1 and one of the following combinations of amino acid substitutions:

[0361] (a) T38R / K / H / W and Q59K / E;

[0362] (b) T38R / K / W and S212G / R / K / P / A / F / W / N / Y / D;

[0363] (c) Q59K / E and S212G / M / R / K / P / A / F / H / W / N / Y / D; or

[0364] (d) T38R / K / H / W, Q59K / E and S212G / M / R / K / P / A / F / H / W / N / Y / D, compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2, wherein said variant polypeptide comprises amino acid residue D or E at position 101 , preferably E at position 101 , referring to the numbering of SEQ ID NO: 2. In one embodiment the variant polypeptide of the present invention preferably has a sequence identity of 82% to 98% to the amino acid sequence according to SEQ ID NO: 1 and one of the following combinations of amino acid substitutions:

[0365] (a) X38R / K / H / W and X59K / E;

[0366] (b) X38R / K / W and X212P / A / G / R / K / F / Y;

[0367] (c) X59K / E and X212P / A / G / R / K / F / Y / M; or

[0368] (d) X38R / K / H / W, X59K / E and X212P / A / G / R / K / F / Y / M, compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2, wherein said variant polypeptide comprises amino acid residue D or E at position 101 , preferably E at position 101 , referring to the numbering of SEQ ID NO: 2.

[0369] In one embodiment the variant polypeptide of the present invention preferably has a sequence identity of 82% to 98% to the amino acid sequence according to SEQ ID NO: 1 and one of the following combinations of amino acid substitutions:

[0370] (a) T38R / K / H / W and Q59K / E;

[0371] (b) T38R / K / W and S212P / A / G / R / K / F / Y;

[0372] (c) Q59K / E and S212P / A / G / R / K / F / Y / M; or

[0373] (d) T38R / K / H / W, Q59K / E and S212P / A / G / R / K / F / Y / M, compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2, wherein said variant polypeptide comprises amino acid residue D or E at position 101 , preferably E at position 101 , referring to the numbering of SEQ ID NO: 2.

[0374] In one embodiment the variant polypeptide of the present invention preferably has a sequence identity of 95.5% to 98% to the amino acid sequence according to SEQ ID NO: 1 and one of the following combinations of amino acid substitutions:

[0375] (a) X38R / K / H / W and X59K / E;

[0376] (b) X38R / K / W and X212G / M / R / K / P / A / F / H / W / N / Y / D;

[0377] (c) X59K / E and X212G / R / K / P / A / F / W / N / Y / D; or

[0378] (d) X38R / K / H / W, X59K / E and X212G / M / R / K / P / A / F / H / W / N / Y / D compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2. In one embodiment the variant polypeptide of the present invention preferably has a sequence identity of 95.5% to 98% to the amino acid sequence according to SEQ ID NO: 1 and one of the following combinations of amino acid substitutions:

[0379] (a) X38W and X212F;

[0380] (b) X38K and X212F;

[0381] (c) X38W and X212P; (d) X38W and X212R;

[0382] (e) X38K and X212P;

[0383] (f) X59K and X212P;

[0384] (g) X38K, X59K, X212P; or

[0385] (h) X38W, X59K, X212P compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2. In one embodiment the variant polypeptide of the present invention preferably has a sequence identity of 95.5% to 98 to the amino acid sequence according to SEQ ID NO: 1 and one of the following combinations of amino acid substitutions:

[0386] (a) T38R / K / H / W and Q59K / E;

[0387] (b) T38R / K / W and S212G / R / K / P / A / F / W / N / Y / D;

[0388] (c) Q59K / E and S212G / R / K / P / A / F / W / N / Y / D; or

[0389] (d) T38R / K / H / W, Q59K / E and S212G / M / R / K / P / A / F / H / W / N / Y / D compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2. In one embodiment the variant polypeptide of the present invention preferably has a sequence identity of 95.5% to 98% to the amino acid sequence according to SEQ ID NO: 1 and one of the following combinations of amino acid substitutions:

[0390] (a) X38R / K / H / W and X59K / E;

[0391] (b) X38R / K / W and X212P / A / G / R / K / F / Y;

[0392] (c) X59K / E and X212P / A / G / R / K / F / Y / M; or

[0393] (d) X38R / K / H / W, X59K / E and X212P / A / G / R / K / F / Y / M compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2. In one embodiment the variant polypeptide of the present invention preferably has a sequence identity of 95.5% to 98% to the amino acid sequence according to SEQ ID NO: 1 and one of the following combinations of amino acid substitutions:

[0394] (a) T38R / K / H / W and Q59K / E;

[0395] (b) T38R / K / W and S212P / A / G / R / K / F / Y;

[0396] (c) Q59K / E and S212P / A / G / R / K / F / Y / M; or

[0397] (d) T38R / K / H / W, Q59K / E and S212P / A / G / R / K / F / Y / M compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2. In one embodiment the variant polypeptide of the present invention preferably has a sequence identity of 95.5% to 98% to the amino acid sequence according to SEQ ID NO: 1 and one of the following combinations of amino acid substitutions:

[0398] (a) X38R / K / H / W and X59K / E; (b) X38R / K / W and X212G / R / K / P / A / F / W / N / Y / D;

[0399] (c) X59K / E and X212G / M / R / K / P / A / F / H / W / N / Y / D; or

[0400] (d) X38R / K / H / W, X59K / E and X212G / M / R / K / P / A / F / H / W / N / Y / D compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2, wherein said variant polypeptide comprises amino acid residue D or E at position 101 , preferably E at position 101 , referring to the numbering of SEQ ID NO: 2.

[0401] In one embodiment the variant polypeptide of the present invention preferably has a sequence identity of 95.5% to 98 to the amino acid sequence according to SEQ ID NO: 1 and one of the following combinations of amino acid substitutions:

[0402] (a) T38R / K / H / W and Q59K / E;

[0403] (b) T38R / K / W and S212G / R / K / P / A / F / W / N / Y / D;

[0404] (c) Q59K / E and S212G / M / R / K / P / A / F / H / W / N / Y / D; or

[0405] (d) T38R / K / H / W, Q59K / E and S212G / M / R / K / P / A / F / H / W / N / Y / D compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2, wherein said variant polypeptide comprises amino acid residue D or E at position 101 , preferably E at position 101 , referring to the numbering of SEQ ID NO: 2.

[0406] In one embodiment the variant polypeptide of the present invention preferably has a sequence identity of 95.5% to 98% to the amino acid sequence according to SEQ ID NO: 1 and one of the following combinations of amino acid substitutions:

[0407] (a) X38R / K / H / W and X59K / E;

[0408] (b) X38R / K / W and X212P / A / G / R / K / F / Y;

[0409] (c) X59K / E and X212P / A / G / R / K / F / Y / M; or

[0410] (d) X38R / K / H / W, X59K / E and X212P / A / G / R / K / F / Y / M compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2, wherein said variant polypeptide comprises amino acid residue D or E at position 101 , preferably E at position 101 , referring to the numbering of SEQ ID NO: 2.

[0411] In one embodiment the variant polypeptide of the present invention preferably has a sequence identity of 95.5% to 98% to the amino acid sequence according to SEQ ID NO: 1 and one of the following combinations of amino acid substitutions:

[0412] (a) T38R / K / H / W and Q59K / E;

[0413] (b) T38R / K / W and S212P / A / G / R / K / F / Y;

[0414] (c) Q59K / E and S212P / A / G / R / K / F / Y / M; or

[0415] (d) T38R / K / H / W, Q59K / E and S212P / A / G / R / K / F / Y / M compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2, wherein said variant polypeptide comprises amino acid residue D or E at position 101 , preferably E at position 101 , referring to the numbering of SEQ ID NO: 2.

[0416] In one embodiment, the variant protease has an amino acid sequence with a sequence identity of at least 95%, at least 95.5%, at least 96%, at least 96.5%, at least 97%, at least 97.5%, at least 98%, at least 98.5%, at least 99% or of 100% to the amino acid sequence according to any one of SEQ ID NO: 65, SEQ ID NO: 66 and SEQ ID NO: 67. In one embodiment, the variant polypeptide of the present invention or the fragment of said polypeptide having protease activity additionally comprises an amino acid substitution at amino residue 210 and / or amino acid residue 271 compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2. In one embodiment, the variant polypeptide of the present invention or the fragment of said polypeptide having protease activity additionally comprises the amino acid substitution X210I / V and / or the amino acid substitution X271 D compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2.

[0417] In one embodiment, the variant polypeptide of the present invention or the fragment of said polypeptide having protease activity additionally comprises the amino acid substitution P210I / V and / or the amino acid substitution E271 D compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2.

[0418] In one embodiment the variant polypeptide of the present invention preferably has a sequence identity of at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 90.5%, at least 91 %, at least 91 .5%, at least 92%, at least 92.5%, at least 93%, at least 93.5%, at least 94%, at least 94.5%, at least 95%, at least 95.1 %, at least 95.2%, at least 95.3%, at least 95.4%, at least 95.5%, at least 95.6%, at least 95.7%, at least 95.8%, at least 95.9% at least 96%, at least 96.1 %, at least 96.2%, at least 96.3%, at least 96.4%, at least 96.5%, at least 96.6%, at least 96.7%, at least 96.8%, at least 96.9%, at least 97%, at least 97.1 %, at least 97.2%, at least 97.3%, at least 97.4%, at least 97.5%, at least 97.6%, at least 97.7%, at least 97.8%, at least 97.9, but less than 100% to the amino acid sequence according to SEQ ID NO: 1 and one of the following combinations of amino acid substitutions:

[0419] (a) X38R / K / H / W, X59K / E and X210 l / V;

[0420] (b) X38R / K / H / W, X210 l / V and X212G / M / R / K / P / A / F / H / W / N / Y / D ;

[0421] (c) X59K / E, X210 l / V and X212G / M / R / K / P / A / F / H / W / N / Y / D;

[0422] (d) X38R / K / H / W, X59K / E, X210 l / V and X212G / M / R / K / P / A / F / H / W / N / Y / D;

[0423] (e) X38R / K / H / W, X59K / E and X271 D;

[0424] (f) X38R / K / H / W, X212G / M / R / K / P / A / F / H / W / N / Y / D and X271 D;

[0425] (g) X59K / E, X212G / M / R / K / P / A / F / H / W / N / Y / D and X271 D;

[0426] (h) X38R / K / H / W, X59K / E, X212G / M / R / K / P / A / F / H / W / N / Y / D and X271 D;

[0427] (I) X38R / K / H / W, X59K / E, X210I / V and X271 D;

[0428] Q) X38R / K / H / W, X210 l / V, X212G / M / R / K / P / A / F / H / W / N / Y / D and X271 D;

[0429] (k) X59K / E, X210I / V, X212G / M / R / K / P / A / F / H / W / N / Y / D and X271 D; or (I) X38R / K / H / W, X59K / E, X21 Ol / V, X212G / M / R / K / P / A / F / H / W / N / Y / D and X271 D compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2. In one embodiment the variant polypeptide of the present invention preferably has a sequence identity of at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 90.5%, at least 91 %, at least 91 .5%, at least 92%, at least 92.5%, at least 93%, at least 93.5%, at least 94%, at least 94.5%, at least 95%, at least 95.1 %, at least 95.2%, at least 95.3%, at least 95.4%, at least 95.5%, at least 95.6%, at least 95.7%, at least 95.8%, at least 95.9% at least 96%, at least 96.1 %, at least 96.2%, at least 96.3%, at least 96.4%, at least 96.5%, at least 96.6%, at least 96.7%, at least 96.8%, at least 96.9%, at least 97%, at least 97.1 %, at least 97.2%, at least 97.3%, at least 97.4%, at least 97.5%, at least 97.6%, at least 97.7%, at least 97.8%, at least 97.9%, but less than 100% to the amino acid sequence according to SEQ ID NO: 1 and one of the following combinations of amino acid substitutions:

[0430] (a) T38R / K / H / W, Q59K / E and P21 Ol / V;

[0431] (b) T38R / K / H / W, P21 Ol / V and S212G / M / R / K / P / A / F / H / W / N / Y / D;

[0432] (c) Q59K / E, P21 Ol / V and S212G / M / R / K / P / A / F / H / W / N / Y / D;

[0433] (d) T38R / K / H / W, Q59K / E, P210I / V and S212G / M / R / K / P / A / F / H / W / N / Y / D;

[0434] (e) T38R / K / H / W, Q59K / E and E271 D;

[0435] (f) T38R / K / H / W, S212G / M / R / K / P / A / F / H / W / N / Y / D and E271 D;

[0436] (g) Q59K / E, S212G / M / R / K / P / A / F / H / W / N / Y / D and E271 D;

[0437] (h) T38R / K / H / W, Q59K / E, S212G / M / R / K / P / A / F / H / W / N / Y / D and E271 D;

[0438] (i) T38R / K / H / W, Q59K / E, P210I / V and E271 D;

[0439] Q) T38R / K / H / W, P210I / V, S212G / M / R / K / P / A / F / H / W / N / Y / D and E271 D;

[0440] (k) Q59K / E, P210I / V, S212G / M / R / K / P / A / F / H / W / N / Y / D and E271 D; or

[0441] (l) T38R / K / H / W, Q59K / E, P210I / V, S212G / M / R / K / P / A / F / H / W / N / Y / D and E271 D compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2. In one embodiment the variant polypeptide of the present invention preferably has a sequence identity of at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 90.5%, at least 91 %, at least 91 .5%, at least 92%, at least 92.5%, at least 93%, at least 93.5%, at least 94%, at least 94.5%, at least 95%, at least 95.1 %, at least 95.2%, at least 95.3%, at least 95.4%, at least 95.5%, at least 95.6%, at least 95.7%, at least 95.8%, at least 95.9% at least 96%, at least 96.1 %, at least 96.2%, at least 96.3%, at least 96.4%, at least 96.5%, at least 96.6%, at least 96.7%, at least 96.8%, at least 96.9%, at least 97%, at least 97.1 %, at least 97.2%, at least 97.3%, at least 97.4%, at least 97.5%, at least 97.6%, at least 97.7%, at least 97.8%, at least 97.9%, but less than 100% to the amino acid sequence according to SEQ ID NO: 1 and one of the following combinations of amino acid substitutions:

[0442] (a) X38R / K / H / W, X59K / E and X210I; (b) X38R / K / H / W, X210I and X212P / A / G / R / K / F / Y / M;

[0443] (c) X59K / E, X21 Ol and X212P / A / G / R / K / F / Y / M;

[0444] (d) X38R / K / H / W, X59K / E, X21 Ol and X212P / A / G / R / K / F / Y / M;

[0445] (e) X38R / K / H / W, X59K / E and X271 D;

[0446] (f) X38R / K / H / W, X212P / A / G / R / K / F / Y / M and X271 D;

[0447] (g) X59K / E, X212P / A / G / R / K / F / Y / M and X271 D;

[0448] (h) X38R / K / H / W, X59K / E, X212P / A / G / R / K / F / Y / M and X271 D;

[0449] (i) X38R / K / H / W, X59K / E, X210I and X271 D;

[0450] G) X38R / K / H / W, X21 Ol, X212P / A / G / R / K / F / Y / M and X271 D;

[0451] (k) X59K / E, X210I, X212P / A / G / R / K / F / Y / M and X271 D; or

[0452] (l) X38R / K / H / W, X59K / E, X210I, X212P / A / G / R / K / F / Y / M and X271 D compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2. In one embodiment the variant polypeptide of the present invention preferably has a sequence identity of at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 90.5%, at least 91 %, at least 91 .5%, at least 92%, at least 92.5%, at least 93%, at least 93.5%, at least 94%, at least 94.5%, at least 95%, at least 95.1 %, at least 95.2%, at least 95.3%, at least 95.4%, at least 95.5%, at least 95.6%, at least 95.7%, at least 95.8%, at least 95.9% at least 96%, at least 96.1 %, at least 96.2%, at least 96.3%, at least 96.4%, at least 96.5%, at least 96.6%, at least 96.7%, at least 96.8%, at least 96.9%, at least 97%, at least 97.1 %, at least 97.2%, at least 97.3%, at least 97.4%, at least 97.5%, at least 97.6%, at least 97.7%, at least 97.8%, at least 97.9%, but less than 100% to the amino acid sequence according to SEQ ID NO: 1 and one of the following combinations of amino acid substitutions:

[0453] (a) T38R / K / H / W, Q59K / E and P210I;

[0454] (b) T38R / K / H / W, P21 Ol and S212P / A / G / R / K / F / Y / M;

[0455] (c) Q59K / E, P210I and S212P / A / G / R / K / F / Y / M;

[0456] (d) T38R / K / H / W, Q59K / E, P210I and S212P / A / G / R / K / F / Y / M;

[0457] (e) T38R / K / H / W, Q59K / E and E271 D;

[0458] (f) T38R / K / H / W, S212P / A / G / R / K / F / Y / M and E271 D;

[0459] (g) Q59K / E, S212P / A / G / R / K / F / Y / M and E271 D;

[0460] (h) T38R / K / H / W, Q59K / E, S212P / A / G / R / K / F / Y / M and E271 D;

[0461] (I) T38R / K / H / W, Q59K / E, P210I and E271 D;

[0462] Q) T38R / K / H / W, P210I, S212P / A / G / R / K / F / Y / M and E271 D;

[0463] (k) Q59K / E, P210I, S212P / A / G / R / K / F / Y / M and E271 D; or

[0464] (l) T38R / K / H / W, Q59K / E, P210I, S212P / A / G / R / K / F / Y / M and E271 D compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2. In one embodiment the variant polypeptide of the present invention preferably has a sequence identity of at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 90.5%, at least 91 %, at least 91 .5%, at least 92%, at least 92.5%, at least 93%, at least 93.5%, at least 94%, at least 94.5%, at least 95%, at least 95.1 %, at least 95.2%, at least 95.3%, at least 95.4%, at least 95.5%, at least 95.6%, at least 95.7%, at least 95.8%, at least 95.9% at least 96%, at least 96.1 %, at least 96.2%, at least 96.3%, at least 96.4%, at least 96.5%, at least 96.6%, at least 96.7%, at least 96.8%, at least 96.9%, at least 97%, at least 97.1 %, at least 97.2%, at least 97.3%, at least 97.4%, at least 97.5%, at least 97.6%, at least 97.7%, at least 97.8%, at least 97.9, but less than 100% to the amino acid sequence according to SEQ ID NO: 1 and one of the following combinations of amino acid substitutions:

[0465] (a) X38R / K / H / W, X59K / E and X21 Ol / V;

[0466] (b) X38R / K / H / W, X210I / V and X212G / M / R / K / P / A / F / H / W / N / Y / D;

[0467] (c) X59K / E, X21 Ol / V and X212G / M / R / K / P / A / F / H / W / N / Y / D;

[0468] (d) X38R / K / H / W, X59K / E, X21 Ol / V and X212G / M / R / K / P / A / F / H / VV / N / Y / D;

[0469] (e) X38R / K / H / W, X59K / E and X271 D;

[0470] (f) X38R / K / H / W, X212G / M / R / K / P / A / F / H / W / N / Y / D and X271 D;

[0471] (g) X59K / E, X212G / M / R / K / P / A / F / H / W / N / Y / D and X271 D;

[0472] (h) X38R / K / H / W, X59K / E, X212G / M / R / K / P / A / F / H / W / N / Y / D and X271 D;

[0473] (I) X38R / K / H / W, X59K / E, X210I / V and X271 D;

[0474] Q) X38R / K / H / W, X21 Ol / V, X212G / M / R / K / P / A / F / H / W / N / Y / D and X271 D;

[0475] (k) X59K / E, X210I / V, X212G / M / R / K / P / A / F / H / W / N / Y / D and X271 D; or

[0476] (l) X38R / K / H / W, X59K / E, X21 Ol / V, X212G / M / R / K / P / A / F / H / W / N / Y / D and X271 D compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2, wherein said variant polypeptide comprises amino acid residue D or E at position 101 , preferably E at position 101 , referring to the numbering of SEQ ID NO: 2.

[0477] In one embodiment the variant polypeptide of the present invention preferably has a sequence identity of at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 90.5%, at least 91 %, at least 91 .5%, at least 92%, at least 92.5%, at least 93%, at least 93.5%, at least 94%, at least 94.5%, at least 95%, at least 95.1 %, at least 95.2%, at least 95.3%, at least 95.4%, at least 95.5%, at least 95.6%, at least 95.7%, at least 95.8%, at least 95.9% at least 96%, at least 96.1 %, at least 96.2%, at least 96.3%, at least 96.4%, at least 96.5%, at least 96.6%, at least 96.7%, at least 96.8%, at least 96.9%, at least 97%, at least 97.1 %, at least 97.2%, at least 97.3%, at least 97.4%, at least 97.5%, at least 97.6%, at least 97.7%, at least 97.8%, at least 97.9%, but less than 100% to the amino acid sequence according to SEQ ID NO: 1 and one of the following combinations of amino acid substitutions:

[0478] (a) T38R / K / H / W, Q59K / E and P21 Ol / V; (b) T38R / K / H / W, P210I / V and S212G / M / R / K / P / A / F / H / W / N / Y / D;

[0479] (c) Q59K / E, P21 Ol / V and S212G / M / R / K / P / A / F / H / W / N / Y / D;

[0480] (d) T38R / K / H / W, Q59K / E, P210I / V and S212G / M / R / K / P / A / F / H / W / N / Y / D;

[0481] (e) T38R / K / H / W, Q59K / E and E271 D;

[0482] (f) T38R / K / H / W, S212G / M / R / K / P / A / F / H / W / N / Y / D and E271 D;

[0483] (g) Q59K / E, S212G / M / R / K / P / A / F / H / W / N / Y / D and E271 D;

[0484] (h) T38R / K / H / W, Q59K / E, S212G / M / R / K / P / A / F / H / W / N / Y / D and E271 D;

[0485] (i) T38R / K / H / W, Q59K / E, P210I / V and E271 D;

[0486] G) T38R / K / H / W, P210I / V, S212G / M / R / K / P / A / F / H / W / N / Y / D and E271 D;

[0487] (k) Q59K / E, P210I / V, S212G / M / R / K / P / A / F / H / W / N / Y / D and E271 D; or

[0488] (l) T38R / K / H / W, Q59K / E, P21 Ol / V, S212G / M / R / K / P / A / F / H / W / N / Y / D and E271 D compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2, wherein said variant polypeptide comprises amino acid residue D or E at position 101 , preferably E at position 101 , referring to the numbering of SEQ ID NO: 2.

[0489] In one embodiment the variant polypeptide of the present invention preferably has a sequence identity of at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 90.5%, at least 91 %, at least 91 .5%, at least 92%, at least 92.5%, at least 93%, at least 93.5%, at least 94%, at least 94.5%, at least 95%, at least 95.1 %, at least 95.2%, at least 95.3%, at least 95.4%, at least 95.5%, at least 95.6%, at least 95.7%, at least 95.8%, at least 95.9% at least 96%, at least 96.1 %, at least 96.2%, at least 96.3%, at least 96.4%, at least 96.5%, at least 96.6%, at least 96.7%, at least 96.8%, at least 96.9%, at least 97%, at least 97.1 %, at least 97.2%, at least 97.3%, at least 97.4%, at least 97.5%, at least 97.6%, at least 97.7%, at least 97.8%, at least 97.9%, but less than 100% to the amino acid sequence according to SEQ ID NO: 1 and one of the following combinations of amino acid substitutions:

[0490] (a) X38R / K / H / W, X59K / E and X210I;

[0491] (b) X38R / K / H / W, X210I and X212P / A / G / R / K / F / Y / M;

[0492] (c) X59K / E, X210I and X212P / A / G / R / K / F / Y / M ;

[0493] (d) X38R / K / H / W, X59K / E, X210I and X212P / A / G / R / K / F / Y / M ;

[0494] (e) X38R / K / H / W, X59K / E and X271 D;

[0495] (f) X38R / K / H / W, X212 P / A / G / R / K / F / Y / M and X271 D;

[0496] (g) X59K / E, X212P / A / G / R / K / F / Y / M and X271 D;

[0497] (h) X38R / K / H / W, X59K / E, X212P / A / G / R / K / F / Y / M and X271 D;

[0498] (I) X38R / K / H / W, X59K / E, X210I and X271 D;

[0499] Q) X38R / K / H / W, X210I, X212P / A / G / R / K / F / Y / M and X271 D;

[0500] (k) X59K / E, X210I, X212P / A / G / R / K / F / Y / M and X271 D; or (I) X38R / K / H / W, X59K / E, X210I, X212P / A / G / R / K / F / Y / M and X271 D compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2, wherein said variant polypeptide comprises amino acid residue D or E at position 101 , preferably E at position 101 , referring to the numbering of SEQ ID NO: 2.

[0501] In one embodiment the variant polypeptide of the present invention preferably has a sequence identity of at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 90.5%, at least 91 %, at least 91 .5%, at least 92%, at least 92.5%, at least 93%, at least 93.5%, at least 94%, at least 94.5%, at least 95%, at least 95.1 %, at least 95.2%, at least 95.3%, at least 95.4%, at least 95.5%, at least 95.6%, at least 95.7%, at least 95.8%, at least 95.9% at least 96%, at least 96.1 %, at least 96.2%, at least 96.3%, at least 96.4%, at least 96.5%, at least 96.6%, at least 96.7%, at least 96.8%, at least 96.9%, at least 97%, at least 97.1 %, at least 97.2%, at least 97.3%, at least 97.4%, at least 97.5%, at least 97.6%, at least 97.7%, at least 97.8%, at least 97.9%, but less than 100% to the amino acid sequence according to SEQ ID NO: 1 and one of the following combinations of amino acid substitutions:

[0502] (a) T38R / K / H / W, Q59K / E and P210I;

[0503] (b) T38R / K / H / W, P210I and S212P / A / G / R / K / F / Y / M;

[0504] (c) Q59K / E, P210I and S212P / A / G / R / K / F / Y / M;

[0505] (d) T38R / K / H / W, Q59K / E, P210I and S212P / A / G / R / K / F / Y / M;

[0506] (e) T38R / K / H / W, Q59K / E and E271 D;

[0507] (f) T38R / K / H / W, S212P / A / G / R / K / F / Y / M and E271 D;

[0508] (g) Q59K / E, S212P / A / G / R / K / F / Y / M and E271 D;

[0509] (h) T38R / K / H / W, Q59K / E, S212P / A / G / R / K / F / Y / M and E271 D;

[0510] (i) T38R / K / H / W, Q59K / E, P210I and E271 D;

[0511] Q) T38R / K / H / W, P210I, S212P / A / G / R / K / F / Y / M and E271 D;

[0512] (k) Q59K / E, P210I, S212P / A / G / R / K / F / Y / M and E271 D; or

[0513] (l) T38R / K / H / W, Q59K / E, P210I, S212P / A / G / R / K / F / Y / M and E271 D compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2, wherein said variant polypeptide comprises amino acid residue D or E at position 101 , preferably E at position 101 , referring to the numbering of SEQ ID NO: 2.

[0514] In one embodiment the variant polypeptide of the present invention preferably has a sequence identity of 82% to 98%, 82.5% to 98%, 83% to 98%, 83.5% to 98%, 84% to 98%, 84.5% to 98%, 85% to 98%, 85.5% to 98%, 86% to 98%, 86.5% to 98%, 87% to 98%, 87.5% to 98%, 88% to 98%, 88.5% to 98%, 89% to 98%, 89.5% to 98%, 90% to 98%, 90.3% to 98%, 90.5% to 98%, 90.7% to 98%, 91 % to 98%, 91 .3% to 98%, 91 .5% to 98%, 91 .7% to 98%, 92% to 98%, 92.3% to 98%, 92.5% to 98%, 92.7% to 98%, 93% to 98%, 93.3% to 98%, 93.5% to 98%, 93.7% to 98%, 94% to 98%, 94.3% to 98%, 94.5% to 98%, 94.7% to 98%, 95% to 98%, 95.1 % to 98%, 95.2% to 98%, 95.3% to 98%, 95.4% to 98%, 95.5% to 98% to the amino acid sequence according to SEQ ID NO: 1 and one of the following combinations of amino acid substitutions:

[0515] (a) X38R / K / H / W, X59K / E and X210I / V;

[0516] (b) X38R / K / H / W, X210I / V and X212G / M / R / K / P / A / F / H / W / N / Y / D;

[0517] (c) X59K / E, X21 Ol / V and X212G / M / R / K / P / A / F / H / W / N / Y / D;

[0518] (d) X38R / K / H / W, X59K / E, X210I / V and X212G / M / R / K / P / A / F / H / W / N / Y / D;

[0519] (e) X38R / K / H / W, X59K / E and X271 D;

[0520] (f) X38R / K / H / W, X212G / M / R / K / P / A / F / H / W / N / Y / D and X271 D;

[0521] (g) X59K / E, X212G / M / R / K / P / A / F / H / W / N / Y / D and X271 D;

[0522] (h) X38R / K / H / W, X59K / E, X212G / M / R / K / P / A / F / H / W / N / Y / D and X271 D;

[0523] (I) X38R / K / H / W, X59K / E, X210I / V and X271 D;

[0524] Q) X38R / K / H / W, X21 Ol / V, X212G / M / R / K / P / A / F / H / W / N / Y / D and X271 D;

[0525] (k) X59K / E, X210I / V, X212G / M / R / K / P / A / F / H / W / N / Y / D and X271 D; or

[0526] (l) X38R / K / H / W, X59K / E, X21 Ol / V, X212G / M / R / K / P / A / F / H / W / N / Y / D and X271 D compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2. In one embodiment the variant polypeptide of the present invention preferably has a sequence identity of 82% to 98%, 82.5% to 98%, 83% to 98%, 83.5% to 98%, 84% to 98%, 84.5% to 98%, 85% to 98%, 85.5% to 98%, 86% to 98%, 86.5% to 98%, 87% to 98%, 87.5% to 98%, 88% to 98%, 88.5% to 98%, 89% to 98%, 89.5% to 98%, 90% to 98%, 90.3% to 98%, 90.5% to 98%, 90.7% to 98%, 91 % to 98%, 91 .3% to 98%, 91 .5% to 98%, 91 .7% to 98%, 92% to 98%, 92.3% to 98%, 92.5% to 98%, 92.7% to 98%, 93% to 98%, 93.3% to 98%, 93.5% to 98%, 93.7% to 98%, 94% to 98%, 94.3% to 98%, 94.5% to 98%, 94.7% to 98%, 95% to 98%, 95.1 % to 98%, 95.2% to 98%, 95.3% to 98%, 95.4% to 98%, 95.5% to 98% to the amino acid sequence according to SEQ ID NO: 1 and one of the following combinations of amino acid substitutions:

[0527] (a) T38R / K / H / W, Q59K / E and P21 Ol / V;

[0528] (b) T38R / K / H / W, P210I / V and S212G / M / R / K / P / A / F / H / W / N / Y / D;

[0529] (c) Q59K / E, P21 Ol / V and S212G / M / R / K / P / A / F / H / W / N / Y / D;

[0530] (d) T38R / K / H / W, Q59K / E, P210I / V and S212G / M / R / K / P / A / F / H / W / N / Y / D;

[0531] (e) T38R / K / H / W, Q59K / E and E271 D;

[0532] (f) T38R / K / H / W, S212G / M / R / K / P / A / F / H / W / N / Y / D and E271 D;

[0533] (g) Q59K / E, S212G / M / R / K / P / A / F / H / W / N / Y / D and E271 D;

[0534] (h) T38R / K / H / W, Q59K / E, S212G / M / R / K / P / A / F / H / W / N / Y / D and E271 D;

[0535] (I) T38R / K / H / W, Q59K / E, P210I / V and E271 D;

[0536] Q) T38R / K / H / W, P210I / V, S212G / M / R / K / P / A / F / H / W / N / Y / D and E271 D;

[0537] (k) Q59K / E, P210I / V, S212G / M / R / K / P / A / F / H / W / N / Y / D and E271 D; or (I) T38R / K / H / W, Q59K / E, P21 Ol / V, S212G / M / R / K / P / A / F / H / W / N / Y / D and E271 D compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2. In one embodiment the variant polypeptide of the present invention preferably has a sequence identity of 82% to 98%, 82.5% to 98%, 83% to 98%, 83.5% to 98%, 84% to 98%, 84.5% to 98%, 85% to 98%, 85.5% to 98%, 86% to 98%, 86.5% to 98%, 87% to 98%, 87.5% to 98%, 88% to 98%, 88.5% to 98%, 89% to 98%, 89.5% to 98%, 90% to 98%, 90.3% to 98%, 90.5% to 98%, 90.7% to 98%, 91 % to 98%, 91 .3% to 98%, 91 .5% to 98%, 91 .7% to 98%, 92% to 98%, 92.3% to 98%, 92.5% to 98%, 92.7% to 98%, 93% to 98%, 93.3% to 98%, 93.5% to 98%, 93.7% to 98%, 94% to 98%, 94.3% to 98%, 94.5% to 98%, 94.7% to 98%, 95% to 98%, 95.1 % to 98%, 95.2% to 98%, 95.3% to 98%, 95.4% to 98%, 95.5% to 98% to the amino acid sequence according to SEQ ID NO: 1 and one of the following combinations of amino acid substitutions:

[0538] (a) X38R / K / H / W, X59K / E and X210I;

[0539] (b) X38R / K / H / W, X210I and X212P / A / G / R / K / F / Y / M;

[0540] (c) X59K / E, X210I and X212P / A / G / R / K / F / Y / M;

[0541] (d) X38R / K / H / W, X59K / E, X210I and X212P / A / G / R / K / F / Y / M;

[0542] (e) X38R / K / H / W, X59K / E and X271 D;

[0543] (f) X38R / K / H / W, X212P / A / G / R / K / F / Y / M and X271 D;

[0544] (g) X59K / E, X212P / A / G / R / K / F / Y / M and X271 D;

[0545] (h) X38R / K / H / W, X59K / E, X212P / A / G / R / K / F / Y / M and X271 D;

[0546] (I) X38R / K / H / W, X59K / E, X210I and X271 D;

[0547] Q) X38R / K / H / W, X210I, X212P / A / G / R / K / F / Y / M and X271 D;

[0548] (k) X59K / E, X210I, X212P / A / G / R / K / F / Y / M and X271 D; or

[0549] (l) X38R / K / H / W, X59K / E, X210I, X212P / A / G / R / K / F / Y / M and X271 D compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2. In one embodiment the variant polypeptide of the present invention preferably has a sequence identity of 82% to 98%, 82.5% to 98%, 83% to 98%, 83.5% to 98%, 84% to 98%, 84.5% to 98%, 85% to 98%, 85.5% to 98%, 86% to 98%, 86.5% to 98%, 87% to 98%, 87.5% to 98%, 88% to 98%, 88.5% to 98%, 89% to 98%, 89.5% to 98%, 90% to 98%, 90.3% to 98%, 90.5% to 98%, 90.7% to 98%, 91 % to 98%, 91 .3% to 98%, 91 .5% to 98%, 91 .7% to 98%, 92% to 98%, 92.3% to 98%, 92.5% to 98%, 92.7% to 98%, 93% to 98%, 93.3% to 98%, 93.5% to 98%, 93.7% to 98%, 94% to 98%, 94.3% to 98%, 94.5% to 98%, 94.7% to 98%, 95% to 98%, 95.1 % to 98%, 95.2% to 98%, 95.3% to 98%, 95.4% to 98%, 95.5% to 98% to the amino acid sequence according to SEQ ID NO: 1 and one of the following combinations of amino acid substitutions:

[0550] (a) T38R / K / H / W, Q59K / E and P210I;

[0551] (b) T38R / K / H / W, P210I and S212P / A / G / R / K / F / Y / M;

[0552] (c) Q59K / E, P210I and S212P / A / G / R / K / F / Y / M; (d) T38R / K / H / W, Q59K / E, P210I and S212P / A / G / R / K / F / Y / M;

[0553] (e) T38R / K / H / W, Q59K / E and E271 D;

[0554] (f) T38R / K / H / W, S212 P / A / G / R / K / F / Y / M and E271 D;

[0555] (g) Q59K / E, S212P / A / G / R / K / F / Y / M and E271 D;

[0556] (h) T38R / K / H / W, Q59K / E, S212P / A / G / R / K / F / Y / M and E271 D;

[0557] (i) T38R / K / H / W, Q59K / E, P210I and E271 D;

[0558] Q) T38R / K / H / W, P210I, S212P / A / G / R / K / F / Y / M and E271 D;

[0559] (k) Q59K / E, P210I, S212P / A / G / R / K / F / Y / M and E271 D; or

[0560] (l) T38R / K / H / W, Q59K / E, P210I, S212P / A / G / R / K / F / Y / M and E271 D compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2. In one embodiment the variant polypeptide of the present invention preferably has a sequence identity of 82% to 98%, 82.5% to 98%, 83% to 98%, 83.5% to 98%, 84% to 98%, 84.5% to 98%, 85% to 98%, 85.5% to 98%, 86% to 98%, 86.5% to 98%, 87% to 98%, 87.5% to 98%, 88% to 98%, 88.5% to 98%, 89% to 98%, 89.5% to 98%, 90% to 98%, 90.3% to 98%, 90.5% to 98%, 90.7% to 98%, 91 % to 98%, 91 .3% to 98%, 91 .5% to 98%, 91 .7% to 98%, 92% to 98%, 92.3% to 98%, 92.5% to 98%, 92.7% to 98%, 93% to 98%, 93.3% to 98%, 93.5% to 98%, 93.7% to 98%, 94% to 98%, 94.3% to 98%, 94.5% to 98%, 94.7% to 98%, 95% to 98%, 95.1 % to 98%, 95.2% to 98%, 95.3% to 98%, 95.4% to 98%, 95.5% to 98% to the amino acid sequence according to SEQ ID NO: 1 and one of the following combinations of amino acid substitutions:

[0561] (a) X38R / K / H / W, X59K / E and X210I / V;

[0562] (b) X38R / K / H / W, X210I / V and X212G / M / R / K / P / A / F / H / W / N / Y / D;

[0563] (c) X59K / E, X210I / V and X212G / M / R / K / P / A / F / H / W / N / Y / D;

[0564] (d) X38R / K / H / W, X59K / E, X210I / V and X212G / M / R / K / P / A / F / H / W / N / Y / D ;

[0565] (e) X38R / K / H / W, X59K / E and X271 D;

[0566] (f) X38R / K / H / W, X212G / M / R / K / P / A / F / H / W / N / Y / D and X271 D;

[0567] (g) X59K / E, X212G / M / R / K / P / A / F / H / W / N / Y / D and X271 D;

[0568] (h) X38R / K / H / W, X59K / E, X212G / M / R / K / P / A / F / H / W / N / Y / D and X271 D;

[0569] (I) X38R / K / H / W, X59K / E, X210I / V and X271 D;

[0570] Q) X38R / K / H / W, X210 l / V, X212G / M / R / K / P / A / F / H / W / N / Y / D and X271 D;

[0571] (k) X59K / E, X210I / V, X212G / M / R / K / P / A / F / H / W / N / Y / D and X271 D; or

[0572] (l) X38R / K / H / W, X59K / E, X210I / V, X212G / M / R / K / P / A / F / H / W / N / Y / D and X271 D compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2, wherein said variant polypeptide comprises amino acid residue D or E at position 101 , preferably E at position 101 , referring to the numbering of SEQ ID NO: 2. In one embodiment the variant polypeptide of the present invention preferably has a sequence identity of 82% to 98%, 82.5% to 98%, 83% to 98%, 83.5% to 98%, 84% to 98%, 84.5% to 98%, 85% to 98%, 85.5% to 98%, 86% to 98%, 86.5% to 98%, 87% to 98%, 87.5% to 98%, 88% to 98%, 88.5% to 98%, 89% to 98%, 89.5% to 98%, 90% to 98%, 90.3% to 98%, 90.5% to 98%, 90.7% to 98%, 91 % to 98%, 91 .3% to 98%, 91 .5% to 98%, 91 .7% to 98%, 92% to 98%, 92.3% to 98%, 92.5% to 98%, 92.7% to 98%, 93% to 98%, 93.3% to 98%, 93.5% to 98%, 93.7% to 98%, 94% to 98%, 94.3% to 98%, 94.5% to 98%, 94.7% to 98%, 95% to 98%, 95.1 % to 98%, 95.2% to 98%, 95.3% to 98%, 95.4% to 98%, 95.5% to 98% to the amino acid sequence according to SEQ ID NO: 1 and one of the following combinations of amino acid substitutions:

[0573] (a) T38R / K / H / W, Q59K / E and P210I / V;

[0574] (b) T38R / K / H / W, P210I / V and S212G / M / R / K / P / A / F / H / W / N / Y / D;

[0575] (c) Q59K / E, P210I / V and S212G / M / R / K / P / A / F / H / W / N / Y / D;

[0576] (d) T38R / K / H / W, Q59K / E, P210I / V and S212G / M / R / K / P / A / F / H / W / N / Y / D;

[0577] (e) T38R / K / H / W, Q59K / E and E271 D;

[0578] (f) T38R / K / H / W, S212G / M / R / K / P / A / F / H / W / N / Y / D and E271 D;

[0579] (g) Q59K / E, S212G / M / R / K / P / A / F / H / W / N / Y / D and E271 D;

[0580] (h) T38R / K / H / W, Q59K / E, S212G / M / R / K / P / A / F / H / W / N / Y / D and E271 D;

[0581] (i) T38R / K / H / W, Q59K / E, P210I / V and E271 D;

[0582] Q) T38R / K / H / W, P210I / V, S212G / M / R / K / P / A / F / H / W / N / Y / D and E271 D;

[0583] (k) Q59K / E, P210I / V, S212G / M / R / K / P / A / F / H / W / N / Y / D and E271 D; or

[0584] (l) T38R / K / H / W, Q59K / E, P210I / V, S212G / M / R / K / P / A / F / H / W / N / Y / D and E271 D compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2, wherein said variant polypeptide comprises amino acid residue D or E at position 101 , preferably E at position 101 , referring to the numbering of SEQ ID NO: 2.

[0585] In one embodiment the variant polypeptide of the present invention preferably has a sequence identity of 82% to 98%, 82.5% to 98%, 83% to 98%, 83.5% to 98%, 84% to 98%, 84.5% to 98%, 85% to 98%, 85.5% to 98%, 86% to 98%, 86.5% to 98%, 87% to 98%, 87.5% to 98%, 88% to 98%, 88.5% to 98%, 89% to 98%, 89.5% to 98%, 90% to 98%, 90.3% to 98%, 90.5% to 98%, 90.7% to 98%, 91 % to 98%, 91 .3% to 98%, 91 .5% to 98%, 91 .7% to 98%, 92% to 98%, 92.3% to 98%, 92.5% to 98%, 92.7% to 98%, 93% to 98%, 93.3% to 98%, 93.5% to 98%, 93.7% to 98%, 94% to 98%, 94.3% to 98%, 94.5% to 98%, 94.7% to 98%, 95% to 98%, 95.1 % to 98%, 95.2% to 98%, 95.3% to 98%, 95.4% to 98%, 95.5% to 98% to the amino acid sequence according to SEQ ID NO: 1 and one of the following combinations of amino acid substitutions:

[0586] (a) X38R / K / H / W, X59K / E and X210I;

[0587] (b) X38R / K / H / W, X210I and X212P / A / G / R / K / F / Y / M;

[0588] (c) X59K / E, X210I and X212P / A / G / R / K / F / Y / M ; (d) X38R / K / H / W, X59K / E, X21 Ol and X212P / A / G / R / K / F / Y / M;

[0589] (e) X38R / K / H / W, X59K / E and X271 D;

[0590] (f) X38R / K / H / W, X212 P / A / G / R / K / F / Y / M and X271 D;

[0591] (g) X59K / E, X212P / A / G / R / K / F / Y / M and X271 D;

[0592] (h) X38R / K / H / W, X59K / E, X212P / A / G / R / K / F / Y / M and X271 D;

[0593] (i) X38R / K / H / W, X59K / E, X210I and X271 D;

[0594] Q) X38R / K / H / W, X21 Ol, X212P / A / G / R / K / F / Y / M and X271 D;

[0595] (k) X59K / E, X210I, X212P / A / G / R / K / F / Y / M and X271 D; or

[0596] (l) X38R / K / H / W, X59K / E, X210I, X212 P / A / G / R / K / F / Y / M and X271 D compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2, wherein said variant polypeptide comprises amino acid residue D or E at position 101 , preferably E at position 101 , referring to the numbering of SEQ ID NO: 2.

[0597] In one embodiment the variant polypeptide of the present invention preferably has a sequence identity of 82% to 98%, 82.5% to 98%, 83% to 98%, 83.5% to 98%, 84% to 98%, 84.5% to 98%, 85% to 98%, 85.5% to 98%, 86% to 98%, 86.5% to 98%, 87% to 98%, 87.5% to 98%, 88% to 98%, 88.5% to 98%, 89% to 98%, 89.5% to 98%, 90% to 98%, 90.3% to 98%, 90.5% to 98%, 90.7% to 98%, 91 % to 98%, 91 .3% to 98%, 91 .5% to 98%, 91 .7% to 98%, 92% to 98%, 92.3% to 98%, 92.5% to 98%, 92.7% to 98%, 93% to 98%, 93.3% to 98%, 93.5% to 98%, 93.7% to 98%, 94% to 98%, 94.3% to 98%, 94.5% to 98%, 94.7% to 98%, 95% to 98%, 95.1 % to 98%, 95.2% to 98%, 95.3% to 98%, 95.4% to 98%, 95.5% to 98% to the amino acid sequence according to SEQ ID NO: 1 and one of the following combinations of amino acid substitutions:

[0598] (a) T38R / K / H / W, Q59K / E and P210I;

[0599] (b) T38R / K / H / W, P210I and S212P / A / G / R / K / F / Y / M;

[0600] (c) Q59K / E, P210I and S212P / A / G / R / K / F / Y / M;

[0601] (d) T38R / K / H / W, Q59K / E, P210I and S212P / A / G / R / K / F / Y / M;

[0602] (e) T38R / K / H / W, Q59K / E and E271 D;

[0603] (f) T38R / K / H / W, S212P / A / G / R / K / F / Y / M and E271 D;

[0604] (g) Q59K / E, S212P / A / G / R / K / F / Y / M and E271 D;

[0605] (h) T38R / K / H / W, Q59K / E, S212P / A / G / R / K / F / Y / M and E271 D;

[0606] (i) T38R / K / H / W, Q59K / E, P210I and E271 D;

[0607] Q) T38R / K / H / W, P210I, S212P / A / G / R / K / F / Y / M and E271 D;

[0608] (k) Q59K / E, P210I, S212P / A / G / R / K / F / Y / M and E271 D; or

[0609] (l) T38R / K / H / W, Q59K / E, P210I, S212P / A / G / R / K / F / Y / M and E271 D compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2, wherein said variant polypeptide comprises amino acid residue D or E at position 101 , preferably E at position 101 , referring to the numbering of SEQ ID NO: 2.

[0610] In one embodiment the variant polypeptide of the present invention preferably has a sequence identity of 82% to 98 to the amino acid sequence according to SEQ ID NO: 1 and one of the following combinations of amino acid substitutions:

[0611] (a) X38R / K / H / W, X59K / E and X210I / V;

[0612] (b) X38R / K / H / W, X210I / V and X212G / M / R / K / P / A / F / H / W / N / Y / D;

[0613] (c) X59K / E, X210 l / V and X212G / M / R / K / P / A / F / H / W / N / Y / D;

[0614] (d) X38R / K / H / W, X59K / E, X210I / V and X212G / M / R / K / P / A / F / H / W / N / Y / D;

[0615] (e) X38R / K / H / W, X59K / E and X271 D;

[0616] (f) X38R / K / H / W, X212G / M / R / K / P / A / F / H / W / N / Y / D and X271 D;

[0617] (g) X59K / E, X212G / M / R / K / P / A / F / H / W / N / Y / D and X271 D;

[0618] (h) X38R / K / H / W, X59K / E, X212G / M / R / K / P / A / F / H / W / N / Y / D and X271 D;

[0619] (I) X38R / K / H / W, X59K / E, X210I / V and X271 D;

[0620] Q) X38R / K / H / W, X210I / V, X212G / M / R / K / P / A / F / H / W / N / Y / D and X271 D;

[0621] (k) X59K / E, X210I / V, X212G / M / R / K / P / A / F / H / W / N / Y / D and X271 D; or

[0622] (l) X38R / K / H / W, X59K / E, X210I / V, X212G / M / R / K / P / A / F / H / W / N / Y / D and X271 D compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2. In one embodiment the variant polypeptide of the present invention preferably has a sequence identity of 82% to 98% to the amino acid sequence according to SEQ ID NO: 1 and one of the following combinations of amino acid substitutions:

[0623] (a) T38R / K / H / W, Q59K / E and P210I / V;

[0624] (b) T38R / K / H / W, P210I / V and S212G / M / R / K / P / A / F / H / W / N / Y / D;

[0625] (c) Q59K / E, P210I / V and S212G / M / R / K / P / A / F / H / W / N / Y / D;

[0626] (d) T38R / K / H / W, Q59K / E, P210I / V and S212G / M / R / K / P / A / F / H / W / N / Y / D;

[0627] (e) T38R / K / H / W, Q59K / E and E271 D;

[0628] (f) T38R / K / H / W, S212G / M / R / K / P / A / F / H / W / N / Y / D and E271 D;

[0629] (g) Q59K / E, S212G / M / R / K / P / A / F / H / W / N / Y / D and E271 D;

[0630] (h) T38R / K / H / W, Q59K / E, S212G / M / R / K / P / A / F / H / W / N / Y / D and E271 D;

[0631] (I) T38R / K / H / W, Q59K / E, P210I / V and E271 D;

[0632] Q) T38R / K / H / W, P210I / V, S212G / M / R / K / P / A / F / H / W / N / Y / D and E271 D;

[0633] (k) Q59K / E, P210I / V, S212G / M / R / K / P / A / F / H / W / N / Y / D and E271 D; or

[0634] (l) T38R / K / H / W, Q59K / E, P210I / V, S212G / M / R / K / P / A / F / H / W / N / Y / D and E271 D compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2. In one embodiment the variant polypeptide of the present invention preferably has a sequence identity of 82% to 98% to the amino acid sequence according to SEQ ID NO: 1 and one of the following combinations of amino acid substitutions:

[0635] (a) X38R / K / H / W, X59K / E and X210I;

[0636] (b) X38R / K / H / W, X210I and X212P / A / G / R / K / F / Y / M;

[0637] (c) X59K / E, X21 Ol and X212P / A / G / R / K / F / Y / M;

[0638] (d) X38R / K / H / W, X59K / E, X21 Ol and X212P / A / G / R / K / F / Y / M;

[0639] (e) X38R / K / H / W, X59K / E and X271 D;

[0640] (f) X38R / K / H / W, X212P / A / G / R / K / F / Y / M and X271 D;

[0641] (g) X59K / E, X212P / A / G / R / K / F / Y / M and X271 D;

[0642] (h) X38R / K / H / W, X59K / E, X212P / A / G / R / K / F / Y / M and X271 D;

[0643] (i) X38R / K / H / W, X59K / E, X210I and X271 D;

[0644] Q) X38R / K / H / W, X21 Ol, X212P / A / G / R / K / F / Y / M and X271 D;

[0645] (k) X59K / E, X210I, X212P / A / G / R / K / F / Y / M and X271 D; or

[0646] (l) X38R / K / H / W, X59K / E, X210I, X212P / A / G / R / K / F / Y / M and X271 D compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2. In one embodiment the variant polypeptide of the present invention preferably has a sequence identity of 82% to 98% to the amino acid sequence according to SEQ ID NO: 1 and one of the following combinations of amino acid substitutions:

[0647] (a) T38R / K / H / W, Q59K / E and P210I;

[0648] (b) T38R / K / H / W, P210I and S212P / A / G / R / K / F / Y / M;

[0649] (c) Q59K / E, P210I and S212P / A / G / R / K / F / Y / M;

[0650] (d) T38R / K / H / W, Q59K / E, P210I and S212P / A / G / R / K / F / Y / M;

[0651] (e) T38R / K / H / W, Q59K / E and E271 D;

[0652] (f) T38R / K / H / W, S212P / A / G / R / K / F / Y / M and E271 D;

[0653] (g) Q59K / E, S212P / A / G / R / K / F / Y / M and E271 D;

[0654] (h) T38R / K / H / W, Q59K / E, S212P / A / G / R / K / F / Y / M and E271 D;

[0655] (i) T38R / K / H / W, Q59K / E, P210I and E271 D;

[0656] Q) T38R / K / H / W, P210I, S212P / A / G / R / K / F / Y / M and E271 D;

[0657] (k) Q59K / E, P210I, S212P / A / G / R / K / F / Y / M and E271 D; or

[0658] (l) T38R / K / H / W, Q59K / E, P21 Ol, S212P / A / G / R / K / F / Y / M and E271 D compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2. In one embodiment the variant polypeptide of the present invention preferably has a sequence identity of 82% to 98 to the amino acid sequence according to SEQ ID NO: 1 and one of the following combinations of amino acid substitutions:

[0659] (a) X38R / K / H / W, X59K / E and X21 Ol / V;

[0660] (b) X38R / K / H / W, X210I / V and X212G / M / R / K / P / A / F / H / W / N / Y / D;

[0661] (c) X59K / E, X21 Ol / V and X212G / M / R / K / P / A / F / H / W / N / Y / D;

[0662] (d) X38R / K / H / W, X59K / E, X21 Ol / V and X212G / M / R / K / P / A / F / H / VV / N / Y / D;

[0663] (e) X38R / K / H / W, X59K / E and X271 D;

[0664] (f) X38R / K / H / W, X212G / M / R / K / P / A / F / H / W / N / Y / D and X271 D;

[0665] (g) X59K / E, X212G / M / R / K / P / A / F / H / W / N / Y / D and X271 D;

[0666] (h) X38R / K / H / W, X59K / E, X212G / M / R / K / P / A / F / H / W / N / Y / D and X271 D;

[0667] (i) X38R / K / H / W, X59K / E, X210I / V and X271 D;

[0668] Q) X38R / K / H / W, X21 Ol / V, X212G / M / R / K / P / A / F / H / W / N / Y / D and X271 D;

[0669] (k) X59K / E, X210I / V, X212G / M / R / K / P / A / F / H / W / N / Y / D and X271 D; or

[0670] (l) X38R / K / H / W, X59K / E, X21 Ol / V, X212G / M / R / K / P / A / F / H / W / N / Y / D and X271 D compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2, wherein said variant polypeptide comprises amino acid residue D or E at position 101 , preferably E at position 101 , referring to the numbering of SEQ ID NO: 2.

[0671] In one embodiment the variant polypeptide of the present invention preferably has a sequence identity of 82% to 98% to the amino acid sequence according to SEQ ID NO: 1 and one of the following combinations of amino acid substitutions:

[0672] (a) T38R / K / H / W, Q59K / E and P21 Ol / V;

[0673] (b) T38R / K / H / W, P210I / V and S212G / M / R / K / P / A / F / H / W / N / Y / D;

[0674] (c) Q59K / E, P21 Ol / V and S212G / M / R / K / P / A / F / H / W / N / Y / D;

[0675] (d) T38R / K / H / W, Q59K / E, P210I / V and S212G / M / R / K / P / A / F / H / W / N / Y / D;

[0676] (e) T38R / K / H / W, Q59K / E and E271 D;

[0677] (f) T38R / K / H / W, S212G / M / R / K / P / A / F / H / W / N / Y / D and E271 D;

[0678] (g) Q59K / E, S212G / M / R / K / P / A / F / H / W / N / Y / D and E271 D;

[0679] (h) T38R / K / H / W, Q59K / E, S212G / M / R / K / P / A / F / H / W / N / Y / D and E271 D;

[0680] (i) T38R / K / H / W, Q59K / E, P210I / V and E271 D;

[0681] Q) T38R / K / H / W, P210I / V, S212G / M / R / K / P / A / F / H / W / N / Y / D and E271 D;

[0682] (k) Q59K / E, P210I / V, S212G / M / R / K / P / A / F / H / W / N / Y / D and E271 D; or

[0683] (l) T38R / K / H / W, Q59K / E, P21 Ol / V, S212G / M / R / K / P / A / F / H / W / N / Y / D and E271 D compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2, wherein said variant polypeptide comprises amino acid residue D or E at position 101 , preferably E at position 101 , referring to the numbering of SEQ ID NO: 2.

[0684] In one embodiment the variant polypeptide of the present invention preferably has a sequence identity of 82% to 98% to the amino acid sequence according to SEQ ID NO: 1 and one of the following combinations of amino acid substitutions:

[0685] (a) X38R / K / H / W, X59K / E and X210I;

[0686] (b) X38R / K / H / W, X210I and X212P / A / G / R / K / F / Y / M;

[0687] (c) X59K / E, X210I and X212P / A / G / R / K / F / Y / M;

[0688] (d) X38R / K / H / W, X59K / E, X210I and X212P / A / G / R / K / F / Y / M;

[0689] (e) X38R / K / H / W, X59K / E and X271 D;

[0690] (f) X38R / K / H / W, X212P / A / G / R / K / F / Y / M and X271 D;

[0691] (g) X59K / E, X212P / A / G / R / K / F / Y / M and X271 D;

[0692] (h) X38R / K / H / W, X59K / E, X212P / A / G / R / K / F / Y / M and X271 D;

[0693] (I) X38R / K / H / W, X59K / E, X210I and X271 D;

[0694] Q) X38R / K / H / W, X210I, X212P / A / G / R / K / F / Y / M and X271 D;

[0695] (k) X59K / E, X210I, X212P / A / G / R / K / F / Y / M and X271 D; or

[0696] (l) X38R / K / H / W, X59K / E, X210I, X212P / A / G / R / K / F / Y / M and X271 D compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2, wherein said variant polypeptide comprises amino acid residue D or E at position 101 , preferably E at position 101 , referring to the numbering of SEQ ID NO: 2.

[0697] In one embodiment the variant polypeptide of the present invention preferably has a sequence identity of 82% to 98% to the amino acid sequence according to SEQ ID NO: 1 and one of the following combinations of amino acid substitutions:

[0698] (a) T38R / K / H / W, Q59K / E and P210I;

[0699] (b) T38R / K / H / W, P210I and S212P / A / G / R / K / F / Y / M;

[0700] (c) Q59K / E, P210I and S212P / A / G / R / K / F / Y / M;

[0701] (d) T38R / K / H / W, Q59K / E, P210I and S212P / A / G / R / K / F / Y / M;

[0702] (e) T38R / K / H / W, Q59K / E and E271 D;

[0703] (f) T38R / K / H / W, S212P / A / G / R / K / F / Y / M and E271 D;

[0704] (g) Q59K / E, S212P / A / G / R / K / F / Y / M and E271 D;

[0705] (h) T38R / K / H / W, Q59K / E, S212P / A / G / R / K / F / Y / M and E271 D;

[0706] (I) T38R / K / H / W, Q59K / E, P210I and E271 D;

[0707] Q) T38R / K / H / W, P210I, S212P / A / G / R / K / F / Y / M and E271 D;

[0708] (k) Q59K / E, P210I, S212P / A / G / R / K / F / Y / M and E271 D; or (I) T38R / K / H / W, Q59K / E, P21 Ol, S212P / A / G / R / K / F / Y / M and E271 D compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2, wherein said variant polypeptide comprises amino acid residue D or E at position 101 , preferably E at position 101 , referring to the numbering of SEQ ID NO: 2.

[0709] In one embodiment the variant polypeptide of the present invention preferably has a sequence identity of 95.5% to 98% to the amino acid sequence according to SEQ ID NO: 1 and one of the following combinations of amino acid substitutions:

[0710] (a) X38R / K / H / W, X59K / E and X210I / V;

[0711] (b) X38R / K / H / W, X210I / V and X212G / M / R / K / P / A / F / H / W / N / Y / D;

[0712] (c) X59K / E, X210 l / V and X212G / M / R / K / P / A / F / H / W / N / Y / D;

[0713] (d) X38R / K / H / W, X59K / E, X210I / V and X212G / M / R / K / P / A / F / H / W / N / Y / D;

[0714] (e) X38R / K / H / W, X59K / E and X271 D;

[0715] (f) X38R / K / H / W, X212G / M / R / K / P / A / F / H / W / N / Y / D and X271 D;

[0716] (g) X59K / E, X212G / M / R / K / P / A / F / H / W / N / Y / D and X271 D;

[0717] (h) X38R / K / H / W, X59K / E, X212G / M / R / K / P / A / F / H / W / N / Y / D and X271 D;

[0718] (I) X38R / K / H / W, X59K / E, X210I / V and X271 D;

[0719] Q) X38R / K / H / W, X210I / V, X212G / M / R / K / P / A / F / H / W / N / Y / D and X271 D;

[0720] (k) X59K / E, X210I / V, X212G / M / R / K / P / A / F / H / W / N / Y / D and X271 D; or

[0721] (l) X38R / K / H / W, X59K / E, X210I / V, X212G / M / R / K / P / A / F / H / W / N / Y / D and X271 D compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2. In one embodiment the variant polypeptide of the present invention preferably has a sequence identity of 95.5% to 98% to the amino acid sequence according to SEQ ID NO: 1 and one of the following combinations of amino acid substitutions:

[0722] (a) T38R / K / H / W, Q59K / E and P210I / V;

[0723] (b) T38R / K / H / W, P210I / V and S212G / M / R / K / P / A / F / H / W / N / Y / D;

[0724] (c) Q59K / E, P210I / V and S212G / M / R / K / P / A / F / H / W / N / Y / D;

[0725] (d) T38R / K / H / W, Q59K / E, P210I / V and S212G / M / R / K / P / A / F / H / W / N / Y / D;

[0726] (e) T38R / K / H / W, Q59K / E and E271 D;

[0727] (f) T38R / K / H / W, S212G / M / R / K / P / A / F / H / W / N / Y / D and E271 D;

[0728] (g) Q59K / E, S212G / M / R / K / P / A / F / H / W / N / Y / D and E271 D;

[0729] (h) T38R / K / H / W, Q59K / E, S212G / M / R / K / P / A / F / H / W / N / Y / D and E271 D;

[0730] (I) T38R / K / H / W, Q59K / E, P210I / V and E271 D;

[0731] Q) T38R / K / H / W, P210I / V, S212G / M / R / K / P / A / F / H / W / N / Y / D and E271 D;

[0732] (k) Q59K / E, P210I / V, S212G / M / R / K / P / A / F / H / W / N / Y / D and E271 D; or (I) T38R / K / H / W, Q59K / E, P21 Ol / V, S212G / M / R / K / P / A / F / H / W / N / Y / D and E271 D compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2. In one embodiment the variant polypeptide of the present invention preferably has a sequence identity of 95.5% to 98 % to the amino acid sequence according to SEQ ID NO: 1 and one of the following combinations of amino acid substitutions:

[0733] (a) X38R / K / H / W, X59K / E and X210I;

[0734] (b) X38R / K / H / W, X210I and X212P / A / G / R / K / F / Y / M;

[0735] (c) X59K / E, X21 Ol and X212P / A / G / R / K / F / Y / M;

[0736] (d) X38R / K / H / W, X59K / E, X21 Ol and X212P / A / G / R / K / F / Y / M;

[0737] (e) X38R / K / H / W, X59K / E and X271 D;

[0738] (f) X38R / K / H / W, X212P / A / G / R / K / F / Y / M and X271 D;

[0739] (g) X59K / E, X212P / A / G / R / K / F / Y / M and X271 D;

[0740] (h) X38R / K / H / W, X59K / E, X212P / A / G / R / K / F / Y / M and X271 D;

[0741] (I) X38R / K / H / W, X59K / E, X210I and X271 D;

[0742] Q) X38R / K / H / W, X21 Ol, X212P / A / G / R / K / F / Y / M and X271 D;

[0743] (k) X59K / E, X210I, X212P / A / G / R / K / F / Y / M and X271 D; or

[0744] (l) X38R / K / H / W, X59K / E, X210I, X212P / A / G / R / K / F / Y / M and X271 D compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2. In one embodiment the variant polypeptide of the present invention preferably has a sequence identity of 95.5% to 98% to the amino acid sequence according to SEQ ID NO: 1 and one of the following combinations of amino acid substitutions:

[0745] (a) T38R / K / H / W, Q59K / E and P210I;

[0746] (b) T38R / K / H / W, P210I and S212P / A / G / R / K / F / Y / M;

[0747] (c) Q59K / E, P210I and S212P / A / G / R / K / F / Y / M;

[0748] (d) T38R / K / H / W, Q59K / E, P210I and S212P / A / G / R / K / F / Y / M;

[0749] (e) T38R / K / H / W, Q59K / E and E271 D;

[0750] (f) T38R / K / H / W, S212P / A / G / R / K / F / Y / M and E271 D;

[0751] (g) Q59K / E, S212P / A / G / R / K / F / Y / M and E271 D;

[0752] (h) T38R / K / H / W, Q59K / E, S212P / A / G / R / K / F / Y / M and E271 D;

[0753] (I) T38R / K / H / W, Q59K / E, P210I and E271 D;

[0754] Q) T38R / K / H / W, P210I, S212P / A / G / R / K / F / Y / M and E271 D;

[0755] (k) Q59K / E, P210I, S212P / A / G / R / K / F / Y / M and E271 D; or

[0756] (l) T38R / K / H / W, Q59K / E, P210I, S212P / A / G / R / K / F / Y / M and E271 D compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2. In one embodiment the variant polypeptide of the present invention preferably has a sequence identity of 95.5% to 98% to the amino acid sequence according to SEQ ID NO: 1 and one of the following combinations of amino acid substitutions:

[0757] (a) X38R / K / H / W, X59K / E and X21 Ol / V;

[0758] (b) X38R / K / H / W, X210I / V and X212G / M / R / K / P / A / F / H / W / N / Y / D;

[0759] (c) X59K / E, X21 Ol / V and X212G / M / R / K / P / A / F / H / W / N / Y / D;

[0760] (d) X38R / K / H / W, X59K / E, X21 Ol / V and X212G / M / R / K / P / A / F / H / VV / N / Y / D;

[0761] (e) X38R / K / H / W, X59K / E and X271 D;

[0762] (f) X38R / K / H / W, X212G / M / R / K / P / A / F / H / W / N / Y / D and X271 D;

[0763] (g) X59K / E, X212G / M / R / K / P / A / F / H / W / N / Y / D and X271 D;

[0764] (h) X38R / K / H / W, X59K / E, X212G / M / R / K / P / A / F / H / W / N / Y / D and X271 D;

[0765] (i) X38R / K / H / W, X59K / E, X210I / V and X271 D;

[0766] Q) X38R / K / H / W, X21 Ol / V, X212G / M / R / K / P / A / F / H / W / N / Y / D and X271 D;

[0767] (k) X59K / E, X210I / V, X212G / M / R / K / P / A / F / H / W / N / Y / D and X271 D; or

[0768] (l) X38R / K / H / W, X59K / E, X21 Ol / V, X212G / M / R / K / P / A / F / H / W / N / Y / D and X271 D compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2, wherein said variant polypeptide comprises amino acid residue D or E at position 101 , preferably E at position 101 , referring to the numbering of SEQ ID NO: 2.

[0769] In one embodiment the variant polypeptide of the present invention preferably has a sequence identity of 95.5% to 98% to the amino acid sequence according to SEQ ID NO: 1 and one of the following combinations of amino acid substitutions:

[0770] (a) T38R / K / H / W, Q59K / E and P21 Ol / V;

[0771] (b) T38R / K / H / W, P210I / V and S212G / M / R / K / P / A / F / H / W / N / Y / D;

[0772] (c) Q59K / E, P21 Ol / V and S212G / M / R / K / P / A / F / H / W / N / Y / D;

[0773] (d) T38R / K / H / W, Q59K / E, P210I / V and S212G / M / R / K / P / A / F / H / W / N / Y / D;

[0774] (e) T38R / K / H / W, Q59K / E and E271 D;

[0775] (f) T38R / K / H / W, S212G / M / R / K / P / A / F / H / W / N / Y / D and E271 D;

[0776] (g) Q59K / E, S212G / M / R / K / P / A / F / H / W / N / Y / D and E271 D;

[0777] (h) T38R / K / H / W, Q59K / E, S212G / M / R / K / P / A / F / H / W / N / Y / D and E271 D;

[0778] (i) T38R / K / H / W, Q59K / E, P210I / V and E271 D;

[0779] Q) T38R / K / H / W, P210I / V, S212G / M / R / K / P / A / F / H / W / N / Y / D and E271 D;

[0780] (k) Q59K / E, P210I / V, S212G / M / R / K / P / A / F / H / W / N / Y / D and E271 D; or

[0781] (l) T38R / K / H / W, Q59K / E, P21 Ol / V, S212G / M / R / K / P / A / F / H / W / N / Y / D and E271 D compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2, wherein said variant polypeptide comprises amino acid residue D or E at position 101 , preferably E at position 101 , referring to the numbering of SEQ ID NO: 2.

[0782] In one embodiment the variant polypeptide of the present invention preferably has a sequence identity of 95.5% to 98 % to the amino acid sequence according to SEQ ID NO: 1 and one of the following combinations of amino acid substitutions:

[0783] (a) X38R / K / H / W, X59K / E and X210I;

[0784] (b) X38R / K / H / W, X210I and X212P / A / G / R / K / F / Y / M;

[0785] (c) X59K / E, X210I and X212P / A / G / R / K / F / Y / M;

[0786] (d) X38R / K / H / W, X59K / E, X210I and X212P / A / G / R / K / F / Y / M;

[0787] (e) X38R / K / H / W, X59K / E and X271 D;

[0788] (f) X38R / K / H / W, X212P / A / G / R / K / F / Y / M and X271 D;

[0789] (g) X59K / E, X212P / A / G / R / K / F / Y / M and X271 D;

[0790] (h) X38R / K / H / W, X59K / E, X212P / A / G / R / K / F / Y / M and X271 D;

[0791] (I) X38R / K / H / W, X59K / E, X210I and X271 D;

[0792] Q) X38R / K / H / W, X210I, X212P / A / G / R / K / F / Y / M and X271 D;

[0793] (k) X59K / E, X210I, X212P / A / G / R / K / F / Y / M and X271 D; or

[0794] (l) X38R / K / H / W, X59K / E, X210I, X212P / A / G / R / K / F / Y / M and X271 D compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2, wherein said variant polypeptide comprises amino acid residue D or E at position 101 , preferably E at position 101 , referring to the numbering of SEQ ID NO: 2.

[0795] In one embodiment the variant polypeptide of the present invention preferably has a sequence identity of 95.5% to 98% to the amino acid sequence according to SEQ ID NO: 1 and one of the following combinations of amino acid substitutions:

[0796] (a) T38R / K / H / W, Q59K / E and P210I;

[0797] (b) T38R / K / H / W, P210I and S212P / A / G / R / K / F / Y / M;

[0798] (c) Q59K / E, P210I and S212P / A / G / R / K / F / Y / M;

[0799] (d) T38R / K / H / W, Q59K / E, P210I and S212P / A / G / R / K / F / Y / M;

[0800] (e) T38R / K / H / W, Q59K / E and E271 D;

[0801] (f) T38R / K / H / W, S212P / A / G / R / K / F / Y / M and E271 D;

[0802] (g) Q59K / E, S212P / A / G / R / K / F / Y / M and E271 D;

[0803] (h) T38R / K / H / W, Q59K / E, S212P / A / G / R / K / F / Y / M and E271 D;

[0804] (I) T38R / K / H / W, Q59K / E, P210I and E271 D;

[0805] Q) T38R / K / H / W, P210I, S212P / A / G / R / K / F / Y / M and E271 D;

[0806] (k) Q59K / E, P210I, S212P / A / G / R / K / F / Y / M and E271 D; or (I) T38R / K / H / W, Q59K / E, P21 Ol, S212P / A / G / R / K / F / Y / M and E271 D compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2, wherein said variant polypeptide comprises amino acid residue D or E at position 101 , preferably E at position 101 , referring to the numbering of SEQ ID NO: 2.

[0807] In one embodiment the variant polypeptide of the present invention or a fragment of said polypeptide having protease activity additionally comprises at least one amino acid substitution at an amino acid residue selected from the group consisting of 9, 18, 21 , 43, 78, 160, 183, 194, 204, 218, 228, 259, 260 and 261 compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2.

[0808] Hence, in one embodiment the present invention relates to a variant polypeptide having protease activity or a fragment of said polypeptide having protease activity, wherein:

[0809] (I) the polypeptide has an amino acid sequence which is at least 82%, but less than 100%, identical to the amino acid sequence as set forth in SEQ ID NO: 1 and

[0810] (ii) the polypeptide or fragment thereof comprises an amino acid substitution at two or more amino acid residues selected from the group consisting of 38, 59 and 212 compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2,

[0811] (ill) the polypeptide or fragment thereof comprises at least one amino acid substitution at an amino acid residue selected from the group consisting of 9, 18, 21 , 43, 78, 160, 183, 194, 204, 218, 228, 259, 260 and 261 compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2, wherein the amino acid substitutions 38F, 38H, 38N, 59A, 59H, 59N, 59S, 212H and 212M are excluded and wherein the fragment of the variant polypeptide comprises 100 to 259 consecutive amino acids of the full-length variant polypeptide.

[0812] In one embodiment the variant polypeptide of the present invention preferably has a sequence identity of 95.5% to 98% to the amino acid sequence according to SEQ ID NO: 1 and one of the following combinations of amino acid substitutions:

[0813] (a) X38R / K / H / W, X59K / E and X210 l / V;

[0814] (b) X38R / K / H / W, X210I / V and X212G / M / R / K / P / A / F / H / W / N / Y / D;

[0815] (c) X59K / E, X210I / V and X212G / M / R / K / P / A / F / H / W / N / Y / D;

[0816] (d) X38R / K / H / W, X59K / E, X210I / V and X212G / M / R / K / P / A / F / H / VV / N / Y / D;

[0817] (e) X38R / K / H / W, X59K / E and X271 D;

[0818] (f) X38R / K / H / W, X212G / M / R / K / P / A / F / H / W / N / Y / D and X271 D;

[0819] (g) X59K / E, X212G / M / R / K / P / A / F / H / W / N / Y / D and X271 D;

[0820] (h) X38R / K / H / W, X59K / E, X212G / M / R / K / P / A / F / H / W / N / Y / D and X271 D;

[0821] (I) X38R / K / H / W, X59K / E, X210I / V and X271 D;

[0822] Q) X38R / K / H / W, X210I / V, X212G / M / R / K / P / A / F / H / W / N / Y / D and X271 D; (k) X59K / E, X210I / V, X212G / M / R / K / P / A / F / H / W / N / Y / D and X271 D; or

[0823] (l) X38R / K / H / W, X59K / E, X21 Ol / V, X212G / M / R / K / P / A / F / H / W / N / Y / D and X271 D compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2, wherein the variant polypeptide further comprises at least one amino acid substitution at an amino acid residue selected from the group consisting of 9, 18, 21 , 43, 78, 160, 183, 194, 204, 218, 228, 259, 260 and 261 compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2.

[0824] In one embodiment the present invention relates to a variant polypeptide having protease activity or a fragment of said polypeptide having protease activity, wherein:

[0825] (I) the polypeptide has an amino acid sequence which is at least 82%, but less than 100%, identical to the amino acid sequence as set forth in SEQ ID NO: 1 and

[0826] (ii) the polypeptide or fragment thereof comprises an amino acid substitution at two or more amino acid residues selected from the group consisting of 38, 59 and 212 compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2,

[0827] (ill) the polypeptide or fragment thereof comprises at least one amino acid substitution at an amino acid residue selected from the group consisting of 9, 18, 21 , 43, 78, 160, 183, 194, 204, 218, 228, 259, 260 and 261 compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2, wherein the amino acid substitutions 38F, 38H, 38N, 59A, 59H, 59N, 59S, 212H and 212M are excluded, wherein the fragment of the variant polypeptide comprises 100 to 259 consecutive amino acids of the full-length variant polypeptide and wherein said variant polypeptide comprises amino acid residue D or E at position 101 , preferably E at position 101 , referring to the numbering of SEQ ID NO: 2.

[0828] In one embodiment the variant polypeptide of the present invention preferably has a sequence identity of 95.5% to 98% to the amino acid sequence according to SEQ ID NO: 1 and one of the following combinations of amino acid substitutions:

[0829] (a) X38R / K / H / W, X59K / E and X210I / V;

[0830] (b) X38R / K / H / W, X210I / V and X212G / M / R / K / P / A / F / H / W / N / Y / D;

[0831] (c) X59K / E, X210I / V and X212G / M / R / K / P / A / F / H / W / N / Y / D;

[0832] (d) X38R / K / H / W, X59K / E, X210I / V and X212G / M / R / K / P / A / F / H / W / N / Y / D;

[0833] (e) X38R / K / H / W, X59K / E and X271 D;

[0834] (f) X38R / K / H / W, X212G / M / R / K / P / A / F / H / W / N / Y / D and X271 D;

[0835] (g) X59K / E, X212G / M / R / K / P / A / F / H / W / N / Y / D and X271 D;

[0836] (h) X38R / K / H / W, X59K / E, X212G / M / R / K / P / A / F / H / W / N / Y / D and X271 D;

[0837] (I) X38R / K / H / W, X59K / E, X210I / V and X271 D;

[0838] Q) X38R / K / H / W, X210I / V, X212G / M / R / K / P / A / F / H / W / N / Y / D and X271 D;

[0839] (k) X59K / E, X210I / V, X212G / M / R / K / P / A / F / H / W / N / Y / D and X271 D; or (I) X38R / K / H / W, X59K / E, X21 Ol / V, X212G / M / R / K / P / A / F / H / W / N / Y / D and X271 D compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2, wherein the variant polypeptide further comprises at least one amino acid substitution at an amino acid residue selected from the group consisting of 9, 18, 21 , 43, 78, 160, 183, 194, 204, 218, 228, 259, 260 and 261 compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2 and wherein said variant polypeptide comprises amino acid residue D or E at position 101 , preferably E at position 101 , referring to the numbering of SEQ ID NO: 2.

[0840] In one embodiment the variant polypeptide of the present invention or a fragment of said polypeptide having protease activity additionally comprises at least one amino acid substitution selected from the group consisting of X9A / Q / E / C / D, X18Q / E / D, X21 l / V / F / W / Y, X43K / R / C / H / D / L / S / W / A / M / Y / Q / F / I, X78N / D / R / W / F / H / K / E / L / Y / M / C / Q, X160H / S / D / E / P, X183D / E / C / Q / A / M, X194E / C / D, X204D / E / C / G, X218T / S / D, X228S, X259E / D, X260D / E / K and X261 L / M / F / E / W / Y / C compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2. Hence, in one embodiment the present invention relates to a variant polypeptide having protease activity or a fragment of said polypeptide having protease activity, wherein:

[0841] (I) the polypeptide has an amino acid sequence which is at least 82%, but less than 100%, identical to the amino acid sequence as set forth in SEQ ID NO: 1 and

[0842] (ii) the polypeptide or fragment thereof comprises an amino acid substitution at two or more amino acid residues selected from the group consisting of 38, 59 and 212 compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2,

[0843] (ill) the polypeptide or fragment thereof comprises at least one amino acid substitution selected from the group consisting of X9A / Q / E / C / D, X18Q / E / D, X21 l / V / F / W / Y, X43K / R / C / H / D / L / S / W / A / M / Y / Q / F / I,

[0844] X78N / D / R / W / F / H / K / E / L / Y / M / C / Q, X160H / S / D / E / P, X183D / E / C / Q / A / M, X194E / C / D, X204D / E / C / G, X218T / S / D, X228S, X259E / D, X260D / E / K and X261 L / M / F / E / W / Y / C compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2, wherein the amino acid substitutions 38F, 38H, 38N, 59A, 59H, 59N, 59S, 212H and 212M are excluded and wherein the fragment of the variant polypeptide comprises 100 to 259 consecutive amino acids of the full-length variant polypeptide.

[0845] In one embodiment the variant polypeptide of the present invention preferably has a sequence identity of 95.5% to 98% to the amino acid sequence according to SEQ ID NO: 1 and one of the following combinations of amino acid substitutions:

[0846] (a) X38R / K / H / W, X59K / E and X210I / V;

[0847] (b) X38R / K / H / W, X210I / V and X212G / M / R / K / P / A / F / H / W / N / Y / D;

[0848] (c) X59K / E, X210I / V and X212G / M / R / K / P / A / F / H / W / N / Y / D;

[0849] (d) X38R / K / H / W, X59K / E, X210I / V and X212G / M / R / K / P / A / F / H / W / N / Y / D; (e) X38R / K / H / W, X59K / E and X271 D;

[0850] (f) X38R / K / H / W, X212G / M / R / K / P / A / F / H / W / N / Y / D and X271 D;

[0851] (g) X59K / E, X212G / M / R / K / P / A / F / H / W / N / Y / D and X271 D;

[0852] (h) X38R / K / H / W, X59K / E, X212G / M / R / K / P / A / F / H / W / N / Y / D and X271 D;

[0853] (i) X38R / K / H / W, X59K / E, X210I / V and X271 D;

[0854] Q) X38R / K / H / W, X21 Ol / V, X212G / M / R / K / P / A / F / H / W / N / Y / D and X271 D;

[0855] (k) X59K / E, X210I / V, X212G / M / R / K / P / A / F / H / W / N / Y / D and X271 D; or

[0856] (l) X38R / K / H / W, X59K / E, X21 Ol / V, X212G / M / R / K / P / A / F / H / W / N / Y / D and X271 D compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2, wherein the variant polypeptide further comprises at least one amino acid substitution selected from the group consisting of X9A / Q / E / C / D, X18Q / E / D, X21 l / V / F / W / Y, X43K / R / C / H / D / L / S / W / A / M / Y / Q / F / I, X78N / D / R / W / F / H / K / E / L / Y / M / C / Q, X160H / S / D / E / P, X183D / E / C / Q / A / M, X194E / C / D, X204D / E / C / G, X218T / S / D, X228S, X259E / D, X260D / E / K and X261 L / M / F / E / W / Y / C compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2.

[0857] In one embodiment the present invention relates to a variant polypeptide having protease activity or a fragment of said polypeptide having protease activity, wherein:

[0858] (i) the polypeptide has an amino acid sequence which is at least 82%, but less than 100%, identical to the amino acid sequence as set forth in SEQ ID NO: 1 and

[0859] (ii) the polypeptide or fragment thereof comprises an amino acid substitution at two or more amino acid residues selected from the group consisting of 38, 59 and 212 compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2,

[0860] (iii) the polypeptide or fragment thereof comprises at least one amino acid substitution selected from the group consisting of X9A / Q / E / C / D, X18Q / E / D, X21 l / V / F / W / Y, X43K / R / C / H / D / L / S / W / A / M / Y / Q / F / I,

[0861] X78N / D / R / W / F / H / K / E / L / Y / M / C / Q, X160H / S / D / E / P, X183D / E / C / Q / A / M, X194E / C / D, X204D / E / C / G, X218T / S / D, X228S, X259E / D, X260D / E / K and X261 L / M / F / E / W / Y / C compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2, wherein the amino acid substitutions 38F, 38H, 38N, 59A, 59H, 59N, 59S, 212H and 212M are excluded, wherein the fragment of the variant polypeptide comprises 100 to 259 consecutive amino acids of the full-length variant polypeptide and wherein said variant polypeptide comprises amino acid residue D or E at position 101 , preferably E at position 101 , referring to the numbering of SEQ ID NO: 2.

[0862] In one embodiment the variant polypeptide of the present invention preferably has a sequence identity of 95.5% to 98% to the amino acid sequence according to SEQ ID NO: 1 and one of the following combinations of amino acid substitutions:

[0863] (a) X38R / K / H / W, X59K / E and X21 Ol / V; (b) X38R / K / H / W, X210I / V and X212G / M / R / K / P / A / F / H / W / N / Y / D;

[0864] (c) X59K / E, X21 Ol / V and X212G / M / R / K / P / A / F / H / W / N / Y / D;

[0865] (d) X38R / K / H / W, X59K / E, X21 Ol / V and X212G / M / R / K / P / A / F / H / W / N / Y / D ;

[0866] (e) X38R / K / H / W, X59K / E and X271 D;

[0867] (f) X38R / K / H / W, X212G / M / R / K / P / A / F / H / W / N / Y / D and X271 D;

[0868] (g) X59K / E, X212G / M / R / K / P / A / F / H / W / N / Y / D and X271 D;

[0869] (h) X38R / K / H / W, X59K / E, X212G / M / R / K / P / A / F / H / W / N / Y / D and X271 D;

[0870] (i) X38R / K / H / W, X59K / E, X210I / V and X271 D;

[0871] G) X38R / K / H / W, X21 Ol / V, X212G / M / R / K / P / A / F / H / W / N / Y / D and X271 D;

[0872] (k) X59K / E, X210I / V, X212G / M / R / K / P / A / F / H / W / N / Y / D and X271 D; or

[0873] (l) X38R / K / H / W, X59K / E, X21 Ol / V, X212G / M / R / K / P / A / F / H / W / N / Y / D and X271 D compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2, wherein the variant polypeptide further comprises at least one amino acid substitution selected from the group consisting of X9A / Q / E / C / D, X18Q / E / D, X21 l / V / F / W / Y, X43K / R / C / H / D / L / S / W / A / M / Y / Q / F / I, X78N / D / R / W / F / H / K / E / L / Y / M / C / Q, X160H / S / D / E / P, X183D / E / C / Q / A / M, X194E / C / D, X204D / E / C / G, X218T / S / D, X228S, X259E / D, X260D / E / K and X261 L / M / F / E / W / Y / C compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2 and wherein said variant polypeptide comprises amino acid residue D or E at position 101 , preferably E at position 101 , referring to the numbering of SEQ ID NO: 2.

[0874] In one embodiment the variant polypeptide of the present invention or a fragment of said polypeptide having protease activity additionally comprises at least one amino acid substitution selected from the group consisting of X9A / Q, X18Q / E / D, X21 V / I, X43K / R, X78N / D, X160H, X183D / E, X194E / D, X204D, X218S / T, X228S, X259D, X260D / E and X261 F / W / Y / L compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2.

[0875] Hence, in one embodiment the present invention relates to a variant polypeptide having protease activity or a fragment of said polypeptide having protease activity, wherein:

[0876] (i) the polypeptide has an amino acid sequence which is at least 82%, but less than 100%, identical to the amino acid sequence as set forth in SEQ ID NO: 1 and

[0877] (ii) the polypeptide or fragment thereof comprises an amino acid substitution at two or more amino acid residues selected from the group consisting of 38, 59 and 212 compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2,

[0878] (iii) the polypeptide or fragment thereof comprises at least one amino acid substitution selected from the group consisting of of X9A / Q, X18Q / E / D, X21 V / I, X43K / R, X78N / D, X160H, X183D / E, X194E / D, X204D, X218S / T, X228S, X259D, X260D / E and X261 F / W / Y / L 261 F / W / Y / L compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2, wherein the amino acid substitutions 38F, 38H, 38N, 59A, 59H, 59N, 59S, 212H and 212M are excluded and wherein the fragment of the variant polypeptide comprises 100 to 259 consecutive amino acids of the full-length variant polypeptide.

[0879] In one embodiment the variant polypeptide of the present invention preferably has a sequence identity of 95.5% to 98% to the amino acid sequence according to SEQ ID NO: 1 and one of the following combinations of amino acid substitutions:

[0880] (a) X38R / K / H / W, X59K / E and X21 Ol / V;

[0881] (b) X38R / K / H / W, X210I / V and X212G / M / R / K / P / A / F / H / W / N / Y / D;

[0882] (c) X59K / E, X21 Ol / V and X212G / M / R / K / P / A / F / H / W / N / Y / D;

[0883] (d) X38R / K / H / W, X59K / E, X21 Ol / V and X212G / M / R / K / P / A / F / H / VV / N / Y / D;

[0884] (e) X38R / K / H / W, X59K / E and X271 D;

[0885] (f) X38R / K / H / W, X212G / M / R / K / P / A / F / H / W / N / Y / D and X271 D;

[0886] (g) X59K / E, X212G / M / R / K / P / A / F / H / W / N / Y / D and X271 D;

[0887] (h) X38R / K / H / W, X59K / E, X212G / M / R / K / P / A / F / H / W / N / Y / D and X271 D;

[0888] (I) X38R / K / H / W, X59K / E, X210I / V and X271 D;

[0889] Q) X38R / K / H / W, X21 Ol / V, X212G / M / R / K / P / A / F / H / W / N / Y / D and X271 D;

[0890] (k) X59K / E, X210I / V, X212G / M / R / K / P / A / F / H / W / N / Y / D and X271 D; or

[0891] (l) X38R / K / H / W, X59K / E, X21 Ol / V, X212G / M / R / K / P / A / F / H / W / N / Y / D and X271 D compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2, wherein the variant polypeptide further comprises at least one amino acid substitution selected from the group consisting of of X9A / Q, X18Q / E / D, X21V / I, X43K / R, X78N / D, X160H, X183D / E, X194E / D, X204D, X218S / T, X228S, X259D, X260D / E and X261 F / W / Y / L compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2.

[0892] In one embodiment the present invention relates to a variant polypeptide having protease activity or a fragment of said polypeptide having protease activity, wherein:

[0893] (I) the polypeptide has an amino acid sequence which is at least 82%, but less than 100%, identical to the amino acid sequence as set forth in SEQ ID NO: 1 and

[0894] (ii) the polypeptide or fragment thereof comprises an amino acid substitution at two or more amino acid residues selected from the group consisting of 38, 59 and 212 compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2,

[0895] (ill) the polypeptide or fragment thereof comprises at least one amino acid substitution selected from the group consisting of of X9A / Q, X18Q / E / D, X21 V / I, X43K / R, X78N / D, X160H, X183D / E, X194E / D, X204D / E, X218S / T, X228S, X259D, X260D / E and X261 F / W / Y / L compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2, wherein the amino acid substitutions 38F, 38H, 38N, 59A, 59H, 59N, 59S, 212H and 212M are excluded, wherein the fragment of the variant polypeptide comprises 100 to 259 consecutive amino acids of the full-length variant polypeptide and wherein said variant polypeptide comprises amino acid residue D or E at position 101 , preferably E at position 101 , referring to the numbering of SEQ ID NO: 2.

[0896] In one embodiment the variant polypeptide of the present invention preferably has a sequence identity of 95.5% to 98% to the amino acid sequence according to SEQ ID NO: 1 and one of the following combinations of amino acid substitutions:

[0897] (a) X38R / K / H / W, X59K / E and X210I / V;

[0898] (b) X38R / K / H / W, X210I / V and X212G / M / R / K / P / A / F / H / W / N / Y / D;

[0899] (c) X59K / E, X210I / V and X212G / M / R / K / P / A / F / H / W / N / Y / D;

[0900] (d) X38R / K / H / W, X59K / E, X210I / V and X212G / M / R / K / P / A / F / H / VV / N / Y / D;

[0901] (e) X38R / K / H / W, X59K / E and X271 D;

[0902] (f) X38R / K / H / W, X212G / M / R / K / P / A / F / H / W / N / Y / D and X271 D;

[0903] (g) X59K / E, X212G / M / R / K / P / A / F / H / W / N / Y / D and X271 D;

[0904] (h) X38R / K / H / W, X59K / E, X212G / M / R / K / P / A / F / H / W / N / Y / D and X271 D;

[0905] (I) X38R / K / H / W, X59K / E, X210I / V and X271 D;

[0906] Q) X38R / K / H / W, X210I / V, X212G / M / R / K / P / A / F / H / W / N / Y / D and X271 D;

[0907] (k) X59K / E, X210I / V, X212G / M / R / K / P / A / F / H / W / N / Y / D and X271 D; or

[0908] (l) X38R / K / H / W, X59K / E, X210I / V, X212G / M / R / K / P / A / F / H / W / N / Y / D and X271 D compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2, wherein the variant polypeptide further comprises at least one amino acid substitution selected from the group consisting of of X9A / Q, X18Q / E / D, X21 V / l, X43K / R, X78N / D, X160H, X183D / E, X194E / D, X204D / E, X218S / T, X228S, X259D, X260D / E and X261 F / W / Y / L compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2 and wherein said variant polypeptide comprises amino acid residue D or E at position 101 , preferably E at position 101 , referring to the numbering of SEQ ID NO: 2.

[0909] In one embodiment the variant polypeptide of the present invention or a fragment of said polypeptide having protease activity additionally comprises at least one amino acid substitution selected from the group consisting of S9A / Q / E / C / D, N18Q / E / D, L21 l / V / F / W / Y, N43K / R / C / H / D / L / S / W / A / M / Y / Q / F / I, S78N / D / R / W / F / H / K / E / L / Y / M / C / Q, S160H / S / D / E / P, N183D / E / C / Q / A / M, A194E / C / D, N204D / E / C / G, N218T / S / D, A228S, S259E / D, T260D / E / K and N261 L / M / F / E / W / Y / C compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2. In one embodiment the variant polypeptide of the present invention or a fragment of said polypeptide having protease activity additionally comprises at least one amino acid substitution selected from the group consisting of S9A / Q, N18Q / E / D, L21 V / I, N43K / R, S78N / D, S160H, N183D / E, A194E / D, N204D / E, N218S / T, A228S, S259D, T260D / E and N261 FI\NI I\_ compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2.

[0910] In one embodiment the variant polypeptide of the present invention or a fragment of said polypeptide having protease activity additionally comprises at least one amino acid substitution at an amino acid residue selected from the group consisting of 24, 25, 103, 109, 144, 182 and 248 compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2.

[0911] Hence, in one embodiment the present invention relates to a variant polypeptide having protease activity or a fragment of said polypeptide having protease activity, wherein:

[0912] (I) the polypeptide has an amino acid sequence which is at least 82%, but less than 100%, identical to the amino acid sequence as set forth in SEQ ID NO: 1 and

[0913] (ii) the polypeptide or fragment thereof comprises an amino acid substitution at two or more amino acid residues selected from the group consisting of 38, 59 and 212 compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2,

[0914] (ill) the polypeptide or fragment thereof comprises at least one amino acid substitution at an amino acid residue selected from the group consisting of 24, 25, 103, 109, 144, 182 and 248 compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2, wherein the amino acid substitutions 38F, 38H, 38N, 59A, 59H, 59N, 59S, 212H and 212M are excluded and wherein the fragment of the variant polypeptide comprises 100 to 259 consecutive amino acids of the full-length variant polypeptide.

[0915] In one embodiment the variant polypeptide of the present invention preferably has a sequence identity of 95.5% to 98% to the amino acid sequence according to SEQ ID NO: 1 and one of the following combinations of amino acid substitutions:

[0916] (a) X38R / K / H / W, X59K / E and X210I / V;

[0917] (b) X38R / K / H / W, X210I / V and X212G / M / R / K / P / A / F / H / W / N / Y / D;

[0918] (c) X59K / E, X210I / V and X212G / M / R / K / P / A / F / H / W / N / Y / D;

[0919] (d) X38R / K / H / W, X59K / E, X210I / V and X212G / M / R / K / P / A / F / H / VV / N / Y / D;

[0920] (e) X38R / K / H / W, X59K / E and X271 D;

[0921] (f) X38R / K / H / W, X212G / M / R / K / P / A / F / H / W / N / Y / D and X271 D;

[0922] (g) X59K / E, X212G / M / R / K / P / A / F / H / W / N / Y / D and X271 D;

[0923] (h) X38R / K / H / W, X59K / E, X212G / M / R / K / P / A / F / H / W / N / Y / D and X271 D;

[0924] (I) X38R / K / H / W, X59K / E, X210I / V and X271 D;

[0925] Q) X38R / K / H / W, X210I / V, X212G / M / R / K / P / A / F / H / W / N / Y / D and X271 D;

[0926] (k) X59K / E, X210I / V, X212G / M / R / K / P / A / F / H / W / N / Y / D and X271 D; or

[0927] (l) X38R / K / H / W, X59K / E, X210I / V, X212G / M / R / K / P / A / F / H / W / N / Y / D and X271 D compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2, wherein the variant polypeptide further comprises at least one amino acid substitution at an amino acid residue selected from the group consisting of 24, 25, 103, 109, 144, 182 and 248 compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2.

[0928] In one embodiment the present invention relates to a variant polypeptide having protease activity or a fragment of said polypeptide having protease activity, wherein:

[0929] (I) the polypeptide has an amino acid sequence which is at least 82%, but less than 100%, identical to the amino acid sequence as set forth in SEQ ID NO: 1 and

[0930] (ii) the polypeptide or fragment thereof comprises an amino acid substitution at two or more amino acid residues selected from the group consisting of 38, 59 and 212 compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2,

[0931] (ill) the polypeptide or fragment thereof comprises at least one amino acid substitution at an amino acid residue selected from the group consisting of 24, 25, 103, 109, 144, 182 and 248 compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2, wherein the amino acid substitutions 38F, 38H, 38N, 59A, 59H, 59N, 59S, 212H and 212M are excluded, wherein the fragment of the variant polypeptide comprises 100 to 259 consecutive amino acids of the full-length variant polypeptide and wherein said variant polypeptide comprises amino acid residue D or E at position 101 , preferably E at position 101 , referring to the numbering of SEQ ID NO: 2.

[0932] In one embodiment the variant polypeptide of the present invention preferably has a sequence identity of 95.5% to 98% to the amino acid sequence according to SEQ ID NO: 1 and one of the following combinations of amino acid substitutions:

[0933] (a) X38R / K / H / W, X59K / E and X210I / V;

[0934] (b) X38R / K / H / W, X210I / V and X212G / M / R / K / P / A / F / H / W / N / Y / D;

[0935] (c) X59K / E, X210I / V and X212G / M / R / K / P / A / F / H / W / N / Y / D;

[0936] (d) X38R / K / H / W, X59K / E, X210I / V and X212G / M / R / K / P / A / F / H / VV / N / Y / D;

[0937] (e) X38R / K / H / W, X59K / E and X271 D;

[0938] (f) X38R / K / H / W, X212G / M / R / K / P / A / F / H / W / N / Y / D and X271 D;

[0939] (g) X59K / E, X212G / M / R / K / P / A / F / H / W / N / Y / D and X271 D;

[0940] (h) X38R / K / H / W, X59K / E, X212G / M / R / K / P / A / F / H / W / N / Y / D and X271 D;

[0941] (I) X38R / K / H / W, X59K / E, X210I / V and X271 D;

[0942] Q) X38R / K / H / W, X210I / V, X212G / M / R / K / P / A / F / H / W / N / Y / D and X271 D;

[0943] (k) X59K / E, X210I / V, X212G / M / R / K / P / A / F / H / W / N / Y / D and X271 D; or

[0944] (l) X38R / K / H / W, X59K / E, X210I / V, X212G / M / R / K / P / A / F / H / W / N / Y / D and X271 D compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2, wherein the variant polypeptide further comprises at least one amino acid substitution at an amino acid residue selected from the group consisting of 24, 25, 103, 109, 144, 182 and 248 compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2 and wherein said variant polypeptide comprises amino acid residue D or E at position 101 , preferably E at position 101 , referring to the numbering of SEQ ID NO: 2.

[0945] In one embodiment the variant polypeptide of the present invention or a fragment of said polypeptide having protease activity additionally comprises at least one amino acid substitution selected from the group consisting of X24K, X25D, X103S, X109K / A, X144N / R, X182K / R / E / C and X248Q / R compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2.

[0946] Hence, in one embodiment the present invention relates to a variant polypeptide having protease activity or a fragment of said polypeptide having protease activity, wherein:

[0947] (I) the polypeptide has an amino acid sequence which is at least 82%, but less than 100%, identical to the amino acid sequence as set forth in SEQ ID NO: 1 and

[0948] (ii) the polypeptide or fragment thereof comprises an amino acid substitution at two or more amino acid residues selected from the group consisting of 38, 59 and 212 compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2,

[0949] (ill) the polypeptide or fragment thereof comprises at least one amino acid substitution selected from the group consisting of X24K, X25D, X103S, X109K / A, X144N / R, X182K / R / E / C and X248Q / R compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2, wherein the amino acid substitutions 38F, 38H, 38N, 59A, 59H, 59N, 59S, 212H and 212M are excluded and wherein the fragment of the variant polypeptide comprises 100 to 259 consecutive amino acids of the full-length variant polypeptide.

[0950] In one embodiment the variant polypeptide of the present invention preferably has a sequence identity of 95.5% to 98% to the amino acid sequence according to SEQ ID NO: 1 and one of the following combinations of amino acid substitutions:

[0951] (a) X38R / K / H / W, X59K / E and X210I / V;

[0952] (b) X38R / K / H / W, X210I / V and X212G / M / R / K / P / A / F / H / W / N / Y / D;

[0953] (c) X59K / E, X210I / V and X212G / M / R / K / P / A / F / H / W / N / Y / D;

[0954] (d) X38R / K / H / W, X59K / E, X210I / V and X212G / M / R / K / P / A / F / H / W / N / Y / D;

[0955] (e) X38R / K / H / W, X59K / E and X271 D;

[0956] (f) X38R / K / H / W, X212G / M / R / K / P / A / F / H / W / N / Y / D and X271 D;

[0957] (g) X59K / E, X212G / M / R / K / P / A / F / H / W / N / Y / D and X271 D;

[0958] (h) X38R / K / H / W, X59K / E, X212G / M / R / K / P / A / F / H / W / N / Y / D and X271 D;

[0959] (I) X38R / K / H / W, X59K / E, X210I / V and X271 D; Q) X38R / K / H / W, X21 Ol / V, X212G / M / R / K / P / A / F / H / W / N / Y / D and X271 D;

[0960] (k) X59K / E, X210I / V, X212G / M / R / K / P / A / F / H / W / N / Y / D and X271 D; or

[0961] (l) X38R / K / H / W, X59K / E, X21 Ol / V, X212G / M / R / K / P / A / F / H / W / N / Y / D and X271 D compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2, wherein the variant polypeptide further comprises at least one amino acid substitution selected from the group consisting of X24K, X25D, X103S, X109K / A, X144N / R, X182K / R / E / C and X248Q / R compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2.

[0962] In one embodiment the present invention relates to a variant polypeptide having protease activity or a fragment of said polypeptide having protease activity, wherein:

[0963] (i) the polypeptide has an amino acid sequence which is at least 82%, but less than 100%, identical to the amino acid sequence as set forth in SEQ ID NO: 1 and

[0964] (ii) the polypeptide or fragment thereof comprises an amino acid substitution at two or more amino acid residues selected from the group consisting of 38, 59 and 212 compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2,

[0965] (iii) the polypeptide or fragment thereof comprises at least one amino acid substitution selected from the group consisting of X24K, X25D, X103S, X109K / A, X144N / R, X182K / R / E / C and X248Q / R compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2, wherein the amino acid substitutions 38F, 38H, 38N, 59A, 59H, 59N, 59S, 212H and 212M are excluded, wherein the fragment of the variant polypeptide comprises 100 to 259 consecutive amino acids of the full-length variant polypeptide and wherein said variant polypeptide comprises amino acid residue D or E at position 101 , preferably E at position 101 , referring to the numbering of SEQ ID NO: 2.

[0966] In one embodiment the variant polypeptide of the present invention preferably has a sequence identity of 95.5% to 98% to the amino acid sequence according to SEQ ID NO: 1 and one of the following combinations of amino acid substitutions:

[0967] (a) X38R / K / H / W, X59K / E and X21 Ol / V;

[0968] (b) X38R / K / H / W, X210I / V and X212G / M / R / K / P / A / F / H / W / N / Y / D;

[0969] (c) X59K / E, X21 Ol / V and X212G / M / R / K / P / A / F / H / W / N / Y / D;

[0970] (d) X38R / K / H / W, X59K / E, X21 Ol / V and X212G / M / R / K / P / A / F / H / W / N / Y / D;

[0971] (e) X38R / K / H / W, X59K / E and X271 D;

[0972] (f) X38R / K / H / W, X212G / M / R / K / P / A / F / H / W / N / Y / D and X271 D;

[0973] (g) X59K / E, X212G / M / R / K / P / A / F / H / W / N / Y / D and X271 D;

[0974] (h) X38R / K / H / W, X59K / E, X212G / M / R / K / P / A / F / H / W / N / Y / D and X271 D;

[0975] (i) X38R / K / H / W, X59K / E, X210I / V and X271 D;

[0976] Q) X38R / K / H / W, X21 Ol / V, X212G / M / R / K / P / A / F / H / W / N / Y / D and X271 D; (k) X59K / E, X210I / V, X212G / M / R / K / P / A / F / H / W / N / Y / D and X271 D; or

[0977] (l) X38R / K / H / W, X59K / E, X21 Ol / V, X212G / M / R / K / P / A / F / H / W / N / Y / D and X271 D compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2, wherein the variant polypeptide further comprises at least one amino acid substitution selected from the group consisting of X24K, X25D, X103S, X109K / A, X144N / R, X182K / R / E / C and X248Q / R compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2 and wherein said variant polypeptide comprises amino acid residue D or E at position 101 , preferably E at position 101 , referring to the numbering of SEQ ID NO: 2.

[0978] In one embodiment the variant polypeptide of the present invention or a fragment of said polypeptide having protease activity additionally comprises at least one amino acid substitution selected from the group consisting of X24K, X25D, X103S, X109K / A, X144N / R, X182K and X248Q compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2.

[0979] Hence, in one embodiment the present invention relates to a variant polypeptide having protease activity or a fragment of said polypeptide having protease activity, wherein:

[0980] (I) the polypeptide has an amino acid sequence which is at least 82%, but less than 100%, identical to the amino acid sequence as set forth in SEQ ID NO: 1 and

[0981] (ii) the polypeptide or fragment thereof comprises an amino acid substitution at two or more amino acid residues selected from the group consisting of 38, 59 and 212 compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2,

[0982] (ill) the polypeptide or fragment thereof comprises at least one amino acid substitution selected from the group consisting of of X24K, X25D, X103S, X109K / A, X144N / R, X182K and X248Q compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2, wherein the amino acid substitutions 38F, 38H, 38N, 59A, 59H, 59N, 59S, 212H and 212M are excluded and wherein the fragment of the variant polypeptide comprises 100 to 259 consecutive amino acids of the full-length variant polypeptide.

[0983] In one embodiment the variant polypeptide of the present invention preferably has a sequence identity of 95.5% to 98% to the amino acid sequence according to SEQ ID NO: 1 and one of the following combinations of amino acid substitutions:

[0984] (a) X38R / K / H / W, X59K / E and X210I / V;

[0985] (b) X38R / K / H / W, X210I / V and X212G / M / R / K / P / A / F / H / W / N / Y / D;

[0986] (c) X59K / E, X210I / V and X212G / M / R / K / P / A / F / H / W / N / Y / D;

[0987] (d) X38R / K / H / W, X59K / E, X210I / V and X212G / M / R / K / P / A / F / H / W / N / Y / D;

[0988] (e) X38R / K / H / W, X59K / E and X271 D;

[0989] (f) X38R / K / H / W, X212G / M / R / K / P / A / F / H / W / N / Y / D and X271 D; (g) X59K / E, X212G / M / R / K / P / A / F / H / W / N / Y / D and X271 D;

[0990] (h) X38R / K / H / W, X59K / E, X212G / M / R / K / P / A / F / H / W / N / Y / D and X271 D;

[0991] (i) X38R / K / H / W, X59K / E, X210I / V and X271 D;

[0992] Q) X38R / K / H / W, X21 Ol / V, X212G / M / R / K / P / A / F / H / W / N / Y / D and X271 D;

[0993] (k) X59K / E, X210I / V, X212G / M / R / K / P / A / F / H / W / N / Y / D and X271 D; or

[0994] (l) X38R / K / H / W, X59K / E, X21 Ol / V, X212G / M / R / K / P / A / F / H / W / N / Y / D and X271 D compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2, wherein the variant polypeptide further comprises at least one amino acid substitution selected from the group consisting of of X24K, X25D, X103S, X109K / A, X144N / R, X182K and X248Q compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2.

[0995] In one embodiment the present invention relates to a variant polypeptide having protease activity or a fragment of said polypeptide having protease activity, wherein:

[0996] (i) the polypeptide has an amino acid sequence which is at least 82%, but less than 100%, identical to the amino acid sequence as set forth in SEQ ID NO: 1 and

[0997] (ii) the polypeptide or fragment thereof comprises an amino acid substitution at two or more amino acid residues selected from the group consisting of 38, 59 and 212 compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2,

[0998] (iii) the polypeptide or fragment thereof comprises at least one amino acid substitution selected from the group consisting of of X24K, X25D, X103S, X109K / A, X144N / R, X182K and X248Q compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2, wherein the amino acid substitutions 38F, 38H, 38N, 59A, 59H, 59N, 59S, 212H and 212M are excluded, wherein the fragment of the variant polypeptide comprises 100 to 259 consecutive amino acids of the full-length variant polypeptide and wherein said variant polypeptide comprises amino acid residue D or E at position 101 , preferably E at position 101 , referring to the numbering of SEQ ID NO: 2.

[0999] In one embodiment the variant polypeptide of the present invention preferably has a sequence identity of 95.5% to 98% to the amino acid sequence according to SEQ ID NO: 1 and one of the following combinations of amino acid substitutions:

[1000] (a) X38R / K / H / W, X59K / E and X21 Ol / V;

[1001] (b) X38R / K / H / W, X210I / V and X212G / M / R / K / P / A / F / H / W / N / Y / D;

[1002] (c) X59K / E, X21 Ol / V and X212G / M / R / K / P / A / F / H / W / N / Y / D;

[1003] (d) X38R / K / H / W, X59K / E, X21 Ol / V and X212G / M / R / K / P / A / F / H / W / N / Y / D;

[1004] (e) X38R / K / H / W, X59K / E and X271 D;

[1005] (f) X38R / K / H / W, X212G / M / R / K / P / A / F / H / W / N / Y / D and X271 D;

[1006] (g) X59K / E, X212G / M / R / K / P / A / F / H / W / N / Y / D and X271 D; (h) X38R / K / H / W, X59K / E, X212G / M / R / K / P / A / F / H / W / N / Y / D and X271 D;

[1007] (i) X38R / K / H / W, X59K / E, X210I / V and X271 D;

[1008] Q) X38R / K / H / W, X21 Ol / V, X212G / M / R / K / P / A / F / H / W / N / Y / D and X271 D;

[1009] (k) X59K / E, X210I / V, X212G / M / R / K / P / A / F / H / W / N / Y / D and X271 D; or

[1010] (l) X38R / K / H / W, X59K / E, X21 Ol / V, X212G / M / R / K / P / A / F / H / W / N / Y / D and X271 D compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2, wherein the variant polypeptide further comprises at least one amino acid substitution selected from the group consisting of of X24K, X25D, X103S, X109K / A, X144N / R, X182K and X248Q compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2 and wherein said variant polypeptide comprises amino acid residue D or E at position 101 , preferably E at position 101 , referring to the numbering of SEQ ID NO: 2. In one embodiment the variant polypeptide of the present invention or a fragment of said polypeptide having protease activity additionally comprises at least one amino acid substitution selected from the group consisting of S24K, G25D, A103S, Q109K / A, S144N / R, Q182K and N248Q compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2.

[1011] In one embodiment the variant polypeptide of the present invention or a fragment of said polypeptide having protease activity additionally comprises at least one amino acid substitution at an amino acid residue selected from the group consisting of 3, 76 and 256 compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2.

[1012] Hence, in one embodiment the present invention relates to a variant polypeptide having protease activity or a fragment of said polypeptide having protease activity, wherein:

[1013] (i) the polypeptide has an amino acid sequence which is at least 82%, but less than 100%, identical to the amino acid sequence as set forth in SEQ ID NO: 1 and

[1014] (ii) the polypeptide or fragment thereof comprises an amino acid substitution at two or more amino acid residues selected from the group consisting of 38, 59 and 212 compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2,

[1015] (iii) the polypeptide or fragment thereof comprises at least one amino acid substitution at an amino acid residue selected from the group consisting of 3, 76 and 256 compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2, wherein the amino acid substitutions 38F, 38H, 38N, 59A, 59H, 59N, 59S, 212H and 212M are excluded and wherein the fragment of the variant polypeptide comprises 100 to 259 consecutive amino acids of the full-length variant polypeptide.

[1016] In one embodiment the variant polypeptide of the present invention preferably has a sequence identity of 95.5% to 98% to the amino acid sequence according to SEQ ID NO: 1 and one of the following combinations of amino acid substitutions: (a) X38R / K / H / W, X59K / E and X21 Ol / V;

[1017] (b) X38R / K / H / W, X210I / V and X212G / M / R / K / P / A / F / H / W / N / Y / D;

[1018] (c) X59K / E, X21 Ol / V and X212G / M / R / K / P / A / F / H / W / N / Y / D;

[1019] (d) X38R / K / H / W, X59K / E, X21 Ol / V and X212G / M / R / K / P / A / F / H / VV / N / Y / D;

[1020] (e) X38R / K / H / W, X59K / E and X271 D;

[1021] (f) X38R / K / H / W, X212G / M / R / K / P / A / F / H / W / N / Y / D and X271 D;

[1022] (g) X59K / E, X212G / M / R / K / P / A / F / H / W / N / Y / D and X271 D;

[1023] (h) X38R / K / H / W, X59K / E, X212G / M / R / K / P / A / F / H / W / N / Y / D and X271 D;

[1024] (i) X38R / K / H / W, X59K / E, X210I / V and X271 D;

[1025] G) X38R / K / H / W, X21 Ol / V, X212G / M / R / K / P / A / F / H / W / N / Y / D and X271 D;

[1026] (k) X59K / E, X210I / V, X212G / M / R / K / P / A / F / H / W / N / Y / D and X271 D; or

[1027] (l) X38R / K / H / W, X59K / E, X21 Ol / V, X212G / M / R / K / P / A / F / H / W / N / Y / D and X271 D compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2, wherein the variant polypeptide further comprises at least one amino acid substitution at an amino acid residue selected from the group consisting of 3, 76 and 256 compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2.

[1028] In one embodiment the present invention relates to a variant polypeptide having protease activity or a fragment of said polypeptide having protease activity, wherein:

[1029] (i) the polypeptide has an amino acid sequence which is at least 82%, but less than 100%, identical to the amino acid sequence as set forth in SEQ ID NO: 1 and

[1030] (ii) the polypeptide or fragment thereof comprises an amino acid substitution at two or more amino acid residues selected from the group consisting of 38, 59 and 212 compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2,

[1031] (iii) the polypeptide or fragment thereof comprises at least one amino acid substitution at an amino acid residue selected from the group consisting of 3, 76 and 256 compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2, wherein the amino acid substitutions 38F, 38H, 38N, 59A, 59H, 59N, 59S, 212H and 212M are excluded, wherein the fragment of the variant polypeptide comprises 100 to 259 consecutive amino acids of the full-length variant polypeptide and wherein said variant polypeptide comprises amino acid residue D or E at position 101 , preferably E at position 101 , referring to the numbering of SEQ ID NO: 2.

[1032] In one embodiment the variant polypeptide of the present invention preferably has a sequence identity of 95.5% to 98% to the amino acid sequence according to SEQ ID NO: 1 and one of the following combinations of amino acid substitutions:

[1033] (a) X38R / K / H / W, X59K / E and X21 Ol / V; (b) X38R / K / H / W, X210I / V and X212G / M / R / K / P / A / F / H / W / N / Y / D;

[1034] (c) X59K / E, X21 Ol / V and X212G / M / R / K / P / A / F / H / W / N / Y / D;

[1035] (d) X38R / K / H / W, X59K / E, X21 Ol / V and X212G / M / R / K / P / A / F / H / W / N / Y / D;

[1036] (e) X38R / K / H / W, X59K / E and X271 D;

[1037] (f) X38R / K / H / W, X212G / M / R / K / P / A / F / H / W / N / Y / D and X271 D;

[1038] (g) X59K / E, X212G / M / R / K / P / A / F / H / W / N / Y / D and X271 D;

[1039] (h) X38R / K / H / W, X59K / E, X212G / M / R / K / P / A / F / H / W / N / Y / D and X271 D;

[1040] (i) X38R / K / H / W, X59K / E, X210I / V and X271 D;

[1041] G) X38R / K / H / W, X21 Ol / V, X212G / M / R / K / P / A / F / H / W / N / Y / D and X271 D;

[1042] (k) X59K / E, X210I / V, X212G / M / R / K / P / A / F / H / W / N / Y / D and X271 D; or

[1043] (l) X38R / K / H / W, X59K / E, X21 Ol / V, X212G / M / R / K / P / A / F / H / W / N / Y / D and X271 D compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2, wherein the variant polypeptide further comprises at least one amino acid substitution at an amino acid residue selected from the group consisting of 3, 76 and 256 compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2 and wherein said variant polypeptide comprises amino acid residue D or E at position 101 , preferably E at position 101 , referring to the numbering of SEQ ID NO: 2.

[1044] In one embodiment the variant polypeptide of the present invention or a fragment of said polypeptide having protease activity additionally comprises at least one amino acid substitution selected from the group consisting of X3T / Q / V, X76D and X256E / T / D / R / P compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2.

[1045] Hence, in one embodiment the present invention relates to a variant polypeptide having protease activity or a fragment of said polypeptide having protease activity, wherein:

[1046] (i) the polypeptide has an amino acid sequence which is at least 82%, but less than 100%, identical to the amino acid sequence as set forth in SEQ ID NO: 1 and

[1047] (ii) the polypeptide or fragment thereof comprises an amino acid substitution at two or more amino acid residues selected from the group consisting of 38, 59 and 212 compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2,

[1048] (iii) the polypeptide or fragment thereof comprises at least one amino acid substitution selected from the group consisting of X3T / Q / V, X76D and X256E / T / D / R / P compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2, wherein the amino acid substitutions 38F, 38H, 38N, 59A, 59H, 59N, 59S, 212H and 212M are excluded and wherein the fragment of the variant polypeptide comprises 100 to 259 consecutive amino acids of the full-length variant polypeptide. In one embodiment the variant polypeptide of the present invention preferably has a sequence identity of 95.5% to 98% to the amino acid sequence according to SEQ ID NO: 1 and one of the following combinations of amino acid substitutions:

[1049] (a) X38R / K / H / W, X59K / E and X21 Ol / V;

[1050] (b) X38R / K / H / W, X210I / V and X212G / M / R / K / P / A / F / H / W / N / Y / D;

[1051] (c) X59K / E, X21 Ol / V and X212G / M / R / K / P / A / F / H / W / N / Y / D;

[1052] (d) X38R / K / H / W, X59K / E, X21 Ol / V and X212G / M / R / K / P / A / F / H / VV / N / Y / D;

[1053] (e) X38R / K / H / W, X59K / E and X271 D;

[1054] (f) X38R / K / H / W, X212G / M / R / K / P / A / F / H / W / N / Y / D and X271 D;

[1055] (g) X59K / E, X212G / M / R / K / P / A / F / H / W / N / Y / D and X271 D;

[1056] (h) X38R / K / H / W, X59K / E, X212G / M / R / K / P / A / F / H / W / N / Y / D and X271 D;

[1057] (i) X38R / K / H / W, X59K / E, X210I / V and X271 D;

[1058] Q) X38R / K / H / W, X21 Ol / V, X212G / M / R / K / P / A / F / H / W / N / Y / D and X271 D;

[1059] (k) X59K / E, X210I / V, X212G / M / R / K / P / A / F / H / W / N / Y / D and X271 D; or

[1060] (l) X38R / K / H / W, X59K / E, X21 Ol / V, X212G / M / R / K / P / A / F / H / W / N / Y / D and X271 D compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2, wherein the variant polypeptide further comprises at least one amino acid substitution selected from the group consisting of X3T / Q / V, X76D and X256E / T / D / R / P compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2.

[1061] In one embodiment the present invention relates to a variant polypeptide having protease activity or a fragment of said polypeptide having protease activity, wherein:

[1062] (i) the polypeptide has an amino acid sequence which is at least 82%, but less than 100%, identical to the amino acid sequence as set forth in SEQ ID NO: 1 and

[1063] (ii) the polypeptide or fragment thereof comprises an amino acid substitution at two or more amino acid residues selected from the group consisting of 38, 59 and 212 compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2,

[1064] (iii) the polypeptide or fragment thereof comprises at least one amino acid substitution selected from the group consisting of X3T / Q / V, X76D and X256E / T / D / R / P compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2, wherein the amino acid substitutions 38F, 38H, 38N, 59A, 59H, 59N, 59S, 212H and 212M are excluded, wherein the fragment of the variant polypeptide comprises 100 to 259 consecutive amino acids of the full-length variant polypeptide and wherein said variant polypeptide comprises amino acid residue D or E at position 101 , preferably E at position 101 , referring to the numbering of SEQ ID NO: 2. In one embodiment the variant polypeptide of the present invention preferably has a sequence identity of 95.5% to 98% to the amino acid sequence according to SEQ ID NO: 1 and one of the following combinations of amino acid substitutions:

[1065] (a) X38R / K / H / W, X59K / E and X21 Ol / V;

[1066] (b) X38R / K / H / W, X210I / V and X212G / M / R / K / P / A / F / H / W / N / Y / D;

[1067] (c) X59K / E, X21 Ol / V and X212G / M / R / K / P / A / F / H / W / N / Y / D;

[1068] (d) X38R / K / H / W, X59K / E, X21 Ol / V and X212G / M / R / K / P / A / F / H / VV / N / Y / D;

[1069] (e) X38R / K / H / W, X59K / E and X271 D;

[1070] (f) X38R / K / H / W, X212G / M / R / K / P / A / F / H / W / N / Y / D and X271 D;

[1071] (g) X59K / E, X212G / M / R / K / P / A / F / H / W / N / Y / D and X271 D;

[1072] (h) X38R / K / H / W, X59K / E, X212G / M / R / K / P / A / F / H / W / N / Y / D and X271 D;

[1073] (i) X38R / K / H / W, X59K / E, X210I / V and X271 D;

[1074] Q) X38R / K / H / W, X21 Ol / V, X212G / M / R / K / P / A / F / H / W / N / Y / D and X271 D;

[1075] (k) X59K / E, X210I / V, X212G / M / R / K / P / A / F / H / W / N / Y / D and X271 D; or

[1076] (l) X38R / K / H / W, X59K / E, X21 Ol / V, X212G / M / R / K / P / A / F / H / W / N / Y / D and X271 D compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2, wherein the variant polypeptide further comprises at least one amino acid substitution selected from the group consisting of X3T / Q / V, X76D and X256E / T / D / R / P compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2 and wherein said variant polypeptide comprises amino acid residue D or E at position 101 , preferably E at position 101 , referring to the numbering of SEQ ID NO: 2.

[1077] In one embodiment the variant polypeptide of the present invention or a fragment of said polypeptide having protease activity additionally comprises at least one amino acid substitution selected from the group consisting of X3T, X76D and X256D compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2.

[1078] Hence, in one embodiment the present invention relates to a variant polypeptide having protease activity or a fragment of said polypeptide having protease activity, wherein:

[1079] (i) the polypeptide has an amino acid sequence which is at least 82%, but less than 100%, identical to the amino acid sequence as set forth in SEQ ID NO: 1 and

[1080] (ii) the polypeptide or fragment thereof comprises an amino acid substitution at two or more amino acid residues selected from the group consisting of 38, 59 and 212 compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2,

[1081] (iii) the polypeptide or fragment thereof comprises at least one amino acid substitution selected from the group consisting of of X3T, X76D and X256D compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2, wherein the amino acid substitutions 38F, 38H, 38N, 59A, 59H, 59N, 59S, 212H and 212M are excluded and wherein the fragment of the variant polypeptide comprises 100 to 259 consecutive amino acids of the full-length variant polypeptide.

[1082] In one embodiment the variant polypeptide of the present invention preferably has a sequence identity of 95.5% to 98% to the amino acid sequence according to SEQ ID NO: 1 and one of the following combinations of amino acid substitutions:

[1083] (a) X38R / K / H / W, X59K / E and X210I / V;

[1084] (b) X38R / K / H / W, X210I / V and X212G / M / R / K / P / A / F / H / W / N / Y / D;

[1085] (c) X59K / E, X21 Ol / V and X212G / M / R / K / P / A / F / H / W / N / Y / D;

[1086] (d) X38R / K / H / W, X59K / E, X21 Ol / V and X212G / M / R / K / P / A / F / H / VV / N / Y / D;

[1087] (e) X38R / K / H / W, X59K / E and X271 D;

[1088] (f) X38R / K / H / W, X212G / M / R / K / P / A / F / H / W / N / Y / D and X271 D;

[1089] (g) X59K / E, X212G / M / R / K / P / A / F / H / W / N / Y / D and X271 D;

[1090] (h) X38R / K / H / W, X59K / E, X212G / M / R / K / P / A / F / H / W / N / Y / D and X271 D;

[1091] (I) X38R / K / H / W, X59K / E, X210I / V and X271 D;

[1092] Q) X38R / K / H / W, X21 Ol / V, X212G / M / R / K / P / A / F / H / W / N / Y / D and X271 D;

[1093] (k) X59K / E, X210I / V, X212G / M / R / K / P / A / F / H / W / N / Y / D and X271 D; or

[1094] (l) X38R / K / H / W, X59K / E, X21 Ol / V, X212G / M / R / K / P / A / F / H / W / N / Y / D and X271 D compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2, wherein the variant polypeptide further comprises at least one amino acid substitution selected from the group consisting of of X3T, X76D and X256D compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2.

[1095] In one embodiment the present invention relates to a variant polypeptide having protease activity or a fragment of said polypeptide having protease activity, wherein:

[1096] (I) the polypeptide has an amino acid sequence which is at least 82%, but less than 100%, identical to the amino acid sequence as set forth in SEQ ID NO: 1 and

[1097] (ii) the polypeptide or fragment thereof comprises an amino acid substitution at two or more amino acid residues selected from the group consisting of 38, 59 and 212 compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2,

[1098] (ill) the polypeptide or fragment thereof comprises at least one amino acid substitution selected from the group consisting of of X3T, X76D and X256D compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2, wherein the amino acid substitutions 38F, 38H, 38N, 59A, 59H, 59N, 59S, 212H and 212M are excluded, wherein the fragment of the variant polypeptide comprises 100 to 259 consecutive amino acids of the full-length variant polypeptide and wherein said variant polypeptide comprises amino acid residue D or E at position 101 , preferably E at position 101 , referring to the numbering of SEQ ID NO: 2.

[1099] In one embodiment the variant polypeptide of the present invention preferably has a sequence identity of 95.5% to 98% to the amino acid sequence according to SEQ ID NO: 1 and one of the following combinations of amino acid substitutions:

[1100] (a) X38R / K / H / W, X59K / E and X21 Ol / V;

[1101] (b) X38R / K / H / W, X210I / V and X212G / M / R / K / P / A / F / H / W / N / Y / D;

[1102] (c) X59K / E, X21 Ol / V and X212G / M / R / K / P / A / F / H / W / N / Y / D;

[1103] (d) X38R / K / H / W, X59K / E, X21 Ol / V and X212G / M / R / K / P / A / F / H / VV / N / Y / D;

[1104] (e) X38R / K / H / W, X59K / E and X271 D;

[1105] (f) X38R / K / W, X212G / M / R / K / P / A / F / H / W / N / Y / D and X271 D;

[1106] (g) X59K / E, X212G / M / R / K / P / A / F / H / W / N / Y / D and X271 D;

[1107] (h) X38R / K / H / W, X59K / E, X212G / M / R / K / P / A / F / H / W / N / Y / D and X271 D;

[1108] (I) X38R / K / H / W, X59K / E, X210I / V and X271 D;

[1109] Q) X38R / K / H / W, X21 Ol / V, X212G / M / R / K / P / A / F / H / W / N / Y / D and X271 D;

[1110] (k) X59K / E, X210I / V, X212G / M / R / K / P / A / F / H / W / N / Y / D and X271 D; or

[1111] (l) X38R / K / H / W, X59K / E, X21 Ol / V, X212G / M / R / K / P / A / F / H / W / N / Y / D and X271 D compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2, wherein the variant polypeptide further comprises at least one amino acid substitution selected from the group consisting of of X3T, X76D and X256D compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2 and wherein said variant polypeptide comprises amino acid residue D or E at position 101 , preferably E at position 101 , referring to the numbering of SEQ ID NO: 2.

[1112] In one embodiment the variant polypeptide of the present invention or a fragment of said polypeptide having protease activity additionally comprises at least one amino acid substitution selected from the group consisting of S3T, N76D and S256D compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2.

[1113] In one embodiment the variant polypeptide of the present invention or a fragment of said polypeptide having protease activity additionally comprises at least one amino acid substitution selected from the group consisting of S3T, N76D and S256D compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2.

[1114] In one embodiment the variant polypeptide of the present invention or a fragment of said polypeptide having protease activity additionally comprises at least one amino acid substitution at an amino acid residue selected from the group consisting of 9, 18, 43, 76, 78, 183, 194, 204, 218, 259 and 261 compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2. Hence, in one embodiment the present invention relates to a variant polypeptide having protease activity or a fragment of said polypeptide having protease activity, wherein:

[1115] (I) the polypeptide has an amino acid sequence which is at least 82%, but less than 100%, identical to the amino acid sequence as set forth in SEQ ID NO: 1 and

[1116] (ii) the polypeptide or fragment thereof comprises an amino acid substitution at two or more amino acid residues selected from the group consisting of 38, 59 and 212 compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2,

[1117] (ill) the polypeptide orfragment thereof comprises at least one amino acid substitution at an amino acid residue selected from the group consisting of 9, 18, 43, 76, 78, 183, 194, 204, 218, 259 and 261 compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2, wherein the amino acid substitutions 38F, 38H, 38N, 59A, 59H, 59N, 59S, 212H and 212M are excluded and wherein the fragment of the variant polypeptide comprises 100 to 259 consecutive amino acids of the full-length variant polypeptide.

[1118] In one embodiment the variant polypeptide of the present invention preferably has a sequence identity of 95.5% to 98% to the amino acid sequence according to SEQ ID NO: 1 and one of the following combinations of amino acid substitutions:

[1119] (a) X38R / K / H / W, X59K / E and X210I / V;

[1120] (b) X38R / K / H / W, X210I / V and X212G / M / R / K / P / A / F / H / W / N / Y / D;

[1121] (c) X59K / E, X210I / V and X212G / M / R / K / P / A / F / H / W / N / Y / D;

[1122] (d) X38R / K / H / W, X59K / E, X210I / V and X212G / M / R / K / P / A / F / H / VV / N / Y / D;

[1123] (e) X38R / K / H / W, X59K / E and X271 D;

[1124] (f) X38R / K / H / W, X212G / M / R / K / P / A / F / H / W / N / Y / D and X271 D;

[1125] (g) X59K / E, X212G / M / R / K / P / A / F / H / W / N / Y / D and X271 D;

[1126] (h) X38R / K / H / W, X59K / E, X212G / M / R / K / P / A / F / H / W / N / Y / D and X271 D;

[1127] (I) X38R / K / H / W, X59K / E, X210I / V and X271 D;

[1128] Q) X38R / K / H / W, X210I / V, X212G / M / R / K / P / A / F / H / W / N / Y / D and X271 D;

[1129] (k) X59K / E, X210I / V, X212G / M / R / K / P / A / F / H / W / N / Y / D and X271 D; or

[1130] (l) X38R / K / H / W, X59K / E, X210I / V, X212G / M / R / K / P / A / F / H / W / N / Y / D and X271 D compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2, wherein the variant polypeptide further comprises at least one amino acid substitution at an amino acid residue selected from the group consisting of 9, 18, 43, 76, 78, 183, 194, 204, 218, 259 and 261 compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2.

[1131] In one embodiment the present invention relates to a variant polypeptide having protease activity or a fragment of said polypeptide having protease activity, wherein: (i) the polypeptide has an amino acid sequence which is at least 82%, but less than 100%, identical to the amino acid sequence as set forth in SEQ ID NO: 1 and

[1132] (ii) the polypeptide or fragment thereof comprises an amino acid substitution at two or more amino acid residues selected from the group consisting of 38, 59 and 212 compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2,

[1133] (ill) the polypeptide orfragment thereof comprises at least one amino acid substitution at an amino acid residue selected from the group consisting of 9, 18, 43, 76, 78, 183, 194, 204, 218, 259 and 261 compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2, wherein the amino acid substitutions 38F, 38H, 38N, 59A, 59H, 59N, 59S, 212H and 212M are excluded, wherein the fragment of the variant polypeptide comprises 100 to 259 consecutive amino acids of the full-length variant polypeptide and wherein said variant polypeptide comprises amino acid residue D or E at position 101 , preferably E at position 101 , referring to the numbering of SEQ ID NO:2.

[1134] In one embodiment the variant polypeptide of the present invention preferably has a sequence identity of 95.5% to 98% to the amino acid sequence according to SEQ ID NO: 1 and one of the following combinations of amino acid substitutions:

[1135] (a) X38R / K / H / W, X59K / E and X210I / V;

[1136] (b) X38R / K / H / W, X210I / V and X212G / M / R / K / P / A / F / H / W / N / Y / D;

[1137] (c) X59K / E, X210I / V and X212G / M / R / K / P / A / F / H / W / N / Y / D;

[1138] (d) X38R / K / H / W, X59K / E, X210I / V and X212G / M / R / K / P / A / F / H / W / N / Y / D ;

[1139] (e) X38R / K / H / W, X59K / E and X271 D;

[1140] (f) X38R / K / H / W, X212G / M / R / K / P / A / F / H / W / N / Y / D and X271 D;

[1141] (g) X59K / E, X212G / M / R / K / P / A / F / H / W / N / Y / D and X271 D;

[1142] (h) X38R / K / H / W, X59K / E, X212G / M / R / K / P / A / F / H / W / N / Y / D and X271 D;

[1143] (I) X38R / K / H / W, X59K / E, X210I / V and X271 D;

[1144] Q) X38R / K / H / W, X210I / V, X212G / M / R / K / P / A / F / H / W / N / Y / D and X271 D;

[1145] (k) X59K / E, X210I / V, X212G / M / R / K / P / A / F / H / W / N / Y / D and X271 D; or

[1146] (l) X38R / K / H / W, X59K / E, X210I / V, X212G / M / R / K / P / A / F / H / W / N / Y / D and X271 D compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2, wherein the variant polypeptide further comprises at least one amino acid substitution at an amino acid residue selected from the group consisting of 9, 18, 43, 76, 78, 183, 194, 204, 218, 259 and 261 compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2 and wherein said variant polypeptide comprises amino acid residue D or E at position 101 , preferably E at position 101 , referring to the numbering of SEQ ID NO: 2. In one embodiment the variant polypeptide of the present invention or a fragment of said polypeptide having protease activity additionally comprises at least one amino acid substitution selected from the group consisting of X9A / Q / E / C / D, X18Q / E / D, X43K / R / C / H / D / L / S / W / A / M / Y / Q / F / I, X76D, X78N / D / R / W / F / H / K / E / L / Y / M / C / Q, X183D / E / C / Q / A / M, X194E / C / D, X204D / E / C / G, X218T / S / D, X259E / D and X261 L / M / F / E / W / Y / C compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2.

[1147] Hence, in one embodiment the present invention relates to a variant polypeptide having protease activity or a fragment of said polypeptide having protease activity, wherein:

[1148] (i) the polypeptide has an amino acid sequence which is at least 82%, but less than 100%, identical to the amino acid sequence as set forth in SEQ ID NO: 1 and

[1149] (ii) the polypeptide or fragment thereof comprises an amino acid substitution at two or more amino acid residues selected from the group consisting of 38, 59 and 212 compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2,

[1150] (iii) the polypeptide or fragment thereof comprises at least one amino acid substitution selected from the group consisting of X9A / Q / E / C / D, X18Q / E / D, X43K / R / C / H / D / L / S / W / A / M / Y / Q / F / I, X76D, X78N / D / R / W / F / H / K / E / L / Y / M / C / Q, X183D / E / C / Q / A / M, X194E / C / D, X204D / E / C / G, X218T / S / D, X259E / D and X261 L / M / F / E / W / Y / C compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2, wherein the amino acid substitutions 38F, 38H, 38N, 59A, 59H, 59N, 59S, 212H and 212M are excluded and wherein the fragment of the variant polypeptide comprises 100 to 259 consecutive amino acids of the full-length variant polypeptide.

[1151] In one embodiment the variant polypeptide of the present invention preferably has a sequence identity of 95.5% to 98% to the amino acid sequence according to SEQ ID NO: 1 and one of the following combinations of amino acid substitutions:

[1152] (a) X38R / K / H / W, X59K / E and X210I / V;

[1153] (b) X38R / K / H / W, X210I / V and X212G / M / R / K / P / A / F / H / W / N / Y / D;

[1154] (c) X59K / E, X210I / V and X212G / M / R / K / P / A / F / H / W / N / Y / D;

[1155] (d) X38R / K / H / W, X59K / E, X210I / V and X212G / M / R / K / P / A / F / H / W / N / Y / D;

[1156] (e) X38R / K / H / W, X59K / E and X271 D;

[1157] (f) X38R / K / H / W, X212G / M / R / K / P / A / F / H / W / N / Y / D and X271 D;

[1158] (g) X59K / E, X212G / M / R / K / P / A / F / H / W / N / Y / D and X271 D;

[1159] (h) X38R / K / H / W, X59K / E, X212G / M / R / K / P / A / F / H / W / N / Y / D and X271 D;

[1160] (i) X38R / K / H / W, X59K / E, X210I / V and X271 D;

[1161] Q) X38R / K / H / W, X210I / V, X212G / M / R / K / P / A / F / H / W / N / Y / D and X271 D;

[1162] (k) X59K / E, X210I / V, X212G / M / R / K / P / A / F / H / W / N / Y / D and X271 D; or

[1163] (l) X38R / K / H / W, X59K / E, X210I / V, X212G / M / R / K / P / A / F / H / W / N / Y / D and X271 D compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2, wherein the variant polypeptide further comprises at least one amino acid substitution selected from the group consisting of X9A / Q / E / C / D, X18Q / E / D, X43K / R / C / H / D / L / S / W / A / M / Y / Q / F / I, X76D, X78N / D / R / W / F / H / K / E / L / Y / M / C / Q, X183D / E / C / Q / A / M, X194E / C / D, X204D / E / C / G, X218T / S / D, X259E / D and X261 L / M / F / E / W / Y / C compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2.

[1164] In one embodiment the present invention relates to a variant polypeptide having protease activity or a fragment of said polypeptide having protease activity, wherein:

[1165] (I) the polypeptide has an amino acid sequence which is at least 82%, but less than 100%, identical to the amino acid sequence as set forth in SEQ ID NO: 1 and

[1166] (ii) the polypeptide or fragment thereof comprises an amino acid substitution at two or more amino acid residues selected from the group consisting of 38, 59 and 212 compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2,

[1167] (ill) the polypeptide or fragment thereof comprises at least one amino acid substitution selected from the group consisting of X9A / Q / E / C / D, X18Q / E / D, X43K / R / C / H / D / L / S / W / A / M / Y / Q / F / I, X76D, X78N / D / R / W / F / H / K / E / L / Y / M / C / Q, X183D / E / C / Q / A / M, X194E / C / D, X204D / E / C / G, X218T / S / D, X259E / D and X261 L / M / F / E / W / Y / C compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2, wherein the amino acid substitutions 38F, 38H, 38N, 59A, 59H, 59N, 59S, 212H and 212M are excluded, wherein the fragment of the variant polypeptide comprises 100 to 259 consecutive amino acids of the full-length variant polypeptide and wherein said variant polypeptide comprises amino acid residue D or E at position 101 , preferably E at position 101 , referring to the numbering of SEQ ID NO:2.

[1168] In one embodiment the variant polypeptide of the present invention preferably has a sequence identity of 95.5% to 98% to the amino acid sequence according to SEQ ID NO: 1 and one of the following combinations of amino acid substitutions:

[1169] (a) X38R / K / H / W, X59K / E and X210I / V;

[1170] (b) X38R / K / H / W, X210I / V and X212G / M / R / K / P / A / F / H / W / N / Y / D;

[1171] (c) X59K / E, X210I / V and X212G / M / R / K / P / A / F / H / W / N / Y / D;

[1172] (d) X38R / K / H / W, X59K / E, X210I / V and X212G / M / R / K / P / A / F / H / W / N / Y / D;

[1173] (e) X38R / K / H / W, X59K / E and X271 D;

[1174] (f) X38R / K / H / W, X212G / M / R / K / P / A / F / H / W / N / Y / D and X271 D;

[1175] (g) X59K / E, X212G / M / R / K / P / A / F / H / W / N / Y / D and X271 D;

[1176] (h) X38R / K / H / W, X59K / E, X212G / M / R / K / P / A / F / H / W / N / Y / D and X271 D;

[1177] (I) X38R / K / H / W, X59K / E, X210I / V and X271 D;

[1178] Q) X38R / K / H / W, X210I / V, X212G / M / R / K / P / A / F / H / W / N / Y / D and X271 D;

[1179] (k) X59K / E, X210I / V, X212G / M / R / K / P / A / F / H / W / N / Y / D and X271 D; or (I) X38R / K / H / W, X59K / E, X21 Ol / V, X212G / M / R / K / P / A / F / H / W / N / Y / D and X271 D compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2, wherein the variant polypeptide further comprises at least one amino acid substitution selected from the group consisting of X9A / Q / E / C / D, X18Q / E / D, X43K / R / C / H / D / L / S / W / A / M / Y / Q / F / I, X76D, X78N / D / R / W / F / H / K / E / L / Y / M / C / Q, X183D / E / C / Q / A / M, X194E / C / D, X204D / E / C / G, X218T / S / D, X259E / D and X261 L / M / F / E / W / Y / C compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2 and wherein said variant polypeptide comprises amino acid residue D or E at position 101 , preferably E at position 101 , referring to the numbering of SEQ ID NO: 2.

[1180] In one embodiment the variant polypeptide of the present invention or a fragment of said polypeptide having protease activity additionally comprises at least one amino acid substitution selected from the group consisting of X9A / Q, X18Q / E / D, X43K / R, X76D, X78N / D, X183D / E, X194E / D, X204D / E, X218S / T, X259D, and X261 F / W / Y / L compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2.

[1181] Hence, in one embodiment the present invention relates to a variant polypeptide having protease activity or a fragment of said polypeptide having protease activity, wherein:

[1182] (I) the polypeptide has an amino acid sequence which is at least 82%, but less than 100%, identical to the amino acid sequence as set forth in SEQ ID NO: 1 and

[1183] (ii) the polypeptide or fragment thereof comprises an amino acid substitution at two or more amino acid residues selected from the group consisting of 38, 59 and 212 compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2,

[1184] (ill) the polypeptide or fragment thereof comprises at least one amino acid substitution selected from the group consisting of X9A / Q, X18Q / E / D, X43K / R, X76D, X78N / D, X183D / E, X194E / D, X204D / E, X218S / T, X259D, and X261 F / W / Y / L compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2, wherein the amino acid substitutions 38F, 38H, 38N, 59A, 59H, 59N, 59S, 212H and 212M are excluded and wherein the fragment of the variant polypeptide comprises 100 to 259 consecutive amino acids of the full-length variant polypeptide.

[1185] In one embodiment the variant polypeptide of the present invention preferably has a sequence identity of 95.5% to 98% to the amino acid sequence according to SEQ ID NO: 1 and one of the following combinations of amino acid substitutions:

[1186] (a) X38R / K / H / W, X59K / E and X210I / V;

[1187] (b) X38R / K / H / W, X210I / V and X212G / M / R / K / P / A / F / H / W / N / Y / D;

[1188] (c) X59K / E, X210I / V and X212G / M / R / K / P / A / F / H / W / N / Y / D;

[1189] (d) X38R / K / H / W, X59K / E, X210I / V and X212G / M / R / K / P / A / F / H / W / N / Y / D;

[1190] (e) X38R / K / H / W, X59K / E and X271 D; (f) X38R / K / H / W, X212G / M / R / K / P / A / F / H / W / N / Y / D and X271 D;

[1191] (g) X59K / E, X212G / M / R / K / P / A / F / H / W / N / Y / D and X271 D;

[1192] (h) X38R / K / H / W, X59K / E, X212G / M / R / K / P / A / F / H / W / N / Y / D and X271 D;

[1193] (i) X38R / K / H / W, X59K / E, X210I / V and X271 D;

[1194] Q) X38R / K / H / W, X21 Ol / V, X212G / M / R / K / P / A / F / H / W / N / Y / D and X271 D;

[1195] (k) X59K / E, X210I / V, X212G / M / R / K / P / A / F / H / W / N / Y / D and X271 D; or

[1196] (l) X38R / K / H / W, X59K / E, X21 Ol / V, X212G / M / R / K / P / A / F / H / W / N / Y / D and X271 D compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2, wherein the variant polypeptide further comprises at least one amino acid substitution selected from the group consisting of X9A / Q, X18Q / E / D, X43K / R, X76D, X78N / D, X183D / E, X194E / D, X204D / E, X218S / T, X259D, and X261 F / W / Y / L compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2. In one embodiment the present invention relates to a variant polypeptide having protease activity or a fragment of said polypeptide having protease activity, wherein:

[1197] (i) the polypeptide has an amino acid sequence which is at least 82%, but less than 100%, identical to the amino acid sequence as set forth in SEQ ID NO: 1 and

[1198] (ii) the polypeptide or fragment thereof comprises an amino acid substitution at two or more amino acid residues selected from the group consisting of 38, 59 and 212 compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2,

[1199] (iii) the polypeptide or fragment thereof comprises at least one amino acid substitution selected from the group consisting of X9A / Q, X18Q / E / D, X43K / R, X76D, X78N / D, X183D / E, X194E / D, X204D / E, X218S / T, X259D, and X261 F / W / Y / L compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2, wherein the amino acid substitutions 38F, 38H, 38N, 59A, 59H, 59N, 59S, 212H and 212M are excluded, wherein the fragment of the variant polypeptide comprises 100 to 259 consecutive amino acids of the full-length variant polypeptide and wherein said variant polypeptide comprises amino acid residue D or E at position 101 , preferably E at position 101 , referring to the numbering of SEQ ID NO:2.

[1200] In one embodiment the variant polypeptide of the present invention preferably has a sequence identity of 95.5% to 98% to the amino acid sequence according to SEQ ID NO: 1 and one of the following combinations of amino acid substitutions:

[1201] (a) X38R / K / H / W, X59K / E and X21 Ol / V;

[1202] (b) X38R / K / H / W, X210I / V and X212G / M / R / K / P / A / F / H / W / N / Y / D;

[1203] (c) X59K / E, X21 Ol / V and X212G / M / R / K / P / A / F / H / W / N / Y / D;

[1204] (d) X38R / K / H / W, X59K / E, X21 Ol / V and X212G / M / R / K / P / A / F / H / W / N / Y / D;

[1205] (e) X38R / K / H / W, X59K / E and X271 D; (f) X38R / K / H / W, X212G / M / R / K / P / A / F / H / W / N / Y / D and X271 D;

[1206] (g) X59K / E, X212G / M / R / K / P / A / F / H / W / N / Y / D and X271 D;

[1207] (h) X38R / K / H / W, X59K / E, X212G / M / R / K / P / A / F / H / W / N / Y / D and X271 D;

[1208] (i) X38R / K / H / W, X59K / E, X210I / V and X271 D;

[1209] Q) X38R / K / H / W, X21 Ol / V, X212G / M / R / K / P / A / F / H / W / N / Y / D and X271 D;

[1210] (k) X59K / E, X210I / V, X212G / M / R / K / P / A / F / H / W / N / Y / D and X271 D; or

[1211] (l) X38R / K / H / W, X59K / E, X21 Ol / V, X212G / M / R / K / P / A / F / H / W / N / Y / D and X271 D compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2, wherein the variant polypeptide further comprises at least one amino acid substitution selected from the group consisting of X9A / Q, X18Q / E / D, X43K / R, X76D, X78N / D, X183D / E, X194E / D, X204D / E, X218S / T, X259D, and X261 F / W / Y / L compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2 and wherein said variant polypeptide comprises amino acid residue D or E at position 101 , preferably E at position 101 , referring to the numbering of SEQ ID NO: 2.

[1212] In one embodiment the variant polypeptide of the present invention or a fragment of said polypeptide having protease activity additionally comprises at least one amino acid substitution selected from the group consisting of S9A / Q / E / C / D, N18Q / E / D, N43K / R / C / H / D / L / S / W / A / M / Y / Q / F / I, N76D, S78N / D / R / W / F / H / K / E / L / Y / M / C / Q, N183D / E / C / Q / A / M, A194E / C / D, N204D / E / C / G, N218T / S / D, S259E / D and N261 L / M / F / E / W / Y / C compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2.

[1213] In one embodiment the variant polypeptide of the present invention or a fragment of said polypeptide having protease activity additionally comprises at least one amino acid substitution selected from the group consisting of S9A / Q, N18Q / E / D, N43K / R, N76D, S78N / D, N183D / E, A194E / D, N204D / E, N218S / T, S259D and N261 F / W / Y / L compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2.

[1214] In one embodiment the variant polypeptide of the present invention or a fragment of said polypeptide having protease activity additionally comprises at least one amino acid substitution at an amino acid residue selected from the group consisting of 18, 21 , 24, 78, 109, 144, 160, 182, 183, 204, 218, 228 and 248 compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2.

[1215] Hence, in one embodiment the present invention relates to a variant polypeptide having protease activity or a fragment of said polypeptide having protease activity, wherein:

[1216] (i) the polypeptide has an amino acid sequence which is at least 82%, but less than 100%, identical to the amino acid sequence as set forth in SEQ ID NO: 1 and

[1217] (ii) the polypeptide or fragment thereof comprises an amino acid substitution at two or more amino acid residues selected from the group consisting of 38, 59 and 212 compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2, (iii) the polypeptide orfragment thereof comprises at least one amino acid substitution at an amino acid residue selected from the group consisting of 18, 21 , 24, 78, 109, 144, 160, 182, 183, 204, 218, 228 and 248 compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2, wherein the amino acid substitutions 38F, 38H, 38N, 59A, 59H, 59N, 59S, 212H and 212M are excluded and wherein the fragment of the variant polypeptide comprises 100 to 259 consecutive amino acids of the full-length variant polypeptide.

[1218] In one embodiment the variant polypeptide of the present invention preferably has a sequence identity of 95.5% to 98% to the amino acid sequence according to SEQ ID NO: 1 and one of the following combinations of amino acid substitutions:

[1219] (a) X38R / K / H / W, X59K / E and X210I / V;

[1220] (b) X38R / K / H / W, X210I / V and X212G / M / R / K / P / A / F / H / W / N / Y / D;

[1221] (c) X59K / E, X210I / V and X212G / M / R / K / P / A / F / H / W / N / Y / D;

[1222] (d) X38R / K / H / W, X59K / E, X210I / V and X212G / M / R / K / P / A / F / H / VV / N / Y / D;

[1223] (e) X38R / K / H / W, X59K / E and X271 D;

[1224] (f) X38R / K / H / W, X212G / M / R / K / P / A / F / H / W / N / Y / D and X271 D;

[1225] (g) X59K / E, X212G / M / R / K / P / A / F / H / W / N / Y / D and X271 D;

[1226] (h) X38R / K / H / W, X59K / E, X212G / M / R / K / P / A / F / H / W / N / Y / D and X271 D;

[1227] (I) X38R / K / H / W, X59K / E, X210I / V and X271 D;

[1228] Q) X38R / K / H / W, X210I / V, X212G / M / R / K / P / A / F / H / W / N / Y / D and X271 D;

[1229] (k) X59K / E, X210I / V, X212G / M / R / K / P / A / F / H / W / N / Y / D and X271 D; or

[1230] (l) X38R / K / H / W, X59K / E, X210I / V, X212G / M / R / K / P / A / F / H / W / N / Y / D and X271 D compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2, wherein the variant polypeptide further comprises at least one amino acid substitution at an amino acid residue selected from the group consisting of 18, 21 , 24, 78, 109, 144, 160, 182, 183, 204, 218, 228 and 248 compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2.

[1231] Hence, in one embodiment the present invention relates to a variant polypeptide having protease activity or a fragment of said polypeptide having protease activity, wherein:

[1232] (I) the polypeptide has an amino acid sequence which is at least 82%, but less than 100%, identical to the amino acid sequence as set forth in SEQ ID NO: 1 and

[1233] (ii) the polypeptide or fragment thereof comprises an amino acid substitution at two or more amino acid residues selected from the group consisting of 38, 59 and 212 compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2, (iii) the polypeptide or fragment thereof comprises at least one amino acid substitution at an amino acid residue selected from the group consisting of 18, 21 , 24, 78, 109, 144, 160, 182, 183, 204, 218, 228 and 248 compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2, wherein the amino acid substitutions 38F, 38H, 38N, 59A, 59H, 59N, 59S, 212H and 212M are excluded, wherein the fragment of the variant polypeptide comprises 100 to 259 consecutive amino acids of the full-length variant polypeptide and wherein said variant polypeptide comprises amino acid residue D or E at position 101 , preferably E at position 101 , referring to the numbering of SEQ ID NO: 2.

[1234] In one embodiment the variant polypeptide of the present invention preferably has a sequence identity of 95.5% to 98% to the amino acid sequence according to SEQ ID NO: 1 and one of the following combinations of amino acid substitutions:

[1235] (a) X38R / K / H / W, X59K / E and X210I / V;

[1236] (b) X38R / K / H / W, X210I / V and X212G / M / R / K / P / A / F / H / W / N / Y / D;

[1237] (c) X59K / E, X210I / V and X212G / M / R / K / P / A / F / H / W / N / Y / D;

[1238] (d) X38R / K / H / W, X59K / E, X210I / V and X212G / M / R / K / P / A / F / H / VV / N / Y / D;

[1239] (e) X38R / K / H / W, X59K / E and X271 D;

[1240] (f) X38R / K / H / W, X212G / M / R / K / P / A / F / H / W / N / Y / D and X271 D;

[1241] (g) X59K / E, X212G / M / R / K / P / A / F / H / W / N / Y / D and X271 D;

[1242] (h) X38R / K / H / W, X59K / E, X212G / M / R / K / P / A / F / H / W / N / Y / D and X271 D;

[1243] (I) X38R / K / H / W, X59K / E, X210I / V and X271 D;

[1244] Q) X38R / K / H / W, X210I / V, X212G / M / R / K / P / A / F / H / W / N / Y / D and X271 D;

[1245] (k) X59K / E, X210I / V, X212G / M / R / K / P / A / F / H / W / N / Y / D and X271 D; or

[1246] (l) X38R / K / H / W, X59K / E, X210I / V, X212G / M / R / K / P / A / F / H / W / N / Y / D and X271 D compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2, wherein the variant polypeptide further comprises at least one amino acid substitution at an amino acid residue selected from the group consisting of 18, 21 , 24, 78, 109, 144, 160, 182, 183, 204, 218, 228 and 248 compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2 and wherein said variant polypeptide comprises amino acid residue D or E at position 101 , preferably E at position 101 , referring to the numbering of SEQ ID NO: 2.

[1247] In one embodiment the variant polypeptide of the present invention or a fragment of said polypeptide having protease activity additionally comprises at least one amino acid substitution selected from the group consisting of X18Q / E / D, X21 l / V / F / W / Y, X24K, X78N / D / R / W / F / H / K / E / L / Y / M / C / Q, X109K / A, X144N / R, X160H / S / D / E / P, X182K / R / E / C, X183D / E / C / Q / A / M, X204D / E / C / G, X218T / S / D, X228S and X248Q / R compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2. Hence, in one embodiment the present invention relates to a variant polypeptide having protease activity or a fragment of said polypeptide having protease activity, wherein:

[1248] (I) the polypeptide has an amino acid sequence which is at least 82%, but less than 100%, identical to the amino acid sequence as set forth in SEQ ID NO: 1 and

[1249] (ii) the polypeptide or fragment thereof comprises an amino acid substitution at two or more amino acid residues selected from the group consisting of 38, 59 and 212 compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2,

[1250] (ill) the polypeptide or fragment thereof comprises at least one amino acid substitution selected from the group consisting of X18Q / E / D, X21 l / V / F / W / Y, X24K, X78N / D / R / W / F / H / K / E / L / Y / M / C / Q, X109K / A, X144N / R, X160H / S / D / E / P, X182K / R / E / C, X183D / E / C / Q / A / M, X204D / E / C / G, X218T / S / D, X228S and X248Q / R compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2, wherein the amino acid substitutions 38F, 38H, 38N, 59A, 59H, 59N, 59S, 212H and 212M are excluded and wherein the fragment of the variant polypeptide comprises 100 to 259 consecutive amino acids of the full-length variant polypeptide.

[1251] In one embodiment the variant polypeptide of the present invention preferably has a sequence identity of 95.5% to 98% to the amino acid sequence according to SEQ ID NO: 1 and one of the following combinations of amino acid substitutions:

[1252] (a) X38R / K / H / W, X59K / E and X210I / V;

[1253] (b) X38R / K / H / W, X210I / V and X212G / M / R / K / P / A / F / H / W / N / Y / D;

[1254] (c) X59K / E, X210I / V and X212G / M / R / K / P / A / F / H / W / N / Y / D;

[1255] (d) X38R / K / H / W, X59K / E, X210I / V and X212G / M / R / K / P / A / F / H / W / N / Y / D ;

[1256] (e) X38R / K / H / W, X59K / E and X271 D;

[1257] (f) X38R / K / H / W, X212G / M / R / K / P / A / F / H / W / N / Y / D and X271 D;

[1258] (g) X59K / E, X212G / M / R / K / P / A / F / H / W / N / Y / D and X271 D;

[1259] (h) X38R / K / H / W, X59K / E, X212G / M / R / K / P / A / F / H / W / N / Y / D and X271 D;

[1260] (I) X38R / K / H / W, X59K / E, X210I / V and X271 D;

[1261] Q) X38R / K / H / W, X210I / V, X212G / M / R / K / P / A / F / H / W / N / Y / D and X271 D;

[1262] (k) X59K / E, X210I / V, X212G / M / R / K / P / A / F / H / W / N / Y / D and X271 D; or

[1263] (l) X38R / K / H / W, X59K / E, X210I / V, X212G / M / R / K / P / A / F / H / W / N / Y / D and X271 D compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2, wherein the variant polypeptide further comprises at least one amino acid substitution selected from the group consisting of X18Q / E / D, X21 l / V / F / W / Y, X24K, X78N / D / R / W / F / H / K / E / L / Y / M / C / Q, X109K / A, X144N / R, X160H / S / D / E / P, X182K / R / E / C, X183D / E / C / Q / A / M, X204D / E / C / G, X218T / S / D, X228S and X248Q / R compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2. In one embodiment the present invention relates to a variant polypeptide having protease activity or a fragment of said polypeptide having protease activity, wherein:

[1264] (i) the polypeptide has an amino acid sequence which is at least 82%, but less than 100%, identical to the amino acid sequence as set forth in SEQ ID NO: 1 and

[1265] (ii) the polypeptide or fragment thereof comprises an amino acid substitution at two or more amino acid residues selected from the group consisting of 38, 59 and 212 compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2,

[1266] (iii) the polypeptide or fragment thereof comprises at least one amino acid substitution selected from the group consisting of X18Q / E / D, X21 l / V / F / W / Y, X24K, X78N / D / R / W / F / H / K / E / L / Y / M / C / Q, X109K / A, X144N / R, X160H / S / D / E / P, X182K / R / E / C, X183D / E / C / Q / A / M, X204D / E / C / G, X218T / S / D, X228S and X248Q / R compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2, wherein the amino acid substitutions 38F, 38H, 38N, 59A, 59H, 59N, 59S, 212H and 212M are excluded, wherein the fragment of the variant polypeptide comprises 100 to 259 consecutive amino acids of the full-length variant polypeptide and wherein said variant polypeptide comprises amino acid residue D or E at position 101 , preferably E at position 101 , referring to the numbering of SEQ ID NO: 2.

[1267] In one embodiment the variant polypeptide of the present invention preferably has a sequence identity of 95.5% to 98% to the amino acid sequence according to SEQ ID NO: 1 and one of the following combinations of amino acid substitutions:

[1268] (a) X38R / K / H / W, X59K / E and X210I / V;

[1269] (b) X38R / K / H / W, X210I / V and X212G / M / R / K / P / A / F / H / W / N / Y / D;

[1270] (c) X59K / E, X210I / V and X212G / M / R / K / P / A / F / H / W / N / Y / D;

[1271] (d) X38R / K / H / W, X59K / E, X210I / V and X212G / M / R / K / P / A / F / H / W / N / Y / D ;

[1272] (e) X38R / K / H / W, X59K / E and X271 D;

[1273] (f) X38R / K / H / W, X212G / M / R / K / P / A / F / H / W / N / Y / D and X271 D;

[1274] (g) X59K / E, X212G / M / R / K / P / A / F / H / W / N / Y / D and X271 D;

[1275] (h) X38R / K / H / W, X59K / E, X212G / M / R / K / P / A / F / H / W / N / Y / D and X271 D;

[1276] (i) X38R / K / H / W, X59K / E, X210I / V and X271 D;

[1277] Q) X38R / K / H / W, X210I / V, X212G / M / R / K / P / A / F / H / W / N / Y / D and X271 D;

[1278] (k) X59K / E, X210I / V, X212G / M / R / K / P / A / F / H / W / N / Y / D and X271 D; or

[1279] (l) X38R / K / H / W, X59K / E, X210I / V, X212G / M / R / K / P / A / F / H / W / N / Y / D and X271 D compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2, wherein the variant polypeptide further comprises at least one amino acid substitution selected from the group consisting of X18Q / E / D, X21 l / V / F / W / Y, X24K, X78N / D / R / W / F / H / K / E / L / Y / M / C / Q, X109K / A, X144N / R, X160H / S / D / E / P, X182K / R / E / C, X183D / E / C / Q / A / M, X204D / E / C / G, X218T / S / D, X228S and X248Q / R compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2, wherein said variant polypeptide comprises amino acid residue D or E at position 101 , preferably E at position 101 , referring to the numbering of SEQ ID NO: 2.

[1280] In one embodiment the variant polypeptide of the present invention or a fragment of said polypeptide having protease activity additionally comprises at least one amino acid substitution selected from the group consisting of X18Q / E / D, X21V / I, X24K, X78N / D, X109K / A, X144N / R, X160H, X182K / R / E, X183D / E, X204D / E, X218S / T, X228S and X248Q compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2. Hence, in one embodiment the present invention relates to a variant polypeptide having protease activity or a fragment of said polypeptide having protease activity, wherein:

[1281] (I) the polypeptide has an amino acid sequence which is at least 82%, but less than 100%, identical to the amino acid sequence as set forth in SEQ ID NO: 1 and

[1282] (ii) the polypeptide or fragment thereof comprises an amino acid substitution at two or more amino acid residues selected from the group consisting of 38, 59 and 212 compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2,

[1283] (ill) the polypeptide or fragment thereof comprises at least one amino acid substitution selected from the group consisting of X18Q / E / D, X21 V / l, X24K, X78N / D, X109K / A, X144N / R, X160H, X182K / R / E, X183D / E, X204D / E, X218S / T, X228S and X248Q compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2, wherein the amino acid substitutions 38F, 38H, 38N, 59A, 59H, 59N, 59S, 212H and 212M are excluded and wherein the fragment of the variant polypeptide comprises 100 to 259 consecutive amino acids of the full-length variant polypeptide.

[1284] In one embodiment the variant polypeptide of the present invention preferably has a sequence identity of 95.5% to 98% to the amino acid sequence according to SEQ ID NO: 1 and one of the following combinations of amino acid substitutions:

[1285] (a) X38R / K / H / W, X59K / E and X210I / V;

[1286] (b) X38R / K / H / W, X210I / V and X212G / M / R / K / P / A / F / H / W / N / Y / D;

[1287] (c) X59K / E, X210I / V and X212G / M / R / K / P / A / F / H / W / N / Y / D;

[1288] (d) X38R / K / H / W, X59K / E, X210I / V and X212G / M / R / K / P / A / F / H / W / N / Y / D;

[1289] (e) X38R / K / H / W, X59K / E and X271 D;

[1290] (f) X38R / K / H / W, X212G / M / R / K / P / A / F / H / W / N / Y / D and X271 D;

[1291] (g) X59K / E, X212G / M / R / K / P / A / F / H / W / N / Y / D and X271 D;

[1292] (h) X38R / K / H / W, X59K / E, X212G / M / R / K / P / A / F / H / W / N / Y / D and X271 D;

[1293] (I) X38R / K / H / W, X59K / E, X210I / V and X271 D; Q) X38R / K / H / W, X21 Ol / V, X212G / M / R / K / P / A / F / H / W / N / Y / D and X271 D;

[1294] (k) X59K / E, X210I / V, X212G / M / R / K / P / A / F / H / W / N / Y / D and X271 D; or

[1295] (l) X38R / K / H / W, X59K / E, X21 Ol / V, X212G / M / R / K / P / A / F / H / W / N / Y / D and X271 D compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2, wherein the variant polypeptide further comprises at least one amino acid substitution selected from the group consisting of X18Q / E / D, X21V / I, X24K, X78N / D, X109K / A, X144N / R, X160H, X182K / R / E, X183D / E, X204D / E, X218S / T, X228S and X248Q compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2.

[1296] In one embodiment the present invention relates to a variant polypeptide having protease activity or a fragment of said polypeptide having protease activity, wherein:

[1297] (i) the polypeptide has an amino acid sequence which is at least 82%, but less than 100%, identical to the amino acid sequence as set forth in SEQ ID NO: 1 and

[1298] (ii) the polypeptide or fragment thereof comprises an amino acid substitution at two or more amino acid residues selected from the group consisting of 38, 59 and 212 compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2,

[1299] (iii) the polypeptide or fragment thereof comprises at least one amino acid substitution selected from the group consisting of X18Q / E / D, X21 V / l, X24K, X78N / D, X109K / A, X144N / R, X160H, X182K / R / E, X183D / E, X204D / E, X218S / T, X228S and X248Q compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2, wherein the amino acid substitutions 38F, 38H, 38N, 59A, 59H, 59N, 59S, 212H and 212M are excluded, wherein the fragment of the variant polypeptide comprises 100 to 259 consecutive amino acids of the full-length variant polypeptide and wherein said variant polypeptide comprises amino acid residue D or E at position 101 , preferably E at position 101 , referring to the numbering of SEQ ID NO: 2.

[1300] In one embodiment the variant polypeptide of the present invention preferably has a sequence identity of 95.5% to 98% to the amino acid sequence according to SEQ ID NO: 1 and one of the following combinations of amino acid substitutions:

[1301] (a) X38R / K / H / W, X59K / E and X21 Ol / V;

[1302] (b) X38R / K / H / W, X210I / V and X212G / M / R / K / P / A / F / H / W / N / Y / D;

[1303] (c) X59K / E, X21 Ol / V and X212G / M / R / K / P / A / F / H / W / N / Y / D;

[1304] (d) X38R / K / H / W, X59K / E, X21 Ol / V and X212G / M / R / K / P / A / F / H / W / N / Y / D;

[1305] (e) X38R / K / H / W, X59K / E and X271 D;

[1306] (f) X38R / K / H / W, X212G / M / R / K / P / A / F / H / W / N / Y / D and X271 D;

[1307] (g) X59K / E, X212G / M / R / K / P / A / F / H / W / N / Y / D and X271 D;

[1308] (h) X38R / K / H / W, X59K / E, X212G / M / R / K / P / A / F / H / W / N / Y / D and X271 D; (i) X38R / K / H / W, X59K / E, X210I / V and X271 D;

[1309] Q) X38R / K / H / W, X21 Ol / V, X212G / M / R / K / P / A / F / H / W / N / Y / D and X271 D;

[1310] (k) X59K / E, X210I / V, X212G / M / R / K / P / A / F / H / W / N / Y / D and X271 D; or

[1311] (l) X38R / K / H / W, X59K / E, X21 Ol / V, X212G / M / R / K / P / A / F / H / W / N / Y / D and X271 D compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2, wherein the variant polypeptide further comprises at least one amino acid substitution selected from the group consisting of X18Q / E / D, X21V / I, X24K, X78N / D, X109K / A, X144N / R, X160H, X182K / R / E, X183D / E, X204D / E, X218S / T, X228S and X248Q compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2, wherein said variant polypeptide comprises amino acid residue D or E at position 101 , preferably E at position 101 , referring to the numbering of SEQ ID NO: 2.

[1312] In one embodiment the variant polypeptide of the present invention or a fragment of said polypeptide having protease activity additionally comprises at least one amino acid substitution selected from the group consisting of N18Q / E / D, L21 l / V / F / W / Y, S24K, S78N / D / R / W / F / H / K / E / L / Y / M / C / Q, Q109K / A, S144N / R, S160H / S / D / E / P, Q182K / R / E / C, N183D / E / C / Q / A / M, N204D / E / C / G, N218T / S / D, A228S and N248Q / R compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2.

[1313] In one embodiment the variant polypeptide of the present invention or a fragment of said polypeptide having protease activity additionally comprises at least one amino acid substitution selected from the group consisting of N18Q / E / D, L21 V / I, S24K, S78N / D, Q109K / A, S144N / R, S160H, Q182K / R / E, N183D / E, N204D / E, N218S / T, A228S and N248Q compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2. In one embodiment the variant polypeptide of the present invention or a fragment of said polypeptide having protease activity additionally comprises at least one amino acid substitution at an amino acid residue selected from the group consisting of 18, 78, 183, 204 and 218 compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2.

[1314] Hence, in one embodiment the present invention relates to a variant polypeptide having protease activity or a fragment of said polypeptide having protease activity, wherein:

[1315] (i) the polypeptide has an amino acid sequence which is at least 82%, but less than 100%, identical to the amino acid sequence as set forth in SEQ ID NO: 1 and

[1316] (ii) the polypeptide or fragment thereof comprises an amino acid substitution at two or more amino acid residues selected from the group consisting of 38, 59 and 212 compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2,

[1317] (iii) the polypeptide or fragment thereof comprises at least one amino acid substitution at an amino acid residue selected from the group consisting of 18, 78, 183, 204 and 218 compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2, wherein the amino acid substitutions 38F, 38H, 38N, 59A, 59H, 59N, 59S, 212H and 212M are excluded and wherein the fragment of the variant polypeptide comprises 100 to 259 consecutive amino acids of the full-length variant polypeptide.

[1318] In one embodiment the variant polypeptide of the present invention preferably has a sequence identity of 95.5% to 98% to the amino acid sequence according to SEQ ID NO: 1 and one of the following combinations of amino acid substitutions:

[1319] (a) X38R / K / H / W, X59K / E and X21 Ol / V;

[1320] (b) X38R / K / H / W, X210I / V and X212G / M / R / K / P / A / F / H / W / N / Y / D;

[1321] (c) X59K / E, X21 Ol / V and X212G / M / R / K / P / A / F / H / W / N / Y / D;

[1322] (d) X38R / K / H / W, X59K / E, X21 Ol / V and X212G / M / R / K / P / A / F / H / VV / N / Y / D;

[1323] (e) X38R / K / H / W, X59K / E and X271 D;

[1324] (f) X38R / K / H / W, X212G / M / R / K / P / A / F / H / W / N / Y / D and X271 D;

[1325] (g) X59K / E, X212G / M / R / K / P / A / F / H / W / N / Y / D and X271 D;

[1326] (h) X38R / K / H / W, X59K / E, X212G / M / R / K / P / A / F / H / W / N / Y / D and X271 D;

[1327] (I) X38R / K / H / W, X59K / E, X210I / V and X271 D;

[1328] Q) X38R / K / H / W, X21 Ol / V, X212G / M / R / K / P / A / F / H / W / N / Y / D and X271 D;

[1329] (k) X59K / E, X210I / V, X212G / M / R / K / P / A / F / H / W / N / Y / D and X271 D; or

[1330] (l) X38R / K / H / W, X59K / E, X21 Ol / V, X212G / M / R / K / P / A / F / H / W / N / Y / D and X271 D compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2, wherein the variant polypeptide further comprises at least one amino acid substitution at an amino acid residue selected from the group consisting of 18, 78, 183, 204 and 218 compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2.

[1331] In one embodiment the present invention relates to a variant polypeptide having protease activity or a fragment of said polypeptide having protease activity, wherein:

[1332] (I) the polypeptide has an amino acid sequence which is at least 82%, but less than 100%, identical to the amino acid sequence as set forth in SEQ ID NO: 1 and

[1333] (ii) the polypeptide or fragment thereof comprises an amino acid substitution at two or more amino acid residues selected from the group consisting of 38, 59 and 212 compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2,

[1334] (ill) the polypeptide or fragment thereof comprises at least one amino acid substitution at an amino acid residue selected from the group consisting of 18, 78, 183, 204 and 218 compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2, wherein the amino acid substitutions 38F, 38H, 38N, 59A, 59H, 59N, 59S, 212H and 212M are excluded, wherein the fragment of the variant polypeptide comprises 100 to 259 consecutive amino acids of the full-length variant polypeptide and wherein said variant polypeptide comprises amino acid residue D or E at position 101 , preferably E at position 101 , referring to the numbering of SEQ ID NO: 2.

[1335] In one embodiment the variant polypeptide of the present invention preferably has a sequence identity of 95.5% to 98% to the amino acid sequence according to SEQ ID NO: 1 and one of the following combinations of amino acid substitutions:

[1336] (a) X38R / K / H / W, X59K / E and X210I / V;

[1337] (b) X38R / K / H / W, X210I / V and X212G / M / R / K / P / A / F / H / W / N / Y / D;

[1338] (c) X59K / E, X210I / V and X212G / M / R / K / P / A / F / H / W / N / Y / D;

[1339] (d) X38R / K / H / W, X59K / E, X210I / V and X212G / M / R / K / P / A / F / H / VV / N / Y / D;

[1340] (e) X38R / K / H / W, X59K / E and X271 D;

[1341] (f) X38R / K / H / W, X212G / M / R / K / P / A / F / H / W / N / Y / D and X271 D;

[1342] (g) X59K / E, X212G / M / R / K / P / A / F / H / W / N / Y / D and X271 D;

[1343] (h) X38R / K / H / W, X59K / E, X212G / M / R / K / P / A / F / H / W / N / Y / D and X271 D;

[1344] (I) X38R / K / H / W, X59K / E, X210I / V and X271 D;

[1345] Q) X38R / K / H / W, X210I / V, X212G / M / R / K / P / A / F / H / W / N / Y / D and X271 D;

[1346] (k) X59K / E, X210I / V, X212G / M / R / K / P / A / F / H / W / N / Y / D and X271 D; or

[1347] (l) X38R / K / H / W, X59K / E, X210I / V, X212G / M / R / K / P / A / F / H / W / N / Y / D and X271 D compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2, wherein the variant polypeptide further comprises at least one amino acid substitution at an amino acid residue selected from the group consisting of 18, 78, 183, 204 and 218 compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2 and wherein said variant polypeptide comprises amino acid residue D or E at position 101 , preferably E at position 101 , referring to the numbering of SEQ ID NO: 2.

[1348] In one embodiment the variant polypeptide of the present invention or a fragment of said polypeptide having protease activity additionally comprises at least one amino acid substitution selected from the group consisting of X18Q / E / D, X78N / D / R / W / F / H / K / E / L / Y / M / C / Q, X183D / E / C / Q / A / M, X204D / E / C / G and X218T / S / D compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2.

[1349] Hence, in one embodiment the present invention relates to a variant polypeptide having protease activity or a fragment of said polypeptide having protease activity, wherein:

[1350] (I) the polypeptide has an amino acid sequence which is at least 82%, but less than 100%, identical to the amino acid sequence as set forth in SEQ ID NO: 1 and (ii) the polypeptide or fragment thereof comprises an amino acid substitution at two or more amino acid residues selected from the group consisting of 38, 59 and 212 compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2,

[1351] (ill) the polypeptide or fragment thereof comprises at least one amino acid substitution selected from the group consisting of X18Q / E / D, X78N / D / R / W / F / H / K / E / L / Y / M / C / Q, X183D / E / C / Q / A / M, X204D / E / C / G and X218T / S / D compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2, wherein the amino acid substitutions 38F, 38H, 38N, 59A, 59H, 59N, 59S, 212H and 212M are excluded and wherein the fragment of the variant polypeptide comprises 100 to 259 consecutive amino acids of the full-length variant polypeptide.

[1352] In one embodiment the variant polypeptide of the present invention preferably has a sequence identity of 95.5% to 98% to the amino acid sequence according to SEQ ID NO: 1 and one of the following combinations of amino acid substitutions:

[1353] (a) X38R / K / H / W, X59K / E and X21 Ol / V;

[1354] (b) X38R / K / H / W, X210I / V and X212G / M / R / K / P / A / F / H / W / N / Y / D;

[1355] (c) X59K / E, X21 Ol / V and X212G / M / R / K / P / A / F / H / W / N / Y / D;

[1356] (d) X38R / K / H / W, X59K / E, X21 Ol / V and X212G / M / R / K / P / A / F / H / W / N / Y / D ;

[1357] (e) X38R / K / H / W, X59K / E and X271 D;

[1358] (f) X38R / K / H / W, X212G / M / R / K / P / A / F / H / W / N / Y / D and X271 D;

[1359] (g) X59K / E, X212G / M / R / K / P / A / F / H / W / N / Y / D and X271 D;

[1360] (h) X38R / K / H / W, X59K / E, X212G / M / R / K / P / A / F / H / W / N / Y / D and X271 D;

[1361] (I) X38R / K / H / W, X59K / E, X210I / V and X271 D;

[1362] Q) X38R / K / H / W, X21 Ol / V, X212G / M / R / K / P / A / F / H / W / N / Y / D and X271 D;

[1363] (k) X59K / E, X210I / V, X212G / M / R / K / P / A / F / H / W / N / Y / D and X271 D; or

[1364] (l) X38R / K / H / W, X59K / E, X21 Ol / V, X212G / M / R / K / P / A / F / H / W / N / Y / D and X271 D compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2, wherein the variant polypeptide further comprises at least one amino acid substitution selected from the group consisting of X18Q / E / D, X78N / D / R / W / F / H / K / E / L / Y / M / C / Q, X183D / E / C / Q / A / M, X204D / E / C / G and X218T / S / D compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2.

[1365] Hence, in one embodiment the present invention relates to a variant polypeptide having protease activity or a fragment of said polypeptide having protease activity, wherein:

[1366] (I) the polypeptide has an amino acid sequence which is at least 82%, but less than 100%, identical to the amino acid sequence as set forth in SEQ ID NO: 1 and (ii) the polypeptide or fragment thereof comprises an amino acid substitution at two or more amino acid residues selected from the group consisting of 38, 59 and 212 compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2,

[1367] (ill) the polypeptide or fragment thereof comprises at least one amino acid substitution selected from the group consisting of X18Q / E / D, X78N / D / R / W / F / H / K / E / L / Y / M / C / Q, X183D / E / C / Q / A / M, X204D / E / C / G and X218T / S / D compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2, wherein the amino acid substitutions 38F, 38H, 38N, 59A, 59H, 59N, 59S, 212H and 212M are excluded, wherein the fragment of the variant polypeptide comprises 100 to 259 consecutive amino acids of the full-length variant polypeptide and wherein said variant polypeptide comprises amino acid residue D or E at position 101 , preferably E at position 101 , referring to the numbering of SEQ ID NO: 2.

[1368] In one embodiment the variant polypeptide of the present invention preferably has a sequence identity of 95.5% to 98% to the amino acid sequence according to SEQ ID NO: 1 and one of the following combinations of amino acid substitutions:

[1369] (a) X38R / K / H / W, X59K / E and X210I / V;

[1370] (b) X38R / K / H / W, X210I / V and X212G / M / R / K / P / A / F / H / W / N / Y / D;

[1371] (c) X59K / E, X210 l / V and X212G / M / R / K / P / A / F / H / W / N / Y / D;

[1372] (d) X38R / K / H / W, X59K / E, X210I / V and X212G / M / R / K / P / A / F / H / W / N / Y / D ;

[1373] (e) X38R / K / H / W, X59K / E and X271 D;

[1374] (f) X38R / K / H / W, X212G / M / R / K / P / A / F / H / W / N / Y / D and X271 D;

[1375] (g) X59K / E, X212G / M / R / K / P / A / F / H / W / N / Y / D and X271 D;

[1376] (h) X38R / K / H / W, X59K / E, X212G / M / R / K / P / A / F / H / W / N / Y / D and X271 D;

[1377] (I) X38R / K / H / W, X59K / E, X210I / V and X271 D;

[1378] Q) X38R / K / H / W, X210I / V, X212G / M / R / K / P / A / F / H / W / N / Y / D and X271 D;

[1379] (k) X59K / E, X210I / V, X212G / M / R / K / P / A / F / H / W / N / Y / D and X271 D; or

[1380] (l) X38R / K / H / W, X59K / E, X210I / V, X212G / M / R / K / P / A / F / H / W / N / Y / D and X271 D compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2, wherein the variant polypeptide further comprises at least one amino acid substitution selected from the group consisting of X18Q / E / D, X78 N / D / R / W / F / H / K / E / L / Y / M / C / Q, X183D / E / C / Q / A / M, X204D / E / C / G and X218T / S / D compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2 and wherein said variant polypeptide comprises amino acid residue D or E at position 101 , preferably E at position 101 , referring to the numbering of SEQ ID NO: 2.

[1381] In one embodiment the variant polypeptide of the present invention or a fragment of said polypeptide having protease activity additionally comprises at least one amino acid substitution selected from the group consisting of X18Q / E / D, X78N / D, X183D / E, X204D and X218S compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2.

[1382] Hence, in one embodiment the present invention relates to a variant polypeptide having protease activity or a fragment of said polypeptide having protease activity, wherein:

[1383] (I) the polypeptide has an amino acid sequence which is at least 82%, but less than 100%, identical to the amino acid sequence as set forth in SEQ ID NO: 1 and

[1384] (ii) the polypeptide or fragment thereof comprises an amino acid substitution at two or more amino acid residues selected from the group consisting of 38, 59 and 212 compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2,

[1385] (ill) the polypeptide or fragment thereof comprises at least one amino acid substitution selected from the group consisting of X18Q / E / D, X78N / D, X183D / E, X204D and X218S compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2, wherein the amino acid substitutions 38F, 38H, 38N, 59A, 59H, 59N, 59S, 212H and 212M are excluded and wherein the fragment of the variant polypeptide comprises 100 to 259 consecutive amino acids of the full-length variant polypeptide.

[1386] In one embodiment the variant polypeptide of the present invention preferably has a sequence identity of 95.5% to 98% to the amino acid sequence according to SEQ ID NO: 1 and one of the following combinations of amino acid substitutions:

[1387] (a) X38R / K / H / W, X59K / E and X210I / V;

[1388] (b) X38R / K / H / W, X210I / V and X212G / M / R / K / P / A / F / H / W / N / Y / D;

[1389] (c) X59K / E, X210I / V and X212G / M / R / K / P / A / F / H / W / N / Y / D;

[1390] (d) X38R / K / H / W, X59K / E, X210I / V and X212G / M / R / K / P / A / F / H / VV / N / Y / D;

[1391] (e) X38R / K / H / W, X59K / E and X271 D;

[1392] (f) X38R / K / H / W, X212G / M / R / K / P / A / F / H / W / N / Y / D and X271 D;

[1393] (g) X59K / E, X212G / M / R / K / P / A / F / H / W / N / Y / D and X271 D;

[1394] (h) X38R / K / H / W, X59K / E, X212G / M / R / K / P / A / F / H / W / N / Y / D and X271 D;

[1395] (I) X38R / K / H / W, X59K / E, X210I / V and X271 D;

[1396] Q) X38R / K / H / W, X210I / V, X212G / M / R / K / P / A / F / H / W / N / Y / D and X271 D;

[1397] (k) X59K / E, X210I / V, X212G / M / R / K / P / A / F / H / W / N / Y / D and X271 D; or

[1398] (l) X38R / K / H / W, X59K / E, X210I / V, X212G / M / R / K / P / A / F / H / W / N / Y / D and X271 D compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2, wherein the variant polypeptide further comprises at least one amino acid substitution selected from the group consisting of X18Q / E / D, X78N / D, X183D / E, X204D and X218S compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2. Hence, in one embodiment the present invention relates to a variant polypeptide having protease activity or a fragment of said polypeptide having protease activity, wherein:

[1399] (I) the polypeptide has an amino acid sequence which is at least 82%, but less than 100%, identical to the amino acid sequence as set forth in SEQ ID NO: 1 and

[1400] (ii) the polypeptide or fragment thereof comprises an amino acid substitution at two or more amino acid residues selected from the group consisting of 38, 59 and 212 compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2,

[1401] (ill) the polypeptide or fragment thereof comprises at least one amino acid substitution selected from the group consisting of X18Q / E / D, X78N / D, X183D / E, X204D and X218S compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2, wherein the amino acid substitutions 38F, 38H, 38N, 59A, 59H, 59N, 59S, 212H and 212M are excluded, wherein the fragment of the variant polypeptide comprises 100 to 259 consecutive amino acids of the full-length variant polypeptide and wherein said variant polypeptide comprises amino acid residue D or E at position 101 , preferably E at position 101 , referring to the numbering of SEQ ID NO: 2.

[1402] In one embodiment the variant polypeptide of the present invention preferably has a sequence identity of 95.5% to 98% to the amino acid sequence according to SEQ ID NO: 1 and one of the following combinations of amino acid substitutions:

[1403] (a) X38R / K / H / W, X59K / E and X210 l / V;

[1404] (b) X38R / K / H / W, X210I / V and X212G / M / R / K / P / A / F / H / W / N / Y / D;

[1405] (c) X59K / E, X210I / V and X212G / M / R / K / P / A / F / H / W / N / Y / D;

[1406] (d) X38R / K / H / W, X59K / E, X210I / V and X212G / M / R / K / P / A / F / H / VV / N / Y / D;

[1407] (e) X38R / K / H / W, X59K / E and X271 D;

[1408] (f) X38R / K / H / W, X212G / M / R / K / P / A / F / H / W / N / Y / D and X271 D;

[1409] (g) X59K / E, X212G / M / R / K / P / A / F / H / W / N / Y / D and X271 D;

[1410] (h) X38R / K / H / W, X59K / E, X212G / M / R / K / P / A / F / H / W / N / Y / D and X271 D;

[1411] (I) X38R / K / H / W, X59K / E, X210I / V and X271 D;

[1412] Q) X38R / K / H / W, X210I / V, X212G / M / R / K / P / A / F / H / W / N / Y / D and X271 D;

[1413] (k) X59K / E, X210I / V, X212G / M / R / K / P / A / F / H / W / N / Y / D and X271 D; or

[1414] (l) X38R / K / H / W, X59K / E, X210I / V, X212G / M / R / K / P / A / F / H / W / N / Y / D and X271 D compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2, wherein the variant polypeptide further comprises at least one amino acid substitution selected from the group consisting of X18Q / E / D, X78N / D, X183D / E, X204D and X218S compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2 and wherein said variant polypeptide comprises amino acid residue D or E at position 101 , preferably E at position 101 , referring to the numbering of SEQ ID NO: 2. In one embodiment the variant polypeptide of the present invention or a fragment of said polypeptide having protease activity additionally comprises at least one amino acid substitution selected from the group consisting of N18Q / E / D, S78N / D / R / W / F / H / K / E / L / Y / M / C / Q, N183D / E / C / Q / A / M, N204D / E / C / G and N218T / S / D compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2.

[1415] In one embodiment the variant polypeptide of the present invention or a fragment of said polypeptide having protease activity additionally comprises at least one amino acid substitution selected from the group consisting of N18Q / E / D, S78N / D, N183D / E, N204D and N218S compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2.

[1416] In one embodiment the variant polypeptide of the present invention or a fragment of said polypeptide having protease activity comprises amino acid substitutions at positions 18, 21 , 24, 25, 38, 78, 103, 109, 144, 160, 182, 183, 194, 204, 210, 212, 218, 228, 248, 260 and 271 compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2 and comprises amino acid residue D or E at position 101 , preferably E at position 101 , referring to the numbering of SEQ ID NO: 2.

[1417] In one embodiment the variant polypeptide of the present invention or a fragment of said polypeptide having protease activity comprises the amino acid substitutions X18Q / E / D, X21V / I, X24K, X25D,X38R / K / H / W, X78N / D, X103S, X109K / A, X144N / R, X160H, X182K / R / E, X183D / E, X194E / D, X204D / E, X210I, X212P / A / G / R / K / F / Y / M, X218S / T, X228S, X248Q / R,248Q / R,X260D / E and X271 D compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2 and comprises amino acid residue D or E at position 101 , preferably E at position 101 , referring to the numbering of SEQ ID NO: 2.

[1418] In one embodiment the variant polypeptide of the present invention or a fragment of said polypeptide having protease activity comprises the amino acid substitutions N18Q / E / D, L21 V / I, S24K, G25D, T38R / K / H / W, S78N / D, A103S, Q109K / A, S144N / R, S160H, Q182K / R / E, N183D / E, A194E / D, N204D / E, P210I, S212P / A / G / R / K / F / Y / M, N218S / T, A228S, N248Q / R,248Q / R,T260D / E and E271 D compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2 and comprises amino acid residue D or E at position 101 , preferably E at position 101 , referring to the numbering of SEQ ID NO: 2.

[1419] Hence, in one embodiment the present invention relates to a variant polypeptide having protease activity or a fragment of said polypeptide having protease activity, wherein:

[1420] (i) the polypeptide has an amino acid sequence which is at least 82%, but less than 100%, identical to the amino acid sequence as set forth in SEQ ID NO: 1 and

[1421] (ii) the polypeptide or fragment thereof comprises amino acid substitutions at positions 18, 21 , 24, 25, 38, 78, 103, 109, 144, 160, 182, 183, 194, 204, 210, 212, 218, 228, 248, 260 and 271 compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2 and comprises amino acid residue D or E at position 101 , preferably E at position 101 , referring to the numbering of SEQ ID NO: 2. Hence, in one embodiment the present invention relates to a variant polypeptide having protease activity or a fragment of said polypeptide having protease activity, wherein:

[1422] (I) the polypeptide has an amino acid sequence which is at least 82%, but less than 100%, identical to the amino acid sequence as set forth in SEQ ID NO: 1 and

[1423] (ii) the polypeptide or fragment thereof comprises the amino acid substitutions X18Q / E / D, X21V / I, X24K, X25D,X38R / K / H / W, X78N / D, X103S, X109K / A, X144N / R, X160H, X182K / R / E, X183D / E, X194E / D, X204D / E, X210I, X212P / A / G / R / K / F / Y / M, X218S / T, X228S, X248Q / R, X260D / E and X271 D compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2 and comprises amino acid residue D or E at position 101 , preferably E at position 101 , referring to the numbering of SEQ ID NO: 2.

[1424] Hence, in one embodiment the present invention relates to a variant polypeptide having protease activity or a fragment of said polypeptide having protease activity, wherein:

[1425] (I) the polypeptide has an amino acid sequence which is at least 82%, but less than 100%, identical to the amino acid sequence as set forth in SEQ ID NO: 1 and

[1426] (ii) the polypeptide or fragment thereof comprises the amino acid substitutions N18Q / E / D, L21V / I, S24K, G25D, T38R / K / H / W, S78N / D, A103S, Q109K / A, S144N / R, S160H;, Q182K / R / E, N183D / E, A194E / D, N204D / E, P210I, S212P / A / G / R / K / F / Y / M, N218S / T, A228S, N248Q / R, T260D / E and E271 D compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2 and comprises amino acid residue D or E at position 101 , preferably E at position 101 , referring to the numbering of SEQ ID NO: 2.

[1427] Hence, in one embodiment the present invention relates to a variant polypeptide having protease activity or a fragment of said polypeptide having protease activity, wherein:

[1428] (I) the polypeptide has an amino acid sequence which is 95.5% to 98% identical to the amino acid sequence as set forth in SEQ ID NO: 1 and

[1429] (ii) the polypeptide or fragment thereof comprises amino acid substitutions at positions 18, 21 , 24, 25, 38, 78, 103, 109, 144, 160, 182, 183, 194, 204, 210, 212, 218, 228, 248, 260 and 271 compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2 and comprises amino acid residue D or E at position 101 , preferably E at position 101 , referring to the numbering of SEQ ID NO: 2.

[1430] Hence, in one embodiment the present invention relates to a variant polypeptide having protease activity or a fragment of said polypeptide having protease activity, wherein:

[1431] (I) the polypeptide has an amino acid sequence which is 95.5% to 98% identical to the amino acid sequence as set forth in SEQ ID NO: 1 and

[1432] (ii) the polypeptide or fragment thereof comprises the amino acid substitutions X18Q / E / D, X21 V / l , X24K, X25D,X38R / K / H / W, X78N / D, X103S, X109K / A, X144N / R, X160H, X182K / R / E, X183D / E, X194E / D, X204D / E, X210I, X212P / A / G / R / K / F / Y / M, X218S / T, X228S, X248Q / R,X260D / E and X271 D compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2 and comprises amino acid residue D or E at position 101 , preferably E at position 101 , referring to the numbering of SEQ ID NO: 2.

[1433] Hence, in one embodiment the present invention relates to a variant polypeptide having protease activity or a fragment of said polypeptide having protease activity, wherein:

[1434] (I) the polypeptide has an amino acid sequence which is 95.5% to 98% identical to the amino acid sequence as set forth in SEQ ID NO: 1 and

[1435] (ii) the polypeptide or fragment thereof comprises the amino acid substitutions N18Q / E / D, L21V / I, S24K, G25D, T38R / K / H / W, S78N / D, A103S, Q109K / A, S144N / R, S160H;, Q182K / R / E, N183D / E, A194E / D, N204D / E, P210I, S212P / A / G / R / K / F / Y / M, N218S / T, A228S, N248Q / R,T260D / E and E271 D compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2 and comprises amino acid residue D or E at position 101 , preferably E at position 101 , referring to the numbering of SEQ ID NO: 2.

[1436] In one embodiment, the amino acid substitution at amino acid residue 9 is X9A and / or the amino acid substitution at amino acid residue 18 is X18Q; and / or the amino acid substitution at amino acid residue 21 is X21 V; and or the amino acid substitution at amino acid residue 38 is X38K / W; and / or the amino acid substitution at amino acid residue 43 is X43K / R; and / or the amino acid substitution at amino acid residue 59 is X59K; and / or the amino acid substitution at amino acid residue 78 is X78N; and / or the amino acid substitution at amino acid residue 109 is X109K; and / or the amino acid substitution at amino acid residue 144 is X144N; and / or the amino acid substitution at amino acid residue 182 is X182K; and / or the amino acid substitution at amino acid residue 183 is X183D; and / or the amino acid substitution at amino acid residue 194 is X194E; and / or the amino acid substitution at amino acid residue 204 is X204D; and / or the amino acid substitution at amino acid residue 210 is X210I; and / or the amino acid substitution at amino acid residue 212 is X212P; and / or the amino acid substitution at amino acid residue 218 is X218S; and / or the amino acid substitution at amino acid residue 248 is X248Q; and / or the amino acid substitution at amino acid residue 259 is X259D; and / or the amino acid substitution at amino acid residue 260 is X260D; and / or the amino acid substitution at amino acid residue 261 is X261 F.

[1437] In one embodiment the variant polypeptide of the present invention or a fragment of said polypeptide having protease activity additionally comprises at least one amino acid substitution at an amino acid residue selected from the group consisting of 8, 108, 133, 137 and 215 compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2.

[1438] Hence, in one embodiment the present invention relates to a variant polypeptide having protease activity or a fragment of said polypeptide having protease activity, wherein:

[1439] (i) the polypeptide has an amino acid sequence which is at least 82%, but less than 100%, identical to the amino acid sequence as set forth in SEQ ID NO: 1 and (ii) the polypeptide or fragment thereof comprises an amino acid substitution at two or more amino acid residues selected from the group consisting of 38, 59 and 212 compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2,

[1440] (ill) the polypeptide or fragment thereof comprises at least one amino acid substitution at an amino acid residue selected from the group consisting of 8, 108, 133, 137 and 215 compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2, wherein the amino acid substitutions 38F, 38H, 38N, 59A, 59H, 59N, 59S, 212H and 212M are excluded and wherein the fragment of the variant polypeptide comprises 100 to 259 consecutive amino acids of the full-length variant polypeptide.

[1441] In one embodiment the variant polypeptide of the present invention preferably has a sequence identity of 95.5% to 98% to the amino acid sequence according to SEQ ID NO: 1 and one of the following combinations of amino acid substitutions:

[1442] (a) X38R / K / H / W, X59K / E and X210I / V;

[1443] (b) X38R / K / H / W, X210I / V and X212G / M / R / K / P / A / F / H / W / N / Y / D;

[1444] (c) X59K / E, X210I / V and X212G / M / R / K / P / A / F / H / W / N / Y / D;

[1445] (d) X38R / K / H / W, X59K / E, X210I / V and X212G / M / R / K / P / A / F / H / VV / N / Y / D;

[1446] (e) X38R / K / H / W, X59K / E and X271 D;

[1447] (f) X38R / K / H / W, X212G / M / R / K / P / A / F / H / W / N / Y / D and X271 D;

[1448] (g) X59K / E, X212G / M / R / K / P / A / F / H / W / N / Y / D and X271 D;

[1449] (h) X38R / K / H / W, X59K / E, X212G / M / R / K / P / A / F / H / W / N / Y / D and X271 D;

[1450] (I) X38R / K / H / W, X59K / E, X210I / V and X271 D;

[1451] Q) X38R / K / H / W, X210 l / V, X212G / M / R / K / P / A / F / H / W / N / Y / D and X271 D;

[1452] (k) X59K / E, X210I / V, X212G / M / R / K / P / A / F / H / W / N / Y / D and X271 D; or

[1453] (l) X38R / K / H / W, X59K / E, X210I / V, X212G / M / R / K / P / A / F / H / W / N / Y / D and X271 D compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2, wherein the variant polypeptide further comprises at least one amino acid substitution at an amino acid residue selected from the group consisting of 8, 108, 133, 137 and 215 compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2.

[1454] In one embodiment the present invention relates to a variant polypeptide having protease activity or a fragment of said polypeptide having protease activity, wherein:

[1455] (I) the polypeptide has an amino acid sequence which is at least 82%, but less than 100%, identical to the amino acid sequence as set forth in SEQ ID NO: 1 and (ii) the polypeptide or fragment thereof comprises an amino acid substitution at two or more amino acid residues selected from the group consisting of 38, 59 and 212 compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2,

[1456] (ill) the polypeptide or fragment thereof comprises at least one amino acid substitution at an amino acid residue selected from the group consisting of 8, 108, 133, 137 and 215 compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2, wherein the amino acid substitutions 38F, 38H, 38N, 59A, 59H, 59N, 59S, 212H and 212M are excluded, wherein the fragment of the variant polypeptide comprises 100 to 259 consecutive amino acids of the full-length variant polypeptide and wherein sai...

Claims

CLAIMS1 . A variant polypeptide having protease activity or a fragment of said polypeptide having protease activity, wherein:(I) the polypeptide has an amino acid sequence which is at least 82%, but less than 100%, identical to the amino acid sequence as set forth in SEQ ID NO: 1 and(ii) the polypeptide or fragment thereof comprises an amino acid substitution at two or more amino acid residues selected from the group consisting of 38, 59 and 212 compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2, wherein the amino acid substitutions 38F, 38H, 38N, 59A, 59H, 59N, 59S, 212H and 212M are excluded and wherein the fragment of the variant polypeptide comprises 100 to 259 consecutive amino acids of the full-length variant polypeptide.

2. The variant polypeptide of claim 1 , wherein:(a) the amino acid substitution at amino acid residue 38 is X38R / KW; and / or(b) the amino acid substitution at amino acid residue 59 is X59K / E; and / or(c) the amino acid substitution at amino acid residue 212 is X212G / R / K / P / A / F / W / N / Y / D compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2.

3. The variant polypeptide of claim 1 or 2, wherein the polypeptide or fragment thereof additionally comprises an amino acid substitution at amino residue 210 and / or amino acid residue 271 compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2.

4. The variant polypeptide of claim 3, wherein:(a) the amino acid substitution at amino acid residue 210 is X21 OIAZ; and / or(b) the amino acid substitution at amino acid residue 271 is X271 D.

5. The variant polypeptide of any one of the preceding claims, wherein the polypeptide or fragment thereof additionally comprises at least one amino acid substitution at an amino acid residue selected from the group consisting of 9, 18, 21, 43, 78, 160, 183, 194, 204, 218, 228, 259, 260 and 261 compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2.

6. The variant polypeptide of claim 5, wherein:(a) the amino acid substitution at amino acid residue 9 is X9A / Q / E / C / D, preferably is X9A / Q; and / or(b) the amino acid substitution at amino acid residue 18 is X18Q / E / D; and / or(c) the amino acid substitution at amino acid residue 21 is X21 l / V / F / W / Y, preferably is X21 V / l; and / or(d) the amino acid substitution at amino acid residue 43 is X43K / R / C / H / D / L / S / W / A / M / Y / Q / F / I , preferably is X43K / R; and / or(e) the amino acid substitution at amino acid residue 78 is X78N / D / R / W / F / H / K / E / L / Y / M / C / Q, preferably is X78N / D; and / or(f) the amino acid substitution at amino acid residue 160 is X160H / S / D / E / P, preferably is X160H, and / or(g) the amino acid substitution at amino acid residue 183 is X183D / E / C / Q / A / M, preferably is X183D / E; and / or(h) the amino acid substitution at amino acid residue 194 is X194E / C / D, preferably is X194E / D; and / or(i) the amino acid substitution at amino acid residue 204 is X204D / E / C / G, preferably is X204D / E; and / or(j) the amino acid substitution at amino acid residue 218 is X218T / S / D, preferably is X218S / T; and / or(k) the amino acid substitution at amino acid residue 228 is X228S; and / or(l) the amino acid substitution at amino acid residue 259 is X259E / D, preferably is X259D; and / or(m) the amino acid substitution at amino acid residue 260 is X260D / E / K, preferably is X260D / E; and / or(n) the amino acid substitution at amino acid residue 261 is X261 L / M / F / E / W / Y / C, preferably is X261 F / W / Y / L.

7. The variant polypeptide of any one of the preceding claims, wherein said polypeptide additionally comprises at least one amino acid substitution at an amino acid residue selected from the group consisting of 24, 25, 103, 109, 144, 182 and 248 compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2.

8. The variant polypeptide of any one of the preceding claims, wherein said polypeptide additionally comprises at least one amino acid substitution at an amino acid residue selected from the group consisting of 3, 76 and 256 compared to the amino acid sequence as set forth in SEQ ID NO: 3 and referring to the numbering of SEQ ID NO: 2.

9. The variant polypeptide of any one of the preceding claims, wherein said polypeptide comprises amino acid residue D or E at position 101, preferably E at position 101, referring to the numbering of SEQ ID NO: 2.

10. The variant polypeptide of any one of the preceding claims, wherein the polypeptide exhibits one or more improved properties compared to the protease according to SEQ ID NO: 3 or 4, preferably wherein the improved properties are selected from:(I) increase in stability,(ii) increase in storage stability, and(ill) increase in storage stability in a detergent composition.

11. A polynucleotide encoding the variant polypeptide of any one of the preceding claims.

12. A composition comprising the variant polypeptide of any one of claims 1 to 10 and at least one additional component, preferably wherein the composition comprises an enzyme stabilizing system, wherein the enzyme stabilizing system preferably comprises at least one compound selected from the group consisting of polyols (preferably, 1,3-propanediol, ethylene glycol, glycerol, 1 ,2-propanediol, or sorbitol), inorganic salts (preferably, CaCI2, MgCI2, or NaCI), short chain (preferably, C1-C3) carboxylic acids or salts thereof (preferably, formic acid, formate (preferably, sodium formate), acetic acid, acetate, or lactate), borate, boric acid, boronic acids (preferably, 4-formyl phenylboronic acid (4-FPBA)), peptide aldehydes (preferably, Z-VAL-H or Z-GAY-H), peptide acetals, and peptide aldehyde hydrosulfite adducts, preferably peptide aldehydes (preferably, Z-VAL-H or Z-GAY-H).

13. The composition of claim 12, wherein the composition comprises one or more additional enzymes different from the variant polypeptide referred to in any of the preceding claims, preferably one or more additional enzymes selected from the group consisting of amylases, second proteases, lipases, cellulases, hemicellulases, mannanases, xylanases, DNAses, dispersins, pectinases, oxidoreductases, and cutinases, preferably selected from amylases, mannanases, cellulase, and lipases, most preferably amylases.

14. The composition of any one of claims 12 or 13, wherein the composition is a detergent composition, preferably a laundry detergent composition or a hard surface cleaning detergent composition, preferably wherein the composition comprises one or more surfactants and / or one or more builder, preferably strong sequestering builder.

15. The composition of any one of claims 12 to 14, wherein the composition further comprises 2-phenoxyethanol and / or 4, 4' -dichloro 2-hydroxydiphenylether, preferably comprising phenoxyethanol in an amount ranging from 2ppm to 5% by weight of the composition; more preferably comprising 0.1 to 2% of phenoxyethanol by weight of the composition and / or preferably comprising 4, 4' -dichloro 2-hydroxydiphenylether in a concentration from 0.001 to 3%, preferably 0.002 to 1%, more preferably 0.01 to 0.6%, each by weight of the composition.

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