Dna enzyme variants

Abstract

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

Description

[0001] This application is a divisional application of Chinese invention patent application filed on October 26, 2018, application number: 201880067842.5, entitled "DNA enzyme variant".

[0002] References to sequence lists

[0003] This application contains a sequence list in computer-readable form, which is incorporated herein by reference. Technical Field

[0004] This invention relates to novel DNase variants that exhibit alterations in one or more properties relative to the parental DNase, including: washing performance, detergent stability, and / or storage stability. The variants of this invention are suitable for use in cleaning processes and detergent compositions, such as laundry compositions and dishwashing compositions (including hand wash and automatic dishwashing compositions). This invention also relates to isolated DNA sequences encoding these variants, expression vectors, host cells, and methods for generating and using the DNase variants of this invention. Background Technology

[0005] For decades, enzymes have been used in detergents, with proteases and amylases being the most commercially relevant for their effectiveness in removing protein- and starch-related stains, respectively. However, most household care stains are complex mixtures of various organic substances. Hard surfaces and fabrics are exposed to skin debris, such as dead cells, sweat and sebum, contaminants, and microorganisms from external sources and from sources such as the human body. Microorganisms can adhere to stains on clothing or surfaces, and such adhered microorganisms can multiply and form biofilms. Biofilms are often embedded within a matrix of extracellular polymeric substances (EPS). Biofilm EPS are polymeric aggregates generally composed of extracellular DNA, proteins, and polysaccharides. Household care stains consist of different organic stains, such as proteins, starches, greases, and DNA. Therefore, stain removal requires different enzyme activities, depending on the specific stain being targeted.

[0006] In order to be useful in the cleaning process, DNA enzymes such as laundry detergent enzymes need to be stable in the detergent composition and compatible with standard detergent components such as surfactants, builders, bleach, etc.

[0007] This invention provides such enzymes suitable for use in detergents, for example, for improving laundry and dishwashing / washing machines. Summary of the Invention

[0008] This invention relates to DNA enzyme variants that are particularly suitable for detergents and cleaning processes, such as laundry and dishwashing.

[0009] One aspect of the invention relates to a DNAse variant comprising two or more substitutions selected from the group consisting of: T1I, T1L, T1V, S13Y, T22P, S25P, S27L, S39P, S42G, S42A, S42T, S57W, S57Y, S57F, S59V, S59I, S59L, V76L, V76I, Q109R, S116D, S116E, T127V, T127I, T127L, S144P, A147H, S167L, S167I, S167V, G175D, and G175E, wherein the positions correspond to the positions in SEQ ID NO:1 (numbered according to SEQ ID NO:1), and wherein the variant corresponds to the polypeptide shown in SEQ ID NO:1 or SEQ ID NO:1. The polypeptide shown in NO:28 has at least 90% sequence identity, and the variant therein has DNase activity. One aspect of the invention relates to a DNase variant comprising two or more of the following mutations compared to the DNase having SEQ ID NO:1: T1I, T1L, T1V, S13Y, T22P, S25P, S27L, S39P, S42G, S42A, S42T, S57W, S57Y, S57F, S59V, S59I, S59L, V76L, V76I, Q109R, S116D, S116E, T127V, T127I, T127L, S144P, A147H, S167L, S167I, S167V, G175D, G175E, wherein the position corresponds to the position in SEQ ID NO:1 (numbered according to SEQ ID NO:1), wherein the variant corresponds to the position in SEQ ID NO:1. The polypeptide shown in NO:1 has at least 80% sequence identity and the variant has DNase activity.

[0010] Another aspect of the invention relates to a method for obtaining a DNA enzyme variant, the method comprising:

[0011] a) Introducing two or more of the following mutations into the parental DNase: T1I, T1L, T1V, S13Y, T22P, S25P, S27L, S39P, S42G, S42A, S42T, S57W, S57Y, S57F, S59V, S59I, S59L, V76L, V76I, Q109R, S116D, S116E, T127V, T127I, T127L, S144P, A147H, S167L, S167I, S167V, G175D, G175E, wherein the positions correspond to the positions in SEQ ID NO:1 (numbered according to SEQ ID NO:1), and wherein the variants have DNase activity.

[0012] b) and recycle the variant.

[0013] A third aspect of the invention relates to the use of the DNA enzyme variant according to the invention for deep cleaning articles, wherein the articles are textiles or hard surfaces.

[0014] sequence

[0015] SEQ ID NO:1: Mature polypeptide obtained from Bacillus cibi

[0016] SEQ ID NO:2: Mature polypeptide obtained from Bacillus Horikoshi

[0017] SEQ ID NO:3: Mature polypeptide obtained from Bacillus sp.-62520

[0018] SEQ ID NO:4: Mature polypeptide obtained from Bacillus Horikoshi

[0019] SEQ ID NO:5: Mature polypeptide obtained from Bacillus Horikoshi

[0020] SEQ ID NO:6: Mature polypeptide obtained from Bacillus species-16840

[0021] SEQ ID NO:7: Mature polypeptide obtained from Bacillus species-16840

[0022] SEQ ID NO:8: Mature polypeptide obtained from Bacillus species-62668

[0023] SEQ ID NO:9: Mature polypeptide obtained from Bacillus species-13395

[0024] SEQ ID NO:10: Mature polypeptide obtained from Bacillus horneckiae

[0025] SEQ ID NO:11: Mature polypeptide obtained from Bacillus species-11238

[0026] SEQ ID NO:12: Mature polypeptide obtained from Bacillus species-62451

[0027] SEQ ID NO:13: Mature polypeptide obtained from Bacillus species-18318

[0028] SEQ ID NO:14: Mature polypeptide obtained from Bacillus idriensis, a strain from the Institute of Pathogenic Biology.

[0029] SEQ ID NO 15 is a mature polypeptide obtained from Bacillus subtilis.

[0030] SEQ ID NO:16: Mature polypeptide obtained from environmental sample J

[0031] SEQ ID NO:17: Mature polypeptide obtained from Bacillus vietnamensis

[0032] SEQ ID NO:18: Mature polypeptide obtained from Bacillus hwajinpoensis

[0033] SEQ ID NO:19: Mature polypeptide obtained from Paenibacillus mucilaginosus

[0034] SEQ ID NO:20: Mature polypeptide obtained from Bacillus indicus

[0035] SEQ ID NO:21: Mature polypeptide obtained from Bacillus marisflavi SEQ ID NO:22: Mature polypeptide obtained from Bacillus luciferensis

[0036] SEQ ID NO:23: Mature polypeptide obtained from Bacillus chrysogenum

[0037] SEQ ID NO:24: Mature polypeptide obtained from Bacillus species SA2-6

[0038] SEQ ID NO:25: Motif [D / M / L][S / T]GYSR[D / N]

[0039] SEQ ID NO:26: motif ASXNRSKG

[0040] SEQ ID NO:27: DNAse variant

[0041] SEQ ID NO:28: DNAse variant

[0042] definition

[0043] The term "auxiliary material" refers to any liquid, solid, or gaseous material selected for a specific type of detergent composition and product form (e.g., liquid, granules, powder, stick, paste, spray, tablet, gel, or foam composition), which is also preferably compatible with the DNase variant enzyme used in the composition. In some respects, granular compositions are in a "compressed" form, while in others, liquid compositions are in a "concentrated" form.

[0044] The term "allelic variant" refers to any of two or more alternative forms of a gene occupying the same chromosomal locus. Allelic variations arise naturally through mutation and can lead to polymorphism within a population. Gene mutations can be silent (encoding a polypeptide with no change) or can encode a polypeptide with a modified amino acid sequence. Allelic variants of a polypeptide are polypeptides encoded by allelic variants of a gene.

[0045] The term "biofilm" refers to any group of microorganisms whose cells adhere to each other on a surface, such as textiles, tableware, or hard surfaces. These adherent cells are often embedded within a matrix of extracellular polymeric substances (EPS). Biofilms (EPS) are polymeric aggregates generally composed of extracellular DNA, proteins, and polysaccharides. Biofilms can form on living or non-living surfaces. Microbial cells growing in biofilms are physiologically different from planktonic cells of the same organism (in contrast, planktonic cells are single cells that can float or float in a liquid medium). Bacteria living in biofilms often exhibit significantly different characteristics from freely floating bacteria of the same species because the dense and protected environment of the membrane allows them to cooperate and interact in different ways. One benefit of this environment is increased resistance to detergents and antibiotics, as the dense extracellular matrix and the outer layer of the cells protect the interior of the community. Bacteria that produce biofilms during laundry washing can be found in the following species: Acinetobacter sp., Aeromicrobium sp., Brevundimonas sp., Microbacterium sp., Micrococcus luteus, Pseudomonas sp., Staphylococcus epidermidis, and Stenotrophomonas sp.

[0046] The term "cDNA" refers to a DNA molecule that can be prepared by reverse transcription from mature, spliced ​​mRNA molecules derived from prokaryotic or eukaryotic cells. cDNA lacks intron sequences that may be present in the corresponding genomic DNA. The initial primary RNA transcript is the precursor of mRNA, which undergoes a series of steps (including splicing) to become mature, spliced ​​mRNA.

[0047] The term "clade" refers to a group of polypeptides grouped together based on homology traits traced back to a common ancestor. A polypeptide clade can be viewed as a phylogenetic tree, and a clade is a group of polypeptides consisting of a common ancestor and all its direct descendants. Polypeptides forming a clade, such as those shown in a phylogenetic tree, often share common properties and are functionally more closely related to other polypeptides not in the clade.

[0048] The term "coding sequence" refers to a polynucleotide that directly identifies the amino acid sequence of its polypeptide product. The boundaries of this coding sequence are generally determined by an open reading frame (ORF), which typically begins with an ATG start codon or alternative start codons (such as GTG and TTG) and ends with a stop codon (such as TAA, TAG, and TGA). The coding sequence can be a polynucleotide from DNA, cDNA, synthetic, or recombinant sources.

[0049] The term "control sequence" refers to all components necessary for the expression of a polynucleotide encoding a variant of the present invention. Each control sequence for a polynucleotide encoding a variant may be native or exogenous, or may be native or exogenous to each other. Such control sequences include, but are not limited to, leader sequences, polyadenylation sequences, propeptide sequences, promoters, signal peptide sequences, and transcription terminators. At a minimum, control sequences include promoters and transcription and translation termination signals. These control sequences may provide adapters for introducing specific restriction enzyme sites that facilitate the linking of these control sequences to the coding regions of polynucleotides encoding variants.

[0050] The term "deep cleaning" refers to the destruction or removal of components of organic matter (such as biofilms and EPS, such as polysaccharides, proteins, DNA, dirt) or other components present in organic matter.

[0051] Unless otherwise specified, the term "detergent composition" includes general-purpose or heavy-duty detergents in granular or powder form, especially cleaning detergents; all-purpose cleaners in liquid, gel, or paste form, especially so-called heavy-duty liquid (HDL) types; liquid de-slip detergents; hand-washing or light-duty dishwashing detergents, especially high-foaming types; machine-washing dishwashing detergents, including various sheet, granular, liquid, and rinsing aid types for home and institutional use; liquid cleaning and disinfecting agents, including antibacterial hand sanitizers, cleaning sticks, soap sticks, mouthwashes, denture cleaners, car or carpet cleaners, and bathroom cleaners; shampoos and conditioners; shower gels and body washes; metal cleaners; and cleaning aids such as bleach additives and "stain remover sticks" or pretreatment types. The terms "detergent composition" and "detergent formulation" are used with respect to mixtures in a washing medium intended for cleaning soiled objects. In some respects, the term "laundry detergent" (e.g., "laundry detergent") is used with respect to the washing of fabrics and / or clothing. In alternative contexts, the term refers to other detergents, such as those used for cleaning tableware, knives, etc. (e.g., "dishwashing detergent"). It is not intended to limit the invention to any particular detergent formulation or composition. The term "detergent composition" is not intended to be limited to compositions containing surfactants. It is intended that, except for variations according to the invention, the term encompasses detergents that may contain: for example, surfactants, builders, chelators or chelating agents, bleaching systems or bleaching components, polymers, fabric conditioners, foaming agents, defoaming agents, dyes, fragrances, darkening inhibitors, optical brighteners, bactericides, fungicides, dirt suspending agents, corrosion inhibitors, enzyme inhibitors or stabilizers, enzyme activators, one or more transferases, hydrolases, oxidoreductases, bluing agents and fluorescent dyes, antioxidants, and solubilizers.

[0052] The terms "DNase," "DNase variant," or "DNase parent" refer to a polypeptide with DNase activity that catalyzes the hydrolytic breakage of phosphodiester bonds in DNA, thereby degrading DNA. DNases belong to the subgroup of esterases (EC-3.1), hydrolases. DNases are classified, for example, as EC3.1.11, EC3.1.12, EC3.1.15, EC3.1.16, EC3.1.21, EC3.1.22, EC 3.1.23, EC 3.1.24, and EC3.1.25, and EC 3.1.21.X, where X = 1, 2, 3, 4, 5, 6, 7, 8, or 9. The terms "DNase" and the expression "polypeptide with DNase activity" are used interchangeably in this application. For the purposes of this invention, DNase activity is determined according to the procedure described in Assay I. In some aspects, the DNase variants of this invention have improved DNase activity compared to parental DNases. In some aspects, the DNAse variants of the present invention have at least 100%, for example at least 110%, at least 120%, at least 130%, at least 140%, at least 150%, at least 160%, at least 170%, at least 180%, at least 190%, or at least 200% DNAse activity compared to the polypeptide shown in SEQ ID NO:1, or have, for example, from 100% to 200% DNAse activity compared to the polypeptide shown in SEQ ID NO:1.

[0053] The term "effective amount of enzyme" refers to the amount of enzyme necessary to achieve the desired enzymatic activity in a particular application, such as in a defined detergent composition. Such an effective amount is readily determined by those skilled in the art and is based on many factors, such as the specific enzyme used, the cleaning application, the specific composition of the detergent composition, and whether a liquid or dry (e.g., granular, strip) composition is required. The term "effective amount of DNAase variant" refers to the amount of the aforementioned DNAase variant that achieves the desired level of enzymatic activity (e.g., in a defined detergent composition).

[0054] The term “expression” includes any step involved in generating variants, including but not limited to: transcription, post-transcriptional modification, translation, post-translational modification, and secretion.

[0055] The term "expression vector" refers to a linear or circular DNA molecule that contains a polynucleotide encoding a variant and is operatively linked to additional nucleotides that provide its expression.

[0056] The term "fabric" encompasses any textile material. Therefore, it is intended that the term cover clothing, as well as fabrics, yarns, fibers, nonwoven materials, natural materials, synthetic materials, and any other textile material.

[0057] The term "high detergent concentration" system includes detergents in which detergent components present in wash water at concentrations greater than approximately 2000 ppm. High detergent concentration systems are generally considered to be used for detergents in Europe.

[0058] The term "host cell" refers to any cell type susceptible to transformation, transfection, transduction, etc., using nucleic acid constructs or expression vectors containing the polynucleotides of this invention. The term "host cell" also encompasses any parental cell progeny that is not entirely identical to the parental cell due to mutations occurring during replication.

[0059] The term "improved property" refers to a characteristic associated with a variant that is improved compared to the parent and / or to a DNase having SEQ ID NO:1, or to a DNase having the same amino acid sequence as the variant but without alterations at two or more of the specified positions. Such improved properties include, but are not limited to, stability, such as detergent stability, and washing performance, such as deep cleaning effect, which may include, but is not limited to, adhesive removal effect. Improved properties also include improved stability in the presence of certain enzymes (e.g., protease stability) or substances (e.g., in the presence of sulfites (e.g., sulfite stability)).

[0060] The term “improved DNase activity” is defined herein as altered DNase activity, for example by increasing the catalytic hydrolysis of phosphodiester bonds in DNA, i.e., the DNase variant exhibits a change in activity relative to (or compared to) the parental DNase activity (e.g., compared to the DNase having SEQ ID NO: 1 or compared to the DNase having the same amino acid sequence but without the alteration at one or more of the specified positions).

[0061] The term "low detergent concentration" system refers to detergents in which detergent components are present in wash water at concentrations of less than about 800 ppm. Asian (e.g., Japanese) detergents are typically considered to be low detergent concentration systems.

[0062] The term "improved washing performance" includes, but is not limited to, the term "deep cleaning effect." Improved properties, such as those compared to the DNAse parent (the DNAse shown in SEQ ID NO:1) or to a DNAse having the same amino acid sequence as the variant but without the alteration at one or more of the specified positions, were used to measure the deep cleaning performance of the DNAse variant according to the invention. Improved performance, such as deep cleaning performance, can be expressed as the reflectance value of a small sample of soiled fabric. After washing and rinsing, the small samples were laid flat and allowed to air dry overnight at room temperature. All washed small samples were evaluated the day after washing. The reflectance of the small samples was evaluated using a Macbeth Color Eye 7000 reflectance spectrophotometer with a very small aperture. Measurements were taken under conditions without UV in the incident light, and reflectance at 460 nm was extracted. A positive reaction indicates that dirt was removed, including dirt adhered to the fabric due to, for example, biofilms or EPS viscous layers.

[0063] The term "isolated polynucleotide" refers to a polynucleotide that has been artificially modified. In some respects, such as as determined by agarose gel electrophoresis, the isolated polynucleotide is at least 1% pure, for example at least 5% pure, at least 10% pure, at least 20% pure, at least 40% pure, at least 60% pure, at least 80% pure, at least 90% pure, and at least 95% pure. The polynucleotide can be genomic, cDNA, RNA, semi-synthetic, synthetically derived, or any combination thereof.

[0064] The term "laundry" encompasses both household and industrial laundry and refers to the process of treating textiles with a solution containing the cleaning or detergent composition of the present invention. The laundry process can be performed, for example, using a household or industrial washing machine or manually.

[0065] The term "mature polypeptide" refers to a polypeptide in its final form after translation and any post-translational modifications (such as N-terminal processing, C-terminal truncation, glycosylation, phosphorylation, etc.). In some aspects, the mature polypeptide is amino acids 1 to 182 of SEQ ID NO: 1. The N-terminus of the mature polypeptide used according to the invention has been experimentally confirmed based on EDMAN N-terminal sequencing data and complete MS data. It is known in the art that host cells can produce mixtures of two or more different mature polypeptides (i.e., with different C-terminal and / or N-terminal amino acids) expressed by the same polynucleotide.

[0066] The term "mature polypeptide coding sequence" refers to a polynucleotide that encodes a mature polypeptide with DNase activity.

[0067] The term "medium detergent concentration" refers to detergents in which detergent components are present in wash water at concentrations between approximately 800 ppm and approximately 2000 ppm. North American detergents are generally considered to be in the medium detergent concentration system.

[0068] The term "nucleic acid construct" refers to a single-stranded or double-stranded nucleic acid molecule isolated from naturally occurring genes, modified to contain nucleic acid fragments in a manner not found in nature, or synthesized. When the nucleic acid construct contains the control sequence required to express the coding sequence of this invention, the term "nucleic acid construct" is synonymous with the term "expression cassette."

[0069] The term "non-textile detergent composition" includes detergent compositions for non-textile surfaces, including but not limited to compositions for cleaning hard surfaces, such as dishwashing detergent compositions (including manual dishwashing compositions), oral compositions, denture compositions, and personal hygiene compositions.

[0070] The term "operably linked" refers to a configuration in which a control sequence is placed in the appropriate position relative to the coding sequence of a polynucleotide, such that the control sequence guides the expression of the coding sequence.

[0071] The terms "parental" DNase, DNase parent, or precursor DNase are used interchangeably. In the context of this invention, "parental DNase" should be understood as a DNase in which at least one alteration is made in the amino acid sequence to produce a DNase variant having an amino acid sequence less than 100% identical to that of the DNase in which the alteration is made, i.e., the parental DNase. Thus, a parent is a DNase having the same amino acid sequence as the variant but without alterations at one or more (or two or more) of these designated positions. It should be understood that the expression "having the same amino acid sequence" in this context refers to 100% sequence identity. In a specific aspect, the DNAseed parent is a DNAseed having at least 60%, at least 61%, at least 62%, at least 63%, at least 64%, at least 65%, at least 66%, at least 67%, at least 68%, at least 69%, at least 70%, at least 72%, at least 73%, at least 74%, at least 75%, at least 80%, at least 81%, at least 82%, at least 83%, at least 84%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, for example at least 99.1%, at least 99.2%, at least 99.3%, at least 99.4%, at least 99.5%, at least 99.6%, or 100% identity with the polypeptide shown in SEQ ID NO:1.

[0072] The term “relevant washing conditions” used in this article refers to the conditions actually used in the household within the detergent segment, specifically washing temperature, time, washing mechanics, detergent concentration, detergent type, and water hardness.

[0073] The term "stability" includes storage stability and stability during use (e.g., during washing), and reflects the stability of the DNase variant according to the invention as a function of time, such as how much activity is retained when the DNase variant is placed in solution, particularly in a detergent solution. This stability is affected by many factors, such as pH, temperature, detergent composition, such as builders, the amount of surfactant, etc. DNase stability can be measured as described in Examples 2-6. The terms "improved stability" or "increased stability" are defined herein as the increased stability of the variant DNase relative to the parent DNase and / or relative to SEQ ID NO:1, as shown in solution. "Improved stability" and "increased stability" include detergent stability. The terms "detergent stability" or "improved detergent stability" can refer to improved stability of DNase activity compared to the parent DNase. DNase stability is measured as described in Examples 2-6.

[0074] The term "substantially pure variant" means a formulation containing, by weight, at most 10%, at most 8%, at most 6%, at most 5%, at most 4%, at most 3%, at most 2%, at most 1%, and at most 0.5% of other polypeptide substances naturally or recombinantly associated with the substantially pure variant. Preferably, the variant is at least 92% pure by weight of the total polypeptide substance present in the formulation, for example, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, at least 99.5%, and 100% pure. The variants of the present invention are preferably in a substantially pure form. This can be achieved, for example, by preparing the variant using well-known recombinant methods or classical purification methods.

[0075] The term "textiles" refers to woven fabrics, as well as short fibers and filaments suitable for conversion into or used as yarns, for weaving, knitting, and non-woven fabrics. This term encompasses yarns made from natural and synthetic (e.g., manufactured) fibers. The term "textile materials" is a general term for fibers, yarn intermediates, yarns, fabrics, and products made from fabrics (e.g., clothing and other articles).

[0076] The term "transcription promoter" is used to refer to the promoter of a DNA region that promotes the transcription of a specific gene. Transcription promoters are typically located near the gene they regulate, on the same strand, and upstream (towards the 5' region of the sense strand).

[0077] The term "transcription terminator" is used to refer to a gene sequence segment that marks the end of a gene or an operon on genomic DNA used for transcription.

[0078] The term "variant" refers to a polypeptide having DNase activity and comprising substitutions at one or more (e.g., several) positions. Substitution means that an amino acid occupying a position is replaced by a different amino acid, deletion means that an amino acid occupying a position is removed, and insertion means that an amino acid is added adjacent to an amino acid occupying a position, for example, 1 to 10 amino acids, preferably 1 to 3 amino acids. The variants of the present invention have at least 20%, for example, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 95%, or at least 100% of the mature polypeptide shown in SEQ ID NO:1, of the DNase activity. The term "DNase variant" refers to a polypeptide having DNase activity and comprising altered (i.e., substitution, insertion, and / or deletion) positions at one or more (or several) positions compared to the parental DNase (e.g., compared to SEQ ID NO:1).

[0079] As used herein, the terms “water hardness” or “degree of hardness” or “dH” or “°dH” refer to German degrees of hardness. It was once defined as 10 mg calcium oxide per liter of water.

[0080] The term "wild-type DNase" refers to DNase expressed by naturally occurring organisms (such as fungi, bacteria, archaea, yeast, plants, or animals found in nature).

[0081] Variant Naming Conventions

[0082] For the purposes of this invention, the polypeptide disclosed in SEQ ID NO:1 is used to determine the corresponding amino acid residues in another DNase. The amino acid sequence of the other DNase is aligned with the polypeptide disclosed in SEQ ID NO:1, and based on this alignment, the amino acid position number corresponding to any amino acid residue in the polypeptide disclosed in SEQ ID NO:1 is determined using, for example, the Needleman-Wunsch algorithm (Needleman and Wunsch, 1970, J. Mol. Biol. [Journal of Molecular Biology] 48:443-453), implemented in the Needleman procedure of the EMBOSS software package (EMBOSS: The European Molecular Biology Open Software Suite, Rice et al., 2000, Trends Genet. [Trends in Genetics] 16:276-277) (preferably version 5.0.0 or later). The parameters used are a vacancy opening penalty of 10, a vacancy extension penalty of 0.5, and an EBLOSUM62 (EMBOSS version of BLOSUM62) substitution matrix.

[0083] The identification of corresponding amino acid residues in another DNase can be determined by comparing multiple polypeptide sequences with their respective default parameters using several computer programs, including but not limited to MUSCLE (multiple sequence comparison by logarithmic prediction; version 3.5 or later; Edgar, 2004, Nucleic Acids Research 32:1792-1797), MAFFT (version 6.857 or later; Katoh and Kuma, 2002, Nucleic Acids Research 30:3059-3066; Katoh et al., 2005, Nucleic Acids Research 33:511-518; Katoh and Toh, 2007, Bioinformatics 23:372-374; Katoh et al., 2009, Methods in Molecular Biology). Biology [Molecular Biology Methods] 537:39-64; Katoh and Toh, 2010, Bioinformatics 26:1899-1900), and EMBOSS EMMA (version 1.83 or later) using ClustalW; Thompson et al., 1994, Nucleic Acids Research 22:4673-4680, using their respective default parameters.

[0084] When another enzyme is dissimilar to the polypeptide of SEQ ID NO:1, making conventional sequence-based comparison methods unable to detect their relationship (Lindahl and Elofsson, 2000, J. Mol. Biol. [Journal of Molecular Biology] 295: 613-615), alternative pairwise sequence comparison algorithms can be used. Higher sensitivity in sequence-based searches can be achieved using search programs that utilize the probabilistic representation (spectrum) of polypeptide families to search a database. For example, the PSI-BLAST program generates multiple spectra through an iterative database search process and can detect distant homologs (Atschul et al., 1997, Nucleic Acids Res. [Nucleic Acids Research] 25: 3389-3402). Even higher sensitivity can be achieved if the polypeptide family or superfamily has one or more representatives in a protein structure database. Programs such as GenTHREADER (Jones, 1999, J. Mol. Biol. [Journal of Molecular Biology] 287:797-815; McGuffin and Jones, 2003, Bioinformatics [Bioinformatics] 19:874-881) utilize information from multiple sources (PSI-BLAST, secondary structure prediction, structural alignment spectra, and solvation potential) as input to a neural network predicting the structural folding of a query sequence. Similarly, the method of Gough et al., 2000, J. Mol. Biol. [Journal of Molecular Biology] 313:903-919 can be used to align sequences of unknown structures with superfamily models existing in the SCOP database. These alignments can then be used to generate homology models of peptides, and the accuracy of such models can be evaluated using various tools developed for this purpose.

[0085] For proteins with known structures, several tools and resources are available for retrieving and generating structure alignments. For example, the SCOP superfamily of proteins has been structurally aligned, and those alignments are accessible and downloadable. Various algorithms, such as distance alignment matrices (Holm and Sander, 1998, Proteins 33:88-96) or combined extensions (Shindyalov and Bourne, 1998, Protein Engineering 11:739-747), can be used to align two or more protein structures, and implementations of these algorithms can also be used to query structure databases containing structures of interest to discover potential structural homologs (e.g., Holm and Park, 2000, Bioinformatics 16:566-567).

[0086] Since different amino acids can be present at a given position depending on the chosen variant parent, amino acid positions are indicated by #1, #2, etc., in the following definitions. In the variants describing the invention, the nomenclature described below has been adapted for ease of reference. Recognized IUPAC single-letter or three-letter amino acid abbreviations are used.

[0087] replace For amino acid substitutions, the following nomenclature is used: original amino acid, position, substituted amino acid. Therefore, the substitution of valine at position #1 with alanine is represented as "Val#1Ala" or "V#1A". Multiple mutations are separated by a plus sign ("+") or a comma (,), for example, "Val#1Ala+"Pro#2Gly" or V#1A,P#2G represents the substitution of valine (V) and proline (P) at positions #1 and #2, respectively, with alanine (A) and glycine (G). If more than one amino acid can be substituted at a given position, these amino acids are listed in parentheses, such as [X] or {X}. Thus, if both Trp and Lys can be substituted for the amino acid occupying position #1, this is represented as X#1{W,K}, X#1[W,K], or X#1[W / K], where X represents an amino acid residue present at the position of the parental DNase (e.g., the DNase shown in SEQ ID NO:1 or a DNase having at least 60% identity with it). In some cases, these variants may be represented as #1{W,K} or X#2P, indicating that the amino acid to be substituted varies depending on the parent. For convenience, when SEQ ID NO:1 is used, the amino acid to be substituted is listed as VA. When NO:1 is used to number substitutions, it is represented by the amino acid (e.g., T1A) at the corresponding position in SEQ ID NO:1. However, those skilled in the art will appreciate that DNase variants containing T1A are not limited to parental DNases having a threonine residue at position 1 corresponding to SEQ ID NO:1. In parental DNases having, for example, asparagine at position 1, those skilled in the art will designate translational mutations as T1A to N1A. In the case where the parental DNase has alanine residue at position 1, those skilled in the art will recognize that the parental DNase has not changed at that position. This also applies to deletions and insertions described below.

[0088] Missing For amino acid deletions, use the following nomenclature: original amino acid, position, * Therefore, the deletion of valine at position #1 is represented as "Val#1*" or "V#1*". Multiple deletions are separated by a plus sign ("+") or a comma, such as "Val#1*+Pro#2*" or "V#1*,P#2*".

[0089] insertThe insertion of additional amino acid residues, such as the insertion of lysine after Val#1, can be represented as Val#1ValLys or V#1VK. Alternatively, the insertion of additional amino acid residues, such as the insertion of lysine after V#1, can be represented as *#1aK. When more than one amino acid residue is inserted, such as the insertion of Lys and Ala after #1, this insertion can be represented as Val#1ValLysGly or V#1VKG. In such cases, the inserted amino acid residues can also be numbered by adding a lowercase letter to the position number of the amino acid residue before the inserted amino acid residue, in this example: *#1aK*#1bG.

[0090] Multiple changes Variants containing multiple changes are separated by a plus sign ("+") or a comma (,), such as "Val#1Trp+Pro#2Gly" or "V#1W,P#2G", which represent that valine and proline at positions #1 and #2 are replaced by tryptophan and glycine, respectively, as described above.

[0091] Different changes: When different changes can be introduced at a single position, these different changes are separated by commas, for example, "Val#1Trp,Lys" or V#1W,K represents the substitution of valine at position #1 with tryptophan or lysine. Therefore, "Val#1Trp,Lys+Pro#2Asp" represents the following variants: "Val#1Trp+Pro#2Asp", "Val#1Lys+Pro#2Asp", or V#1W,K+P#2D. In the context of this invention, the term "substitution group" is a variant containing more than one mutation compared to the parent or reference enzyme. For example, the substitution group T1I+S13Y (compared to SEQ ID NO:1) is a variant of SEQ ID NO:1 that contains two mutations, T1I+S13Y, compared to SEQ ID NO:1.

[0092] Specific nomenclature used for clades

[0093] For the purposes of this invention, the designation [IV] or [I / V] indicates that the amino acid at that position can be isoleucine (Ile, I) or valine (Val, V). Similarly, for other combinations as described herein, the designations [LVI] and [L / V / I] indicate that the amino acid at that position can be leucine (Leu, L), valine (Val, V), or isoleucine (Ile, I), and so on. Unless further limited otherwise, amino acid X is defined such that it can be any of the 20 naturally occurring amino acids. Detailed Implementation

[0094] This invention provides a novel DNase preferably obtained from Bacillus species (particularly Bacillus foodae). The DNase of this invention contains at least 60% sequence identity with the polypeptide having SEQ ID NO:1, and includes changes at at least two amino acid positions compared to the DNase having SEQ ID NO:1. In some aspects, the DNase variants of this invention comprise an amino acid sequence containing at least two amino acid substitutions made at positions equivalent to those in SEQ ID NO:1. This invention also relates to methods for producing DNase variants. Compared to the parental DNase or compared to SEQ ID NO:1, the DNase variants of this invention have at least one improved property. These properties include, but are not limited to, stability such as stability in detergents, storage stability, stability during washing and thermal stability, washing performance, particularly deep cleaning performance, increased expression levels, and reduced odor.

[0095] DNA enzyme variants

[0096] Some aspects of the present invention relate to a DNase variant of SEQ ID NO:1 or a DNase variant having at least 60% identity with it, and to a method for generating a DNase variant of SEQ ID NO:1 or a DNase having at least 60% identity with it.

[0097] The present invention relates to a DNAse that includes variations at two or more positions selected from a list consisting of the following: positions 1, 13, 22, 25, 27, 39, 42, 57, 59, 76, 109, 116, 127, 144, 147, 167 and 175, wherein each position corresponds to a position of the polypeptide of SEQ ID NO:1, wherein said variant has DNAse activity.

[0098] In some respects, the variant has at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, such as at least 96%, at least 97%, at least 98%, or at least 99%, but less than 100% sequence identity with the mature parental DNase (e.g., the DNase shown in SEQ ID NO:1).

[0099] In some respects, the variant has DNase activity, wherein the variant has at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, such as at least 96%, at least 97%, at least 98%, or at least 99%, but less than 100% sequence identity with the polypeptide shown in SEQ ID NO:1.

[0100] In some preferred aspects, the change is a substitution, and the DNase variants of the present invention comprise two or more substitutions (compared to SEQ ID NO:1), wherein the substitutions are selected from the group consisting of: T1I, T1L, T1V, S13Y, T22P, S25P, S27L, S39P, S42G, S42A, S42T, S57W, S57Y, S57F, S59V, S59I, S59L, V76L, V76I, Q109R, S116D, S116E, T127V, T127I, T127L, S144P, A147H, S167L, S167I, S167V, G175D, and G175E, wherein the variants have DNase activity, and wherein each position corresponds to the polypeptide shown in SEQ ID NO:1 (numbered according to SEQ ID NO:1).

[0101] In some aspects, the number of substitutions in the variants of the invention is 2-20, for example 2-10 and 2-5, such as 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 substitutions. In one aspect, the total number of substitutions compared to SEQ ID NO:1 is 3-20, for example at least 5 to 20, at least 10 to 20, or for example at least 15 to 20 substitutions, compared to a DNAseed containing the amino acid sequence shown in SEQ ID NO:1.

[0102] The present invention further relates to variants comprising the DNase parent polypeptide of SEQ ID NO:1, wherein the variant contains changes at at least two positions selected from the positions of SEQ ID NO:1, corresponding to the positions of SEQ ID NO:1: 1, 13, 22, 25, 27, 39, 42, 57, 59, 76, 109, 116, 127, 144, 147, 167, and 175, respectively, and wherein the variant has at least 60%, at least 70%, at least 80%, at least 90%, or at least 95% of the same amino acid sequence as SEQ ID NO:1, wherein the variant has DNase activity, and wherein the variant has at least one improved property compared to the DNase of SEQ ID NO:1.

[0103] In one aspect, the present invention relates to a DNA enzyme variant comprising substitutions at positions corresponding to positions 1 and 13 of SEQ ID NO:1, wherein the variant has DNA enzyme activity.

[0104] In one aspect, the present invention relates to a DNA enzyme variant comprising substitutions at positions corresponding to positions 1 and 22 of SEQ ID NO:1, wherein the variant has DNA enzyme activity.

[0105] In one aspect, the present invention relates to a DNA enzyme variant comprising substitutions at positions corresponding to positions 1 and 25 of SEQ ID NO:1, wherein the variant has DNA enzyme activity.

[0106] In one aspect, the present invention relates to a DNA enzyme variant comprising substitutions at positions corresponding to positions 1 and 27 of SEQ ID NO:1, wherein the variant has DNA enzyme activity.

[0107] In one aspect, the present invention relates to a DNA enzyme variant comprising substitutions at positions corresponding to positions 1 and 39 of SEQ ID NO:1, wherein the variant has DNA enzyme activity.

[0108] In one aspect, the present invention relates to a DNA enzyme variant comprising substitutions at positions corresponding to positions 1 and 42 of SEQ ID NO:1, wherein the variant has DNA enzyme activity.

[0109] In one aspect, the present invention relates to a DNA enzyme variant comprising substitutions at positions corresponding to positions 1 and 57 of SEQ ID NO:1, wherein the variant has DNA enzyme activity.

[0110] In one aspect, the present invention relates to a DNA enzyme variant comprising substitutions at positions corresponding to positions 1 and 59 of SEQ ID NO:1, wherein the variant has DNA enzyme activity.

[0111] In one aspect, the present invention relates to a DNA enzyme variant comprising substitutions at positions corresponding to positions 1 and 76 of SEQ ID NO:1, wherein the variant has DNA enzyme activity.

[0112] In one aspect, the present invention relates to a DNA enzyme variant comprising a substitution at positions corresponding to positions 1 and 109 of SEQ ID NO:1, wherein the variant has DNA enzyme activity.

[0113] In one aspect, the present invention relates to a DNA enzyme variant comprising a substitution at positions corresponding to positions 1 and 116 of SEQ ID NO:1, wherein the variant has DNA enzyme activity.

[0114] In one aspect, the present invention relates to a DNA enzyme variant comprising substitutions at positions corresponding to positions 1 and 127 of SEQ ID NO:1, wherein the variant has DNA enzyme activity.

[0115] In one aspect, the present invention relates to a DNA enzyme variant comprising substitutions at positions corresponding to positions 1 and 144 of SEQ ID NO:1, wherein the variant has DNA enzyme activity.

[0116] In one aspect, the present invention relates to a DNA enzyme variant comprising a substitution at positions corresponding to positions 1 and 147 of SEQ ID NO:1, wherein the variant has DNA enzyme activity.

[0117] In one aspect, the present invention relates to a DNA enzyme variant comprising a substitution at positions corresponding to positions 1 and 167 of SEQ ID NO:1, wherein the variant has DNA enzyme activity.

[0118] In one aspect, the present invention relates to a DNA enzyme variant comprising a substitution at positions corresponding to positions 1 and 175 of SEQ ID NO:1, wherein the variant has DNA enzyme activity.

[0119] In one aspect, the present invention relates to a DNA enzyme variant comprising substitutions at positions corresponding to positions 13 and 22 of SEQ ID NO:1, wherein the variant has DNA enzyme activity.

[0120] In one aspect, the present invention relates to a DNA enzyme variant comprising substitutions at positions corresponding to positions 13 and 25 of SEQ ID NO:1, wherein the variant has DNA enzyme activity.

[0121] In one aspect, the present invention relates to a DNA enzyme variant comprising substitutions at positions corresponding to positions 13 and 27 of SEQ ID NO:1, wherein the variant has DNA enzyme activity.

[0122] In one aspect, the present invention relates to a DNA enzyme variant comprising substitutions at positions corresponding to positions 13 and 39 of SEQ ID NO:1, wherein the variant has DNA enzyme activity.

[0123] In one aspect, the present invention relates to a DNA enzyme variant comprising substitutions at positions corresponding to positions 13 and 42 of SEQ ID NO:1, wherein the variant has DNA enzyme activity.

[0124] In one aspect, the present invention relates to a DNA enzyme variant comprising substitutions at positions corresponding to positions 13 and 57 of SEQ ID NO:1, wherein the variant has DNA enzyme activity.

[0125] In one aspect, the present invention relates to a DNA enzyme variant comprising substitutions at positions corresponding to positions 13 and 59 of SEQ ID NO:1, wherein the variant has DNA enzyme activity.

[0126] In one aspect, the present invention relates to a DNA enzyme variant comprising substitutions at positions corresponding to positions 13 and 76 of SEQ ID NO:1, wherein the variant has DNA enzyme activity.

[0127] In one aspect, the present invention relates to a DNA enzyme variant comprising substitutions at positions corresponding to positions 13 and 109 of SEQ ID NO:1, wherein the variant has DNA enzyme activity.

[0128] In one aspect, the present invention relates to a DNA enzyme variant comprising substitutions at positions corresponding to positions 13 and 116 of SEQ ID NO:1, wherein the variant has DNA enzyme activity.

[0129] In one aspect, the present invention relates to a DNA enzyme variant comprising substitutions at positions corresponding to positions 13 and 127 of SEQ ID NO:1, wherein the variant has DNA enzyme activity.

[0130] In one aspect, the present invention relates to a DNA enzyme variant comprising substitutions at positions corresponding to positions 13 and 144 of SEQ ID NO:1, wherein the variant has DNA enzyme activity.

[0131] In one aspect, the present invention relates to a DNA enzyme variant comprising substitutions at positions corresponding to positions 13 and 147 of SEQ ID NO:1, wherein the variant has DNA enzyme activity.

[0132] In one aspect, the present invention relates to a DNA enzyme variant comprising substitutions at positions corresponding to positions 13 and 167 of SEQ ID NO:1, wherein the variant has DNA enzyme activity.

[0133] In one aspect, the present invention relates to a DNA enzyme variant comprising substitutions at positions corresponding to positions 13 and 175 of SEQ ID NO:1, wherein the variant has DNA enzyme activity.

[0134] In one aspect, the present invention relates to a DNA enzyme variant comprising substitutions at positions corresponding to positions 22 and 25 of SEQ ID NO:1, wherein the variant has DNA enzyme activity.

[0135] In one aspect, the present invention relates to a DNA enzyme variant comprising substitutions at positions corresponding to positions 22 and 27 of SEQ ID NO:1, wherein the variant has DNA enzyme activity.

[0136] In one aspect, the present invention relates to a DNA enzyme variant comprising substitutions at positions corresponding to positions 22 and 39 of SEQ ID NO:1, wherein the variant has DNA enzyme activity.

[0137] In one aspect, the present invention relates to a DNA enzyme variant comprising substitutions at positions corresponding to positions 22 and 42 of SEQ ID NO:1, wherein the variant has DNA enzyme activity.

[0138] In one aspect, the present invention relates to a DNA enzyme variant comprising substitutions at positions corresponding to positions 22 and 57 of SEQ ID NO:1, wherein the variant has DNA enzyme activity.

[0139] In one aspect, the present invention relates to a DNA enzyme variant comprising substitutions at positions corresponding to positions 22 and 59 of SEQ ID NO:1, wherein the variant has DNA enzyme activity.

[0140] In one aspect, the present invention relates to a DNA enzyme variant comprising substitutions at positions corresponding to positions 22 and 76 of SEQ ID NO:1, wherein the variant has DNA enzyme activity.

[0141] In one aspect, the present invention relates to a DNA enzyme variant comprising substitutions at positions corresponding to positions 22 and 109 of SEQ ID NO:1, wherein the variant has DNA enzyme activity.

[0142] In one aspect, the present invention relates to a DNA enzyme variant comprising substitutions at positions corresponding to positions 22 and 116 of SEQ ID NO:1, wherein the variant has DNA enzyme activity.

[0143] In one aspect, the present invention relates to a DNA enzyme variant comprising substitutions at positions corresponding to positions 22 and 127 of SEQ ID NO:1, wherein the variant has DNA enzyme activity.

[0144] In one aspect, the present invention relates to a DNA enzyme variant comprising substitutions at positions corresponding to positions 22 and 144 of SEQ ID NO:1, wherein the variant has DNA enzyme activity.

[0145] In one aspect, the present invention relates to a DNA enzyme variant comprising substitutions at positions corresponding to positions 22 and 147 of SEQ ID NO:1, wherein the variant has DNA enzyme activity.

[0146] In one aspect, the present invention relates to a DNA enzyme variant comprising substitutions at positions corresponding to positions 22 and 167 of SEQ ID NO:1, wherein the variant has DNA enzyme activity.

[0147] In one aspect, the present invention relates to a DNA enzyme variant comprising substitutions at positions corresponding to positions 22 and 175 of SEQ ID NO:1, wherein the variant has DNA enzyme activity.

[0148] In one aspect, the present invention relates to a DNA enzyme variant comprising substitutions at positions corresponding to positions 25 and 27 of SEQ ID NO:1, wherein the variant has DNA enzyme activity.

[0149] In one aspect, the present invention relates to a DNA enzyme variant comprising substitutions at positions corresponding to positions 25 and 39 of SEQ ID NO:1, wherein the variant has DNA enzyme activity.

[0150] In one aspect, the present invention relates to a DNA enzyme variant comprising substitutions at positions corresponding to positions 25 and 42 of SEQ ID NO:1, wherein the variant has DNA enzyme activity.

[0151] In one aspect, the present invention relates to a DNA enzyme variant comprising substitutions at positions corresponding to positions 25 and 57 of SEQ ID NO:1, wherein the variant has DNA enzyme activity.

[0152] In one aspect, the present invention relates to a DNA enzyme variant comprising substitutions at positions corresponding to positions 25 and 59 of SEQ ID NO:1, wherein the variant has DNA enzyme activity.

[0153] In one aspect, the present invention relates to a DNA enzyme variant comprising substitutions at positions corresponding to positions 25 and 76 of SEQ ID NO:1, wherein the variant has DNA enzyme activity.

[0154] In one aspect, the present invention relates to a DNA enzyme variant comprising substitutions at positions corresponding to positions 25 and 109 of SEQ ID NO:1, wherein the variant has DNA enzyme activity.

[0155] In one aspect, the present invention relates to a DNA enzyme variant comprising substitutions at positions corresponding to positions 25 and 116 of SEQ ID NO:1, wherein the variant has DNA enzyme activity.

[0156] In one aspect, the present invention relates to a DNA enzyme variant comprising substitutions at positions corresponding to positions 25 and 127 of SEQ ID NO:1, wherein the variant has DNA enzyme activity.

[0157] In one aspect, the present invention relates to a DNA enzyme variant comprising substitutions at positions corresponding to positions 25 and 144 of SEQ ID NO:1, wherein the variant has DNA enzyme activity.

[0158] In one aspect, the present invention relates to a DNA enzyme variant comprising substitutions at positions corresponding to positions 25 and 147 of SEQ ID NO:1, wherein the variant has DNA enzyme activity.

[0159] In one aspect, the present invention relates to a DNA enzyme variant comprising substitutions at positions corresponding to positions 25 and 167 of SEQ ID NO:1, wherein the variant has DNA enzyme activity.

[0160] In one aspect, the present invention relates to a DNA enzyme variant comprising a substitution at positions corresponding to positions 25 and 175 of SEQ ID NO:1, wherein the variant has DNA enzyme activity.

[0161] In one aspect, the present invention relates to a DNA enzyme variant comprising substitutions at positions corresponding to positions 27 and 39 of SEQ ID NO:1, wherein the variant has DNA enzyme activity.

[0162] In one aspect, the present invention relates to a DNA enzyme variant comprising substitutions at positions corresponding to positions 27 and 42 of SEQ ID NO:1, wherein the variant has DNA enzyme activity.

[0163] In one aspect, the present invention relates to a DNA enzyme variant comprising a substitution at positions corresponding to positions 27 and 57 of SEQ ID NO:1, wherein the variant has DNA enzyme activity.

[0164] In one aspect, the present invention relates to a DNA enzyme variant comprising substitutions at positions corresponding to positions 27 and 59 of SEQ ID NO:1, wherein the variant has DNA enzyme activity.

[0165] In one aspect, the present invention relates to a DNA enzyme variant comprising substitutions at positions corresponding to positions 27 and 76 of SEQ ID NO:1, wherein the variant has DNA enzyme activity.

[0166] In one aspect, the present invention relates to a DNA enzyme variant comprising substitutions at positions corresponding to positions 27 and 109 of SEQ ID NO:1, wherein the variant has DNA enzyme activity.

[0167] In one aspect, the present invention relates to a DNA enzyme variant comprising substitutions at positions corresponding to positions 27 and 116 of SEQ ID NO:1, wherein the variant has DNA enzyme activity.

[0168] In one aspect, the present invention relates to a DNA enzyme variant comprising a substitution at positions corresponding to positions 27 and 127 of SEQ ID NO:1, wherein the variant has DNA enzyme activity.

[0169] In one aspect, the present invention relates to a DNA enzyme variant comprising substitutions at positions corresponding to positions 27 and 144 of SEQ ID NO:1, wherein the variant has DNA enzyme activity.

[0170] In one aspect, the present invention relates to a DNA enzyme variant comprising substitutions at positions corresponding to positions 27 and 147 of SEQ ID NO:1, wherein the variant has DNA enzyme activity.

[0171] In one aspect, the present invention relates to a DNA enzyme variant comprising substitutions at positions corresponding to positions 27 and 167 of SEQ ID NO:1, wherein the variant has DNA enzyme activity.

[0172] In one aspect, the present invention relates to a DNA enzyme variant comprising substitutions at positions corresponding to positions 27 and 175 of SEQ ID NO:1, wherein the variant has DNA enzyme activity.

[0173] In one aspect, the present invention relates to a DNA enzyme variant comprising substitutions at positions corresponding to positions 39 and 42 of SEQ ID NO:1, wherein the variant has DNA enzyme activity.

[0174] In one aspect, the present invention relates to a DNA enzyme variant comprising substitutions at positions corresponding to positions 39 and 57 of SEQ ID NO:1, wherein the variant has DNA enzyme activity.

[0175] In one aspect, the present invention relates to a DNA enzyme variant comprising a substitution at positions corresponding to positions 39 and 59 of SEQ ID NO:1, wherein the variant has DNA enzyme activity.

[0176] In one aspect, the present invention relates to a DNA enzyme variant comprising substitutions at positions corresponding to positions 39 and 76 of SEQ ID NO:1, wherein the variant has DNA enzyme activity.

[0177] In one aspect, the present invention relates to a DNA enzyme variant comprising substitutions at positions corresponding to positions 39 and 109 of SEQ ID NO:1, wherein the variant has DNA enzyme activity.

[0178] In one aspect, the present invention relates to a DNA enzyme variant comprising substitutions at positions corresponding to positions 39 and 116 of SEQ ID NO:1, wherein the variant has DNA enzyme activity.

[0179] In one aspect, the present invention relates to a DNA enzyme variant comprising substitutions at positions corresponding to positions 39 and 127 of SEQ ID NO:1, wherein the variant has DNA enzyme activity.

[0180] In one aspect, the present invention relates to a DNA enzyme variant comprising substitutions at positions corresponding to positions 39 and 144 of SEQ ID NO:1, wherein the variant has DNA enzyme activity.

[0181] In one aspect, the present invention relates to a DNA enzyme variant comprising substitutions at positions corresponding to positions 39 and 147 of SEQ ID NO:1, wherein the variant has DNA enzyme activity.

[0182] In one aspect, the present invention relates to a DNA enzyme variant comprising substitutions at positions corresponding to positions 39 and 167 of SEQ ID NO:1, wherein the variant has DNA enzyme activity.

[0183] In one aspect, the present invention relates to a DNA enzyme variant comprising substitutions at positions corresponding to positions 39 and 175 of SEQ ID NO:1, wherein the variant has DNA enzyme activity.

[0184] In one aspect, the present invention relates to a DNA enzyme variant comprising substitutions at positions corresponding to positions 42 and 57 of SEQ ID NO:1, wherein the variant has DNA enzyme activity.

[0185] In one aspect, the present invention relates to a DNA enzyme variant comprising substitutions at positions corresponding to positions 42 and 59 of SEQ ID NO:1, wherein the variant has DNA enzyme activity.

[0186] In one aspect, the present invention relates to a DNA enzyme variant comprising substitutions at positions corresponding to positions 42 and 76 of SEQ ID NO:1, wherein the variant has DNA enzyme activity.

[0187] In one aspect, the present invention relates to a DNA enzyme variant comprising substitutions at positions corresponding to positions 42 and 76 of SEQ ID NO:1, wherein the variant has DNA enzyme activity.

[0188] In one aspect, the present invention relates to a DNA enzyme variant comprising substitutions at positions corresponding to positions 42 and 116 of SEQ ID NO:1, wherein the variant has DNA enzyme activity.

[0189] In one aspect, the present invention relates to a DNA enzyme variant comprising substitutions at positions corresponding to positions 42 and 127 of SEQ ID NO:1, wherein the variant has DNA enzyme activity.

[0190] In one aspect, the present invention relates to a DNA enzyme variant comprising substitutions at positions corresponding to positions 42 and 144 of SEQ ID NO:1, wherein the variant has DNA enzyme activity.

[0191] In one aspect, the present invention relates to a DNA enzyme variant comprising substitutions at positions corresponding to positions 42 and 147 of SEQ ID NO:1, wherein the variant has DNA enzyme activity.

[0192] In one aspect, the present invention relates to a DNA enzyme variant comprising substitutions at positions corresponding to positions 42 and 167 of SEQ ID NO:1, wherein the variant has DNA enzyme activity.

[0193] In one aspect, the present invention relates to a DNA enzyme variant comprising substitutions at positions corresponding to positions 42 and 175 of SEQ ID NO:1, wherein the variant has DNA enzyme activity.

[0194] In one aspect, the present invention relates to a DNA enzyme variant comprising a substitution at positions corresponding to positions 57 and 59 of SEQ ID NO:1, wherein the variant has DNA enzyme activity.

[0195] In one aspect, the present invention relates to a DNA enzyme variant comprising substitutions at positions corresponding to positions 57 and 76 of SEQ ID NO:1, wherein the variant has DNA enzyme activity.

[0196] In one aspect, the present invention relates to a DNA enzyme variant comprising substitutions at positions corresponding to positions 57 and 109 of SEQ ID NO:1, wherein the variant has DNA enzyme activity.

[0197] In one aspect, the present invention relates to a DNA enzyme variant comprising substitutions at positions corresponding to positions 57 and 116 of SEQ ID NO:1, wherein the variant has DNA enzyme activity.

[0198] In one aspect, the present invention relates to a DNA enzyme variant comprising a substitution at positions corresponding to positions 57 and 127 of SEQ ID NO:1, wherein the variant has DNA enzyme activity.

[0199] In one aspect, the present invention relates to a DNA enzyme variant comprising substitutions at positions corresponding to positions 57 and 144 of SEQ ID NO:1, wherein the variant has DNA enzyme activity.

[0200] In one aspect, the present invention relates to a DNA enzyme variant comprising a substitution at positions corresponding to positions 57 and 147 of SEQ ID NO:1, wherein the variant has DNA enzyme activity.

[0201] In one aspect, the present invention relates to a DNA enzyme variant comprising substitutions at positions corresponding to positions 57 and 167 of SEQ ID NO:1, wherein the variant has DNA enzyme activity.

[0202] In one aspect, the present invention relates to a DNA enzyme variant comprising substitutions at positions corresponding to positions 57 and 175 of SEQ ID NO:1, wherein the variant has DNA enzyme activity.

[0203] In one aspect, the present invention relates to a DNA enzyme variant comprising substitutions at positions corresponding to positions 59 and 76 of SEQ ID NO:1, wherein the variant has DNA enzyme activity.

[0204] In one aspect, the present invention relates to a DNA enzyme variant comprising substitutions at positions corresponding to positions 59 and 109 of SEQ ID NO:1, wherein the variant has DNA enzyme activity.

[0205] In one aspect, the present invention relates to a DNA enzyme variant comprising substitutions at positions corresponding to positions 59 and 116 of SEQ ID NO:1, wherein the variant has DNA enzyme activity.

[0206] In one aspect, the present invention relates to a DNA enzyme variant comprising substitutions at positions corresponding to positions 59 and 127 of SEQ ID NO:1, wherein the variant has DNA enzyme activity.

[0207] In one aspect, the present invention relates to a DNA enzyme variant comprising substitutions at positions corresponding to positions 59 and 144 of SEQ ID NO:1, wherein the variant has DNA enzyme activity.

[0208] In one aspect, the present invention relates to a DNA enzyme variant comprising substitutions at positions corresponding to positions 59 and 147 of SEQ ID NO:1, wherein the variant has DNA enzyme activity.

[0209] In one aspect, the present invention relates to a DNA enzyme variant comprising substitutions at positions corresponding to positions 59 and 167 of SEQ ID NO:1, wherein the variant has DNA enzyme activity.

[0210] In one aspect, the present invention relates to a DNA enzyme variant comprising substitutions at positions corresponding to positions 59 and 175 of SEQ ID NO:1, wherein the variant has DNA enzyme activity.

[0211] In one aspect, the present invention relates to a DNA enzyme variant comprising substitutions at positions corresponding to positions 76 and 109 of SEQ ID NO:1, wherein the variant has DNA enzyme activity.

[0212] In one aspect, the present invention relates to a DNA enzyme variant comprising substitutions at positions corresponding to positions 76 and 116 of SEQ ID NO:1, wherein the variant has DNA enzyme activity.

[0213] In one aspect, the present invention relates to a DNA enzyme variant comprising substitutions at positions corresponding to positions 76 and 127 of SEQ ID NO:1, wherein the variant has DNA enzyme activity.

[0214] In one aspect, the present invention relates to a DNA enzyme variant comprising substitutions at positions corresponding to positions 76 and 144 of SEQ ID NO:1, wherein the variant has DNA enzyme activity.

[0215] In one aspect, the present invention relates to a DNA enzyme variant comprising substitutions at positions corresponding to positions 76 and 147 of SEQ ID NO:1, wherein the variant has DNA enzyme activity.

[0216] In one aspect, the present invention relates to a DNA enzyme variant comprising substitutions at positions corresponding to positions 76 and 167 of SEQ ID NO:1, wherein the variant has DNA enzyme activity.

[0217] In one aspect, the present invention relates to a DNA enzyme variant comprising substitutions at positions corresponding to positions 76 and 175 of SEQ ID NO:1, wherein the variant has DNA enzyme activity.

[0218] In one aspect, the present invention relates to a DNA enzyme variant comprising substitutions at positions corresponding to positions 109 and 116 of SEQ ID NO:1, wherein the variant has DNA enzyme activity.

[0219] In one aspect, the present invention relates to a DNA enzyme variant comprising substitutions at positions corresponding to positions 109 and 127 of SEQ ID NO:1, wherein the variant has DNA enzyme activity.

[0220] In one aspect, the present invention relates to a DNA enzyme variant comprising substitutions at positions corresponding to positions 109 and 144 of SEQ ID NO:1, wherein the variant has DNA enzyme activity.

[0221] In one aspect, the present invention relates to a DNA enzyme variant comprising substitutions at positions corresponding to positions 109 and 147 of SEQ ID NO:1, wherein the variant has DNA enzyme activity.

[0222] In one aspect, the present invention relates to a DNA enzyme variant comprising substitutions at positions corresponding to positions 109 and 167 of SEQ ID NO:1, wherein the variant has DNA enzyme activity.

[0223] In one aspect, the present invention relates to a DNA enzyme variant comprising substitutions at positions corresponding to positions 109 and 175 of SEQ ID NO:1, wherein the variant has DNA enzyme activity.

[0224] In one aspect, the present invention relates to a DNA enzyme variant comprising substitutions at positions corresponding to positions 116 and 127 of SEQ ID NO:1, wherein the variant has DNA enzyme activity.

[0225] In one aspect, the present invention relates to a DNA enzyme variant comprising substitutions at positions corresponding to positions 116 and 144 of SEQ ID NO:1, wherein the variant has DNA enzyme activity.

[0226] In one aspect, the present invention relates to a DNA enzyme variant comprising substitutions at positions corresponding to positions 116 and 147 of SEQ ID NO:1, wherein the variant has DNA enzyme activity.

[0227] In one aspect, the present invention relates to a DNA enzyme variant comprising substitutions at positions corresponding to positions 116 and 167 of SEQ ID NO:1, wherein the variant has DNA enzyme activity.

[0228] In one aspect, the present invention relates to a DNA enzyme variant comprising substitutions at positions corresponding to positions 116 and 175 of SEQ ID NO:1, wherein the variant has DNA enzyme activity.

[0229] In one aspect, the present invention relates to a DNA enzyme variant comprising substitutions at positions corresponding to positions 144 and 147 of SEQ ID NO:1, wherein the variant has DNA enzyme activity.

[0230] In one aspect, the present invention relates to a DNA enzyme variant comprising substitutions at positions corresponding to positions 144 and 167 of SEQ ID NO:1, wherein the variant has DNA enzyme activity.

[0231] In one aspect, the present invention relates to a DNA enzyme variant comprising substitutions at positions corresponding to positions 144 and 175 of SEQ ID NO:1, wherein the variant has DNA enzyme activity.

[0232] In one aspect, the present invention relates to a DNA enzyme variant comprising a substitution at positions corresponding to positions 147 and 167 of SEQ ID NO:1, wherein the variant has DNA enzyme activity.

[0233] In one aspect, the present invention relates to a DNA enzyme variant comprising substitutions at positions corresponding to positions 147 and 175 of SEQ ID NO:1, wherein the variant has DNA enzyme activity.

[0234] In one aspect, the present invention relates to a DNA enzyme variant comprising substitutions at positions corresponding to positions 167 and 175 of SEQ ID NO:1, wherein the variant has DNA enzyme activity.

[0235] The present invention further relates to DNAse variants comprising combinations of substitutions as described below.

[0236] In one aspect, the present invention relates to a DNAse variant comprising a substituted T1I+S13Y, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNAse activity.

[0237] In one aspect, the present invention relates to a DNAse variant comprising a substitution for T1I+T22P, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNAse activity.

[0238] In one aspect, the present invention relates to a DNAse variant comprising a substituted T1I+S39P, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNAse activity.

[0239] In one aspect, the present invention relates to a DNAse variant comprising a substituted T1I+S42G, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNAse activity.

[0240] In one aspect, the present invention relates to a DNase variant comprising a substituted T1I+S42A, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNase activity.

[0241] In one aspect, the present invention relates to a DNAse variant comprising a substituted T1I+S42T, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNAse activity.

[0242] In one aspect, the present invention relates to a DNAse variant comprising a substituted T1I+S57W, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNAse activity.

[0243] In one aspect, the present invention relates to a DNAse variant comprising a substituted T1I+S57Y, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNAse activity.

[0244] In one aspect, the present invention relates to a DNAse variant comprising a substituted T1I+S57F, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNAse activity.

[0245] In one aspect, the present invention relates to a DNAse variant comprising a substituted T1I+S59V, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNAse activity.

[0246] In one aspect, the present invention relates to a DNAse variant comprising a substituted T1I+S59I, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNAse activity.

[0247] In one aspect, the present invention relates to a DNase variant comprising a substituted T1I+S59L, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNase activity.

[0248] In one aspect, the present invention relates to a DNase variant comprising a substituted T1I+V76L, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNase activity.

[0249] In one aspect, the present invention relates to a DNase variant comprising a substituted T1I+V76I, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNase activity.

[0250] In one aspect, the present invention relates to a DNAse variant comprising a substituted T1I+Q109R, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNAse activity.

[0251] In one aspect, the present invention relates to a DNAse variant comprising a substituted T1I+S116D, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNAse activity.

[0252] In one aspect, the present invention relates to a DNAse variant comprising a substituted T1I+S116E, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNAse activity.

[0253] In one aspect, the present invention relates to a DNAse variant comprising a substitution for T1I+T127V, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNAse activity.

[0254] In one aspect, the present invention relates to a DNase variant comprising a substituted T1I+T127I, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNase activity.

[0255] In one aspect, the present invention relates to a DNase variant comprising a substituted T1I+T127L, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNase activity.

[0256] In one aspect, the present invention relates to a DNase variant comprising a substituted T1I+S144P, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNase activity.

[0257] In one aspect, the present invention relates to a DNase variant comprising a substituted T1I+A147H, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNase activity.

[0258] In one aspect, the present invention relates to a DNase variant comprising a substituted T1I+S167L, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNase activity.

[0259] In one aspect, the present invention relates to a DNase variant comprising a substituted T1I+S167I, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNase activity.

[0260] In one aspect, the present invention relates to a DNAse variant comprising a substituted T1I+S167V, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNAse activity.

[0261] In one aspect, the present invention relates to a DNAse variant comprising a substituted T1I+G175D, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNAse activity.

[0262] In one aspect, the present invention relates to a DNAse variant comprising a substituted T1I+G175E, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNAse activity.

[0263] In one aspect, the present invention relates to a DNase variant comprising a substituted T1L+T22P, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNase activity.

[0264] In one aspect, the present invention relates to a DNAse variant comprising a substituted T1L+S13Y, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNAse activity.

[0265] In one aspect, the present invention relates to a DNase variant comprising a substituted T1L+T22P, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNase activity.

[0266] In one aspect, the present invention relates to a DNAse variant comprising a substituted T1L+S25P, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNAse activity.

[0267] In one aspect, the present invention relates to a DNase variant comprising a substituted T1L+S27L, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNase activity.

[0268] In one aspect, the present invention relates to a DNAse variant comprising a substituted T1L+S39P, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNAse activity.

[0269] In one aspect, the present invention relates to a DNAse variant comprising a substituted T1L+S42G, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNAse activity.

[0270] In one aspect, the present invention relates to a DNAse variant comprising a substituted T1L+S42A, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNAse activity.

[0271] In one aspect, the present invention relates to a DNAse variant comprising a substituted T1L+S42T, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNAse activity.

[0272] In one aspect, the present invention relates to a DNAse variant comprising a substituted T1L+S57W, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNAse activity.

[0273] In one aspect, the present invention relates to a DNAse variant comprising a substituted T1L+S57Y, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNAse activity.

[0274] In one aspect, the present invention relates to a DNAse variant comprising a substituted T1L+S57F, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNAse activity.

[0275] In one aspect, the present invention relates to a DNAse variant comprising a substituted T1L+S59V, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNAse activity.

[0276] In one aspect, the present invention relates to a DNAse variant comprising a substituted T1L+S59I, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNAse activity.

[0277] In one aspect, the present invention relates to a DNAse variant comprising a substitution for T1L+S59L, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNAse activity.

[0278] In one aspect, the present invention relates to a DNase variant comprising a substituted T1L+V76L, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNase activity.

[0279] In one aspect, the present invention relates to a DNase variant comprising a substituted T1L+V76I, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNase activity.

[0280] In one aspect, the present invention relates to a DNAse variant comprising a substituted T1L+Q109R, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNAse activity.

[0281] In one aspect, the present invention relates to a DNAse variant comprising a substituted T1L+S116D, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNAse activity.

[0282] In one aspect, the present invention relates to a DNAse variant comprising a substituted T1L+S116E, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNAse activity.

[0283] In one aspect, the present invention relates to a DNAse variant comprising a substitution for T1L+T127V, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNAse activity.

[0284] In one aspect, the present invention relates to a DNAse variant comprising a substituted T1L+T127I, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNAse activity.

[0285] In one aspect, the present invention relates to a DNase variant comprising a substitution for T1L+T127L, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNase activity.

[0286] In one aspect, the present invention relates to a DNAse variant comprising a substituted T1L+S144P, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNAse activity.

[0287] In one aspect, the present invention relates to a DNase variant comprising a substituted T1L+A147H, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNase activity.

[0288] In one aspect, the present invention relates to a DNase variant comprising a substituted T1L+S167L, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNase activity.

[0289] In one aspect, the present invention relates to a DNAse variant comprising a substituted T1L+S167I, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNAse activity.

[0290] In one aspect, the present invention relates to a DNase variant comprising a substituted T1L+S167V, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNase activity.

[0291] In one aspect, the present invention relates to a DNAse variant comprising a substituted T1L+G175D, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNAse activity.

[0292] In one aspect, the present invention relates to a DNAse variant comprising a substituted T1L+G175E, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNAse activity.

[0293] In one aspect, the present invention relates to a DNAse variant comprising a substituted T1V+S13Y, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNAse activity.

[0294] In one aspect, the present invention relates to a DNAse variant comprising a substitution for T1V+T22P, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNAse activity.

[0295] In one aspect, the present invention relates to a DNAse variant comprising a substituted T1V+S25P, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNAse activity.

[0296] In one aspect, the present invention relates to a DNAse variant comprising a substituted T1V+S27L, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNAse activity.

[0297] In one aspect, the present invention relates to a DNAse variant comprising a substituted T1V+S39P, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNAse activity.

[0298] In one aspect, the present invention relates to a DNAse variant comprising a substituted T1V+S42G, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNAse activity.

[0299] In one aspect, the present invention relates to a DNase variant comprising a substituted T1V+S42A, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNase activity.

[0300] In one aspect, the present invention relates to a DNase variant comprising a substituted T1V+S42T, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNase activity.

[0301] In one aspect, the present invention relates to a DNAse variant comprising a substituted T1V+S57W, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNAse activity.

[0302] In one aspect, the present invention relates to a DNAse variant comprising a substituted T1V+S57Y, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNAse activity.

[0303] In one aspect, the present invention relates to a DNAse variant comprising a substituted T1V+S57F, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNAse activity.

[0304] In one aspect, the present invention relates to a DNAse variant comprising a substitution for T1V+S59V, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNAse activity.

[0305] In one aspect, the present invention relates to a DNAse variant comprising a substituted T1V+S59I, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNAse activity.

[0306] In one aspect, the present invention relates to a DNAse variant comprising a substituted T1V+S59L, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNAse activity.

[0307] In one aspect, the present invention relates to a DNase variant comprising a substituted T1V+V76L, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNase activity.

[0308] In one aspect, the present invention relates to a DNase variant comprising a substituted T1V+V76I, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNase activity.

[0309] In one aspect, the present invention relates to a DNAse variant comprising a substituted T1V+Q109R, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNAse activity.

[0310] In one aspect, the present invention relates to a DNAse variant comprising a substituted T1V+S116D, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNAse activity.

[0311] In one aspect, the present invention relates to a DNAse variant comprising a substituted T1V+S116E, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNAse activity.

[0312] In one aspect, the present invention relates to a DNAse variant comprising a substitution for T1V+T127V, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNAse activity.

[0313] In one aspect, the present invention relates to a DNAse variant comprising a substituted T1V+T127I, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNAse activity.

[0314] In one aspect, the present invention relates to a DNase variant comprising a substituted T1V+T127L, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNase activity.

[0315] In one aspect, the present invention relates to a DNAse variant comprising a substituted T1V+S144P, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNAse activity.

[0316] In one aspect, the present invention relates to a DNAse variant comprising a substituted T1V+A147H, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNAse activity.

[0317] In one aspect, the present invention relates to a DNase variant comprising a substituted T1V+S167L, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNase activity.

[0318] In one aspect, the present invention relates to a DNase variant comprising a substituted T1V+S167I, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNase activity.

[0319] In one aspect, the present invention relates to a DNAse variant comprising a substitution for T1V+S167V, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNAse activity.

[0320] In one aspect, the present invention relates to a DNAse variant comprising a substituted T1V+G175D, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNAse activity.

[0321] In one aspect, the present invention relates to a DNAse variant comprising a substituted T1V+G175E, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNAse activity.

[0322] In one aspect, the present invention relates to a DNase variant comprising a substituted S13Y+T22P, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNase activity.

[0323] In one aspect, the present invention relates to a DNAse variant comprising a substitution for S13Y+S25P, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNAse activity.

[0324] In one aspect, the present invention relates to a DNase variant comprising a substitution for S13Y+S27L, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNase activity.

[0325] In one aspect, the present invention relates to a DNAse variant comprising a substitution for S13Y+S39P, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNAse activity.

[0326] In one aspect, the present invention relates to a DNAse variant comprising a substitution for S13Y+S42G, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNAse activity.

[0327] In one aspect, the present invention relates to a DNase variant comprising a substitution for S13Y+S42A, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNase activity.

[0328] In one aspect, the present invention relates to a DNase variant comprising a substitution for S13Y+S42T, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNase activity.

[0329] In one aspect, the present invention relates to a DNAse variant comprising a substitution for S13Y+S57W, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNAse activity.

[0330] In one aspect, the present invention relates to a DNase variant comprising a substitution for S13Y+S57Y, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNase activity.

[0331] In one aspect, the present invention relates to a DNAse variant comprising a substitution for S13Y+S57F, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNAse activity.

[0332] In one aspect, the present invention relates to a DNAse variant comprising a substitution for S13Y+S59V, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNAse activity.

[0333] In one aspect, the present invention relates to a DNase variant comprising a substituted S13Y+S59I, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNase activity.

[0334] In one aspect, the present invention relates to a DNAse variant comprising a substitution for S13Y+S59L, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNAse activity.

[0335] In one aspect, the present invention relates to a DNase variant comprising a substituted S13Y+V76L, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNase activity.

[0336] In one aspect, the present invention relates to a DNase variant comprising a substituted S13Y+V76I, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNase activity.

[0337] In one aspect, the present invention relates to a DNAse variant comprising a substituted S13Y+Q109R, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNAse activity.

[0338] In one aspect, the present invention relates to a DNAse variant comprising a substitution for S13Y+S116D, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNAse activity.

[0339] In one aspect, the present invention relates to a DNAse variant comprising a substitution for S13Y+S116E, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNAse activity.

[0340] In one aspect, the present invention relates to a DNAse variant comprising a substitution for S13Y+T127V, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNAse activity.

[0341] In one aspect, the present invention relates to a DNase variant comprising a substituted S13Y+T127I, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNase activity.

[0342] In one aspect, the present invention relates to a DNase variant comprising a substituted S13Y+T127L, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNase activity.

[0343] In one aspect, the present invention relates to a DNase variant comprising a substitution for S13Y+S144P, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNase activity.

[0344] In one aspect, the present invention relates to a DNase variant comprising a substituted S13Y+A147H, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNase activity.

[0345] In one aspect, the present invention relates to a DNase variant comprising a substituted S13Y+S167L, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNase activity.

[0346] In one aspect, the present invention relates to a DNase variant comprising a substituted S13Y+S167I, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNase activity.

[0347] In one aspect, the present invention relates to a DNase variant comprising a substitution for S13Y+S167V, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNase activity.

[0348] In one aspect, the present invention relates to a DNAse variant comprising a substituted S13Y+G175D, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNAse activity.

[0349] In one aspect, the present invention relates to a DNase variant comprising a substituted S13Y+G175E, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNase activity.

[0350] In one aspect, the present invention relates to a DNase variant comprising a substituted T22P+S25P, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNase activity.

[0351] In one aspect, the present invention relates to a DNase variant comprising a substituted T22P+S27L, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNase activity.

[0352] In one aspect, the present invention relates to a DNase variant comprising a substituted T22P+S39P, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNase activity.

[0353] In one aspect, the present invention relates to a DNAse variant comprising a substituted T22P+S42G, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNAse activity.

[0354] In one aspect, the present invention relates to a DNase variant comprising a substituted T22P+S42A, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNase activity.

[0355] In one aspect, the present invention relates to a DNase variant comprising a substituted T22P+S42T, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNase activity.

[0356] In one aspect, the present invention relates to a DNase variant comprising a substituted T22P+S57W, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNase activity.

[0357] In one aspect, the present invention relates to a DNase variant comprising a substituted T22P+S57Y, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNase activity.

[0358] In one aspect, the present invention relates to a DNAse variant comprising a substituted T22P+S57F, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNAse activity.

[0359] In one aspect, the present invention relates to a DNAse variant comprising a substituted T22P+S59V, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNAse activity.

[0360] In one aspect, the present invention relates to a DNase variant comprising a substituted T22P+S59I, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNase activity.

[0361] In one aspect, the present invention relates to a DNase variant comprising a substituted T22P+S59L, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNase activity.

[0362] In one aspect, the present invention relates to a DNase variant comprising a substituted T22P+V76L, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNase activity.

[0363] In one aspect, the present invention relates to a DNase variant comprising a substituted T22P+V76I, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNase activity.

[0364] In one aspect, the present invention relates to a DNAse variant comprising a substituted T22P+Q109R, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNAse activity.

[0365] In one aspect, the present invention relates to a DNase variant comprising a substituted T22P+S116D, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNase activity.

[0366] In one aspect, the present invention relates to a DNase variant comprising a substituted T22P+S116E, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNase activity.

[0367] In one aspect, the present invention relates to a DNase variant comprising a substituted T22P+T127V, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNase activity.

[0368] In one aspect, the present invention relates to a DNase variant comprising a substituted T22P+T127I, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNase activity.

[0369] In one aspect, the present invention relates to a DNase variant comprising a substituted T22P+T127L, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNase activity.

[0370] In one aspect, the present invention relates to a DNase variant comprising a substituted T22P+S144P, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNase activity.

[0371] In one aspect, the present invention relates to a DNase variant comprising a substituted T22P+A147H, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNase activity.

[0372] In one aspect, the present invention relates to a DNase variant comprising a substituted T22P+S167L, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNase activity.

[0373] In one aspect, the present invention relates to a DNase variant comprising a substituted T22P+S167I, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNase activity.

[0374] In one aspect, the present invention relates to a DNase variant comprising a substituted T22P+S167V, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNase activity.

[0375] In one aspect, the present invention relates to a DNAse variant comprising a substituted T22P+G175D, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNAse activity.

[0376] In one aspect, the present invention relates to a DNase variant comprising a substituted T22P+G175E, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNase activity.

[0377] In one aspect, the present invention relates to a DNase variant comprising a substitution for S25P+S27L, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNase activity.

[0378] In one aspect, the present invention relates to a DNAse variant comprising a substitution for S25P+S39P, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNAse activity.

[0379] In one aspect, the present invention relates to a DNAse variant comprising a substitution for S25P+S42G, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNAse activity.

[0380] In one aspect, the present invention relates to a DNase variant comprising a substitution for S25P+S42A, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNase activity.

[0381] In one aspect, the present invention relates to a DNAse variant comprising a substitution for S25P+S42T, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNAse activity.

[0382] In one aspect, the present invention relates to a DNAse variant comprising a substitution for S25P+S57W, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNAse activity.

[0383] In one aspect, the present invention relates to a DNase variant comprising a substitution for S25P+S57Y, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNase activity.

[0384] In one aspect, the present invention relates to a DNAse variant comprising a substitution for S25P+S57F, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNAse activity.

[0385] In one aspect, the present invention relates to a DNAse variant comprising a substitution for S25P+S59V, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNAse activity.

[0386] In one aspect, the present invention relates to a DNase variant comprising a substitution for S25P+S59I, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNase activity.

[0387] In one aspect, the present invention relates to a DNase variant comprising a substitution for S25P+S59L, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNase activity.

[0388] In one aspect, the present invention relates to a DNase variant comprising a substituted S25P+V76L, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNase activity.

[0389] In one aspect, the present invention relates to a DNase variant comprising a substituted S25P+V76I, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNase activity.

[0390] In one aspect, the present invention relates to a DNAse variant comprising a substituted S25P+Q109R, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNAse activity.

[0391] In one aspect, the present invention relates to a DNAse variant comprising a substitution for S25P+S116D, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNAse activity.

[0392] In one aspect, the present invention relates to a DNAse variant comprising a substitution for S25P+S116E, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNAse activity.

[0393] In one aspect, the present invention relates to a DNAse variant comprising a substitution for S25P+T127V, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNAse activity.

[0394] In one aspect, the present invention relates to a DNase variant comprising a substituted S25P+T127I, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNase activity.

[0395] In one aspect, the present invention relates to a DNase variant comprising a substituted S25P+T127L, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNase activity.

[0396] In one aspect, the present invention relates to a DNase variant comprising a substitution for S25P+S144P, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNase activity.

[0397] In one aspect, the present invention relates to a DNase variant comprising a substituted S25P+A147H, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNase activity.

[0398] In one aspect, the present invention relates to a DNase variant comprising a substitution for S25P+S167L, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNase activity.

[0399] In one aspect, the present invention relates to a DNase variant comprising a substituted S25P+S167I, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNase activity.

[0400] In one aspect, the present invention relates to a DNAse variant comprising a substitution for S25P+S167V, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNAse activity.

[0401] In one aspect, the present invention relates to a DNase variant comprising a substituted S25P+G175D, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNase activity.

[0402] In one aspect, the present invention relates to a DNase variant comprising a substituted S25P+G175E, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNase activity.

[0403] In one aspect, the present invention relates to a DNAse variant comprising a substitution for S39P+S42G, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNAse activity.

[0404] In one aspect, the present invention relates to a DNAse variant comprising a substitution for S39P+S42A, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNAse activity.

[0405] In one aspect, the present invention relates to a DNAse variant comprising a substitution for S39P+S42T, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNAse activity.

[0406] In one aspect, the present invention relates to a DNAse variant comprising a substitution for S39P+S57W, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNAse activity.

[0407] In one aspect, the present invention relates to a DNase variant comprising a substitution for S39P+S57Y, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNase activity.

[0408] In one aspect, the present invention relates to a DNAse variant comprising a substitution for S39P+S57F, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNAse activity.

[0409] In one aspect, the present invention relates to a DNAse variant comprising a substitution for S39P+S59V, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNAse activity.

[0410] In one aspect, the present invention relates to a DNase variant comprising a substitution for S39P+S59I, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNase activity.

[0411] In one aspect, the present invention relates to a DNAse variant comprising a substitution for S39P+S59L, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNAse activity.

[0412] In one aspect, the present invention relates to a DNase variant comprising a substituted S39P+V76L, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNase activity.

[0413] In one aspect, the present invention relates to a DNase variant comprising a substituted S39P+V76I, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNase activity.

[0414] In one aspect, the present invention relates to a DNAse variant comprising a substituted S39P+Q109R, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNAse activity.

[0415] In one aspect, the present invention relates to a DNAse variant comprising a substitution for S39P+S116D, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNAse activity.

[0416] In one aspect, the present invention relates to a DNAse variant comprising a substitution for S39P+S116E, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNAse activity.

[0417] In one aspect, the present invention relates to a DNAse variant comprising a substitution for S39P+T127V, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNAse activity.

[0418] In one aspect, the present invention relates to a DNase variant comprising a substituted S39P+T127I, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNase activity.

[0419] In one aspect, the present invention relates to a DNase variant comprising a substituted S39P+T127L, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNase activity.

[0420] In one aspect, the present invention relates to a DNase variant comprising replacing S39P+S144P, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNase activity.

[0421] In one aspect, the present invention relates to a DNase variant comprising a substituted S39P+A147H, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNase activity.

[0422] In one aspect, the present invention relates to a DNase variant comprising a substitution for S39P+S167L, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNase activity.

[0423] In one aspect, the present invention relates to a DNase variant comprising a substitution for S39P+S167I, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNase activity.

[0424] In one aspect, the present invention relates to a DNAse variant comprising a substitution for S39P+S167V, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNAse activity.

[0425] In one aspect, the present invention relates to a DNAse variant comprising a substituted S39P+G175D, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNAse activity.

[0426] In one aspect, the present invention relates to a DNAse variant comprising a substituted S39P+G175E, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNAse activity.

[0427] In one aspect, the present invention relates to a DNAse variant comprising a substitution for S42G+S57W, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNAse activity.

[0428] In one aspect, the present invention relates to a DNAse variant comprising a substitution for S42G+S57Y, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNAse activity.

[0429] In one aspect, the present invention relates to a DNAse variant comprising a substitution for S42G+S57F, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNAse activity.

[0430] In one aspect, the present invention relates to a DNAse variant comprising a substitution for S42G+S59V, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNAse activity.

[0431] In one aspect, the present invention relates to a DNAse variant comprising a substitution for S42G+S59I, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNAse activity.

[0432] In one aspect, the present invention relates to a DNase variant comprising a substitution for S42G+S59L, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNase activity.

[0433] In one aspect, the present invention relates to a DNase variant comprising a substituted S42G+V76L, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNase activity.

[0434] In one aspect, the present invention relates to a DNase variant comprising a substituted S42G+V76I, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNase activity.

[0435] In one aspect, the present invention relates to a DNAse variant comprising a substituted S42G+Q109R, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNAse activity.

[0436] In one aspect, the present invention relates to a DNAse variant comprising a substitution for S42G+S116D, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNAse activity.

[0437] In one aspect, the present invention relates to a DNAse variant comprising a substitution for S42G+S116E, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNAse activity.

[0438] In one aspect, the present invention relates to a DNase variant comprising a substituted S42G+T127V, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNase activity.

[0439] In one aspect, the present invention relates to a DNase variant comprising a substituted S42G+T127I, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNase activity.

[0440] In one aspect, the present invention relates to a DNase variant comprising a substituted S42G+T127L, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNase activity.

[0441] In one aspect, the present invention relates to a DNase variant comprising a substitution for S42G+S144P, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNase activity.

[0442] In one aspect, the present invention relates to a DNase variant comprising a substituted S42G+A147H, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNase activity.

[0443] In one aspect, the present invention relates to a DNase variant comprising a substitution for S42G+S167L, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNase activity.

[0444] In one aspect, the present invention relates to a DNase variant comprising a substitution for S42G+S167I, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNase activity.

[0445] In one aspect, the present invention relates to a DNAse variant comprising a substitution for S42G+S167V, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNAse activity.

[0446] In one aspect, the present invention relates to a DNase variant comprising a substituted S42G+G175D, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNase activity.

[0447] In one aspect, the present invention relates to a DNAse variant comprising a substituted S42G+G175E, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNAse activity.

[0448] In one aspect, the present invention relates to a DNase variant comprising a substitution for S42A+S57W, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNase activity.

[0449] In one aspect, the present invention relates to a DNase variant comprising a substitution for S42A+S57Y, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNase activity.

[0450] In one aspect, the present invention relates to a DNAse variant comprising a substitution for S42A+S57F, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNAse activity.

[0451] In one aspect, the present invention relates to a DNAse variant comprising a substitution for S42A+S59V, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNAse activity.

[0452] In one aspect, the present invention relates to a DNase variant comprising a substitution for S42A+S59I, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNase activity.

[0453] In one aspect, the present invention relates to a DNAse variant comprising a substitution for S42A+S59L, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNAse activity.

[0454] In one aspect, the present invention relates to a DNase variant comprising a substituted S42A+V76L, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNase activity.

[0455] In one aspect, the present invention relates to a DNase variant comprising a substituted S42A+V76I, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNase activity.

[0456] In one aspect, the present invention relates to a DNAse variant comprising a substituted S42A+Q109R, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNAse activity.

[0457] In one aspect, the present invention relates to a DNAse variant comprising a substitution for S42A+S116D, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNAse activity.

[0458] In one aspect, the present invention relates to a DNAse variant comprising a substitution for S42A+S116E, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNAse activity.

[0459] In one aspect, the present invention relates to a DNase variant comprising a substituted S42A+T127V, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNase activity.

[0460] In one aspect, the present invention relates to a DNase variant comprising a substituted S42A+T127I, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNase activity.

[0461] In one aspect, the present invention relates to a DNase variant comprising a substituted S42A+T127L, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNase activity.

[0462] In one aspect, the present invention relates to a DNase variant comprising a substitution for S42A+S144P, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNase activity.

[0463] In one aspect, the present invention relates to a DNase variant comprising a substitution for S42A+A147H, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNase activity.

[0464] In one aspect, the present invention relates to a DNase variant comprising a substitution for S42A+S167L, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNase activity.

[0465] In one aspect, the present invention relates to a DNase variant comprising a substitution for S42A+S167I, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNase activity.

[0466] In one aspect, the present invention relates to a DNase variant comprising a substitution for S42A+S167V, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNase activity.

[0467] In one aspect, the present invention relates to a DNase variant comprising a substituted S42A+G175D, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNase activity.

[0468] In one aspect, the present invention relates to a DNase variant comprising a substituted S42A+G175E, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNase activity.

[0469] In one aspect, the present invention relates to a DNAse variant comprising a substitution for S42T+S57W, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNAse activity.

[0470] In one aspect, the present invention relates to a DNAse variant comprising a substitution for S42T+S57Y, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNAse activity.

[0471] In one aspect, the present invention relates to a DNAse variant comprising a substitution for S42T+S57F, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNAse activity.

[0472] In one aspect, the present invention relates to a DNAse variant comprising a substitution for S42T+S59V, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNAse activity.

[0473] In one aspect, the present invention relates to a DNAse variant comprising a substitution for S42T+S59I, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNAse activity.

[0474] In one aspect, the present invention relates to a DNAse variant comprising a substitution for S42T+S59L, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNAse activity.

[0475] In one aspect, the present invention relates to a DNase variant comprising a substituted S42T+V76L, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNase activity.

[0476] In one aspect, the present invention relates to a DNase variant comprising a substituted S42T+V76I, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNase activity.

[0477] In one aspect, the present invention relates to a DNAse variant comprising a substituted S42T+Q109R, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNAse activity.

[0478] In one aspect, the present invention relates to a DNAse variant comprising a substitution for S42T+S116D, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNAse activity.

[0479] In one aspect, the present invention relates to a DNAse variant comprising a substitution for S42T+S116E, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNAse activity.

[0480] In one aspect, the present invention relates to a DNAse variant comprising a substitution for S42T+T127V, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNAse activity.

[0481] In one aspect, the present invention relates to a DNase variant comprising a substituted S42T+T127I, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNase activity.

[0482] In one aspect, the present invention relates to a DNase variant comprising a substituted S42T+T127L, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNase activity.

[0483] In one aspect, the present invention relates to a DNase variant comprising a substitution for S42T+S144P, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNase activity.

[0484] In one aspect, the present invention relates to a DNase variant comprising a substituted S42T+A147H, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNase activity.

[0485] In one aspect, the present invention relates to a DNase variant comprising a substitution for S42T+S167L, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNase activity.

[0486] In one aspect, the present invention relates to a DNase variant comprising a substituted S42T+S167I, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNase activity.

[0487] In one aspect, the present invention relates to a DNAse variant comprising a substitution for S42T+S167V, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNAse activity.

[0488] In one aspect, the present invention relates to a DNAse variant comprising a substituted S42T+G175D, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNAse activity.

[0489] In one aspect, the present invention relates to a DNAse variant comprising a substituted S42T+G175E, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNAse activity.

[0490] In one aspect, the present invention relates to a DNAse variant comprising a substitution for S57W+S59V, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNAse activity.

[0491] In one aspect, the present invention relates to a DNAse variant comprising a substitution for S57W+S59I, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNAse activity.

[0492] In one aspect, the present invention relates to a DNase variant comprising replacing S57W+S59L, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNase activity.

[0493] In one aspect, the present invention relates to a DNase variant comprising a substituted S57W+V76L, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNase activity.

[0494] In one aspect, the present invention relates to a DNase variant comprising a substituted S57W+V76I, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNase activity.

[0495] In one aspect, the present invention relates to a DNAse variant comprising a substituted S57W+Q109R, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNAse activity.

[0496] In one aspect, the present invention relates to a DNAse variant comprising a substitution for S57W+S116D, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNAse activity.

[0497] In one aspect, the present invention relates to a DNAse variant comprising a substitution for S57W+S116E, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNAse activity.

[0498] In one aspect, the present invention relates to a DNAse variant comprising a substitution for S57W+T127V, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNAse activity.

[0499] In one aspect, the present invention relates to a DNase variant comprising a substituted S57W+T127I, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNase activity.

[0500] In one aspect, the present invention relates to a DNase variant comprising a substituted S57W+T127L, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNase activity.

[0501] In one aspect, the present invention relates to a DNAse variant comprising a substitution for S57W+S144P, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNAse activity.

[0502] In one aspect, the present invention relates to a DNase variant comprising a substituted S57W+A147H, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNase activity.

[0503] In one aspect, the present invention relates to a DNase variant comprising a substitution for S57W+S167L, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNase activity.

[0504] In one aspect, the present invention relates to a DNase variant comprising a substitution for S57W+S167I, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNase activity.

[0505] In one aspect, the present invention relates to a DNAse variant comprising a substitution for S57W+S167V, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNAse activity.

[0506] In one aspect, the present invention relates to a DNAse variant comprising a substituted S57W+G175D, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNAse activity.

[0507] In one aspect, the present invention relates to a DNAse variant comprising a substituted S57W+G175E, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNAse activity.

[0508] In one aspect, the present invention relates to a DNAse variant comprising a substitution for S57Y+S59V, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNAse activity.

[0509] In one aspect, the present invention relates to a DNase variant comprising a substitution for S57Y+S59I, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNase activity.

[0510] In one aspect, the present invention relates to a DNase variant comprising a substitution for S57Y+S59L, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNase activity.

[0511] In one aspect, the present invention relates to a DNase variant comprising a substituted S57Y+V76L, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNase activity.

[0512] In one aspect, the present invention relates to a DNase variant comprising a substituted S57Y+V76I, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNase activity.

[0513] In one aspect, the present invention relates to a DNAse variant comprising a substituted S57Y+Q109R, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNAse activity.

[0514] In one aspect, the present invention relates to a DNase variant comprising a substitution for S57Y+S116D, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNase activity.

[0515] In one aspect, the present invention relates to a DNase variant comprising a substitution for S57Y+S116E, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNase activity.

[0516] In one aspect, the present invention relates to a DNase variant comprising a substitution for S57Y+T127V, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNase activity.

[0517] In one aspect, the present invention relates to a DNase variant comprising a substituted S57Y+T127I, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNase activity.

[0518] In one aspect, the present invention relates to a DNase variant comprising a substituted S57Y+T127L, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNase activity.

[0519] In one aspect, the present invention relates to a DNase variant comprising a substitution for S57Y+S144P, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNase activity.

[0520] In one aspect, the present invention relates to a DNase variant comprising a substituted S57Y+A147H, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNase activity.

[0521] In one aspect, the present invention relates to a DNase variant comprising a substitution for S57Y+S167L, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNase activity.

[0522] In one aspect, the present invention relates to a DNase variant comprising a substitution for S57Y+S167I, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNase activity.

[0523] In one aspect, the present invention relates to a DNase variant comprising a substitution for S57Y+S167V, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNase activity.

[0524] In one aspect, the present invention relates to a DNAse variant comprising a substituted S57Y+G175D, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNAse activity.

[0525] In one aspect, the present invention relates to a DNase variant comprising a substituted S57Y+G175E, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNase activity.

[0526] In one aspect, the present invention relates to a DNAse variant comprising a substitution for S57F+S59V, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNAse activity.

[0527] In one aspect, the present invention relates to a DNAse variant comprising a substitution for S57F+S59I, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNAse activity.

[0528] In one aspect, the present invention relates to a DNAse variant comprising a substitution for S57F+S59L, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNAse activity.

[0529] In one aspect, the present invention relates to a DNase variant comprising a substituted S57F+V76L, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNase activity.

[0530] In one aspect, the present invention relates to a DNase variant comprising a substituted S57F+V76I, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNase activity.

[0531] In one aspect, the present invention relates to a DNAse variant comprising a substituted S57F+Q109R, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNAse activity.

[0532] In one aspect, the present invention relates to a DNAse variant comprising a substituted S57F+Q109R, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNAse activity.

[0533] In one aspect, the present invention relates to a DNAse variant comprising a substitution for S57F+S116D, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNAse activity.

[0534] In one aspect, the present invention relates to a DNAse variant comprising a substitution for S57F+S116E, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNAse activity.

[0535] In one aspect, the present invention relates to a DNAse variant comprising a substitution for S57F+T127V, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNAse activity.

[0536] In one aspect, the present invention relates to a DNase variant comprising a substituted S57F+T127I, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNase activity.

[0537] In one aspect, the present invention relates to a DNase variant comprising a substituted S57F+T127L, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNase activity.

[0538] In one aspect, the present invention relates to a DNAse variant comprising a substitution for S57F+S144P, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNAse activity.

[0539] In one aspect, the present invention relates to a DNase variant comprising a substituted S57F+A147H, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNase activity.

[0540] In one aspect, the present invention relates to a DNase variant comprising a substitution for S57F+S167L, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNase activity.

[0541] In one aspect, the present invention relates to a DNAse variant comprising a substitution for S57F+S167I, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNAse activity.

[0542] In one aspect, the present invention relates to a DNAse variant comprising a substitution for S57F+S167V, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNAse activity.

[0543] In one aspect, the present invention relates to a DNAse variant comprising a substituted S57F+G175D, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNAse activity.

[0544] In one aspect, the present invention relates to a DNAse variant comprising a substituted S57F+G175E, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNAse activity.

[0545] In one aspect, the present invention relates to a DNase variant comprising a substituted S59V+V76L, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNase activity.

[0546] In one aspect, the present invention relates to a DNase variant comprising a substituted S59V+V76I, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNase activity.

[0547] In one aspect, the present invention relates to a DNAse variant comprising a substituted S59V+Q109R, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNAse activity.

[0548] In one aspect, the present invention relates to a DNAse variant comprising a substitution for S59V+S116D, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNAse activity.

[0549] In one aspect, the present invention relates to a DNAse variant comprising a substitution for S59V+S116E, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNAse activity.

[0550] In one aspect, the present invention relates to a DNAse variant comprising a substitution for S59V+T127V, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNAse activity.

[0551] In one aspect, the present invention relates to a DNase variant comprising a substituted S59V+T127I, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNase activity.

[0552] In one aspect, the present invention relates to a DNAse variant comprising a substituted S59V+T127L, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNAse activity.

[0553] In one aspect, the present invention relates to a DNase variant comprising a substitution for S59V+S144P, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNase activity.

[0554] In one aspect, the present invention relates to a DNase variant comprising a substituted S59V+A147H, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNase activity.

[0555] In one aspect, the present invention relates to a DNase variant comprising a substitution for S59V+S167L, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNase activity.

[0556] In one aspect, the present invention relates to a DNase variant comprising a substitution for S59V+S167I, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNase activity.

[0557] In one aspect, the present invention relates to a DNase variant comprising replacing S59V+S167V, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNase activity.

[0558] In one aspect, the present invention relates to a DNAse variant comprising a substituted S59V+G175D, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNAse activity.

[0559] In one aspect, the present invention relates to a DNAse variant comprising a substituted S59V+G175E, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNAse activity.

[0560] In one aspect, the present invention relates to a DNase variant comprising a substituted S59I+V76L, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNase activity.

[0561] In one aspect, the present invention relates to a DNase variant comprising a substituted S59I+V76I, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNase activity.

[0562] In one aspect, the present invention relates to a DNAse variant comprising a substituted S59I+Q109R, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNAse activity.

[0563] In one aspect, the present invention relates to a DNAse variant comprising a substitution for S59I+S116D, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNAse activity.

[0564] In one aspect, the present invention relates to a DNAse variant comprising a substitution for S59I+S116E, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNAse activity.

[0565] In one aspect, the present invention relates to a DNAse variant comprising a substitution for S59I+T127V, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNAse activity.

[0566] In one aspect, the present invention relates to a DNase variant comprising a substituted S59I+T127I, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNase activity.

[0567] In one aspect, the present invention relates to a DNase variant comprising a substituted S59I+T127L, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNase activity.

[0568] In one aspect, the present invention relates to a DNase variant comprising a substitution for S59I+S144P, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNase activity.

[0569] In one aspect, the present invention relates to a DNase variant comprising a substituted S59I+A147H, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNase activity.

[0570] In one aspect, the present invention relates to a DNase variant comprising a substitution for S59I+S167L, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNase activity.

[0571] In one aspect, the present invention relates to a DNase variant comprising a substitution for S59I+S167I, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNase activity.

[0572] In one aspect, the present invention relates to a DNAse variant comprising a substitution for S59I+S167V, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNAse activity.

[0573] In one aspect, the present invention relates to a DNAse variant comprising a substituted S59I+G175D, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNAse activity.

[0574] In one aspect, the present invention relates to a DNAse variant comprising a substituted S59I+G175E, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNAse activity.

[0575] In one aspect, the present invention relates to a DNase variant comprising a substituted S59L+V76L, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNase activity.

[0576] In one aspect, the present invention relates to a DNase variant comprising a substituted S59L+V76I, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNase activity.

[0577] In one aspect, the present invention relates to a DNAse variant comprising a substituted S59L+Q109R, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNAse activity.

[0578] In one aspect, the present invention relates to a DNAse variant comprising a substitution for S59L+S116D, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNAse activity.

[0579] In one aspect, the present invention relates to a DNAse variant comprising a substitution for S59L+S116E, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNAse activity.

[0580] In one aspect, the present invention relates to a DNAse variant comprising a substitution for S59L+T127V, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNAse activity.

[0581] In one aspect, the present invention relates to a DNase variant comprising a substituted S59L+T127I, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNase activity.

[0582] In one aspect, the present invention relates to a DNase variant comprising a substituted S59L+T127L, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNase activity.

[0583] In one aspect, the present invention relates to a DNase variant comprising a substitution for S59L+S144P, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNase activity.

[0584] In one aspect, the present invention relates to a DNase variant comprising a substituted S59L+A147H, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNase activity.

[0585] In one aspect, the present invention relates to a DNase variant comprising replacing S59L+S167L, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNase activity.

[0586] In one aspect, the present invention relates to a DNase variant comprising a substituted S59L+S167I, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNase activity.

[0587] In one aspect, the present invention relates to a DNAse variant comprising a substitution for S59L+S167V, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNAse activity.

[0588] In one aspect, the present invention relates to a DNAse variant comprising a substituted S59L+G175D, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNAse activity.

[0589] In one aspect, the present invention relates to a DNAse variant comprising a substituted S59L+G175E, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNAse activity.

[0590] In one aspect, the present invention relates to a DNAse variant comprising a substituted V76L+Q109R, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNAse activity.

[0591] In one aspect, the present invention relates to a DNase variant comprising a substituted V76L+S116D, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNase activity.

[0592] In one aspect, the present invention relates to a DNase variant comprising a substituted V76L+S116E, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNase activity.

[0593] In one aspect, the present invention relates to a DNase variant comprising a substituted V76L+T127V, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNase activity.

[0594] In one aspect, the present invention relates to a DNase variant comprising a substituted V76L+T127I, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNase activity.

[0595] In one aspect, the present invention relates to a DNase variant comprising a substituted V76L+T127L, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNase activity.

[0596] In one aspect, the present invention relates to a DNase variant comprising a substituted V76L+S144P, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNase activity.

[0597] In one aspect, the present invention relates to a DNase variant comprising a substituted V76L+A147H, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNase activity.

[0598] In one aspect, the present invention relates to a DNase variant comprising a substituted V76L+S167L, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNase activity.

[0599] In one aspect, the present invention relates to a DNase variant comprising a substituted V76L+S167I, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNase activity.

[0600] In one aspect, the present invention relates to a DNase variant comprising a substituted V76L+S167V, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNase activity.

[0601] In one aspect, the present invention relates to a DNase variant comprising a substituted V76L+G175D, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNase activity.

[0602] In one aspect, the present invention relates to a DNase variant comprising a substituted V76L+G175E, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNase activity.

[0603] In one aspect, the present invention relates to a DNAse variant comprising a substituted V76I+Q109R, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNAse activity.

[0604] In one aspect, the present invention relates to a DNase variant comprising a substituted V76I+S116D, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNase activity.

[0605] In one aspect, the present invention relates to a DNase variant comprising a substituted V76I+S116E, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNase activity.

[0606] In one aspect, the present invention relates to a DNase variant comprising a substituted V76I+T127V, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNase activity.

[0607] In one aspect, the present invention relates to a DNase variant comprising a substituted V76I+T127I, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNase activity.

[0608] In one aspect, the present invention relates to a DNase variant comprising a substituted V76I+T127L, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNase activity.

[0609] In one aspect, the present invention relates to a DNase variant comprising a substituted V76I+S144P, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNase activity.

[0610] In one aspect, the present invention relates to a DNase variant comprising a substituted V76I+A147H, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNase activity.

[0611] In one aspect, the present invention relates to a DNase variant comprising a substituted V76I+S167L, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNase activity.

[0612] In one aspect, the present invention relates to a DNase variant comprising a substituted V76I+S167I, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNase activity.

[0613] In one aspect, the present invention relates to a DNase variant comprising a substituted V76I+S167V, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNase activity.

[0614] In one aspect, the present invention relates to a DNAse variant comprising a substituted V76I+G175D, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNAse activity.

[0615] In one aspect, the present invention relates to a DNase variant comprising a substituted V76I+G175E, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNase activity.

[0616] In one aspect, the present invention relates to a DNAse variant comprising a substituted Q109R+S116D, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNAse activity.

[0617] In one aspect, the present invention relates to a DNAse variant comprising a substituted Q109R+S116E, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNAse activity.

[0618] In one aspect, the present invention relates to a DNAse variant comprising a substituted Q109R+T127V, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNAse activity.

[0619] In one aspect, the present invention relates to a DNase variant comprising a substituted Q109R+T127I, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNase activity.

[0620] In one aspect, the present invention relates to a DNase variant comprising a substituted Q109R+T127L, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNase activity.

[0621] In one aspect, the present invention relates to a DNase variant comprising a substituted Q109R+S144P, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNase activity.

[0622] In one aspect, the present invention relates to a DNase variant comprising a substituted Q109R+A147H, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNase activity.

[0623] In one aspect, the present invention relates to a DNase variant comprising a substituted Q109R+S167L, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNase activity.

[0624] In one aspect, the present invention relates to a DNase variant comprising a substituted Q109R+S167I, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNase activity.

[0625] In one aspect, the present invention relates to a DNAse variant comprising a substituted Q109R+S167V, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNAse activity.

[0626] In one aspect, the present invention relates to a DNAse variant comprising a substituted Q109R+G175D, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNAse activity.

[0627] In one aspect, the present invention relates to a DNAse variant comprising a substituted Q109R+G175E, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNAse activity.

[0628] In one aspect, the present invention relates to a DNase variant comprising a substituted S116D+T127V, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNase activity.

[0629] In one aspect, the present invention relates to a DNase variant comprising a substituted S116D+T127I, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNase activity.

[0630] In one aspect, the present invention relates to a DNase variant comprising a substituted S116D+T127L, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNase activity.

[0631] In one aspect, the present invention relates to a DNase variant comprising a substituted S116D+S144P, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNase activity.

[0632] In one aspect, the present invention relates to a DNase variant comprising a substituted S116D+A147H, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNase activity.

[0633] In one aspect, the present invention relates to a DNase variant comprising a substituted S116D+S167L, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNase activity.

[0634] In one aspect, the present invention relates to a DNase variant comprising a substituted S116D+S167I, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNase activity.

[0635] In one aspect, the present invention relates to a DNase variant comprising a substitution for S116D+S167V, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNase activity.

[0636] In one aspect, the present invention relates to a DNase variant comprising a substituted S116D+G175D, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNase activity.

[0637] In one aspect, the present invention relates to a DNase variant comprising a substituted S116D+G175E, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNase activity.

[0638] In one aspect, the present invention relates to a DNase variant comprising a substituted S116E+T127V, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNase activity.

[0639] In one aspect, the present invention relates to a DNase variant comprising a substituted S116E+T127I, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNase activity.

[0640] In one aspect, the present invention relates to a DNase variant comprising a substituted S116E+T127L, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNase activity.

[0641] In one aspect, the present invention relates to a DNase variant comprising a substituted S116E+S144P, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNase activity.

[0642] In one aspect, the present invention relates to a DNase variant comprising a substituted S116E+A147H, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNase activity.

[0643] In one aspect, the present invention relates to a DNase variant comprising a substitution for S116E+S167L, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNase activity.

[0644] In one aspect, the present invention relates to a DNase variant comprising a substitution for S116E+S167I, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNase activity.

[0645] In one aspect, the present invention relates to a DNase variant comprising a substitution for S116E+S167V, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNase activity.

[0646] In one aspect, the present invention relates to a DNase variant comprising a substituted S116E+G175D, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNase activity.

[0647] In one aspect, the present invention relates to a DNase variant comprising a substituted S116E+G175E, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNase activity.

[0648] In one aspect, the present invention relates to a DNase variant comprising a substituted T127V+S144P, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNase activity.

[0649] In one aspect, the present invention relates to a DNase variant comprising a substituted T127V+A147H, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNase activity.

[0650] In one aspect, the present invention relates to a DNase variant comprising a substituted T127V+S167L, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNase activity.

[0651] In one aspect, the present invention relates to a DNase variant comprising a substituted T127V+S167I, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNase activity.

[0652] In one aspect, the present invention relates to a DNase variant comprising a substitution for T127V+S167V, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNase activity.

[0653] In one aspect, the present invention relates to a DNAse variant comprising a substituted T127V+G175D, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNAse activity.

[0654] In one aspect, the present invention relates to a DNase variant comprising a substituted T127V+G175E, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNase activity.

[0655] In one aspect, the present invention relates to a DNase variant comprising a substituted T127I+S144P, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNase activity.

[0656] In one aspect, the present invention relates to a DNase variant comprising a substituted T127I+A147H, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNase activity.

[0657] In one aspect, the present invention relates to a DNase variant comprising a substituted T127I+S167L, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNase activity.

[0658] In one aspect, the present invention relates to a DNase variant comprising a substituted T127I+S167I, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNase activity.

[0659] In one aspect, the present invention relates to a DNase variant comprising a substituted T127I+S167V, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNase activity.

[0660] In one aspect, the present invention relates to a DNase variant comprising a substituted T127I+G175D, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNase activity.

[0661] In one aspect, the present invention relates to a DNase variant comprising a substituted T127I+G175E, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNase activity.

[0662] In one aspect, the present invention relates to a DNase variant comprising a substituted T127L+S144P, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNase activity.

[0663] In one aspect, the present invention relates to a DNase variant comprising a substituted T127L+A147H, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNase activity.

[0664] In one aspect, the present invention relates to a DNase variant comprising a substituted T127L+S167L, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNase activity.

[0665] In one aspect, the present invention relates to a DNase variant comprising a substituted T127L+S167I, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNase activity.

[0666] In one aspect, the present invention relates to a DNase variant comprising a substituted T127L+S167V, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNase activity.

[0667] In one aspect, the present invention relates to a DNase variant comprising a substituted T127L+G175D, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNase activity.

[0668] In one aspect, the present invention relates to a DNase variant comprising a substituted T127L+G175E, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNase activity.

[0669] In one aspect, the present invention relates to a DNase variant comprising a substituted S144P+A147H, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNase activity.

[0670] In one aspect, the present invention relates to a DNase variant comprising a substituted S144P+S167L, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNase activity.

[0671] In one aspect, the present invention relates to a DNase variant comprising a substituted S144P+S167I, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNase activity.

[0672] In one aspect, the present invention relates to a DNase variant comprising a substitution for S144P+S167V, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNase activity.

[0673] In one aspect, the present invention relates to a DNase variant comprising a substituted S144P+G175D, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNase activity.

[0674] In one aspect, the present invention relates to a DNase variant comprising a substituted S144P+G175E, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNase activity.

[0675] In one aspect, the present invention relates to a DNase variant comprising a substituted A147H+S167L, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNase activity.

[0676] In one aspect, the present invention relates to a DNase variant comprising a substituted A147H+S167I, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNase activity.

[0677] In one aspect, the present invention relates to a DNase variant comprising a substituted A147H+S167V, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNase activity.

[0678] In one aspect, the present invention relates to a DNase variant comprising a substituted A147H+G175D, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNase activity.

[0679] In one aspect, the present invention relates to a DNase variant comprising a substituted A147H+G175E, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNase activity.

[0680] In one aspect, the present invention relates to a DNase variant comprising a substituted S167L+G175D, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNase activity.

[0681] In one aspect, the present invention relates to a DNase variant comprising a substituted S167L+G175E, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNase activity.

[0682] In one aspect, the present invention relates to a DNase variant comprising a substituted S167I+G175D, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNase activity.

[0683] In one aspect, the present invention relates to a DNase variant comprising a substituted S167I+G175E, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNase activity.

[0684] In one aspect, the present invention relates to a DNase variant comprising a substituted S167V+G175D, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNase activity.

[0685] In one aspect, the present invention relates to a DNase variant comprising a substituted S167V+G175E, wherein the position corresponds to the position of SEQ ID NO:1, and wherein the variant has DNase activity.

[0686] In some preferred aspects, the DNAse variant has at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, for example at least 96%, at least 97%, at least 98%, or at least 99%, but less than 100% sequence identity with the mature parental DNAse.

[0687] In some preferred aspects, the DNAse variant has at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, such as at least 96%, at least 97%, at least 98%, or at least 99%, but less than 100% sequence identity with the polypeptide shown in SEQ ID NO:1.

[0688] In some aspects, the improved properties are increased stability, such as improved detergent stability, improved washing stability, and improved thermal stability. Some aspects of the invention relate to DNase variants having an improvement factor greater than 1 when the DNase variant is tested for the property of interest in a relevant assay, wherein the property of the reference DNase is assigned a value of 1. In some aspects, the property is stability, such as storage stability. Some aspects of the invention relate to DNase variants having an improvement factor greater than 1 when the DNase variant is tested for the property of interest in the assays described in Examples 2, 3, 4, 5, or 6, wherein the property of the reference DNase is assigned a value of 1. In some aspects, the property is stability, such as storage stability.

[0689] In some aspects, the improved properties are increased stability, such as improved detergent stability, improved washing stability, and improved thermal stability. Some aspects of the invention relate to DNase variants having an improvement factor greater than 1 when the DNase variant is tested for the property of interest in a relevant assay, wherein the property of the reference DNase is assigned a value of 1. In some aspects, said property is stability, such as storage stability.

[0690] In some respects, the improved property is improved detergent stability.

[0691] In some respects, the improved property is improved protease stability.

[0692] In some respects, the improved property is the improved stability in the presence of sulfites.

[0693] As shown in Examples 3, 4, and 5, the variants according to the invention are improved when the residual activity ratio is defined as being higher than 1 compared to a reference DNA enzyme under the measurement conditions.

[0694]

[0695] In some aspects, in at least one of assays A, B, or C, the variant is improved compared to a reference DNase, wherein assay A measures improved protease stability, assay B measures improved stability in the presence of sulfite, and assay C measures improved detergent stability, as shown in Examples 3, 4, and 5. In some aspects, in at least two of assays A, B, or C, the variant is improved compared to a reference DNase, wherein assay A measures improved protease stability (Example 4), assay B measures improved stability in the presence of sulfite (Example 5), and assay C (Example 3) measures improved detergent stability, as shown in Examples 3, 4, and 5. In some aspects, in all three of assays A, B, or C, the variant is improved compared to a reference DNase, wherein assay A measures improved protease stability, assay B measures improved stability in the presence of sulfite, and assay C measures improved detergent stability, as shown in Examples 3, 4, and 5.

[0696] In some respects, the variants according to the invention have improved stability relative to a reference DNA enzyme, as measured by a residual activity ratio (RAR) greater than 1.0.

[0697] The residual activity ratio (RAR) of the DNAse variants of the present invention can be calculated as RA. 变体 / RA 参考 As shown in Examples 3, 4 and 5, the improved variants were identified as variants with a residual activity ratio (RAR) greater than 1.0 compared to the reference DNAse.

[0698] In some respects, the variants according to the invention have improved stability relative to a reference DNA enzyme, as measured by a half-life improvement factor (HIF) greater than 1.0.

[0699] The half-life improvement factor (HIF) of the DNAse variant of the present invention can be calculated as T. 1 / 2变体 / T 1 / 2参考As shown in Examples 2 or 6, the improved variants were identified as variants with a half-life improvement factor HIF greater than 1.0 compared to a reference DNase. Productive combinations of mutations according to the invention are those combinations that result in an improvement factor (e.g., HIF or RAR) greater than 1, or, in other words, combinations of mutations that result in variants with at least one improved property compared to the starting molecule (i.e., precursor, reference parent polypeptide, etc.). In a particularly preferred aspect, combinations of mutations according to the invention produce DNase variants with improved stability, wherein HIF > 1.0. In some aspects, variants according to the invention are combined with a reference DNase (e.g., SEQ ID NO: 1 or SEQ ID NO: 1). NO:28) has a half-life improvement factor (HIF) of at least the following values: 1.1; 1.2; 1.3; 1.4; 1.5; 1.6; 1.7; 1.8; 1.9; 2.0; 2.1; 2.2; 2.3; 2.4; 2.5; 2.6; 2.7, 2.8; 2.9; 3.0, 3.1; 3.2; 3.3; 3.4; 3.5, 3.6, 3.7, 3.8, 3.9; 4.0, 4.1; 4.2; 4.3; 4.4; 4.5, 4.6, 4.7, 4.8, 4.9; 5.0, 5.1; 5.2; 5.3; 5.4; 5.5, 5.6, 5.7, 5.8, 5.9; 3. 0, 6.1; 6.2; 6.3; 6.4; 6.5, 6.6, 6.7, 6.8, 6.9; 7.0, 7.1; 7.2; 7.3; 7.4; 7.5, 7.6, 7.7, 7.8, 7.9; 8.0, 8.1; 8.2; 8.3; 8.4; 8.5, 8.6, 8.7, 8.8, 8.9; 9.0, 9.1; 9.2; 9.3; 9.4; 9.5, 9.6, 9.7, 9.8, 9.9; 10.0, 10.1; 10.2; 10.3; 10.4; 10.5, 10.6, 10.7, 10.8, 10.9; 12, 15, 16, 20, 25 or 30.

[0700] In a particularly preferred aspect, the combination of mutations according to the invention produces a DNase variant with improved stability, wherein RAR > 1.0. In some aspects, the variants according to the invention are combined with a reference DNase (e.g., SEQ ID NO: 1 or SEQ ID NO: 1). NO:28) has a residual activity ratio (RAR) of at least: 1.1; 1.2; 1.3; 1.4; 1.5; 1.6; 1.7; 1.8; 1.9; 2.0; 2.1; 2.2; 2.3; 2.4; 2.5; 2.6; 2.7, 2.8; 2.9; 3.0, 3.1; 3.2; 3.3; 3.4; 3.5, 3.6, 3.7, 3.8, 3.9; 4.0, 4.1; 4.2; 4.3; 4.4; 4.5, 4.6, 4.7, 4.8, 4.9; 5.0, 5.1; 5.2; 5.3; 5.4; 5.5, 5.6, 5.7, 5.8, 5.9; 3.0 6.1; 6.2; 6.3; 6.4; 6.5, 6.6, 6.7, 6.8, 6.9; 7.0, 7.1; 7.2; 7.3; 7.4; 7.5, 7.6, 7.7, 7.8, 7.9; 8.0, 8.1; 8.2; 8.3; 8.4; 8.5, 8.6, 8.7, 8.8, 8.9; 9.0, 9.1; 9.2; 9.3; 9.4; 9.5, 9.6, 9.7, 9.8, 9.9; 10.0, 10.1; 10.2; 10.3; 10.4; 10.5, 10.6, 10.7, 10.8, 10.9; 12, 15, 16, 20, 25 or 30.

[0701] A preferred embodiment relates to a DNase variant with improved stability, wherein HIF > 1.0 compared to SEQ ID NO:1 or SEQ ID NO:28. A preferred embodiment relates to a DNase variant with improved stability, wherein the half-life improvement factor is at least 1.5 compared to SEQ ID NO:1 or SEQ ID NO:28 when measured as described in Examples 2-6.

[0702] A preferred embodiment relates to a DNase variant with improved stability, wherein the RAR > 1.0 compared to SEQ ID NO:1 or SEQ ID NO:28. A preferred embodiment relates to a DNase variant with improved stability, wherein the residual activity ratio (RAR) is at least 1.5 compared to SEQ ID NO:1 or SEQ ID NO:28 when measured as described in Examples 3, 4, 5, or 7.

[0703] A preferred embodiment relates to a DNase variant with improved stability, wherein HIF > 1.0 compared to SEQ ID NO:1 or SEQ ID NO:28. A preferred embodiment relates to a DNase variant with improved stability, wherein the half-life improvement factor (HIF) is at least 1.2 (e.g., 1.5) compared to SEQ ID NO:1 or SEQ ID NO:28, wherein HIF is calculated as: T 1 / 2DNA酶变体 / T 1 / 2参考 and the half-life (T) of DNase variants and DNase references (e.g., SEQ ID NO:1 or SEQ ID NO:28). 1 / 2 The residual activity (RA) of each DNAse variant and reference DNAse (e.g., SEQ ID NO:1 or SEQ ID NO:28) was calculated as: slope (stressed sample, 48.5 °C for 20 min) / slope (unstressed sample, 21 °C for 20 min).

[0704] A preferred embodiment relates to a DNAse variant with improved stability, wherein the residual activity ratio (RAR) compared to SEQ ID NO:1 or SEQ ID NO:28 is >1.0. A preferred embodiment relates to a DNAse variant with improved stability, wherein the residual activity ratio (RAR) compared to SEQ ID NO:1 or SEQ ID NO:28 is at least 1.2, for example 1.5, wherein the RAR is calculated as:

[0705]

[0706] As described in Measurement A in the Examples section.

[0707] A preferred aspect of the invention relates to a DNAse variant comprising, compared to the DNAse having SEQ ID NO:1, two or more substitutions selected from the group consisting of: T1I, T1L, T1V, S13Y, T22P, S25P, S27L, S39P, S42G, S42A, S42T, S57W, S57Y, S57F, S59V, S59I, S59L, V76L, V76I, Q109R, S116D, S116E, T127V, T127I, T127L, S144P, A147H, S167L, S167I, S167V, G175D, and G175E, wherein the positions correspond to the positions in SEQ ID NO:1 (numbered according to SEQ ID NO:1), and wherein the variant corresponds to the polypeptide shown in SEQ ID NO:1 or SEQ ID NO:1. The polypeptide shown in ID NO:28 has at least 90% sequence identity, wherein the variant has DNase activity, and wherein the DNase variant has improved stability compared to a DNase having the polypeptide shown in SEQ ID NO:1 or the polypeptide shown in SEQ ID NO:28, wherein the improved stability is measured with an improvement factor (e.g., RAR or HIF) > 1.0.

[0708] In some preferred aspects, the DNAse variants of the present invention contain variations at two or more positions selected from the group consisting of: 1, 13, 22, 25, 27, 39, 42, 57, 59, 76, 109, 116, 127, 144, 147, 167, and 175, wherein the variants have at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, for example at least 96%, at least 97%, at least 98%, or at least 99%, but less than 100% sequence identity with the polypeptide shown in SEQ ID NO:1, wherein each position corresponds to a position of the polypeptide shown in SEQ ID NO:1 (numbered according to SEQ ID NO:1), and wherein the variants have the same sequence identity as the parental DNAse (e.g., containing the DNAse variants with the same sequence identity as SEQ ID NO:1). Compared to the DNA enzyme of the polypeptide with the amino acid sequence shown in IDNO:1, it has at least one improved property, preferably improved stability, wherein the stability is tested as described in Example 1.

[0709] In some preferred aspects, the DNAse variants of the present invention comprise alterations (compared to SEQ ID NO:1) at two or more locations, said alterations being selected from the group consisting of: T1I, T1L, T1V, S13Y, T22P, S25P, S27L, S39P, S42G, S42A, S42T, S57W, S57Y, S57F, S59V, S59I, S59L, V76L, V76I, Q109R, S116D, S116E, T127V, T127I, T127L, S144P, A147H, S167L, S167I, S167V, G175D, and G175E, said variants being compared to SEQ ID NO:1. The polypeptide shown in NO:1 has at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, for example at least 96%, at least 97%, at least 98%, or at least 99%, but less than 100% sequence identity, wherein each position corresponds to a position of the polypeptide shown in SEQ ID NO:1 (numbered according to SEQ ID NO:1), and wherein said variant has at least one improved property compared to the parental DNase (e.g., a DNase containing a polypeptide having the amino acid sequence shown in SEQ ID NO:1), preferably said improved property being improved stability, wherein stability is tested as described in Examples 2 to 6.

[0710] In some preferred aspects, the DNAse variants of the present invention comprise one or more substitution groups selected from the following group (with SEQ ID NO: ... Compared to NO:1, this group consists of the following: T1I+S13Y, T1I+T22P, T1I+S25P, T1I+S27L, T1I+S39P, T1I+S42G, T1I+S42A, T1I+S42T, T1I+S57W, T1I+S57Y, T1I+S57F, T1I+S59V, T1I+S59I, T1I+S59L, T1I+V76L, T1I+V76I, T1I+Q109R, T1I+S116D, T1I+S116E, T1I+T127V, T1I+T127I, T1I+T127L, T1I+S144P, T1I+A147H , T1I+S167L, T1I+S167I, T1I+S167V, T1I+G175D, T1I+G175E, T1L+S13Y, T1L+T22P, T1L+S25P, T1L+S27L, T1L+S39P, T1L+S42G, T1L+S42A, T1L+S4 2T, T1L+S57W, T1L+S57Y, T1L+S57F, T1L+S59V, T1L+S59I, T1L+S59L, T1L +V76L, T1L+V76I, T1L+Q109R, T1L+S116D, T1L+S116E, T1L+T127V, T1L+T 127I, T1L+T127L, T1L+S144P, T1L+A147H, T1L+S167L, T1L+S167I, T1L+ S167V, T1L+G175D, T1L+G175E, T1V+S13Y, T1V+T22P, T1V+S25P, T1V+S27 L, T1V+S39P, T1V+S42G, T1V+S42A, T1V+S42T, T1V+S57W, T1V+S57Y, T1V+ S57F, T1V+S59V, T1V+S59I, T1V+S59L, T1V+V76L, T1V+V76I, T1V+Q109R, T1V+S116D, T1V+S116E, T1V+T127V, T1V+T127I, T1V+T127L, T1V+S144P, T1V+A147H, T1V+S167L, T1V+S167I, T1V+S167V, T1V+G175D, T1V+G175E, S13Y+T22P, S13Y+S25P, S13Y+S27L, S13Y+S39P, S13Y+S42G, S13Y+S42A, S13Y+S42T, S13Y+S57W, S13Y+S57Y, S13Y+S57F, S13Y+S59V, S13Y+S59I,S13Y+S59L、S13Y+V76L、S13Y+V76I、S13Y+Q109R、S13Y+S116D、S13Y+S116E 、S13Y+T127V、S13Y+T127I、S13Y+T127L、S13Y+S144P、S13Y+A147H、S13Y+S 167L、S13Y+S167I、S13Y+S167V、S13Y+G175D、S13Y+G175E、T22P+S25P、T22 P+S27L、T22P+S39P、T22P+S42G、T22P+S42A、T22P+S42T、T22P+S57W、T22P+ S57Y, T22P+S57F, T22P+S59V, T22P+S59I, T22P+S59L, T22P+V76L, T22P+V76I, T22P+Q109R, T22P+S116D, T22P+S116E, T22P+T127V, T22P+T127I, T22P +T127L、T22P+S144P、T22P+A147H、T22P+S167L、T22P+S167I、T22P+S167V、 T22P+G175D、T22P+G175E、S25P+S27L、S25P+S39P、S25P+S42G、S25P+S42A、 S25P+S42T、S25P+S57W、S25P+S57Y、S25P+S57F、S25P+S59V、S25P+S59I、S2 5P+S59L、S25P+V76L、S25P+V76I、S25P+Q109R、S25P+S116D、S25P+S116E、S 25P+T127V、S25P+T127I、S25P+T127L、S25P+S144P、S25P+A147H、S25P+S16 7L、S25P+S167I、S25P+S167V、S25P+G175D、S25P+G175E、S27L+S39P、S27L+ S42G, S27L+S42A, S27L+S42T, S27L+S57W, S27L+S57Y, S27L+S57F, S27L+S59V, S27L+S59I, S27L+S59L, S27L+V76L, S27L+V76I, S27L+Q109R, S27L+S11 6D, S27L+S116E, S27L+T127V, S27L+T127I, S27L+T127L, S27L+S144P, S27L+A147H, S27L+S167L, S27L+S167I, S27L+S167V, S27L+G175D, S27L+G175ES39P+S42G、S39P+S42A、S39P+S42T、S39P+S57W、S39P+S57Y、S39P+S57F、S39P+S59V、S39P+S59I、S39P+S59L、S39P+V76L、S39P+V76I、S39P+Q109R、S39P+S116D、S39P+S116E、S39P+T127V、S39P+T127I、S39P+T127L、S39P+S144P、S39P+A147H、S39P+S167L、S39P+S167I、S39P+S167V、S39P+G175D、S39P+G175E、S42G+S57W、S42G+S57Y、S42G+S57F、S42G+S59V、S42G+S59I、S42G+S59L、S42G+V76L、S42G+V76I、S42G+Q109R、S42G+S116D、S42G+S116E、S42G+T127V、S42G+T127I、S42G+T127L、S42G+S144P、S42G+A147H、S42G+S167L、S42G+S167I、S42G+S167V、S42G+G175D、S42G+G175E、S42A+S57W、S42A+S57Y、S42A+S57F、S42A+S59V、S42A+S59I、S42A+S59L、S42A+V76L、S42A+V76I、S42A+Q109R、S42A+S116D、S42A+S116E、S42A+T127V、S42A+T127I、S42A+T127L、S42A+S144P、S42A+A147H、S42A+S167L、S42A+S167I、S42A+S167V、S42A+G175D、S42A+G175E、S42T+S57W、S42T+S57Y、S42T+S57F、S42T+S59V、S42T+S59I、S42T+S59L、S42T+V76L、S42T+V76I、S42T+Q109R、S42T+S116D、S42T+S116E、S42T+T127V、S42T+T127I、S42T+T127L、S42T+S144P、S42T+A147H、S42T+S167L、S42T+S167I、S42T+S167V、S42T+G175D、S42T+G175E、S57W+S59V、S57W+S59I、S57W+S59L、S57W+V76L、S57W+V76I、S57W+Q109R、S57W+S116D、S57W+S116E、S57W+T127V、S57W+T127I、S57W+T127L、S57W+S144P、S57W+A147H、S57W+S167L、S57W+S167I、S57W+S167V、S57W+G175D、S57W+G175E、S57Y+S59V、S57Y+S59I、S57Y+S59L、S57Y+V76L、S57Y+V76I、S57Y+Q109R、S57Y+S116D、S57Y+S116E、S57Y+T127V、S57Y+T127I、S57Y+T127L、S57Y+S144P、S57Y+A147H、S57Y+S167L、S57Y+S167I、S57Y+S167V、S57Y+G175D、S57Y+G175E、S57F+S59V、S57F+S59I、S57F+S59L、S57F+V76L、S57F+V76I、S57F+Q109R、S57F+S116D、S57F+S116E、S57F+T127V、S57F+T127I、S57F+T127L、S57F+S144P、S57F+A147H、S57F+S167L、S57F+S167I、S57F+S167V、S57F+G175D、S57F+G175E、S59V+V76L、S59V+V76I、S59V+Q109R、S59V+S116D、S59V+S116E、S59V+T127V、S59V+T127I、S59V+T127L、S59V+S144P、S59V+A147H、S59V+S167L、S59V+S167I、S59V+S167V、S59V+G175D、S59V+G175E、S59I+V76L、S59I+V76I、S59I+Q109R、S59I+S116D、S59I+S116E、S59I+T127V、S59I+T127I、S59I+T127L、S59I+S144P、S59I+A147H、S59I+S167L、S59I+S167I、S59I+S167V、S59I+G175D、S59I+G175E、S59L+V76L、S59L+V76I、S59L+Q109R、S59L+S116D、S59L+S116E、S59L+T127V、S59L+T127I、S59L+T127L、S59L+S144P、S59L+A147H、S59L+S167L、S59L+S167I、S59L+S167V、S59L+G175D、S59L+G175E、V76L+Q109R、V76L+S116D、V76L+S116E、V76L+T127V、V76L+T127I、V76L+T127L、V76L+S144P、V76L+A147H、V76L+S167L、V76L+S167I、V76L+S167V、V76L+G175D、V76L+G175E、V76I+Q109R、V76I+S116D、V76I+S116E、V76I+T127V、V76I+T127I、V76I+T127L、V76I+S144P、V76I+A147H、V76I+S167L、V76I+S167I、V76I+S167V、V76I+G175D、V76I+G175E、Q109R+S116D、Q109R+S116E、Q109R+T127V、Q109R+T127I、Q109R+T127L、Q109R+S144P、Q109R+A147H、Q109R+S167L、Q109R+S167I、Q109R+S167V、Q109R+G175D、Q109R+G175E、S116D+T127V、S116D+T127I、S116D+T127L、S116D+S144P、S116D+A147H、S116D+S167L、S116D+S167I、S116D+S167V、S116D+G175D、S116D+G175E、S116E+T127V、S116E+T127I、S116E+T127L、S116E+S144P、S116E+A147H、S116E+S167L、S116E+S167I、S116E+S167V、S116E+G175D、S116E+G175E、T127V+S144P、T127V+A147H、T127V+S167L、T127V+S167I、T127V+S167V、T127V+G175D、T127V+G175E、T127I+S144P、T127I+A147H、T127I+S167L、T127I+S167I、T127I+S167V、T127I+G175D、T127I+G175E、T127L+S144P、T127L+A147H、T127L+S167L、T127L+S167I、T127L+S167V、T127L+G175D、T127L+G175E、S144P+A147H、S144P+S167L、S144P+S167I、S144P+S167V、S144P+G175D、S144P+G175E、A147H+S167L, A147H+S167I, A147H+S167V, A147H+G175D, A147H+G175E, S167L+G175D, S167L+G175E, S167I+G175D, S167I+G175E, S167V+G175D, and S167V+G175E, wherein said variants have at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, for example at least 96%, at least 97%, at least 98%, or at least 99%, but less than 100% sequence identity with the polypeptide shown in SEQ ID NO:1, wherein each position corresponds to a position of the polypeptide shown in SEQ ID NO:1 (according to SEQ ID NO:1). (Numbered as NO:1), and said variant has at least one improved property compared to the parental DNase (e.g., a DNase comprising a polypeptide having the amino acid sequence shown in SEQ ID NO:1), preferably said improved property being improved stability, wherein stability is tested as described in Examples 2 to 6.

[0711] In some respects, the DNA enzyme variants of the present invention have improved stability, as measured by the half-life improvement factor HIF, compared to the parent, or to a DNA enzyme having the polypeptide sequence shown in SEQ ID NO:1, or to a DNA enzyme having the polypeptide sequence shown in SEQ ID NO:28, or to a DNA enzyme having the same amino acid sequence as the DNA enzyme variant but without alteration at two or more specified positions.

[0712] In some respects, the DNA enzyme variants of the present invention have improved stability, such as as measured by residual activity ratio (RAR), compared to the parent, or to a DNA enzyme having the polypeptide sequence shown in SEQ ID NO:1, or to a DNA enzyme having the polypeptide sequence shown in SEQ ID NO:28, or to a DNA enzyme having the same amino acid sequence as the DNA enzyme variant but without alteration at two or more designated positions.

[0713] In some preferred aspects, the DNase variants of the present invention contain changes at two or more positions selected from the group consisting of: 1, 13, 22, 25, 27, 39, 42, 57, 59, 76, 109, 116, 127, 144, 147, 167, and 175, wherein each position corresponds to a position of the polypeptide of SEQ ID NO:1, and wherein each change provides a DNase variant having an increased stability as measured by a half-life modifier HIF, which is compatible with a parental DNase (e.g., containing the polypeptide of SEQ ID NO:1). The variant has at least 1.05, for example 1.08, for example 1.1, for example 1.15, for example 1.2, for example 1.25, for example 1.3, for example 1.4, for example 1.5, for example 1.6, for example 1.7, for example 1.8, for example 1.9, for example 2, for example 3, for example 4, for example 5 or for example at least 10 compared to the DNA enzyme of the polypeptide shown in SEQ ID NO:1, wherein the variant has at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, for example at least 96%, at least 97%, at least 98%, or at least 99%, but less than 100% sequence identity with the polypeptide shown in SEQ ID NO:1, wherein each position corresponds to a position of the polypeptide shown in SEQ ID NO:1 (numbered according to SEQ ID NO:1).

[0714] In some preferred aspects, the DNAse variants of the present invention contain two or more substitutions (compared to SEQ). IDNO:1), wherein the substitution is selected from the group consisting of: T1I, T1L, T1V, S13Y, T22P, S25P, S27L, S39P, S42G, S42A, S42T, S57W, S57Y, S57F, S59V, S59I, S59L, V76L, V76I, Q109R, S116D, S116E, T127V, T127I, T127L, S144P, A147H, S167L, S167I, S167V, G175D, and G175E, wherein each substitution provides a DNase variant having increased stability as measured by the half-life modifier HIF, the half-life modifier HIF being compatible with the parental DNase (e.g., containing the SEQ ID NO:1). The variant has at least 1.05, for example 1.08, for example 1.1, for example 1.15, for example 1.2, for example 1.25, for example 1.3, for example 1.4, for example 1.5, for example 1.6, for example 1.7, for example 1.8, for example 1.9, for example 2, for example 3, for example 4, for example 5 or for example at least 10 compared to the DNA enzyme of the polypeptide shown in SEQ ID NO:1, wherein the variant has at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, for example at least 96%, at least 97%, at least 98%, or at least 99%, but less than 100% sequence identity with the polypeptide shown in SEQ ID NO:1, wherein each position corresponds to a position of the polypeptide shown in SEQ ID NO:1 (numbered according to SEQ ID NO:1).

[0715] In some preferred aspects, the DNase variants of the present invention comprise two or more substitutions (compared to SEQ ID NO:1), wherein said substitutions are selected from the group consisting of: T1I, T1V, S13Y, T22P, S25P, S27L, S39P, S42G, S42A, S42T, S57W, S57F, S59V, S59I, S59L, V76L, Q109R, S116D, S116E, T127V, T127I, T127L, S144P, A147H, S167L, S167I, S167V, G175D, and G175E, wherein each substitution provides a DNase variant having increased stability as measured by a half-life improvement factor HIF, said half-life improvement factor HIF being compatible with a parental DNase (e.g., comprising SEQ ID NO:1). The variant has at least 1.05, for example 1.08, for example 1.1, for example 1.15, for example 1.2, for example 1.25, for example 1.3, for example 1.4, for example 1.5, for example 1.6, for example 1.7, for example 1.8, for example 1.9, for example 2, for example 3, for example 4, for example 5 or for example at least 10 compared to the DNA enzyme of the polypeptide shown in SEQ ID NO:1, wherein the variant has at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, for example at least 96%, at least 97%, at least 98%, or at least 99%, but less than 100% sequence identity with the polypeptide shown in SEQ ID NO:1, wherein each position corresponds to a position of the polypeptide shown in SEQ ID NO:1 (numbered according to SEQ ID NO:1).

[0716] In some preferred aspects, the DNase variants of the present invention comprise two or more substitutions (compared to SEQ ID NO:1), wherein said substitutions are selected from the group consisting of: T1I, T1V, S13Y, T22P, S25P, S27L, S39P, S42G, S42A, S42T, S57W, S57F, S59V, S59I, S59L, V76L, Q109R, S116D, S116E, T127V, T127I, T127L, S144P, S167L, S167I, S167V, G175D, and G175E, wherein each substitution provides a DNase variant having increased stability as measured by a half-life improvement factor HIF, said half-life improvement factor HIF being compatible with a parental DNase (e.g., comprising SEQ ID NO:1). The variant has at least 1.05, for example 1.08, for example 1.1, for example 1.15, for example 1.2, for example 1.25, for example 1.3, for example 1.4, for example 1.5, for example 1.6, for example 1.7, for example 1.8, for example 1.9, for example 2, for example 3, for example 4, for example 5 or for example at least 10 compared to the DNA enzyme of the polypeptide shown in SEQ ID NO:1, wherein the variant has at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, for example at least 96%, at least 97%, at least 98%, or at least 99%, but less than 100% sequence identity with the polypeptide shown in SEQ ID NO:1, wherein each position corresponds to a position of the polypeptide shown in SEQ ID NO:1 (numbered according to SEQ ID NO:1).

[0717] In some preferred aspects, the DNAse variants of the present invention comprise one or more substitution groups (compared to SEQ ID NO 1), wherein said one or more substitution groups are selected from the group consisting of: T1I+S13Y, T1I+T22P, T1I+S25P, T1I+S27L, T1I+S39P, T1I+S42G, T1I+S42A, T1I+S42T, T1I+S57W, T1I+S57Y, T1I+S57F, T1I+S59V, T1I+S59I, T1I+S59L, T1I+V76L, T1I+V76I, T1I+Q109R, T1I+S116D, T1I+S116E, T1I+T127V, T1I+T127I, T1I+T127L, T1I+ S144P, T1I+A147H, T1I+S167L, T1I+S167I, T1I+S167V, T1I+G175D, T1I +G175E, T1L+S13Y, T1L+T22P, T1L+S25P, T1L+S27L, T1L+S39P, T1L+S42G , T1L+S42A, T1L+S42T, T1L+S57W, T1L+S57Y, T1L+S57F, T1L+S59V, T1L+ S59I, T1L+S59L, T1L+V76L, T1L+V76I, T1L+Q109R, T1L+S116D, T1L+S116 E. T1L+T127V, T1L+T127I, T1L+T127L, T1L+S144P, T1L+A147H, T1L+S16 7L, T1L+S167I, T1L+S167V, T1L+G175D, T1L+G175E, T1V+S13Y, T1V+T22P , T1V+S25P, T1V+S27L, T1V+S39P, T1V+S42G, T1V+S42A, T1V+S42T, T1V+ S57W, T1V+S57Y, T1V+S57F, T1V+S59V, T1V+S59I, T1V+S59L, T1V+V76L, T 1V+V76I, T1V+Q109R, T1V+S116D, T1V+S116E, T1V+T127V, T1V+T127I, T 1V+T127L, T1V+S144P, T1V+A147H, T1V+S167L, T1V+S167I, T1V+S167V, T 1V+G175D, T1V+G175E, S13Y+T22P, S13Y+S25P, S13Y+S27L, S13Y+S39P, S13Y+S42G, S13Y+S42A, S13Y+S42T, S13Y+S57W, S13Y+S57Y, S13Y+S57F,S13Y+S59V、S13Y+S59I、S13Y+S59L、S13Y+V76L、S13Y+V76I、S13Y+Q109R、S 13Y+S116D、S13Y+S116E、S13Y+T127V、S13Y+T127I、S13Y+T127L、S13Y+S144 P、S13Y+A147H、S13Y+S167L、S13Y+S167I、S13Y+S167V、S13Y+G175D、S13Y+ G175E、T22P+S25P、T22P+S27L、T22P+S39P、T22P+S42G、T22P+S42A、T22P+S4 2T, T22P+S57W, T22P+S57Y, T22P+S57F, T22P+S59V, T22P+S59I, T22P+S59L, T22P+V76L, T22P+V76I, T22P+Q109R, T22P+S116D, T22P+S116E, T22P+T127 V、T22P+T127I、T22P+T127L、T22P+S144P、T22P+A147H、T22P+S167L、T22P+ S167I、T22P+S167V、T22P+G175D、T22P+G175E、S25P+S27L、S25P+S39P、S25P +S42G、S25P+S42A、S25P+S42T、S25P+S57W、S25P+S57Y、S25P+S57F、S25P+S 59V、S25P+S59I、S25P+S59L、S25P+V76L、S25P+V76I、S25P+Q109R、S25P+S11 6D、S25P+S116E、S25P+T127V、S25P+T127I、S25P+T127L、S25P+S144P、S25P +A147H、S25P+S167L、S25P+S167I、S25P+S167V、S25P+G175D、S25P+G175E、S 27L+S39P, S27L+S42G, S27L+S42A, S27L+S42T, S27L+S57W, S27L+S57Y, S27L+S57F, S27L+S59V, S27L+S59I, S27L+S59L, S27L+V76L, S27L+V76I, S27L+Q 109R, S27L+S116D, S27L+S116E, S27L+T127V, S27L+T127I, S27L+T127L, S27L+S144P, S27L+A147H, S27L+S167L, S27L+S167I, S27L+S167V, S27L+G175DS27L+G175E、S39P+S42G、S39P+S42A、S39P+S42T、S39P+S57W、S39P+S57Y、S39P+S57F、S39P+S59V、S39P+S59I、S39P+S59L、S39P+V76L、S39P+V76I、S39P+Q109R、S39P+S116D、S39P+S116E、S39P+T127V、S39P+T127I、S39P+T127L、S39P+S144P、S39P+A147H、S39P+S167L、S39P+S167I、S39P+S167V、S39P+G175D、S39P+G175E、S42G+S57W、S42G+S57Y、S42G+S57F、S42G+S59V、S42G+S59I、S42G+S59L、S42G+V76L、S42G+V76I、S42G+Q109R、S42G+S116D、S42G+S116E、S42G+T127V、S42G+T127I、S42G+T127L、S42G+S144P、S42G+A147H、S42G+S167L、S42G+S167I、S42G+S167V、S42G+G175D、S42G+G175E、S42A+S57W、S42A+S57Y、S42A+S57F、S42A+S59V、S42A+S59I、S42A+S59L、S42A+V76L、S42A+V76I、S42A+Q109R、S42A+S116D、S42A+S116E、S42A+T127V、S42A+T127I、S42A+T127L、S42A+S144P、S42A+A147H、S42A+S167L、S42A+S167I、S42A+S167V、S42A+G175D、S42A+G175E、S42T+S57W、S42T+S57Y、S42T+S57F、S42T+S59V、S42T+S59I、S42T+S59L、S42T+V76L、S42T+V76I、S42T+Q109R、S42T+S116D、S42T+S116E、S42T+T127V、S42T+T127I、S42T+T127L、S42T+S144P、S42T+A147H、S42T+S167L、S42T+S167I、S42T+S167V、S42T+G175D、S42T+G175E、S57W+S59V、S57W+S59I、S57W+S59L、S57W+V76L、S57W+V76I、S57W+Q109R、S57W+S116D、S57W+S116E、S57W+T127V、S57W+T127I、S57W+T127L、S57W+S144P、S57W+A147H、S57W+S167L、S57W+S167I、S57W+S167V、S57W+G175D、S57W+G175E、S57Y+S59V、S57Y+S59I、S57Y+S59L、S57Y+V76L、S57Y+V76I、S57Y+Q109R、S57Y+S116D、S57Y+S116E、S57Y+T127V、S57Y+T127I、S57Y+T127L、S57Y+S144P、S57Y+A147H、S57Y+S167L、S57Y+S167I、S57Y+S167V、S57Y+G175D、S57Y+G175E、S57F+S59V、S57F+S59I、S57F+S59L、S57F+V76L、S57F+V76I、S57F+Q109R、S57F+S116D、S57F+S116E、S57F+T127V、S57F+T127I、S57F+T127L、S57F+S144P、S57F+A147H、S57F+S167L、S57F+S167I、S57F+S167V、S57F+G175D、S57F+G175E、S59V+V76L、S59V+V76I、S59V+Q109R、S59V+S116D、S59V+S116E、S59V+T127V、S59V+T127I、S59V+T127L、S59V+S144P、S59V+A147H、S59V+S167L、S59V+S167I、S59V+S167V、S59V+G175D、S59V+G175E、S59I+V76L、S59I+V76I、S59I+Q109R、S59I+S116D、S59I+S116E、S59I+T127V、S59I+T127I、S59I+T127L、S59I+S144P、S59I+A147H、S59I+S167L、S59I+S167I、S59I+S167V、S59I+G175D、S59I+G175E、S59L+V76L、S59L+V76I、S59L+Q109R、S59L+S116D、S59L+S116E、S59L+T127V、S59L+T127I、S59L+T127L、S59L+S144P、S59L+A147H、S59L+S167L、S59L+S167I、S59L+S167V、S59L+G175D、S59L+G175E、V76L+Q109R、V76L+S116D、V76L+S116E、V76L+T127V、V76L+T127I、V76L+T127L、V76L+S144P、V76L+A147H、V76L+S167L、V76L+S167I、V76L+S167V、V76L+G175D、V76L+G175E、V76I+Q109R、V76I+S116D、V76I+S116E、V76I+T127V、V76I+T127I、V76I+T127L、V76I+S144P、V76I+A147H、V76I+S167L、V76I+S167I、V76I+S167V、V76I+G175D、V76I+G175E、Q109R+S116D、Q109R+S116E、Q109R+T127V、Q109R+T127I、Q109R+T127L、Q109R+S144P、Q109R+A147H、Q109R+S167L、Q109R+S167I、Q109R+S167V、Q109R+G175D、Q109R+G175E、S116D+T127V、S116D+T127I、S116D+T127L、S116D+S144P、S116D+A147H、S116D+S167L、S116D+S167I、S116D+S167V、S116D+G175D、S116D+G175E、S116E+T127V、S116E+T127I、S116E+T127L、S116E+S144P、S116E+A147H、S116E+S167L、S116E+S167I、S116E+S167V、S116E+G175D、S116E+G175E、T127V+S144P、T127V+A147H、T127V+S167L、T127V+S167I、T127V+S167V、T127V+G175D、T127V+G175E、T127I+S144P、T127I+A147H、T127I+S167L、T127I+S167I、T127I+S167V、T127I+G175D、T127I+G175E、T127L+S144P、T127L+A147H、T127L+S167L、T127L+S167I、T127L+S167V、T127L+G175D、T127L+G175E、S144P+A147H、S144P+S167L、S144P+S167I、S144P+S167V、S144P+G175D、S144P+G175E, A147H+S167L, A147H+S167I, A147H+S167V, A147H+G175D, A147H+G175E, S167L+G175D, S167L+G175E, S167I+G175D, S167I+G175E, S167V+G175D, and S167V+G175E, wherein each substitution provides a DNase variant having increased stability as measured by a half-life modifier HIF, the half-life modifier HIF being compatible with the parental DNase (e.g., containing SEQ ID NO). The variant has at least 1.05, for example 1.08, for example 1.1, for example 1.15, for example 1.2, for example 1.25, for example 1.3, for example 1.4, for example 1.5, for example 1.6, for example 1.7, for example 1.8, for example 1.9, for example 2, for example 3, for example 4, for example 5, or for example at least 10 compared to the DNA enzyme of the polypeptide shown in SEQ ID NO:1, wherein said variant has at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, for example at least 96%, at least 97%, at least 98%, or at least 99%, but less than 100% sequence identity with the polypeptide shown in SEQ ID NO:1, wherein each position corresponds to a position of the polypeptide shown in SEQ ID NO:1 (numbered according to SEQ ID NO:1).

[0718] In some preferred embodiments of the present invention, the DNAse variants of the present invention comprise one or more of the following substitution groups: T1I+T22P, T1I+S39P, T1I+S42G, T1I+S57W, T1I+S59V, T1I+V76L, T1I+S116D, T1I+T127V, T1I+S167L, T1I+G175D, T22P+S42G, T22P+V76L, T22P+S116D, T22P+T127V, T22P+A147H, T22P+G 175D, S25P+S39P, S25P+S42G, S25P+S59V, S25P+V76L, S25P+S116D, S25P+T127V, S25P+S167L, S39P+S42G, S39P+S57 W, S39P+S59V, S39P+V76L, S39P+S116D, S39P+T127V, S39P+S144P, S39P+A147H, S39P+S167L, S42G+S57W, S42G+S59V , S42G+V76L, S42G+S116D, S42G+T127V, S42G+S144P, S42G+A147H, S42G+S167L, S42G+G175D, S57W+S59V, S57W+S116 D. S57W+T127V, S57W+A147H, S59V+V76L, S59V+S116D, S59V+A147H, S59V+S167L, S59V+G175D, V76L+S116D, V76L+T1 27V, V76L+S144P, V76L+A147H, V76L+S167L, V76L+G175D, S116D+T127V, S116D+S144P, S116D+A147H, S116D+S167L, S116D+G175D, T127V+S144P, T127V+A147H, T127V+S167L, T127V+G175D, S144P+A147H, S144P+S167L or S167L+G175D.

[0719] In some preferred embodiments of the present invention, the DNase variants of the present invention comprise one or more of the following substitution groups: T1I+T22P, T1I+S39P, T1I+S42G, T1I+S57W, T1I+S59V, T1I+V76L, T1I+S116D, T1I+T127V, T1I+S167L, T1I+G175D, T22P+S42G, T22P+V76L, T22P+S116D, T22P+T127V, T22P+A147H, T22 P+G175D, S25P+S39P, S25P+S42G, S25P+S59V, S25P+V76L, S25P+S116D, S25P+T127V, S25P+S167L, S39P+S42G, S39 P+S57W, S39P+S59V, S39P+V76L, S39P+S116D, S39P+T127V, S39P+S144P, S39P+A147H, S39P+S167L, S42G+S57W, S42 G+S59V, S42G+V76L, S42G+S116D, S42G+T127V, S42G+S144P, S42G+S167L, S42G+G175D, S57W+S59V, S57W+S116D, S 57W+T127V, S57W+A147H, S59V+V76L, S59V+S116D, S59V+A147H, S59V+S167L, S59V+G175D, V76L+S116D, V76L+T127 V, V76L+S144P, V76L+A147H, V76L+S167L, V76L+G175D, S116D+T127V, S116D+S144P, S116D+A147H, S116D+S167L, S116D+G175D, T127V+S144P, T127V+A147H, T127V+S167L, T127V+G175D, S144P+A147H, S144P+S167L or S167L+G175D.

[0720] In some preferred aspects, the DNase variants of the present invention comprise one or more substitution groups (compared to SEQ ID NO 1), wherein said one or more substitution groups are selected from the group consisting of: T1I+T22P, T1I+S39P, T1I+S42G, T1I+S57W, T1I+S59V, T1I+V76L, T1I+S116D, T1I+T127V, T1I+S167L, T1I+G175D, T22P+S42G, T22P+V76L, T22P+S116D, T22P+T127V, T22P+A147H, T22P+G175D, S25P+S39P, S25P+S42G, S25P+S59V, S25P+V76L, S25P+S116D, S25P+T127V, S25P+S167L, S39P+S42G, S39P+S57W, S39P+S59V, S39P+V76L, S39P+S116D, S39P+T127V, S39P+S144P, S39P+A147H, S39P+S167L, S42G+S57W, S42G+S59V, S42G+V76L, S42G+S116D, S42G+T127V, S42G+S144 P, S42G+A147H, S42G+S167L, S42G+G175D, S57W+S59V, S57W+S116D, S57W+T127V, S57W+A147H, S59V+V76L, S59V+S116D, S59V+ A147H, S59V+S167L, S59V+G175D, V76L+S116D, V76L+T127V, V76L+S144P, V76L+A147H, V76L+S167L, V76L+G175D, S116D+T127 V, S116D+S144P, S116D+A147H, S116D+S167L, S116D+G175D, T127V+S144P, T127V+A147H, T127V+S167L, T127V+G175D, S144P+A147H, S144P+S167L, and S167L+G175D, wherein each substitution provides a DNase variant having increased stability as measured by a half-life modifier HIF, the half-life modifier HIF being compatible with the parental DNase (e.g., containing SEQ ID NO(S116D+S144P), S116D+A147H, T127V+S167L, T127V+G175D), wherein each substitution provides a DNase variant having increased stability as measured by a half-life modifier HIF, the half-life modifier HIF being compatible with the parental DNase (e.g., containing SEQ ID NO(S116D+S144P), S116D+A147H, S116D+S167L, and S167L+G175D). The amino acid sequence of the polypeptide (DNA enzyme) shown in ID NO:1 is at least 1.05, for example 1.08, for example 1.1, for example 1.15, for example 1.2, for example 1.25, for example 1.3, for example 1.4, for example 1.5, for example 1.6, for example 1.7, for example 1.8, for example 1.9, such as 2, 3, 4, 5, or at least 10, wherein the DNase variant has at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, for example at least 96%, at least 97%, at least 98%, or at least 99%, but less than 100% sequence identity with the polypeptide shown in SEQ ID NO:1, wherein each position corresponds to a position of the polypeptide shown in SEQ ID NO:1 (numbered according to SEQ ID NO:1).

[0721] In some preferred aspects, the DNAse variants of the present invention comprise one or more substitution groups (compared to SEQ ID NO 1), wherein said one or more substitution groups are selected from the group consisting of: T1I+T22P, T1I+S39P, T1I+S42G, T1I+S57W, T1I+S59V, T1I+V76L, T1I+S116D, T1I+T127V, T1I+S167L, T1I+G175D, T22P+S42G, T22P+V76L, T22P+S116D, T22P+T127V, T22P+A147H, T22P+G175D, S25P+S39P, S25P+S4 2G, S25P+S59V, S25P+V76L, S25P+S116D, S25P+T127V, S25P+S167L, S39P+S42G, S39P+S57W, S39P+S59V, S39P+V76L, S39P+S1 16D, S39P+T127V, S39P+S144P, S39P+A147H, S39P+S167L, S42G+S57W, S42G+S59V, S42G+V76L, S42G+S116D, S42G+T127V, S42 G+S144P, S42G+S167L, S42G+G175D, S57W+S59V, S57W+S116D, S57W+T127V, S57W+A147H, S59V+V76L, S59V+S116D, S59V+A147 S 116D+S144P, S116D+A147H, S116D+S167L, S116D+G175D, T127V+S144P, T127V+A147H, T127V+S167L, T127V+G175D, S144P+A147H, S144P+S167L, and S167L+G175D, wherein each substitution provides a DNase variant having increased stability as measured by a half-life modifier HIF, the half-life modifier HIF being compatible with the parental DNase (e.g., containing SEQ ID NO(S)). The ratio of the amino acid sequence of the polypeptide (DNA enzyme) shown in IDNO:1 is at least 1.05, for example 1.08, for example 1.1, for example 1.15, for example 1.2, for example 1.25, for example 1.3, for example 1.4, for example 1.5, for example 1.6, for example 1.7, for example 1.8, for example 1.9, such as 2, 3, 4, 5, or at least 10, wherein the DNase variant has at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, for example at least 96%, at least 97%, at least 98%, or at least 99%, but less than 100% sequence identity with the polypeptide shown in SEQ ID NO:1, wherein each position corresponds to a position of the polypeptide shown in SEQ ID NO:1 (numbered according to SEQ ID NO:1).

[0722] In a preferred embodiment, the combination of mutations contains a synergistic effect, i.e., the half-life improvement factor (HIF) of the combination of two mutations is greater than the sum of the half-life improvement factors (HIF) of each mutation alone.

[0723] In some preferred embodiments of the present invention, the DNAse variants of the present invention comprise one or more of the following substitution groups: T1I+S39P, T1I+S42G, T22P+S116D, T22P+T127V, S25P+S39P, S25P+S42G, S25P+S59V, S25P+V76L, S25P+S116D, S25P+T127V, S25P+S167L, S39P+S42G, S39P+S57W, S39P+S59V, S39P+V76L, S39P+S116D, S39P+T127V, S39P+A147H, S39P+S167L, S42G+S57W, S42G+S59V, S 42G+V76L, S42G+S116D, S42G+T127V, S42G+S144P, S42G+A147H, S42G+S167L, S42G+G175D, S57W+S59V, S57W+S116D, S57W+T127V, S59V+V76L, S59V+S116D, S59V+A147H, V76L+S116D, V76L+T127V, S116D+S144P, S116D+A147H, S116D+S167L, T127V+S144P, T127V+A147H, T127V+S167L, T127V+G175D or S144P+A147H.

[0724] In some preferred embodiments of the present invention, the DNAse variants of the present invention comprise one or more of the following substitution groups: T1I+S39P, T1I+S42G, T22P+S116D, T22P+T127V, S25P+S39P, S25P+S42G, S25P+S59V, S25P+V76L, S25P+S116D, S25P+T127V, S25P+S167L, S39P+S42G, S39P+S57W, S39P+S59V, S39P+V76L, S39P+S116D, S39P+T127V, S39P+A147H, S39P+S167L, S42G+S57W, S42G+ S59V, S42G+V76L, S42G+S116D, S42G+T127V, S42G+S144P, S42G+S167L, S42G +G175D, S57W+S59V, S57W+S116D, S57W+T127V, S59V+V76L, S59V+S116D, S59 V+A147H, V76L+S116D, V76L+T127V, S116D+S144P, S116D+A147H, S116D+S167L, T127V+S144P, T127V+A147H, T127V+S167L, T127V+G175D or S144P+A147H.

[0725] In some preferred aspects, the DNAse variants of the present invention comprise one or more substitution groups (compared to SEQ ID NO 1), wherein said one or more substitution groups are selected from the group consisting of: T1I+S39P, T1I+S42G, T22P+S116D, T22P+T127V, S25P+S39P, S25P+S42G, S25P+S59V, S25P+V76L, S25P+S116D, S25P+T127V, S25P+S167L, S3 9P+S42G, S39P+S57W, S39P+S59V, S39P+V76L, S39P+S116D, S39P+T127V, S39P+A147H, S 39P+S167L, S42G+S57W, S42G+S59V, S42G+V76L, S42G+S116D, S42G+T127V, S42G+S144P, S42G+A147H, S42G+S167L, S42G+G175D, S57W+S59V, S57W+S116D, S57W+T127V, S59V+V7 6L, S59V+S116D, S59V+A147H, V76L+S116D, V76L+T127V, S116D+S144P, S116D+A147H, S1 16D+S167L, T127V+S144P, T127V+A147H, T127V+S167L, T127V+G175D, and S144P+A147H, wherein each substitution provides a DNase variant having increased stability as measured by a half-life modifier HIF, the half-life modifier HIF being compatible with the parental DNase (e.g., containing SEQ ID NO(S167L)). The variant has at least 1.05, for example 1.08, for example 1.1, for example 1.15, for example 1.2, for example 1.25, for example 1.3, for example 1.4, for example 1.5, for example 1.6, for example 1.7, for example 1.8, for example 1.9, for example 2, for example 3, for example 4, for example 5 or for example at least 10 compared to the DNA enzyme of the polypeptide shown in SEQ ID NO:1, wherein the variant has at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, for example at least 96%, at least 97%, at least 98%, or at least 99%, but less than 100% sequence identity with the polypeptide shown in SEQ ID NO:1, wherein each position corresponds to a position of the polypeptide shown in SEQ ID NO:1 (numbered according to SEQ ID NO:1).

[0726] In some preferred aspects, the DNAse variants of the present invention comprise one or more substitution groups (compared to SEQ ID NO 1), wherein said one or more substitution groups are selected from the group consisting of: T1I+S39P, T1I+S42G, T22P+S116D, T22P+T127V, S25P+S39P, S25P+S42G, S25P+S59V, S25P+V76L, S25P+S116D, S25P+T127V, S25P+S167L , S39P+S42G, S39P+S57W, S39P+S59V, S39P+V76L, S39P+S116D, S39P+T127V, S39P+A14 7H, S39P+S167L, S42G+S57W, S42G+S59V, S42G+V76L, S42G+S116D, S42G+T127V, S42G+ S144P, S42G+S167L, S42G+G175D, S57W+S59V, S57W+S116D, S57W+T127V, S59V+V76L, S 59V+S116D, S59V+A147H, V76L+S116D, V76L+T127V, S116D+S144P, S116D+A147H, S116 D+S167L, T127V+S144P, T127V+A147H, T127V+S167L, T127V+G175D, and S144P+A147H, wherein each substitution provides a DNase variant having increased stability as measured by a half-life modifier HIF, the half-life modifier HIF being compatible with the parental DNase (e.g., containing SEQ ID NO(S167L)). The variant has at least 1.05, for example 1.08, for example 1.1, for example 1.15, for example 1.2, for example 1.25, for example 1.3, for example 1.4, for example 1.5, for example 1.6, for example 1.7, for example 1.8, for example 1.9, for example 2, for example 3, for example 4, for example 5 or for example at least 10 compared to the DNA enzyme of the polypeptide shown in SEQ ID NO:1, wherein the variant has at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, for example at least 96%, at least 97%, at least 98%, or at least 99%, but less than 100% sequence identity with the polypeptide shown in SEQ ID NO:1, wherein each position corresponds to a position of the polypeptide shown in SEQ ID NO:1 (numbered according to SEQ ID NO:1).

[0727] Variants of the invention preferably comprise the conserved motifs [D / M / L][S / T]GYSR[D / N] (SEQ ID NO:25) or ASXNRSKG (SEQ ID NO:26), which are shared by DNases in the GYS-clade described below. The ASXNRSKG motif corresponds to positions 125 to 133 of SEQ ID NO:1, and the [D / M / L][S / T]GYSR[D / N] motif corresponds to positions 26 to 32 of SEQ ID NO:1. As explained in the "Definition," a clade comprises a group of polypeptides grouped together based on homology characteristics traced back to a common ancestor. Polypeptides forming a group of clades, such as those shown in a phylogenetic tree, often share common properties and are more closely related to other polypeptides not in the clade.

[0728] Therefore, a preferred aspect of the invention relates to a variant of a DNAse parent, wherein the variant comprises one or both of the motif [D / M / L][S / T]GYSR[D / N] (SEQ ID NO:25) or ASXNRSKG (SEQ ID NO:26), and wherein the variant comprises changes at two or more positions selected from a list of the following: positions 1, 13, 22, 25, 27, 39, 42, 57, 59, 76, 109, 116, 127, 144, 147, 167, and 175, wherein each position corresponds to a position of the polypeptide in SEQ ID NO:1, wherein each change provides a DNAse variant having at least one improved property compared to the polypeptide shown in SEQ ID NO:1, wherein the variant corresponds to the polypeptide in SEQ ID NO:1. The polypeptide shown in NO:1 has at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, for example at least 96%, at least 97%, at least 98%, or at least 99%, but less than 100% sequence identity, and wherein each position corresponds to a position of the polypeptide shown in SEQ ID NO:1 (numbered according to SEQ ID NO:1).

[0729] A preferred aspect of the invention relates to a variant of a DNase parent, said variant comprising two or more substitutions (compared to SEQ ID NO:1), said substitutions being selected from the group consisting of: T1I, T1L, T1V, S13Y, T22P, S25P, S27L, S39P, S42G, S42A, S42T, S57W, S57Y, S57F, S59V, S59I, S59L, V76L, V76I, Q109R, S116D, S116E, T127V, T127I, T127L, S144P, A147H, S167L, S167I, S167V, G175D, and G175E, wherein each substitution provides a DNase variant having at least one improved property compared to the polypeptide shown in SEQ ID NO:1, said variant being compared to SEQ ID NO:1. The polypeptide shown in NO:1 has at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, for example at least 96%, at least 97%, at least 98%, or at least 99%, but less than 100% sequence identity, and wherein each position corresponds to a position of the polypeptide shown in SEQ ID NO:1 (numbered according to SEQ ID NO:1).

[0730] A preferred aspect of the invention relates to a variant of a DNAse parent, wherein the variant comprises one or both of the motifs [D / M / L][S / T]GYSR[D / N] (SEQ ID NO:25) or ASXNRSKG (SEQ ID NO:26), and wherein the variant comprises one or more substitution groups (compared to SEQ ID NO:26). NO:1), the substitution group is selected from the group consisting of: T1I+S13Y, T1I+T22P, T1I+S25P, T1I+S27L, T1I+S39P, T1I+S42G, T1I+S42A, T1I+S42T, T1I+S57W, T1I+S57Y, T1I+S57F, T1I+S59V, T1I+S59I, T1I+S59L, T1I+V76L, T1I+V76I, T1I+Q109R, T1I+S116D, T1I+S116E, T1I+T127V, T1I+T127 I, T1I+T127L, T1I+S144P, T1I+A147H, T1I+S167L, T1I+S167I, T1I+S167V, T1I+G175D, T1I+G175E, T1L+S13Y, T1L+T22P, T1L+S25P , T1L+S27L, T1L+S39P, T1L+S42G, T1L+S42A, T1L+S42T, T1L+S57W, T1L+S57Y, T1L+S57F, T1L+S59V, T1L+S59I, T1L+S59L, T1L+V76L, T1L+V76I, T1L+Q109R, T1L+S116D, T1L+S116E, T1L+T127V, T1L+T127I, T1L+T127L, T1L+S144P, T1L+A147H, T1L+S167L, T1L+S167I , T1L+S167V, T1L+G175D, T1L+G175E, T1V+S13Y, T1V+T22P, T1V+S25P, T1V+S27L, T1V+S39P, T1V+S42G, T1V+S42A, T1V+S42T, T1V+S 57W, T1V+S57Y, T1V+S57F, T1V+S59V, T1V+S59I, T1V+S59L, T1V+V76L, T1V+V76I, T1V+Q109R, T1V+S116D, T1V+S116E, T1V+T127V, T 1V+T127I, T1V+T127L, T1V+S144P, T1V+A147H, T1V+S167L, T1V+S167I, T1V+S167V, T1V+G175D, T1V+G175E, S13Y+T22P, S13Y+S25P,S13Y+S27L、S13Y+S39P、S13Y+S42G、S13Y+S42A、S13Y+S42T、S13Y+S57W、S13Y+S57Y、S13Y+S57F、S13Y+S59V、S13Y+S59I、S13Y+S59L、S13Y+V76L、S13Y+V76I、S13Y+Q109R、S13Y+S116D、S13Y+S116E、S13Y+T127V、S13Y+T127I、S13Y+T127L、S13Y+S144P、S13Y+A147H、S13Y+S167L、S13Y+S167I、S13Y+S167V、S13Y+G175D、S13Y+G175E、T22P+S25P、T22P+S27L、T22P+S39P、T22P+S42G、T22P+S42A、T22P+S42T、T22P+S57W、T22P+S57Y、T22P+S57F、T22P+S59V、T22P+S59I、T22P+S59L、T22P+V76L、T22P+V76I、T22P+Q109R、T22P+S116D、T22P+S116E、T22P+T127V、T22P+T127I、T22P+T127L、T22P+S144P、T22P+A147H、T22P+S167L、T22P+S167I、T22P+S167V、T22P+G175D、T22P+G175E、S25P+S27L、S25P+S39P、S25P+S42G、S25P+S42A、S25P+S42T、S25P+S57W、S25P+S57Y、S25P+S57F、S25P+S59V、S25P+S59I、S25P+S59L、S25P+V76L、S25P+V76I、S25P+Q109R、S25P+S116D、S25P+S116E、S25P+T127V、S25P+T127I、S25P+T127L、S25P+S144P、S25P+A147H、S25P+S167L、S25P+S167I、S25P+S167V、S25P+G175D、S25P+G175E、S27L+S39P、S27L+S42G、S27L+S42A、S27L+S42T、S27L+S57W、S27L+S57Y、S27L+S57F、S27L+S59V、S27L+S59I、S27L+S59L、S27L+V76L、S27L+V76I、S27L+Q109R、S27L+S116D、S27L+S116E、S27L+T127V、S27L+T127I、S27L+T127L、S27L+S144P、S27L+A147H、S27L+S167L、S27L+S167I、S27L+S167V、S27L+G175D、S27L+G175E、S39P+S42G、S39P+S42A、S39P+S42T、S39P+S57W、S39P+S57Y、S39P+S57F、S39P+S59V、S39P+S59I、S39P+S59L、S39P+V76L、S39P+V76I、S39P+Q109R、S39P+S116D、S39P+S116E、S39P+T127V、S39P+T127I、S39P+T127L、S39P+S144P、S39P+A147H、S39P+S167L、S39P+S167I、S39P+S167V、S39P+G175D、S39P+G175E、S42G+S57W、S42G+S57Y、S42G+S57F、S42G+S59V、S42G+S59I、S42G+S59L、S42G+V76L、S42G+V76I、S42G+Q109R、S42G+S116D、S42G+S116E、S42G+T127V、S42G+T127I、S42G+T127L、S42G+S144P、S42G+A147H、S42G+S167L、S42G+S167I、S42G+S167V、S42G+G175D、S42G+G175E、S42A+S57W、S42A+S57Y、S42A+S57F、S42A+S59V、S42A+S59I、S42A+S59L、S42A+V76L、S42A+V76I、S42A+Q109R、S42A+S116D、S42A+S116E、S42A+T127V、S42A+T127I、S42A+T127L、S42A+S144P、S42A+A147H、S42A+S167L、S42A+S167I、S42A+S167V、S42A+G175D、S42A+G175E、S42T+S57W、S42T+S57Y、S42T+S57F、S42T+S59V、S42T+S59I、S42T+S59L、S42T+V76L、S42T+V76I、S42T+Q109R、S42T+S116D、S42T+S116E、S42T+T127V、S42T+T127I、S42T+T127L、S42T+S144P、S42T+A147H、S42T+S167L、S42T+S167I、S42T+S167V、S42T+G175D、S42T+G175E、S57W+S59V、S57W+S59I、S57W+S59L、S57W+V76L、S57W+V76I、S57W+Q109R、S57W+S116D、S57W+S116E、S57W+T127V、S57W+T127I、S57W+T127L、S57W+S144P、S57W+A147H、S57W+S167L、S57W+S167I、S57W+S167V、S57W+G175D、S57W+G175E、S57Y+S59V、S57Y+S59I、S57Y+S59L、S57Y+V76L、S57Y+V76I、S57Y+Q109R、S57Y+S116D、S57Y+S116E、S57Y+T127V、S57Y+T127I、S57Y+T127L、S57Y+S144P、S57Y+A147H、S57Y+S167L、S57Y+S167I、S57Y+S167V、S57Y+G175D、S57Y+G175E、S57F+S59V、S57F+S59I、S57F+S59L、S57F+V76L、S57F+V76I、S57F+Q109R、S57F+S116D、S57F+S116E、S57F+T127V、S57F+T127I、S57F+T127L、S57F+S144P、S57F+A147H、S57F+S167L、S57F+S167I、S57F+S167V、S57F+G175D、S57F+G175E、S59V+V76L、S59V+V76I、S59V+Q109R、S59V+S116D、S59V+S116E、S59V+T127V、S59V+T127I、S59V+T127L、S59V+S144P、S59V+A147H、S59V+S167L、S59V+S167I、S59V+S167V、S59V+G175D、S59V+G175E、S59I+V76L、S59I+V76I、S59I+Q109R、S59I+S116D、S59I+S116E、S59I+T127V、S59I+T127I、S59I+T127L、S59I+S144P、S59I+A147H、S59I+S167L、S59I+S167I、S59I+S167V、S59I+G175D、S59I+G175E、S59L+V76L、S59L+V76I、S59L+Q109R、S59L+S116D、S59L+S116E、S59L+T127V、S59L+T127I、S59L+T127L、S59L+S144P、S59L+A147H、S59L+S167L、S59L+S167I、S59L+S167V、S59L+G175D、S59L+G175E、V76L+Q109R、V76L+S116D、V76L+S116E、V76L+T127V、V76L+T127I、V76L+T127L、V76L+S144P、V76L+A147H、V76L+S167L、V76L+S167I、V76L+S167V、V76L+G175D、V76L+G175E、V76I+Q109R、V76I+S116D、V76I+S116E、V76I+T127V、V76I+T127I、V76I+T127L、V76I+S144P、V76I+A147H、V76I+S167L、V76I+S167I、V76I+S167V、V76I+G175D、V76I+G175E、Q109R+S116D、Q109R+S116E、Q109R+T127V、Q109R+T127I、Q109R+T127L、Q109R+S144P、Q109R+A147H、Q109R+S167L、Q109R+S167I、Q109R+S167V、Q109R+G175D、Q109R+G175E、S116D+T127V、S116D+T127I、S116D+T127L、S116D+S144P、S116D+A147H、S116D+S167L、S116D+S167I、S116D+S167V、S116D+G175D、S116D+G175E、S116E+T127V、S116E+T127I、S116E+T127L、S116E+S144P、S116E+A147H、S116E+S167L、S116E+S167I、S116E+S167V、S116E+G175D、S116E+G175E、T127V+S144P、T127V+A147H、T127V+S167L、T127V+S167I、T127V+S167V、T127V+G175D、T127V+G175E、T127I+S144P、T127I+A147H、T127I+S167L、T127I+S167I、T127I+S167V、T127I+G175D、T127I+G175E、T127L+S144P、T127L+A147H、T127L+S167L、T127L+S167I、T127L+S167V、T127L+G175D, T127L+G175E, S144P+A147H, S144P+S167L, S144P+S167I, S144P+S167V, S144P+G175D, S144P+G175E, A147H+S167L, A147H+S167I, A147H+S167V, A147H+G175D, A147H+G175E, S167L+G175D, S167L+G175E, S167I+G175D, S167I+G175E, S167V+G175D, and S167V+G175E, wherein each substitution provides a DNase variant, said DNase variant being associated with SEQ ID Compared to the polypeptide shown in SEQ ID NO:1, the variant has at least one improved property, wherein the variant has at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, for example at least 96%, at least 97%, at least 98%, or at least 99%, but less than 100% sequence identity with the polypeptide shown in SEQ ID NO:1, and wherein each position corresponds to a position of the polypeptide shown in SEQ ID NO:1 (numbered according to SEQ ID NO:1).

[0731] A preferred aspect of the invention relates to a variant of a DNase parent, wherein the variant comprises one or both of the motif [D / M / L][S / T]GYSR[D / N] (SEQ ID NO:25) or ASXNRSKG (SEQ ID NO:26), and wherein the variant comprises changes at two or more positions selected from a list consisting of: 1, 13, 22, 25, 27, 39, 42, 57, 59, 76, 109, 116, 127, 144, 147, 167, and 175, wherein each position corresponds to a position of the polypeptide in SEQ ID NO:1, wherein each substitution provides a DNase variant having increased stability as measured by a half-life modifier HIF, the half-life modifier HIF being associated with the parent DNase (e.g., comprising a SEQ ID NO:26). The variant has at least 1.05, for example 1.08, for example 1.1, for example 1.15, for example 1.2, for example 1.25, for example 1.3, for example 1.4, for example 1.5, for example 1.6, for example 1.7, for example 1.8, for example 1.9, for example 2, for example 3, for example 4, for example 5 or for example at least 10 compared to the DNA enzyme of the polypeptide shown in SEQ ID NO:1, wherein the variant has at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, for example at least 96%, at least 97%, at least 98%, or at least 99%, but less than 100% sequence identity with the polypeptide shown in SEQ ID NO:1.

[0732] A preferred aspect of the invention relates to a variant of a DNAse parent, said variant comprising one or both of the motif [D / M / L][S / T]GYSR[D / N] (SEQ ID NO:25) or ASXNRSKG (SEQ ID NO:26), and said variant comprising two or more substitutions selected from the group consisting of: T1V, S13Y, S27L, S39P, S42A, S42T, S57W, S57F, S59V, S59I, S59L, S116E, T127I, T127L, S144P, S167L, S167I, S167V, G175D, and G175E, said position corresponding to the position in SEQ ID NO:1 (according to SEQ ID NO:26). (Numbered NO:1), wherein each substitution provides a DNase variant having an increase in stability as measured by a half-life improvement factor HIF, the half-life improvement factor HIF being at least 1.05, e.g., 1.08, e.g., 1.1, e.g., 1.15, e.g., 1.2, e.g., 1.25, e.g., 1.3, e.g., 1.4, e.g., 1.5, e.g., 1.6, e.g., 1.7, e.g., 1.8, e.g., 1.9, e.g., 2, e.g., 3, e.g., 4, e.g., 5, or e.g., at least 10 compared to a parental DNase (e.g., a DNase comprising a polypeptide having the amino acid sequence shown in SEQ ID NO:1), wherein the variant has at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, e.g., at least 96%, at least 97%, at least 98%, or at least 99%, but less than 100% sequence identity with the polypeptide shown in SEQ ID NO:1.

[0733] A preferred aspect of the invention relates to variants of the DNAse parent, wherein said variants comprise two or more substitutions (as per SEQ ID). Compared to NO:1), wherein the substitution is selected from the group consisting of: T1I, T1L, T1V, S13Y, T22P, S25P, S27L, S39P, S42G, S42A, S42T, S57W, S57Y, S57F, S59V, S59I, S59L, V76L, V76I, Q109R, S116D, S116E, T127V, T127I, T127L, S144P, A147H, S167L, S167I, S167V, G175D, and G175E, wherein each substitution provides a DNase variant having increased stability as measured by a half-life improvement factor HIF, the half-life improvement factor HIF being compatible with the parental DNase (e.g., containing SEQ ID NO:1). The variant has at least 1.05, for example 1.08, for example 1.1, for example 1.15, for example 1.2, for example 1.25, for example 1.3, for example 1.4, for example 1.5, for example 1.6, for example 1.7, for example 1.8, for example 1.9, for example 2, for example 3, for example 4, for example 5 or for example at least 10 compared to the DNA enzyme of the polypeptide shown in SEQ ID NO:1, wherein the variant has at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, for example at least 96%, at least 97%, at least 98%, or at least 99%, but less than 100% sequence identity with the polypeptide shown in SEQ ID NO:1, wherein each position corresponds to a position of the polypeptide shown in SEQ ID NO:1 (numbered according to SEQ ID NO:1).

[0734] A preferred aspect of the invention relates to a variant of a DNAse parent, wherein the variant comprises one or both of the motifs [D / M / L][S / T]GYSR[D / N] (SEQ ID NO:25) or ASXNRSKG (SEQ ID NO:26), and wherein the variant comprises one or more substitution groups (as in SEQ ID NO:26). Compared to NO:1, the substitution group is selected from the group consisting of: T1I+S13Y, T1I+T22P, T1I+S25P, T1I+S27L, T1I+S39P, T1I+S42G, T1I+S42A, T1I+S42T, T1I+S57W, T1I+S57Y, T1I+S57F, T1I+S59V, T1I+S59I, T1I+S59L, T1I+V76L, T1I+V76I, T1I+Q109R, T1I+S116D, T1I+S116E, T1I+T127V, T1I+T12 7I, T1I+T127L, T1I+S144P, T1I+A147H, T1I+S167L, T1I+S167I, T1I+S167V, T1I+G175D, T1I+G175E, T1L+S13Y, T1L+T22P, T1L+S25 P, T1L+S27L, T1L+S39P, T1L+S42G, T1L+S42A, T1L+S42T, T1L+S57W, T1L+S57Y, T1L+S57F, T1L+S59V, T1L+S59I, T1L+S59L, T1L+V76L , T1L+V76I, T1L+Q109R, T1L+S116D, T1L+S116E, T1L+T127V, T1L+T127I, T1L+T127L, T1L+S144P, T1L+A147H, T1L+S167L, T1L+S167 I. T1L+S167V, T1L+G175D, T1L+G175E, T1V+S13Y, T1V+T22P, T1V+S25P, T1V+S27L, T1V+S39P, T1V+S42G, T1V+S42A, T1V+S42T, T1V+S 57W, T1V+S57Y, T1V+S57F, T1V+S59V, T1V+S59I, T1V+S59L, T1V+V76L, T1V+V76I, T1V+Q109R, T1V+S116D, T1V+S116E, T1V+T127V, T 1V+T127I, T1V+T127L, T1V+S144P, T1V+A147H, T1V+S167L, T1V+S167I, T1V+S167V, T1V+G175D, T1V+G175E, S13Y+T22P, S13Y+S25P,S13Y+S27L、S13Y+S39P、S13Y+S42G、S13Y+S42A、S13Y+S42T、S13Y+S57W、S13Y+S57Y、S13Y+S57F、S13Y+S59V、S13Y+S59I、S13Y+S59L、S13Y+V76L、S13Y+V76I、S13Y+Q109R、S13Y+S116D、S13Y+S116E、S13Y+T127V、S13Y+T127I、S13Y+T127L、S13Y+S144P、S13Y+A147H、S13Y+S167L、S13Y+S167I、S13Y+S167V、S13Y+G175D、S13Y+G175E、T22P+S25P、T22P+S27L、T22P+S39P、T22P+S42G、T22P+S42A、T22P+S42T、T22P+S57W、T22P+S57Y、T22P+S57F、T22P+S59V、T22P+S59I、T22P+S59L、T22P+V76L、T22P+V76I、T22P+Q109R、T22P+S116D、T22P+S116E、T22P+T127V、T22P+T127I、T22P+T127L、T22P+S144P、T22P+A147H、T22P+S167L、T22P+S167I、T22P+S167V、T22P+G175D、T22P+G175E、S25P+S27L、S25P+S39P、S25P+S42G、S25P+S42A、S25P+S42T、S25P+S57W、S25P+S57Y、S25P+S57F、S25P+S59V、S25P+S59I、S25P+S59L、S25P+V76L、S25P+V76I、S25P+Q109R、S25P+S116D、S25P+S116E、S25P+T127V、S25P+T127I、S25P+T127L、S25P+S144P、S25P+A147H、S25P+S167L、S25P+S167I、S25P+S167V、S25P+G175D、S25P+G175E、S27L+S39P、S27L+S42G、S27L+S42A、S27L+S42T、S27L+S57W、S27L+S57Y、S27L+S57F、S27L+S59V、S27L+S59I、S27L+S59L、S27L+V76L、S27L+V76I、S27L+Q109R、S27L+S116D、S27L+S116E、S27L+T127V、S27L+T127I、S27L+T127L、S27L+S144P、S27L+A147H、S27L+S167L、S27L+S167I、S27L+S167V、S27L+G175D、S27L+G175E、S39P+S42G、S39P+S42A、S39P+S42T、S39P+S57W、S39P+S57Y、S39P+S57F、S39P+S59V、S39P+S59I、S39P+S59L、S39P+V76L、S39P+V76I、S39P+Q109R、S39P+S116D、S39P+S116E、S39P+T127V、S39P+T127I、S39P+T127L、S39P+S144P、S39P+A147H、S39P+S167L、S39P+S167I、S39P+S167V、S39P+G175D、S39P+G175E、S42G+S57W、S42G+S57Y、S42G+S57F、S42G+S59V、S42G+S59I、S42G+S59L、S42G+V76L、S42G+V76I、S42G+Q109R、S42G+S116D、S42G+S116E、S42G+T127V、S42G+T127I、S42G+T127L、S42G+S144P、S42G+A147H、S42G+S167L、S42G+S167I、S42G+S167V、S42G+G175D、S42G+G175E、S42A+S57W、S42A+S57Y、S42A+S57F、S42A+S59V、S42A+S59I、S42A+S59L、S42A+V76L、S42A+V76I、S42A+Q109R、S42A+S116D、S42A+S116E、S42A+T127V、S42A+T127I、S42A+T127L、S42A+S144P、S42A+A147H、S42A+S167L、S42A+S167I、S42A+S167V、S42A+G175D、S42A+G175E、S42T+S57W、S42T+S57Y、S42T+S57F、S42T+S59V、S42T+S59I、S42T+S59L、S42T+V76L、S42T+V76I、S42T+Q109R、S42T+S116D、S42T+S116E、S42T+T127V、S42T+T127I、S42T+T127L、S42T+S144P、S42T+A147H、S42T+S167L、S42T+S167I、S42T+S167V、S42T+G175D、S42T+G175E、S57W+S59V、S57W+S59I、S57W+S59L、S57W+V76L、S57W+V76I、S57W+Q109R、S57W+S116D、S57W+S116E、S57W+T127V、S57W+T127I、S57W+T127L、S57W+S144P、S57W+A147H、S57W+S167L、S57W+S167I、S57W+S167V、S57W+G175D、S57W+G175E、S57Y+S59V、S57Y+S59I、S57Y+S59L、S57Y+V76L、S57Y+V76I、S57Y+Q109R、S57Y+S116D、S57Y+S116E、S57Y+T127V、S57Y+T127I、S57Y+T127L、S57Y+S144P、S57Y+A147H、S57Y+S167L、S57Y+S167I、S57Y+S167V、S57Y+G175D、S57Y+G175E、S57F+S59V、S57F+S59I、S57F+S59L、S57F+V76L、S57F+V76I、S57F+Q109R、S57F+S116D、S57F+S116E、S57F+T127V、S57F+T127I、S57F+T127L、S57F+S144P、S57F+A147H、S57F+S167L、S57F+S167I、S57F+S167V、S57F+G175D、S57F+G175E、S59V+V76L、S59V+V76I、S59V+Q109R、S59V+S116D、S59V+S116E、S59V+T127V、S59V+T127I、S59V+T127L、S59V+S144P、S59V+A147H、S59V+S167L、S59V+S167I、S59V+S167V、S59V+G175D、S59V+G175E、S59I+V76L、S59I+V76I、S59I+Q109R、S59I+S116D、S59I+S116E、S59I+T127V、S59I+T127I、S59I+T127L、S59I+S144P、S59I+A147H、S59I+S167L、S59I+S167I、S59I+S167V、S59I+G175D、S59I+G175E、S59L+V76L、S59L+V76I、S59L+Q109R、S59L+S116D、S59L+S116E、S59L+T127V、S59L+T127I、S59L+T127L、S59L+S144P、S59L+A147H、S59L+S167L、S59L+S167I、S59L+S167V、S59L+G175D、S59L+G175E、V76L+Q109R、V76L+S116D、V76L+S116E、V76L+T127V、V76L+T127I、V76L+T127L、V76L+S144P、V76L+A147H、V76L+S167L、V76L+S167I、V76L+S167V、V76L+G175D、V76L+G175E、V76I+Q109R、V76I+S116D、V76I+S116E、V76I+T127V、V76I+T127I、V76I+T127L、V76I+S144P、V76I+A147H、V76I+S167L、V76I+S167I、V76I+S167V、V76I+G175D、V76I+G175E、Q109R+S116D、Q109R+S116E、Q109R+T127V、Q109R+T127I、Q109R+T127L、Q109R+S144P、Q109R+A147H、Q109R+S167L、Q109R+S167I、Q109R+S167V、Q109R+G175D、Q109R+G175E、S116D+T127V、S116D+T127I、S116D+T127L、S116D+S144P、S116D+A147H、S116D+S167L、S116D+S167I、S116D+S167V、S116D+G175D、S116D+G175E、S116E+T127V、S116E+T127I、S116E+T127L、S116E+S144P、S116E+A147H、S116E+S167L、S116E+S167I、S116E+S167V、S116E+G175D、S116E+G175E、T127V+S144P、T127V+A147H、T127V+S167L、T127V+S167I、T127V+S167V、T127V+G175D、T127V+G175E、T127I+S144P、T127I+A147H、T127I+S167L、T127I+S167I、T127I+S167V、T127I+G175D、T127I+G175E、T127L+S144P、T127L+A147H、T127L+S167L、T127L+S167I、T127L+S167V、T127L+G175D, T127L+G175E, S144P+A147H, S144P+S167L, S144P+S167I, S144P+S167V, S14 4P+G175D, S144P+G175E, A147H+S167L, A147H+S167I, A147H+S167V, A147H+G175D, A147H+G 175E, S167L+G175D, S167L+G175E, S167I+G175D, S167I+G175E, S167V+G175D, and S167V+G175E, wherein each substitution provides a DNase variant having increased stability as measured by a half-life modifier HIF, the half-life modifier HIF being compatible with the parental DNase (e.g., containing SEQ ID NO(S)). The variant has at least 1.05, for example 1.08, for example 1.1, for example 1.15, for example 1.2, for example 1.25, for example 1.3, for example 1.4, for example 1.5, for example 1.6, for example 1.7, for example 1.8, for example 1.9, for example 2, for example 3, for example 4, for example 5, or for example at least 10 compared to the DNA enzyme of the polypeptide shown in SEQ ID NO:1, wherein said variant has at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, for example at least 96%, at least 97%, at least 98%, or at least 99%, but less than 100% sequence identity with the polypeptide shown in SEQ ID NO:1, wherein each position corresponds to a position of the polypeptide shown in SEQ ID NO:1 (numbered according to SEQ ID NO:1).

[0735] Preferably, the DNase variants of the present invention comprise one or more of the following substitution groups: T1I+T22P, T1I+S39P, T1I+S42G, T1I+S57W, T1I+S59V, T1I+V76L, T1I+S116D, T1I+T127V, T1I+S167L, T1I+G175D, T22P+S42G, T22P+V76L, T22P+S116D, T22P+T127V, T22P+A147H, T22P+G175D, S 25P+S39P, S25P+S42G, S25P+S59V, S25P+V76L, S25P+S116D, S25P+T127V, S25P+S167L, S39P+S42G, S39P+S57W, S39 P+S59V, S39P+V76L, S39P+S116D, S39P+T127V, S39P+S144P, S39P+A147H, S39P+S167L, S42G+S57W, S42G+S59V, S42 G+V76L, S42G+S116D, S42G+T127V, S42G+S144P, S42G+A147H, S42G+S167L, S42G+G175D, S57W+S59V, S57W+S116D, S 57W+T127V, S57W+A147H, S59V+V76L, S59V+S116D, S59V+A147H, S59V+S167L, S59V+G175D, V76L+S116D, V76L+T127 V, V76L+S144P, V76L+A147H, V76L+S167L, V76L+G175D, S116D+T127V, S116D+S144P, S116D+A147H, S116D+S167L, S116D+G175D, T127V+S144P, T127V+A147H, T127V+S167L, T127V+G175D, S144P+A147H, S144P+S167L or S167L+G175D.

[0736] Preferably, the DNase variants of the present invention comprise one or more of the following substitution groups: T1I+T22P, T1I+S39P, T1I+S42G, T1I+S57W, T1I+S59V, T1I+V76L, T1I+S116D, T1I+T127V, T1I+S167L, T1I+G175D, T22P+S42G, T22P+V76L, T22P+S116D, T22P+T127V, T22P+A147H, T22P+G175D , S25P+S39P, S25P+S42G, S25P+S59V, S25P+V76L, S25P+S116D, S25P+T127V, S25P+S167L, S39P+S42G, S39P+S57W , S39P+S59V, S39P+V76L, S39P+S116D, S39P+T127V, S39P+S144P, S39P+A147H, S39P+S167L, S42G+S57W, S42G+S59 V, S42G+V76L, S42G+S116D, S42G+T127V, S42G+S144P, S42G+S167L, S42G+G175D, S57W+S59V, S57W+S116D, S57W+ T127V, S57W+A147H, S59V+V76L, S59V+S116D, S59V+A147H, S59V+S167L, S59V+G175D, V76L+S116D, V76L+T127V, V 76L+S144P, V76L+A147H, V76L+S167L, V76L+G175D, S116D+T127V, S116D+S144P, S116D+A147H, S116D+S167L, S116D+G175D, T127V+S144P, T127V+A147H, T127V+S167L, T127V+G175D, S144P+A147H, S144P+S167L or S167L+G175D.

[0737] A preferred aspect of the invention relates to a variant of a DNAse parent, wherein the variant comprises one or both of the motifs [D / M / L][S / T]GYSR[D / N] (SEQ ID NO:25) or ASXNRSKG (SEQ ID NO:26), and wherein the variant comprises one or more substitution groups (as in SEQ ID NO:26). Compared to NO:1), the substitution group is selected from the group consisting of: T1I+T22P, T1I+S39P, T1I+S42G, T1I+S57W, T1I+S59V, T1I+V76L, T1I+S116D, T1I+T127V, T1I+S167L, T1I+G175D, T22P+S42G, T22P+V76L, T22P+S116D, T22P+T127V, T22P+A147H, T22P+G175D, S25P+S39P, S25P+S42G, S25 P+S59V, S25P+V76L, S25P+S116D, S25P+T127V, S25P+S167L, S39P+S42G, S39P+S57W, S39P+S59V, S39P+V76L, S39P+S116D, S39 P+T127V, S39P+S144P, S39P+A147H, S39P+S167L, S42G+S57W, S42G+S59V, S42G+V76L, S42G+S116D, S42G+T127V, S42G+S144P, S42G+A147H, S42G+S167L, S42G+G175D, S57W+S59V, S57W+S116D, S57W+T127V, S57W+A147H, S59V+V76L, S59V+S116D, S59V+A 147H, S59V+S167L, S59V+G175D, V76L+S116D, V76L+T127V, V76L+S144P, V76L+A147H, V76L+S167L, V76L+G175D, S116D+T127V The substitutions are S116D+S144P, S116D+A147H, S116D+S167L, S116D+G175D, T127V+S144P, T127V+A147H, T127V+S167L, T127V+G175D, S144P+A147H, S144P+S167L, and S167L+G175D, wherein each substitution provides a DNase variant having an increased stability as measured by a half-life improvement factor HIF, which is at least 1.05, for example 1, compared to a parental DNase (e.g., a DNase comprising a polypeptide having the amino acid sequence shown in SEQ ID NO:1).08, for example 1.1, for example 1.15, for example 1.2, for example 1.25, for example 1.3, for example 1.4, for example 1.5, for example 1.6, for example 1.7, for example 1.8, for example 1.9, for example 2, for example 3, for example 4, for example 5, or for example at least 10, wherein said variants have at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, for example at least 96%, at least 97%, at least 98%, or at least 99%, but less than 100% sequence identity with the polypeptide shown in SEQ ID NO:1, wherein each position corresponds to a position of the polypeptide shown in SEQ ID NO:1 (numbered according to SEQ ID NO:1).

[0738] A preferred aspect of the invention relates to a variant of a DNAse parent, wherein the variant comprises one or both of the motifs [D / M / L][S / T]GYSR[D / N] (SEQ ID NO:25) or ASXNRSKG (SEQ ID NO:26), and wherein the variant comprises one or more substitution groups (as in SEQ ID NO:26). Compared to NO:1, the substitution group is selected from the group consisting of: T1I+T22P, T1I+S39P, T1I+S42G, T1I+S57W, T1I+S59V, T1I+V76L, T1I+S116D, T1I+T127V, T1I+S167L, T1I+G175D, T22P+S42G, T22P+V76L, T22P+S116D, T22P+T127V, T22P+A147H, T22P+G175D, S25P+S39P, S25P+S42G. S25P+S59V, S25P+V76L, S25P+S116D, S25P+T127V, S25P+S167L, S39P+S42G, S39P+S57W, S39P+S59V, S39P+V76L, S39P+S116 D. S39P+T127V, S39P+S144P, S39P+A147H, S39P+S167L, S42G+S57W, S42G+S59V, S42G+V76L, S42G+S116D, S42G+T127V, S42G+ S144P, S42G+S167L, S42G+G175D, S57W+S59V, S57W+S116D, S57W+T127V, S57W+A147H, S59V+V76L, S59V+S116D, S59V+A147H , S59V+S167L, S59V+G175D, V76L+S116D, V76L+T127V, V76L+S144P, V76L+A147H, V76L+S167L, V76L+G175D, S116D+T127V, S1 16D+S144P, S116D+A147H, S116D+S167L, S116D+G175D, T127V+S144P, T127V+A147H, T127V+S167L, T127V+G175D, S144P+A147H, S144P+S167L, and S167L+G175D, wherein each substitution provides a DNase variant having an increased stability as measured by a half-life improvement factor HIF, which is at least 1.05, for example 1.08, for example 1.1, for example 1, compared to a parental DNase (e.g., a DNase comprising a polypeptide having the amino acid sequence shown in SEQ ID NO:1).15, for example 1.2, for example 1.25, for example 1.3, for example 1.4, for example 1.5, for example 1.6, for example 1.7, for example 1.8, for example 1.9, for example 2, for example 3, for example 4, for example 5, or for example at least 10, wherein said variants have at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, for example at least 96%, at least 97%, at least 98%, or at least 99%, but less than 100% sequence identity with the polypeptide shown in SEQ ID NO:1, wherein each position corresponds to a position of the polypeptide shown in SEQ ID NO:1 (numbered according to SEQ ID NO:1).

[0739] Preferably, the DNAse variants of the present invention comprise one or more of the following substitution groups: T1I+S39P, T1I+S42G, T22P+S116D, T22P+T127V, S25P+S39P, S25P+S42G, S25P+S59V, S25P+V76L, S25P+S116D, S25P+T127V, S25P+S167L, S39P+S42G, S39P+S57W, S39P+S59V, S39P+V76L, S39P+S116D, S39P+T127V, S39P+A147H, S39P+S167L, S42G+S57W, S42G+S59V, S42G+ V76L, S42G+S116D, S42G+T127V, S42G+S144P, S42G+A147H, S42G+S167L, S42 G+G175D, S57W+S59V, S57W+S116D, S57W+T127V, S59V+V76L, S59V+S116D, S5 9V+A147H, V76L+S116D, V76L+T127V, S116D+S144P, S116D+A147H, S116D+S1 67L, T127V+S144P, T127V+A147H, T127V+S167L, T127V+G175D or S144P+A147H.

[0740] Preferably, the DNase variants of the present invention comprise one or more of the following substitution groups: T1I+S39P, T1I+S42G, T22P+S116D, T22P+T127V, S25P+S39P, S25P+S42G, S25P+S59V, S25P+V76L, S25P+S116D, S25P+T127V, S25P+S167L, S39P+S42G, S39P+S57W, S39P+S59V, S39P+V76L, S39P+S116D, S39P+T127V, S39P+A147H, S39P+S167L, S42G+S57W, S42G+S59V. S42G+V76L, S42G+S116D, S42G+T127V, S42G+S144P, S42G+S167L, S42G+G1 75D, S57W+S59V, S57W+S116D, S57W+T127V, S59V+V76L, S59V+S116D, S59V+ A147H, V76L+S116D, V76L+T127V, S116D+S144P, S116D+A147H, S116D+S167L, T127V+S144P, T127V+A147H, T127V+S167L, T127V+G175D or S144P+A147H.

[0741] A preferred aspect of the invention relates to a variant of a DNAse parent, wherein the variant comprises one or both of the motif [D / M / L][S / T]GYSR[D / N] (SEQ ID NO:25) or ASXNRSKG (SEQ ID NO:26), and wherein the variant comprises one or more substitution groups (compared to SEQ ID NO:1) selected from the group consisting of: T1I+S39P, T1I+S42G, T22P+S116D, T22P+T127V, S25P+S39P, S25P+S42G, S25P+S59V, S25P+V76L, S25P+S116D, S25P+T127V, S25P+S167L, S39P+ S42G, S39P+S57W, S39P+S59V, S39P+V76L, S39P+S116D, S39P+T127V, S39P+A147H, S39P +S167L, S42G+S57W, S42G+S59V, S42G+V76L, S42G+S116D, S42G+T127V, S42G+S144P, S4 2G+A147H, S42G+S167L, S42G+G175D, S57W+S59V, S57W+S116D, S57W+T127V, S59V+V76L , S59V+S116D, S59V+A147H, V76L+S116D, V76L+T127V, S116D+S144P, S116D+A147H, S11 6D+S167L, T127V+S144P, T127V+A147H, T127V+S167L, T127V+G175D, and S144P+A147H, wherein each substitution provides a DNase variant having increased stability as measured by a half-life modifier HIF, the half-life modifier HIF being compatible with the parental DNase (e.g., containing SEQ ID NO(S167L)). The variant has at least 1.05, for example 1.08, for example 1.1, for example 1.15, for example 1.2, for example 1.25, for example 1.3, for example 1.4, for example 1.5, for example 1.6, for example 1.7, for example 1.8, for example 1.9, for example 2, for example 3, for example 4, for example 5 or for example at least 10 compared to the DNA enzyme of the polypeptide shown in SEQ ID NO:1, wherein the variant has at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, for example at least 96%, at least 97%, at least 98%, or at least 99%, but less than 100% sequence identity with the polypeptide shown in SEQ ID NO:1, wherein each position corresponds to a position of the polypeptide shown in SEQ ID NO:1 (numbered according to SEQ ID NO:1).

[0742] A preferred aspect of the invention relates to variants of a DNAse parent, wherein the variant comprises one or both of the motif [D / M / L][S / T]GYSR[D / N] (SEQ ID NO:25) or ASXNRSKG (SEQ ID NO:26), and wherein the variant comprises one or more substitution groups (compared to SEQ ID NO:1) selected from the group consisting of: T1I+S39P, T1I+S42G, T22P+S116D, T22P+T127V, S25P+S39P, S25P+S42G, S25P+S59V, S25P+V76L, S25P+S116D, S25P+T127V, S25P+S167L. S39P+S42G, S39P+S57W, S39P+S59V, S39P+V76L, S39P+S116D, S39P+T127V, S39P+A147 H. S39P+S167L, S42G+S57W, S42G+S59V, S42G+V76L, S42G+S116D, S42G+T127V, S42G+S 144P, S42G+S167L, S42G+G175D, S57W+S59V, S57W+S116D, S57W+T127V, S59V+V76L, S5 9V+S116D, S59V+A147H, V76L+S116D, V76L+T127V, S116D+S144P, S116D+A147H, S116D +S167L, T127V+S144P, T127V+A147H, T127V+S167L, T127V+G175D, and S144P+A147H, wherein each substitution provides a DNase variant having increased stability as measured by a half-life modifier HIF, the half-life modifier HIF being compatible with the parental DNase (e.g., containing SEQ ID NO(S167L), T127V+S144P, T127V+A147H), and S127V+S167L, T127V+G175D, and S144P+A147H). The variant has at least 1.05, for example 1.08, for example 1.1, for example 1.15, for example 1.2, for example 1.25, for example 1.3, for example 1.4, for example 1.5, for example 1.6, for example 1.7, for example 1.8, for example 1.9, for example 2, for example 3, for example 4, for example 5 or for example at least 10 compared to the DNA enzyme of the polypeptide shown in SEQ ID NO:1, wherein the variant has at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, for example at least 96%, at least 97%, at least 98%, or at least 99%, but less than 100% sequence identity with the polypeptide shown in SEQ ID NO:1, wherein each position corresponds to a position of the polypeptide shown in SEQ ID NO:1 (numbered according to SEQ ID NO:1).

[0743] Variants according to the invention may include additional mutations to the mutations listed above. These additional modifications should preferably not significantly alter the improved properties of the variant DNase.

[0744] Adding any one of the mutations T1I, T1L, T1V, S13Y, T22P, S25P, S27L, S39P, S42G, S42A, S42T, S57W, S57Y, S57F, S59V, S59I, S59L, V76L, V76I, Q109R, S116D, S116E, T127V, T127I, T127L, S144P, A147H, S167L, S167I, S167V, G175D, or G175E has been shown to improve stability. Adding more than one mutation to the parent molecule or starting molecule (e.g., SEQ ID NO:1 or a polypeptide having at least 80% sequence identity with it) has also been shown to provide additional, even synergistic, stability, see, for example, Examples 2 through 6. In a preferred embodiment, a variant of the invention comprises at least three substitutions selected from the group consisting of: T1I, S13Y, T22P, S25P, S27L, S39P, S42G, S57W, S59V, V76L, Q109R, S116D, T127V, S144P, A147H, S167L, and G175D.

[0745] In one embodiment, the DNAse variant is selected from the group consisting of:

[0746] T1I+S13Y+T22P、T1I+S13Y+S25P、T1I+S13Y+S27L、T1I+S13Y+S39P、T1I+S13Y+S42G、T1I+S13Y+S57W、T1I+S13Y+S59V、T1I+S13Y+V76L、T1I+S13Y+Q109R、T1I+S13Y+S116D、T1I+S13Y+T127V、T1I+S13Y+S144P、T1I+S13Y+A147H、T1I+S13Y+S167L、T1I+S13Y+G175D、T1I+T22P+S25P、T1I+T22P+S27L、T1I+T22P+S39P、T1I+T22P+S42G、T1I+T22P+S57W、T1I+T22P+S59V、T1I+T22P+V76L、T1I+T22P+Q109R、T1I+T22P+S116D、T1I+T22P+T127V、T1I+T22P+S144P、T1I+T22P+A147H、T1I+T22P+S167L、T1I+T22P+G175D、T1I+S25P+S27L、T1I+S25P+S39P、T1I+S25P+S42G、T1I+S25P+S57W、T1I+S25P+S59V、T1I+S25P+V76L、T1I+S25P+Q109R、T1I+S25P+S116D、T1I+S25P+T127V、T1I+S25P+S144P、T1I+S25P+A147H、T1I+S25P+S167L、T1I+S25P+G175D、T1I+S27L+S39P、T1I+S27L+S42G、T1I+S27L+S57W、T1I+S27L+S59V、T1I+S27L+V76L、T1I+S27L+Q109R、T1I+S27L+S116D、T1I+S27L+T127V、T1I+S27L+S144P、T1I+S27L+A147H、T1I+S27L+S167L、T1I+S27L+G175D、T1I+S39P+S42G、T1I+S39P+S57W、T1I+S39P+S59V、T1I+S39P+V76L、T1I+S39P+Q109R、T1I+S39P+S116D、T1I+S39P+T127V、T1I+S39P+S144P、T1I+S39P+A147H、T1I+S39P+S167L、T1I+S39P+G175D、T1I+S42G+S57W、T1I+S42G+S59V、T1I+S42G+V76L、T1I+S42G+Q109R、T1I+S42G+S116D、T1I+S42G+T127V、T1I+S42G+S144P、T1I+S42G+A147H、T1I+S42G+S167L、T1I+S42G+G175D、T1I+S57W+S59V、T1I+S57W+V76L、T1I+S57W+Q109R、T1I+S57W+S116D、T1I+S57W+T127V、T1I+S57W+S144P、T1I+S57W+A147H、T1I+S57W+S167L、T1I+S57W+G175D、T1I+S59V+V76L、T1I+S59V+Q109R、T1I+S59V+S116D、T1I+S59V+T127V、T1I+S59V+S144P、T1I+S59V+A147H、T1I+S59V+S167L、T1I+S59V+G175D、T1I+V76L+Q109R、T1I+V76L+S116D、T1I+V76L+T127V、T1I+V76L+S144P、T1I+V76L+A147H、T1I+V76L+S167L、T1I+V76L+G175D、T1I+Q109R+S116D、T1I+Q109R+T127V、T1I+Q109R+S144P、T1I+Q109R+A147H、T1I+Q109R+S167L、T1I+Q109R+G175D、T1I+S116D+T127V、T1I+S116D+S144P、T1I+S116D+A147H、T1I+S116D+S167L、T1I+S116D+G175D、T1I+T127V+S144P、T1I+T127V+A147H、T1I+T127V+S167L、T1I+T127V+G175D、T1I+S144P+A147H、T1I+S144P+S167L、T1I+S144P+G175D、T1I+A147H+S167L、T1I+A147H+G175D、T1I+S167L+G175D、S13Y+T22P+S25P、S13Y+T22P+S27L、S13Y+T22P+S39P、S13Y+T22P+S42G、S13Y+T22P+S57W、S13Y+T22P+S59V、S13Y+T22P+V76L、S13Y+T22P+Q109R、S13Y+T22P+S116D、S13Y+T22P+T127V、S13Y+T22P+S144P、S13Y+T22P+A147H、S13Y+T22P+S167L、S13Y+T22P+G175D、S13Y+S25P+S27L、S13Y+S25P+S39P、S13Y+S25P+S42G、S13Y+S25P+S57W、S13Y+S25P+S59V、S13Y+S25P+V76L、S13Y+S25P+Q109R、S13Y+S25P+S116D、S13Y+S25P+T127V、S13Y+S25P+S144P、S13Y+S25P+A147H、S13Y+S25P+S167L、S13Y+S25P+G175D、S13Y+S27L+S39P、S13Y+S27L+S42G、S13Y+S27L+S57W、S13Y+S27L+S59V、S13Y+S27L+V76L、S13Y+S27L+Q109R、S13Y+S27L+S116D、S13Y+S27L+T127V、S13Y+S27L+S144P、S13Y+S27L+A147H、S13Y+S27L+S167L、S13Y+S27L+G175D、S13Y+S39P+S42G、S13Y+S39P+S57W、S13Y+S39P+S59V、S13Y+S39P+V76L、S13Y+S39P+Q109R、S13Y+S39P+S116D、S13Y+S39P+T127V、S13Y+S39P+S144P、S13Y+S39P+A147H、S13Y+S39P+S167L、S13Y+S39P+G175D、S13Y+S42G+S57W、S13Y+S42G+S59V、S13Y+S42G+V76L、S13Y+S42G+Q109R、S13Y+S42G+S116D、S13Y+S42G+T127V、S13Y+S42G+S144P、S13Y+S42G+A147H、S13Y+S42G+S167L、S13Y+S42G+G175D、S13Y+S57W+S59V、S13Y+S57W+V76L、S13Y+S57W+Q109R、S13Y+S57W+S116D、S13Y+S57W+T127V、S13Y+S57W+S144P、S13Y+S57W+A147H、S13Y+S57W+S167L、S13Y+S57W+G175D、S13Y+S59V+V76L、S13Y+S59V+Q109R、S13Y+S59V+S116D、S13Y+S59V+T127V、S13Y+S59V+S144P、S13Y+S59V+A147H、S13Y+S59V+S167L、S13Y+S59V+G175D、S13Y+V76L+Q109R、S13Y+V76L+S116D、S13Y+V76L+T127V、S13Y+V76L+S144P、S13Y+V76L+A147H、S13Y+V76L+S167L、S13Y+V76L+G175D、S13Y+Q109R+S116D、S13Y+Q109R+T127V、S13Y+Q109R+S144P、S13Y+Q109R+A147H、S13Y+Q109R+S167L、S13Y+Q109R+G175D、S13Y+S116D+T127V、S13Y+S116D+S144P、S13Y+S116D+A147H、S13Y+S116D+S167L、S13Y+S116D+G175D、S13Y+T127V+S144P、S13Y+T127V+A147H、S13Y+T127V+S167L、S13Y+T127V+G175D、S13Y+S144P+A147H、S13Y+S144P+S167L、S13Y+S144P+G175D、S13Y+A147H+S167L、S13Y+A147H+G175D、S13Y+S167L+G175D、T22P+S25P+S27L、T22P+S25P+S39P、T22P+S25P+S42G、T22P+S25P+S57W、T22P+S25P+S59V、T22P+S25P+V76L、T22P+S25P+Q109R、T22P+S25P+S116D、T22P+S25P+T127V、T22P+S25P+S144P、T22P+S25P+A147H、T22P+S25P+S167L、T22P+S25P+G175D、T22P+S27L+S39P、T22P+S27L+S42G、T22P+S27L+S57W、T22P+S27L+S59V、T22P+S27L+V76L、T22P+S27L+Q109R、T22P+S27L+S116D、T22P+S27L+T127V、T22P+S27L+S144P、T22P+S27L+A147H、T22P+S27L+S167L、T22P+S27L+G175D、T22P+S39P+S42G、T22P+S39P+S57W、T22P+S39P+S59V、T22P+S39P+V76L、T22P+S39P+Q109R、T22P+S39P+S116D、T22P+S39P+T127V、T22P+S39P+S144P、T22P+S39P+A147H、T22P+S39P+S167L、T22P+S39P+G175D、T22P+S42G+S57W、T22P+S42G+S59V、T22P+S42G+V76L、T22P+S42G+Q109R、T22P+S42G+S116D、T22P+S42G+T127V、T22P+S42G+S144P、T22P+S42G+A147H、T22P+S42G+S167L、T22P+S42G+G175D、T22P+S57W+S59V、T22P+S57W+V76L、T22P+S57W+Q109R、T22P+S57W+S116D、T22P+S57W+T127V、T22P+S57W+S144P、T22P+S57W+A147H、T22P+S57W+S167L、T22P+S57W+G175D、T22P+S59V+V76L、T22P+S59V+Q109R、T22P+S59V+S116D、T22P+S59V+T127V、T22P+S59V+S144P、T22P+S59V+A147H、T22P+S59V+S167L、T22P+S59V+G175D、T22P+V76L+Q109R、T22P+V76L+S116D、T22P+V76L+T127V、T22P+V76L+S144P、T22P+V76L+A147H、T22P+V76L+S167L、T22P+V76L+G175D、T22P+Q109R+S116D、T22P+Q109R+T127V、T22P+Q109R+S144P、T22P+Q109R+A147H、T22P+Q109R+S167L、T22P+Q109R+G175D、T22P+S116D+T127V、T22P+S116D+S144P、T22P+S116D+A147H、T22P+S116D+S167L、T22P+S116D+G175D、T22P+T127V+S144P、T22P+T127V+A147H、T22P+T127V+S167L、T22P+T127V+G175D、T22P+S144P+A147H、T22P+S144P+S167L、T22P+S144P+G175D、T22P+A147H+S167L、T22P+A147H+G175D、T22P+S167L+G175D、S25P+S27L+S39P、S25P+S27L+S42G、S25P+S27L+S57W、S25P+S27L+S59V、S25P+S27L+V76L、S25P+S27L+Q109R、S25P+S27L+S116D、S25P+S27L+T127V、S25P+S27L+S144P、S25P+S27L+A147H、S25P+S27L+S167L、S25P+S27L+G175D、S25P+S39P+S42G、S25P+S39P+S57W、S25P+S39P+S59V、S25P+S39P+V76L、S25P+S39P+Q109R、S25P+S39P+S116D、S25P+S39P+T127V、S25P+S39P+S144P、S25P+S39P+A147H、S25P+S39P+S167L、S25P+S39P+G175D、S25P+S42G+S57W、S25P+S42G+S59V、S25P+S42G+V76L、S25P+S42G+Q109R、S25P+S42G+S116D、S25P+S42G+T127V、S25P+S42G+S144P、S25P+S42G+A147H、S25P+S42G+S167L、S25P+S42G+G175D、S25P+S57W+S59V、S25P+S57W+V76L、S25P+S57W+Q109R、S25P+S57W+S116D、S25P+S57W+T127V、S25P+S57W+S144P、S25P+S57W+A147H、S25P+S57W+S167L、S25P+S57W+G175D、S25P+S59V+V76L、S25P+S59V+Q109R、S25P+S59V+S116D、S25P+S59V+T127V、S25P+S59V+S144P、S25P+S59V+A147H、S25P+S59V+S167L、S25P+S59V+G175D、S25P+V76L+Q109R、S25P+V76L+S116D、S25P+V76L+T127V、S25P+V76L+S144P、S25P+V76L+A147H、S25P+V76L+S167L、S25P+V76L+G175D、S25P+Q109R+S116D、S25P+Q109R+T127V、S25P+Q109R+S144P、S25P+Q109R+A147H、S25P+Q109R+S167L、S25P+Q109R+G175D、S25P+S116D+T127V、S25P+S116D+S144P、S25P+S116D+A147H、S25P+S116D+S167L、S25P+S116D+G175D、S25P+T127V+ S144P、S25P+T127V+A147H、S25P+T127V+S167L、S25P+T127V+G175D、S25P+ S144P+A147H、S25P+S144P+S167L、S25P+S144P+G175D、S25P+A147H+S167L 、S25P+A147H+G175D、S25P+S167L+G175D、S27L+S39P+S42G、S27L+S39P+S57 W、S27L+S39P+S59V、S27L+S39P+V76L、S27L+S39P+Q109R、S27L+S39P+S116 D、S27L+S39P+T127V、S27L+S39P+S144P、S27L+S39P+A147H、S27L+S39P+S16 7L, S27L+S39P+G175D, S27L+S42G+S57W, S27L+S42G+S59V, S27L+S42G+V76L, S27L+S42G+Q109R, S27L+S42G+S116D, S27L+S42G+T127V, S27L+S42G+S14 4P, S27L+S42G+A147H, S27L+S42G+S167L, S27L+S42G+G175D, S27L+S57W+S59V, S27L+S57W+V76L, S27L+S57W+Q109R, S27L+S57W+S116D, S27L+S57W+T1 27V, S27L+S57W+S144P, S27L+S57W+A147H, S27L+S57W+S167L, S27L+S57W+G175D, S27L+S59V+V76L, S27L+S59V+Q109R, S27L+S59V+S116D, S27L+S59V+ T127V, S27L+S59V+S144P, S27L+S59V+A147H, S27L+S59V+S167L, S27L+S59V+G175D, S27L+V76L+Q109R, S27L+V76L+S116D, S27L+V76L+T127V, S27L+V7 6L+S144P、S27L+V76L+A147H、S27L+V76L+S167L、S27L+V76L+G175D、S27L+ Q109R+S116D、S27L+Q109R+T127V、S27L+Q109R+S144P、S27L+Q109R+A147H、S27L+Q109R+S167L、S27L+Q109R+G175D、S27L+S116D+T127V、S27L+S116D+S144P、S27L+S116D+A147H、S27L+S116D+S167L、S27L+S116D+G175D、S27L+T127V+S144P、S27L+T127V+A147H、S27L+T127V+S167L、S27L+T127V+G175D、S27L+S144P+A147H、S27L+S144P+S167L、S27L+S144P+G175D、S27L+A147H+S167L、S27L+A147H+G175D、S27L+S167L+G175D、S39P+S42G+S57W、S39P+S42G+S59V、S39P+S42G+V76L、S39P+S42G+Q109R、S39P+S42G+S116D、S39P+S42G+T127V、S39P+S42G+S144P、S39P+S42G+A147H、S39P+S42G+S167L、S39P+S42G+G175D、S39P+S57W+S59V、S39P+S57W+V76L、S39P+S57W+Q109R、S39P+S57W+S116D、S39P+S57W+T127V、S39P+S57W+S144P、S39P+S57W+A147H、S39P+S57W+S167L、S39P+S57W+G175D、S39P+S59V+V76L、S39P+S59V+Q109R、S39P+S59V+S116D、S39P+S59V+T127V、S39P+S59V+S144P、S39P+S59V+A147H、S39P+S59V+S167L、S39P+S59V+G175D、S39P+V76L+Q109R、S39P+V76L+S116D、S39P+V76L+T127V、S39P+V76L+S144P、S39P+V76L+A147H、S39P+V76L+S167L、S39P+V76L+G175D、S39P+Q109R+S116D、S39P+Q109R+T127V、S39P+Q109R+S144P、S39P+Q109R+A147H、S39P+Q109R+S167L、S39P+Q109R+G175D、S39P+S116D+T127V、S39P+S116D+S144P、S39P+S116D+A147H、S39P+S116D+S167L、S39P+S116D+G175D、S39P+T127V+S144P、S39P+T127V+A147H、S39P+T127V+S167L、S39P+T127V+G175D、S39P+S144P+A147H、S39P+S144P+S167L、S39P+S144P+G175D、S39P+A147H+S167L、S39P+A147H+G175D、S39P+S167L+G175D、S42G+S57W+S59V、S42G+S57W+V76L、S42G+S57W+Q109R、S42G+S57W+S116D、S42G+S57W+T127V、S42G+S57W+S144P、S42G+S57W+A147H、S42G+S57W+S167L、S42G+S57W+G175D、S42G+S59V+V76L、S42G+S59V+Q109R、S42G+S59V+S116D、S42G+S59V+T127V、S42G+S59V+S144P、S42G+S59V+A147H、S42G+S59V+S167L、S42G+S59V+G175D、S42G+V76L+Q109R、S42G+V76L+S116D、S42G+V76L+T127V、S42G+V76L+S144P、S42G+V76L+A147H、S42G+V76L+S167L、S42G+V76L+G175D、S42G+Q109R+S116D、S42G+Q109R+T127V、S42G+Q109R+S144P、S42G+Q109R+A147H、S42G+Q109R+S167L、S42G+Q109R+G175D、S42G+S116D+T127V、S42G+S116D+S144P、S42G+S116D+A147H、S42G+S116D+S167L、S42G+S116D+G175D、S42G+T127V+S144P、S42G+T127V+A147H、S42G+T127V+S167L、S42G+T127V+G175D、S42G+S144P+A147H、S42G+S144P+S167L、S42G+S144P+G175D、S42G+A147H+S167L、S42G+A147H+G175D、S42G+S167L+G175D、S57W+S59V+V76L、S57W+S59V+Q109R、S57W+S59V+S116D、S57W+S59V+T127V、S57W+S59V+S144P、S57W+S59V+A147H、S57W+S59V+S167L、S57W+S59V+G175D、S57W+V76L+Q109R、S57W+V76L+S116D、S57W+V76L+T127V、S57W+V76L+S144P、S57W+V76L+A147H、S57W+V76L+S167L、S57W+V76L+G175D、S57W+Q109R+S116D、S57W+Q109R+T127V、S57W+Q109R+S144P、S57W+Q109R+A147H、S57W+Q109R+S167L、S57W+Q109R+G175D、S57W+S116D+T127V、S57W+S116D+S144P、S57W+S116D+A147H、S57W+S116D+S167L、S57W+S116D+G175D、S57W+T127V+S144P、S57W+T127V+A147H、S57W+T127V+S167L、S57W+T127V+G175D、S57W+S144P+A147H、S57W+S144P+S167L、S57W+S144P+G175D、S57W+A147H+S167L、S57W+A147H+G175D、S57W+S167L+G175D、S59V+V76L+Q109R、S59V+V76L+S116D、S59V+V76L+T127V、S59V+V76L+S144P、S59V+V76L+A147H、S59V+V76L+S167L、S59V+V76L+G175D、S59V+Q109R+S116D、S59V+Q109R+T127V、S59V+Q109R+S144P、S59V+Q109R+A147H、S59V+Q109R+S167L、S59V+Q109R+G175D、S59V+S116D+T127V、S59V+S116D+S144P、S59V+S116D+A147H、S59V+S116D+S167L、S59V+S116D+G175D、S59V+T127V+S144P、S59V+T127V+A147H、S59V+T127V+S167L、S59V+T127V+G175D、S59V+S144P+A147H、S59V+S144P+S167L、S59V+S144P+G175D、S59V+A147H+S167L、S59V+A147H+G175D、S59V+S167L+G175D、V76L+Q109R+S116D、V76L+Q109R+T127V、V76L+Q109R+S144P、V76L+Q109R+A147H、V76L+Q109R+S167L、V76L+Q109R+G175D、V76L+S116D+T127V、V76L+S116D+S144P、V76L+S116D+A147H、V76L+S116D+S167L、V76L+S116D+G175D、V76L+T127V+S144P、V76L+T127V+A147H、V76L+T127V+S167L、V76L+T127V+G175D、V76L+S144P+A147H、V76L+S144P+S167L、V76L+S144P+G175D、V76L+A147H+S167L、V76L+A147H+G175D、V76L+S167L+G175D、Q109R+S116D+T127V、Q109R+S116D+S144P、Q109R+S116D+A147H、Q109R+S116D+S167L、Q109R+S116D+G175D、Q109R+T127V+S144P、Q109R+T127V+A147H、Q109R+T127V+S167L、Q109R+T127V+G175D、Q109R+S144P+A147H、Q109R+S144P+S167L、Q109R+S144P+G175D、Q109R+A147H+S167L、Q109R+A147H+G175D、Q109R+S167L+G175D、S116D+T127V+S144P、S116D+T127V+A147H、S116D+T127V+S167L、S116D+T127V+G175D、S116D+S144P+A147H、S116D+S144P+S167L、S116D+S144P+G175D、S116D+A147H+S167L、S116D+A147H+G175D、S116D+S167L+G175D、T127V+S144P+A147H、T127V+S144P+S167L、T127V+S144P+G175D、T127V+A147H+S167L、T127V+A147H+G175D、T127V+S167L+G175D、S144P+A147H+S167L、S144P+A147H+G175D、S144P+S167L+G175D and A147H+S167L+G175D.

[0747] In one embodiment, the two or more mutations are selected from any of the following: T1I, S13Y, T22P, S25P, S27L, S39P, S42G, S57W, S59V, V76L, Q109R, S116, T127V, S144P, A147H, S167L, or G175D. In one embodiment, the two or more mutations are selected from any of the following: T1I, S13Y, T22P, S25P, S27L, S39P, S42G, S57W, S59V, V76L, Q109R, S116D, T127V, S144P, A147H, S167L, or G175D. Preferably, the variant contains more than 10 mutations from the following mutations, preferably 11, 12, 13, 14, 15, 16, or all of the following mutations: T1I, S13Y, T22P, S25P, S27L, S39P, S42G, S57W, S59V, V76L, Q109R, S116D, T127V, S144P, A147H, S167L, or G175D.

[0748] In a particularly preferred embodiment, the DNA enzyme variant is selected from any of the following:

[0749] T1I+S13Y+T22P+S25P+S27L+S39P+S42G+S57W+S59V+V76L+Q109R+S116D+T127V+S144P;

[0750] T1I+S13Y+T22P+S25P+S27L+S39P+S42G+S57W+S59V+V76L+Q109R+S116D+T127V+A147H;

[0751] T1I+S13Y+T22P+S25P+S27L+S39P+S42G+S57W+S59V+V76L+Q109R+S116D+T127V+S167L;

[0752] T1I+S13Y+T22P+S25P+S27L+S39P+S42G+S57W+S59V+V76L+Q109R+S116D+T127V+G175D;

[0753] T1I+S13Y+T22P+S25P+S27L+S39P+S42G+S57W+S59V+V76L+Q109R+S116D+S144P+A147H;

[0754] T1I+S13Y+T22P+S25P+S27L+S39P+S42G+S57W+S59V+V76L+Q109R+S116D+S144P+S167L;

[0755] T1I+S13Y+T22P+S25P+S27L+S39P+S42G+S57W+S59V+V76L+Q109R+S116D+S144P+G175D;

[0756] T1I+S13Y+T22P+S25P+S27L+S39P+S42G+S57W+S59V+V76L+Q109R+S116D+A147H+S167L;

[0757] T1I+S13Y+T22P+S25P+S27L+S39P+S42G+S57W+S59V+V76L+Q109R+S116D+A147H+G175D;

[0758] T1I+S13Y+T22P+S25P+S27L+S39P+S42G+S57W+S59V+V76L+Q109R+S116D+S167L+G175D;

[0759] T1I+S13Y+T22P+S25P+S27L+S39P+S42G+S57W+S59V+V76L+Q109R+T127V+S144P+A147H;

[0760] T1I+S13Y+T22P+S25P+S27L+S39P+S42G+S57W+S59V+V76L+Q109R+T127V+S144P+S167L;

[0761] T1I+S13Y+T22P+S25P+S27L+S39P+S42G+S57W+S59V+V76L+Q109R+T127V+S144P+G175D;

[0762] T1I+S13Y+T22P+S25P+S27L+S39P+S42G+S57W+S59V+V76L+Q109R+T127V+A147H+S167L;

[0763] T1I+S13Y+T22P+S25P+S27L+S39P+S42G+S57W+S59V+V76L+Q109R+T127V+A147H+G175D;

[0764] T1I+S13Y+T22P+S25P+S27L+S39P+S42G+S57W+S59V+V76L+Q109R+T127V+S167L+G175D;

[0765] T1I+S13Y+T22P+S25P+S27L+S39P+S42G+S57W+S59V+V76L+Q109R+S144P+A147H+S167L;

[0766] T1I+S13Y+T22P+S25P+S27L+S39P+S42G+S57W+S59V+V76L+Q109R+S144P+A147H+G175D;

[0767] T1I+S13Y+T22P+S25P+S27L+S39P+S42G+S57W+S59V+V76L+Q109R+S144P+S167L+G175D;

[0768] T1I+S13Y+T22P+S25P+S27L+S39P+S42G+S57W+S59V+V76L+Q109R+A147H+S167L+G175D;

[0769] T1I+S13Y+T22P+S25P+S27L+S39P+S42G+S57W+S59V+V76L+S116D+T127V+S144P+A147H;

[0770] T1I+S13Y+T22P+S25P+S27L+S39P+S42G+S57W+S59V+V76L+S116D+T127V+S144P+S167L;

[0771] T1I+S13Y+T22P+S25P+S27L+S39P+S42G+S57W+S59V+V76L+S116D+T127V+S144P+G175D;

[0772] T1I+S13Y+T22P+S25P+S27L+S39P+S42G+S57W+S59V+V76L+S116D+T127V+A147H+S167L;

[0773] T1I+S13Y+T22P+S25P+S27L+S39P+S42G+S57W+S59V+V76L+S116D+T127V+A147H+G175D;

[0774] T1I+S13Y+T22P+S25P+S27L+S39P+S42G+S57W+S59V+V76L+S116D+T127V+S167L+G175D;

[0775] T1I+S13Y+T22P+S25P+S27L+S39P+S42G+S57W+S59V+V76L+S116D+S144P+A147H+S167L;

[0776] T1I+S13Y+T22P+S25P+S27L+S39P+S42G+S57W+S59V+V76L+S116D+S144P+A147H+G175D;

[0777] T1I+S13Y+T22P+S25P+S27L+S39P+S42G+S57W+S59V+V76L+S116D+S144P+S167L+G175D;

[0778] T1I+S13Y+T22P+S25P+S27L+S39P+S42G+S57W+S59V+V76L+S116D+A147H+S167L+G175D;

[0779] T1I+S13Y+T22P+S25P+S27L+S39P+S42G+S57W+S59V+V76L+T127V+S144P+A147H+S167L;

[0780] T1I+S13Y+T22P+S25P+S27L+S39P+S42G+S57W+S59V+V76L+T127V+S144P+A147H+G175D;

[0781] T1I+S13Y+T22P+S25P+S27L+S39P+S42G+S57W+S59V+V76L+T127V+S144P+S167L+G175D;

[0782] T1I+S13Y+T22P+S25P+S27L+S39P+S42G+S57W+S59V+V76L+T127V+A147H+S167L+G175D;

[0783] T1I+S13Y+T22P+S25P+S27L+S39P+S42G+S57W+S59V+V76L+S144P+A147H+S167L+G175D;

[0784] T1I+S13Y+T22P+S25P+S27L+S39P+S42G+S57W+S59V+Q109R+S116D+T127V+S144P+A147H;

[0785] T1I+S13Y+T22P+S25P+S27L+S39P+S42G+S57W+S59V+Q109R+S116D+T127V+S144P+S167L

[0786] T1I+S13Y+T22P+S25P+S27L+S39P+S42G+S57W+S59V+Q109R+S116D+T127V+S144P+G175D;

[0787] T1I+S13Y+T22P+S25P+S27L+S39P+S42G+S57W+S59V+Q109R+S116D+T127V+A147H+S167L

[0788] T1I+S13Y+T22P+S25P+S27L+S39P+S42G+S57W+S59V+Q109R+S116D+T127V+A147H+G175D;

[0789] T1I+S13Y+T22P+S25P+S27L+S39P+S42G+S57W+S59V+Q109R+S116D+T127V+S167L+G175D;

[0790] T1I+S13Y+T22P+S25P+S27L+S39P+S42G+S57W+S59V+Q109R+S116D+S144P+A147H+S167;

[0791] T1I+S13Y+T22P+S25P+S27L+S39P+S42G+S57W+S59V+Q109R+S116D+S144P+A147H+G175D;

[0792] T1I+S13Y+T22P+S25P+S27L+S39P+S42G+S57W+S59V+Q109R+S116D+S144P+S167L+G175D;

[0793] T1I+S13Y+T22P+S25P+S27L+S39P+S42G+S57W+S59V+Q109R+S116D+A147H+S167L+G175D;

[0794] T1I+S13Y+T22P+S25P+S27L+S39P+S42G+S57W+S59V+Q109R+T127V+S144P+A147H+S167;

[0795] T1I+S13Y+T22P+S25P+S27L+S39P+S42G+S57W+S59V+Q109R+T127V+S144P+A147H+G175D;

[0796] T1I+S13Y+T22P+S25P+S27L+S39P+S42G+S57W+S59V+Q109R+T127V+S144P+S167L+G175;

[0797] T1I+S13Y+T22P+S25P+S27L+S39P+S42G+S57W+S59V+Q109R+T127V+A147H+S167L+G175D;

[0798] T1I+S13Y+T22P+S25P+S27L+S39P+S42G+S57W+S59V+Q109R+S144P+A147H+S167L+G175D;

[0799] T1I+S13Y+T22P+S25P+S27L+S39P+S42G+S57W+S59V+S116D+T127V+S144P+A147H+S167L

[0800] T1I+S13Y+T22P+S25P+S27L+S39P+S42G+S57W+S59V+S116D+T127V+S144P+A147H+G175D;

[0801] T1I+S13Y+T22P+S25P+S27L+S39P+S42G+S57W+S59V+S116D+T127V+S144P+S167L+G175D;

[0802] T1I+S13Y+T22P+S25P+S27L+S39P+S42G+S57W+S59V+S116D+T127V+A147H+S167L+G175D;

[0803] T1I+S13Y+T22P+S25P+S27L+S39P+S42G+S57W+S59V+S116D+S144P+A147H+S167L+G175D;

[0804] T1I+S13Y+T22P+S25P+S27L+S39P+S42G+S57W+S59V+T127V+S144P+A147H+S167L+G175D;

[0805] T1I+S13Y+T22P+S25P+S27L+S39P+S42G+S57W+V76L+Q109R+S116D+T127V+S144P+A147H;

[0806] T1I+S13Y+T22P+S25P+S27L+S39P+S42G+S57W+V76L+Q109R+S116D+T127V+S144P+S167L;

[0807] T1I+S13Y+T22P+S25P+S27L+S39P+S42G+S57W+V76L+Q109R+S116D+T127V+S144P+G175D;

[0808] T1I+S13Y+T22P+S25P+S27L+S39P+S42G+S57W+V76L+Q109R+S116D+T127V+A147H+S167L

[0809] T1I+S13Y+T22P+S25P+S27L+S39P+S42G+S57W+V76L+Q109R+S116D+T127V+A147H+G175D;

[0810] T1I+S13Y+T22P+S25P+S27L+S39P+S42G+S57W+V76L+Q109R+S116D+T127V+S167L+G175D;

[0811] T1I+S13Y+T22P+S25P+S27L+S39P+S42G+S57W+V76L+Q109R+S116D+S144P+A147H+S167L

[0812] T1I+S13Y+T22P+S25P+S27L+S39P+S42G+S57W+V76L+Q109R+S116D+S144P+A147H+G175D;

[0813] T1I+S13Y+T22P+S25P+S27L+S39P+S42G+S57W+V76L+Q109R+S116D+S144P+S167L+G175D;

[0814] T1I+S13Y+T22P+S25P+S27L+S39P+S42G+S57W+V76L+Q109R+S116D+A147H+S167L+G175D;

[0815] T1I+S13Y+T22P+S25P+S27L+S39P+S42G+S57W+V76L+Q109R+T127V+S144P+A147H+S167L

[0816] T1I+S13Y+T22P+S25P+S27L+S39P+S42G+S57W+V76L+Q109R+T127V+S144P+A147H+G175D;

[0817] T1I+S13Y+T22P+S25P+S27L+S39P+S42G+S57W+V76L+Q109R+T127V+S144P+S167L+G175D;

[0818] T1I+S13Y+T22P+S25P+S27L+S39P+S42G+S57W+V76L+Q109R+T127V+A147H+S167L+G175D;

[0819] T1I+S13Y+T22P+S25P+S27L+S39P+S42G+S57W+V76L+Q109R+S144P+A147H+S167L+G175D;

[0820] T1I+S13Y+T22P+S25P+S27L+S39P+S42G+S57W+V76L+S116D+T127V+S144P+A147H+S167L;

[0821] T1I+S13Y+T22P+S25P+S27L+S39P+S42G+S57W+V76L+S116D+T127V+S144P+A147H+G175D;

[0822] T1I+S13Y+T22P+S25P+S27L+S39P+S42G+S57W+V76L+S116D+T127V+S144P+S167L+G175D;

[0823] T1I+S13Y+T22P+S25P+S27L+S39P+S42G+S57W+V76L+S116D+T127V+A147H+S167L+G175D;

[0824] T1I+S13Y+T22P+S25P+S27L+S39P+S42G+S57W+V76L+S116D+S144P+A147H+S167L+G175D;

[0825] T1I+S13Y+T22P+S25P+S27L+S39P+S42G+S57W+V76L+T127V+S144P+A147H+S167L+G175D;

[0826] T1I+S13Y+T22P+S25P+S27L+S39P+S42G+S57W+Q109R+S116D+T127V+S144P+A147H+S167L;

[0827] T1I+S13Y+T22P+S25P+S27L+S39P+S42G+S57W+Q109R+S116D+T127V+S144P+A147H+G175D;

[0828] T1I+S13Y+T22P+S25P+S27L+S39P+S42G+S57W+Q109R+S116D+T127V+S144P+S167L+G175D;

[0829] T1I+S13Y+T22P+S25P+S27L+S39P+S42G+S57W+Q109R+S116D+T127V+A147H+S167L+G175D;

[0830] T1I+S13Y+T22P+S25P+S27L+S39P+S42G+S57W+Q109R+S116D+S144P+A147H+S167L+G175D;

[0831] T1I+S13Y+T22P+S25P+S27L+S39P+S42G+S57W+Q109R+T127V+S144P+A147H+S167L+G175D;

[0832] T1I+S13Y+T22P+S25P+S27L+S39P+S42G+S57W+S116D+T127V+S144P+A147H+S167L+G175D;

[0833] T1I+S13Y+T22P+S25P+S27L+S39P+S42G+S59V+V76L+Q109R+S116D+T127V+S144P+A147H;

[0834] T1I+S13Y+T22P+S25P+S27L+S39P+S42G+S59V+V76L+Q109R+S116D+T127V+S144P+S167L;

[0835] T1I+S13Y+T22P+S25P+S27L+S39P+S42G+S59V+V76L+Q109R+S116D+T127V+S144P+G175D;

[0836] T1I+S13Y+T22P+S25P+S27L+S39P+S42G+S59V+V76L+Q109R+S116D+T127V+A147H+S167L;

[0837] T1I+S13Y+T22P+S25P+S27L+S39P+S42G+S59V+V76L+Q109R+S116D+T127V+A147H+G175D;

[0838] T1I+S13Y+T22P+S25P+S27L+S39P+S42G+S59V+V76L+Q109R+S116D+T127V+S167L+G175D;

[0839] T1I+S13Y+T22P+S25P+S27L+S39P+S42G+S59V+V76L+Q109R+S116D+S144P+A147H+S167L;

[0840] T1I+S13Y+T22P+S25P+S27L+S39P+S42G+S59V+V76L+Q109R+S116D+S144P+A147H+G175D;

[0841] T1I+S13Y+T22P+S25P+S27L+S39P+S42G+S59V+V76L+Q109R+S116D+S144P+S167L+G175D;

[0842] T1I+S13Y+T22P+S25P+S27L+S39P+S42G+S59V+V76L+Q109R+S116D+A147H+S167L+G175D

[0843] T1I+S13Y+T22P+S25P+S27L+S39P+S42G+S59V+V76L+Q109R+T127V+S144P+A147H+S167L;

[0844] T1I+S13Y+T22P+S25P+S27L+S39P+S42G+S59V+V76L+Q109R+T127V+S144P+A147H+G175D;

[0845] T1I+S13Y+T22P+S25P+S27L+S39P+S42G+S59V+V76L+Q109R+T127V+S144P+S167L+G175D;

[0846] T1I+S13Y+T22P+S25P+S27L+S39P+S42G+S59V+V76L+Q109R+T127V+A147H+S167L+G175D;

[0847] T1I+S13Y+T22P+S25P+S27L+S39P+S42G+S59V+V76L+Q109R+S144P+A147H+S167L+G175D;

[0848] T1I+S13Y+T22P+S25P+S27L+S39P+S42G+S59V+V76L+S116D+T127V+S144P+A147H+S167L;

[0849] T1I+S13Y+T22P+S25P+S27L+S39P+S42G+S59V+V76L+S116D+T127V+S144P+A147H+G175D;

[0850] T1I+S13Y+T22P+S25P+S27L+S39P+S42G+S59V+V76L+S116D+T127V+S144P+S167L+G175D;

[0851] T1I+S13Y+T22P+S25P+S27L+S39P+S42G+S59V+V76L+S116D+T127V+A147H+S167L+G175D;

[0852] T1I+S13Y+T22P+S25P+S27L+S39P+S42G+S59V+V76L+S116D+S144P+A147H+S167L+G175D;

[0853] T1I+S13Y+T22P+S25P+S27L+S39P+S42G+S59V+V76L+T127V+S144P+A147H+S167L+G175D;

[0854] T1I+S13Y+T22P+S25P+S27L+S39P+S42G+S59V+Q109R+S116D+T127V+S144P+A147H+S167L;

[0855] T1I+S13Y+T22P+S25P+S27L+S39P+S42G+S59V+Q109R+S116D+T127V+S144P+A147H+G175D;

[0856] T1I+S13Y+T22P+S25P+S27L+S39P+S42G+S59V+Q109R+S116D+T127V+S144P+S167L+G175D;

[0857] T1I+S13Y+T22P+S25P+S27L+S39P+S42G+S59V+Q109R+S116D+T127V+A147H+S167L+G175D;

[0858] T1I+S13Y+T22P+S25P+S27L+S39P+S42G+S59V+Q109R+S116D+S144P+A147H+S167L+G175D;

[0859] T1I+S13Y+T22P+S25P+S27L+S39P+S42G+S59V+Q109R+T127V+S144P+A147H+S167L+G175D;

[0860] T1I+S13Y+T22P+S25P+S27L+S39P+S42G+S59V+S116D+T127V+S144P+A147H+S167L+G175D;

[0861] T1I+S13Y+T22P+S25P+S27L+S39P+S42G+V76L+Q109R+S116D+T127V+S144P+A147H+S167L;

[0862] T1I+S13Y+T22P+S25P+S27L+S39P+S42G+V76L+Q109R+S116D+T127V+S144P+A147H+G175D;

[0863] T1I+S13Y+T22P+S25P+S27L+S39P+S42G+V76L+Q109R+S116D+T127V+S144P+S167L+G175D;

[0864] T1I+S13Y+T22P+S25P+S27L+S39P+S42G+V76L+Q109R+S116D+T127V+A147H+S167L+G175D;

[0865] T1I+S13Y+T22P+S25P+S27L+S39P+S42G+V76L+Q109R+S116D+S144P+A147H+S167L+G175D;

[0866] T1I+S13Y+T22P+S25P+S27L+S39P+S42G+V76L+Q109R+T127V+S144P+A147H+S167L+G175D;

[0867] T1I+S13Y+T22P+S25P+S27L+S39P+S42G+V76L+S116D+T127V+S144P+A147H+S167L+G175D;

[0868] T1I+S13Y+T22P+S25P+S27L+S39P+S42G+Q109R+S116D+T127V+S144P+A147H+S167L+G175D;

[0869] T1I+S13Y+T22P+S25P+S27L+S39P+S57W+S59V+V76L+Q109R+S116D+T127V+S144P+A147H;

[0870] T1I+S13Y+T22P+S25P+S27L+S39P+S57W+S59V+V76L+Q109R+S116D+T127V+S144P+S167L;

[0871] T1I+S13Y+T22P+S25P+S27L+S39P+S57W+S59V+V76L+Q109R+S116D+T127V+S144P+G175D;

[0872] T1I+S13Y+T22P+S25P+S27L+S39P+S57W+S59V+V76L+Q109R+S116D+T127V+A147H+S167L;

[0873] T1I+S13Y+T22P+S25P+S27L+S39P+S57W+S59V+V76L+Q109R+S116D+T127V+A147H+G175D;

[0874] T1I+S13Y+T22P+S25P+S27L+S39P+S57W+S59V+V76L+Q109R+S116D+T127V+S167L+G175D;

[0875] T1I+S13Y+T22P+S25P+S27L+S39P+S57W+S59V+V76L+Q109R+S116D+S144P+A147H+S167L;

[0876] T1I+S13Y+T22P+S25P+S27L+S39P+S57W+S59V+V76L+Q109R+S116D+S144P+A147H+G175D;

[0877] T1I+S13Y+T22P+S25P+S27L+S39P+S57W+S59V+V76L+Q109R+S116D+S144P+S167L+G175D;

[0878] T1I+S13Y+T22P+S25P+S27L+S39P+S57W+S59V+V76L+Q109R+S116D+A147H+S167L+G175D;

[0879] T1I+S13Y+T22P+S25P+S27L+S39P+S57W+S59V+V76L+Q109R+T127V+S144P+A147H+S167L;

[0880] T1I+S13Y+T22P+S25P+S27L+S39P+S57W+S59V+V76L+Q109R+T127V+S144P+A147H+G175D;

[0881] T1I+S13Y+T22P+S25P+S27L+S39P+S57W+S59V+V76L+Q109R+T127V+S144P+S167L+G175D;

[0882] T1I+S13Y+T22P+S25P+S27L+S39P+S57W+S59V+V76L+Q109R+T127V+A147H+S167L+G175D;

[0883] T1I+S13Y+T22P+S25P+S27L+S39P+S57W+S59V+V76L+Q109R+S144P+A147H+S167L+G175D;

[0884] T1I+S13Y+T22P+S25P+S27L+S39P+S57W+S59V+V76L+S116D+T127V+S144P+A147H+S167L;

[0885] T1I+S13Y+T22P+S25P+S27L+S39P+S57W+S59V+V76L+S116D+T127V+S144P+A147H+G175D;

[0886] T1I+S13Y+T22P+S25P+S27L+S39P+S57W+S59V+V76L+S116D+T127V+S144P+S167L+G175D;

[0887] T1I+S13Y+T22P+S25P+S27L+S39P+S57W+S59V+V76L+S116D+T127V+A147H+S167L+G175D;

[0888] T1I+S13Y+T22P+S25P+S27L+S39P+S57W+S59V+V76L+S116D+S144P+A147H+S167L+G175D;

[0889] T1I+S13Y+T22P+S25P+S27L+S39P+S57W+S59V+V76L+T127V+S144P+A147H+S167L+G175D;

[0890] T1I+S13Y+T22P+S25P+S27L+S39P+S57W+S59V+Q109R+S116D+T127V+S144P+A147H+S167L;

[0891] T1I+S13Y+T22P+S25P+S27L+S39P+S57W+S59V+Q109R+S116D+T127V+S144P+A147H+G175D;

[0892] T1I+S13Y+T22P+S25P+S27L+S39P+S57W+S59V+Q109R+S116D+T127V+S144P+S167L+G175D;

[0893] T1I+S13Y+T22P+S25P+S27L+S39P+S57W+S59V+Q109R+S116D+T127V+A147H+S167L+G175D;

[0894] T1I+S13Y+T22P+S25P+S27L+S39P+S57W+S59V+Q109R+S116D+S144P+A147H+S167L+G175D;

[0895] T1I+S13Y+T22P+S25P+S27L+S39P+S57W+S59V+Q109R+T127V+S144P+A147H+S167L+G175D;

[0896] T1I+S13Y+T22P+S25P+S27L+S39P+S57W+S59V+S116D+T127V+S144P+A147H+S167L+G175D;

[0897] T1I+S13Y+T22P+S25P+S27L+S39P+S57W+V76L+Q109R+S116D+T127V+S144P+A147H+S167L;

[0898] T1I+S13Y+T22P+S25P+S27L+S39P+S57W+V76L+Q109R+S116D+T127V+S144P+A147H+G175D;

[0899] T1I+S13Y+T22P+S25P+S27L+S39P+S57W+V76L+Q109R+S116D+T127V+S144P+S167L+G175D;

[0900] T1I+S13Y+T22P+S25P+S27L+S39P+S57W+V76L+Q109R+S116D+T127V+A147H+S167L+G175D;

[0901] T1I+S13Y+T22P+S25P+S27L+S39P+S57W+V76L+Q109R+S116D+S144P+A147H+S167L+G175D;

[0902] T1I+S13Y+T22P+S25P+S27L+S39P+S57W+V76L+Q109R+T127V+S144P+A147H+S167L+G175D;

[0903] T1I+S13Y+T22P+S25P+S27L+S39P+S57W+V76L+S116D+T127V+S144P+A147H+S167L+G175D;

[0904] T1I+S13Y+T22P+S25P+S27L+S39P+S57W+Q109R+S116D+T127V+S144P+A147H+S167L+G175D;

[0905] T1I+S13Y+T22P+S25P+S27L+S39P+S59V+V76L+Q109R+S116D+T127V+S144P+A147H+S167L;

[0906] T1I+S13Y+T22P+S25P+S27L+S39P+S59V+V76L+Q109R+S116D+T127V+S144P+A147H+G175D;

[0907] T1I+S13Y+T22P+S25P+S27L+S39P+S59V+V76L+Q109R+S116D+T127V+S144P+S167L+G175D;

[0908] T1I+S13Y+T22P+S25P+S27L+S39P+S59V+V76L+Q109R+S116D+T127V+A147H+S167L+G175D;

[0909] T1I+S13Y+T22P+S25P+S27L+S39P+S59V+V76L+Q109R+S116D+S144P+A147H+S167L+G175D;

[0910] T1I+S13Y+T22P+S25P+S27L+S39P+S59V+V76L+Q109R+T127V+S144P+A147H+S167L+G175D;

[0911] T1I+S13Y+T22P+S25P+S27L+S39P+S59V+V76L+S116D+T127V+S144P+A147H+S167L+G175D;

[0912] T1I+S13Y+T22P+S25P+S27L+S39P+S59V+Q109R+S116D+T127V+S144P+A147H+S167L+G175D;

[0913] T1I+S13Y+T22P+S25P+S27L+S39P+V76L+Q109R+S116D+T127V+S144P+A147H+S167L+G175D;

[0914] T1I+S13Y+T22P+S25P+S27L+S42G+S57W+S59V+V76L+Q109R+S116D+T127V+S144P+A147;

[0915] T1I+S13Y+T22P+S25P+S27L+S42G+S57W+S59V+V76L+Q109R+S116D+T127V+S144P+S167;

[0916] T1I+S13Y+T22P+S25P+S27L+S42G+S57W+S59V+V76L+Q109R+S116D+T127V+S144P+G175D;

[0917] T1I+S13Y+T22P+S25P+S27L+S42G+S57W+S59V+V76L+Q109R+S116D+T127V+A147H+S167;

[0918] T1I+S13Y+T22P+S25P+S27L+S42G+S57W+S59V+V76L+Q109R+S116D+T127V+A147H+G175D;

[0919] T1I+S13Y+T22P+S25P+S27L+S42G+S57W+S59V+V76L+Q109R+S116D+T127V+S167L+G175;

[0920] T1I+S13Y+T22P+S25P+S27L+S42G+S57W+S59V+V76L+Q109R+S116D+S144P+A147H+S167;

[0921] T1I+S13Y+T22P+S25P+S27L+S42G+S57W+S59V+V76L+Q109R+S116D+S144P+A147H+G175D;

[0922] T1I+S13Y+T22P+S25P+S27L+S42G+S57W+S59V+V76L+Q109R+S116D+S144P+S167L+G175;

[0923] T1I+S13Y+T22P+S25P+S27L+S42G+S57W+S59V+V76L+Q109R+S116D+A147H+S167L+G175D;

[0924] T1I+S13Y+T22P+S25P+S27L+S42G+S57W+S59V+V76L+Q109R+T127V+S144P+A147H+S167;

[0925] T1I+S13Y+T22P+S25P+S27L+S42G+S57W+S59V+V76L+Q109R+T127V+S144P+A147H+G175D;

[0926] T1I+S13Y+T22P+S25P+S27L+S42G+S57W+S59V+V76L+Q109R+T127V+S144P+S167L+G175D;

[0927] T1I+S13Y+T22P+S25P+S27L+S42G+S57W+S59V+V76L+Q109R+T127V+A147H+S167L+G175D;

[0928] T1I+S13Y+T22P+S25P+S27L+S42G+S57W+S59V+V76L+Q109R+S144P+A147H+S167L+G175D;

[0929] T1I+S13Y+T22P+S25P+S27L+S42G+S57W+S59V+V76L+S116D+T127V+S144P+A147H+S167L;

[0930] T1I+S13Y+T22P+S25P+S27L+S42G+S57W+S59V+V76L+S116D+T127V+S144P+A147H+G175D;

[0931] T1I+S13Y+T22P+S25P+S27L+S42G+S57W+S59V+V76L+S116D+T127V+S144P+S167L+G175D;

[0932] T1I+S13Y+T22P+S25P+S27L+S42G+S57W+S59V+V76L+S116D+T127V+A147H+S167L+G175D;

[0933] T1I+S13Y+T22P+S25P+S27L+S42G+S57W+S59V+V76L+S116D+S144P+A147H+S167L+G175D;

[0934] T1I+S13Y+T22P+S25P+S27L+S42G+S57W+S59V+V76L+T127V+S144P+A147H+S167L+G175D;

[0935] T1I+S13Y+T22P+S25P+S27L+S42G+S57W+S59V+Q109R+S116D+T127V+S144P+A147H+S167L;

[0936] T1I+S13Y+T22P+S25P+S27L+S42G+S57W+S59V+Q109R+S116D+T127V+S144P+A147H+G175D;

[0937] T1I+S13Y+T22P+S25P+S27L+S42G+S57W+S59V+Q109R+S116D+T127V+S144P+S167L+G175D;

[0938] T1I+S13Y+T22P+S25P+S27L+S42G+S57W+S59V+Q109R+S116D+T127V+A147H+S167L+G175D;

[0939] T1I+S13Y+T22P+S25P+S27L+S42G+S57W+S59V+Q109R+S116D+S144P+A147H+S167L+G175D;

[0940] T1I+S13Y+T22P+S25P+S27L+S42G+S57W+S59V+Q109R+T127V+S144P+A147H+S167L+G175D;

[0941] T1I+S13Y+T22P+S25P+S27L+S42G+S57W+S59V+S116D+T127V+S144P+A147H+S167L+G175D;

[0942] T1I+S13Y+T22P+S25P+S27L+S42G+S57W+V76L+Q109R+S116D+T127V+S144P+A147H+S167L;

[0943] T1I+S13Y+T22P+S25P+S27L+S42G+S57W+V76L+Q109R+S116D+T127V+S144P+A147H+G175D;

[0944] T1I+S13Y+T22P+S25P+S27L+S42G+S57W+V76L+Q109R+S116D+T127V+S144P+S167L+G175D;

[0945] T1I+S13Y+T22P+S25P+S27L+S42G+S57W+V76L+Q109R+S116D+T127V+A147H+S167L+G175D;

[0946] T1I+S13Y+T22P+S25P+S27L+S42G+S57W+V76L+Q109R+S116D+S144P+A147H+S167L+G175D;

[0947] T1I+S13Y+T22P+S25P+S27L+S42G+S57W+V76L+Q109R+T127V+S144P+A147H+S167L+G175D;

[0948] T1I+S13Y+T22P+S25P+S27L+S42G+S57W+V76L+S116D+T127V+S144P+A147H+S167L+G175D;

[0949] T1I+S13Y+T22P+S25P+S27L+S42G+S57W+Q109R+S116D+T127V+S144P+A147H+S167L+G175D;

[0950] T1I+S13Y+T22P+S25P+S27L+S42G+S59V+V76L+Q109R+S116D+T127V+S144P+A147H+S167L;

[0951] T1I+S13Y+T22P+S25P+S27L+S42G+S59V+V76L+Q109R+S116D+T127V+S144P+A147H+G175D;

[0952] T1I+S13Y+T22P+S25P+S27L+S42G+S59V+V76L+Q109R+S116D+T127V+S144P+S167L+G175D;

[0953] T1I+S13Y+T22P+S25P+S27L+S42G+S59V+V76L+Q109R+S116D+T127V+A147H+S167L+G175D;

[0954] T1I+S13Y+T22P+S25P+S27L+S42G+S59V+V76L+Q109R+S116D+S144P+A147H+S167L+G175D;

[0955] T1I+S13Y+T22P+S25P+S27L+S42G+S59V+V76L+Q109R+T127V+S144P+A147H+S167L+G175D;

[0956] T1I+S13Y+T22P+S25P+S27L+S42G+S59V+V76L+S116D+T127V+S144P+A147H+S167L+G175D;

[0957] T1I+S13Y+T22P+S25P+S27L+S42G+S59V+Q109R+S116D+T127V+S144P+A147H+S167L+G175D;

[0958] T1I+S13Y+T22P+S25P+S27L+S42G+V76L+Q109R+S116D+T127V+S144P+A147H+S167L+G175D;

[0959] T1I+S13Y+T22P+S25P+S27L+S57W+S59V+V76L+Q109R+S116D+T127V+S144P+A147H+S167L;

[0960] T1I+S13Y+T22P+S25P+S27L+S57W+S59V+V76L+Q109R+S116D+T127V+S144P+A147H+G175D;

[0961] T1I+S13Y+T22P+S25P+S27L+S57W+S59V+V76L+Q109R+S116D+T127V+S144P+S167L+G175D;

[0962] T1I+S13Y+T22P+S25P+S27L+S57W+S59V+V76L+Q109R+S116D+T127V+A147H+S167L+G175D;

[0963] T1I+S13Y+T22P+S25P+S27L+S57W+S59V+V76L+Q109R+S116D+S144P+A147H+S167L+G175D;

[0964] T1I+S13Y+T22P+S25P+S27L+S57W+S59V+V76L+Q109R+T127V+S144P+A147H+S167L+G175D;

[0965] T1I+S13Y+T22P+S25P+S27L+S57W+S59V+V76L+S116D+T127V+S144P+A147H+S167L+G175D;

[0966] T1I+S13Y+T22P+S25P+S27L+S57W+S59V+Q109R+S116D+T127V+S144P+A147H+S167L+G175D;

[0967] T1I+S13Y+T22P+S25P+S27L+S57W+V76L+Q109R+S116D+T127V+S144P+A147H+S167L+G175D;

[0968] T1I+S13Y+T22P+S25P+S27L+S59V+V76L+Q109R+S116D+T127V+S144P+A147H+S167L+G175D;

[0969] T1I+S13Y+T22P+S25P+S39P+S42G+S57W+S59V+V76L+Q109R+S116D+T127V+S144P+A147H;

[0970] T1I+S13Y+T22P+S25P+S39P+S42G+S57W+S59V+V76L+Q109R+S116D+T127V+S144P+S167L;

[0971] T1I+S13Y+T22P+S25P+S39P+S42G+S57W+S59V+V76L+Q109R+S116D+T127V+S144P+G175D;

[0972] T1I+S13Y+T22P+S25P+S39P+S42G+S57W+S59V+V76L+Q109R+S116D+T127V+A147H+S167L;

[0973] T1I+S13Y+T22P+S25P+S39P+S42G+S57W+S59V+V76L+Q109R+S116D+T127V+A147H+G175D;

[0974] T1I+S13Y+T22P+S25P+S39P+S42G+S57W+S59V+V76L+Q109R+S116D+T127V+S167L+G175D;

[0975] T1I+S13Y+T22P+S25P+S39P+S42G+S57W+S59V+V76L+Q109R+S116D+S144P+A147H+S167;

[0976] T1I+S13Y+T22P+S25P+S39P+S42G+S57W+S59V+V76L+Q109R+S116D+S144P+A147H+G175D;

[0977] T1I+S13Y+T22P+S25P+S39P+S42G+S57W+S59V+V76L+Q109R+S116D+S144P+S167L+G175D;

[0978] T1I+S13Y+T22P+S25P+S39P+S42G+S57W+S59V+V76L+Q109R+S116D+A147H+S167L+G175D;

[0979] T1I+S13Y+T22P+S25P+S39P+S42G+S57W+S59V+V76L+Q109R+T127V+S144P+A147H+S167;

[0980] T1I+S13Y+T22P+S25P+S39P+S42G+S57W+S59V+V76L+Q109R+T127V+S144P+A147H+G175D;

[0981] T1I+S13Y+T22P+S25P+S39P+S42G+S57W+S59V+V76L+Q109R+T127V+S144P+S167L+G175;

[0982] T1I+S13Y+T22P+S25P+S39P+S42G+S57W+S59V+V76L+Q109R+T127V+A147H+S167L+G175D;

[0983] T1I+S13Y+T22P+S25P+S39P+S42G+S57W+S59V+V76L+Q109R+S144P+A147H+S167L+G175D;

[0984] T1I+S13Y+T22P+S25P+S39P+S42G+S57W+S59V+V76L+S116D+T127V+S144P+A147H+S167;

[0985] T1I+S13Y+T22P+S25P+S39P+S42G+S57W+S59V+V76L+S116D+T127V+S144P+A147H+G175D;

[0986] T1I+S13Y+T22P+S25P+S39P+S42G+S57W+S59V+V76L+S116D+T127V+S144P+S167L+G175D;

[0987] T1I+S13Y+T22P+S25P+S39P+S42G+S57W+S59V+V76L+S116D+T127V+A147H+S167L+G175D;

[0988] T1I+S13Y+T22P+S25P+S39P+S42G+S57W+S59V+V76L+S116D+S144P+A147H+S167L+G175D;

[0989] T1I+S13Y+T22P+S25P+S39P+S42G+S57W+S59V+V76L+T127V+S144P+A147H+S167L+G175D;

[0990] T1I+S13Y+T22P+S25P+S39P+S42G+S57W+S59V+Q109R+S116D+T127V+S144P+A147H+S167L;

[0991] T1I+S13Y+T22P+S25P+S39P+S42G+S57W+S59V+Q109R+S116D+T127V+S144P+A147H+G175D;

[0992] T1I+S13Y+T22P+S25P+S39P+S42G+S57W+S59V+Q109R+S116D+T127V+S144P+S167L+G175D;

[0993] T1I+S13Y+T22P+S25P+S39P+S42G+S57W+S59V+Q109R+S116D+T127V+A147H+S167L+G175D;

[0994] T1I+S13Y+T22P+S25P+S39P+S42G+S57W+S59V+Q109R+S116D+S144P+A147H+S167L+G175D;

[0995] T1I+S13Y+T22P+S25P+S39P+S42G+S57W+S59V+Q109R+T127V+S144P+A147H+S167L+G175D;

[0996] T1I+S13Y+T22P+S25P+S39P+S42G+S57W+S59V+S116D+T127V+S144P+A147H+S167L+G175D;

[0997] T1I+S13Y+T22P+S25P+S39P+S42G+S57W+V76L+Q109R+S116D+T127V+S144P+A147H+S167L;

[0998] T1I+S13Y+T22P+S25P+S39P+S42G+S57W+V76L+Q109R+S116D+T127V+S144P+A147H+G175D;

[0999] T1I+S13Y+T22P+S25P+S39P+S42G+S57W+V76L+Q109R+S116D+T127V+S144P+S167L+G175D;

[1000] T1I+S13Y+T22P+S25P+S39P+S42G+S57W+V76L+Q109R+S116D+T127V+A147H+S167L+G175D;

[1001] T1I+S13Y+T22P+S25P+S39P+S42G+S57W+V76L+Q109R+S116D+S144P+A147H+S167L+G175D;

[1002] T1I+S13Y+T22P+S25P+S39P+S42G+S57W+V76L+Q109R+T127V+S144P+A147H+S167L+G175D;

[1003] T1I+S13Y+T22P+S25P+S39P+S42G+S57W+V76L+S116D+T127V+S144P+A147H+S167L+G175D;

[1004] T1I+S13Y+T22P+S25P+S39P+S42G+S57W+Q109R+S116D+T127V+S144P+A147H+S167L+G175D;

[1005] T1I+S13Y+T22P+S25P+S39P+S42G+S59V+V76L+Q109R+S116D+T127V+S144P+A147H+S167L;

[1006] T1I+S13Y+T22P+S25P+S39P+S42G+S59V+V76L+Q109R+S116D+T127V+S144P+A147H+G175D;

[1007] T1I+S13Y+T22P+S25P+S39P+S42G+S59V+V76L+Q109R+S116D+T127V+S144P+S167L+G175D;

[1008] T1I+S13Y+T22P+S25P+S39P+S42G+S59V+V76L+Q109R+S116D+T127V+A147H+S167L+G175D;

[1009] T1I+S13Y+T22P+S25P+S39P+S42G+S59V+V76L+Q109R+S116D+S144P+A147H+S167L+G175D;

[1010] T1I+S13Y+T22P+S25P+S39P+S42G+S59V+V76L+Q109R+T127V+S144P+A147H+S167L+G175D;

[1011] T1I+S13Y+T22P+S25P+S39P+S42G+S59V+V76L+S116D+T127V+S144P+A147H+S167L+G175D;

[1012] T1I+S13Y+T22P+S25P+S39P+S42G+S59V+Q109R+S116D+T127V+S144P+A147H+S167L+G175D;

[1013] T1I+S13Y+T22P+S25P+S39P+S42G+V76L+Q109R+S116D+T127V+S144P+A147H+S167L+G175D;

[1014] T1I+S13Y+T22P+S25P+S39P+S57W+S59V+V76L+Q109R+S116D+T127V+S144P+A147H+S167L;

[1015] T1I+S13Y+T22P+S25P+S39P+S57W+S59V+V76L+Q109R+S116D+T127V+S144P+A147H+G175D;

[1016] T1I+S13Y+T22P+S25P+S39P+S57W+S59V+V76L+Q109R+S116D+T127V+S144P+S167L+G175D;

[1017] T1I+S13Y+T22P+S25P+S39P+S57W+S59V+V76L+Q109R+S116D+T127V+A147H+S167L+G175D;

[1018] T1I+S13Y+T22P+S25P+S39P+S57W+S59V+V76L+Q109R+S116D+S144P+A147H+S167L+G175D;

[1019] T1I+S13Y+T22P+S25P+S39P+S57W+S59V+V76L+Q109R+T127V+S144P+A147H+S167L+G175D;

[1020] T1I+S13Y+T22P+S25P+S39P+S57W+S59V+V76L+S116D+T127V+S144P+A147H+S167L+G175D;

[1021] T1I+S13Y+T22P+S25P+S39P+S57W+S59V+Q109R+S116D+T127V+S144P+A147H+S167L+G175D;

[1022] T1I+S13Y+T22P+S25P+S39P+S57W+V76L+Q109R+S116D+T127V+S144P+A147H+S167L+G175D;

[1023] T1I+S13Y+T22P+S25P+S39P+S59V+V76L+Q109R+S116D+T127V+S144P+A147H+S167L+G175D;

[1024] T1I+S13Y+T22P+S25P+S42G+S57W+S59V+V76L+Q109R+S116D+T127V+S144P+A147H+S167L;

[1025] T1I+S13Y+T22P+S25P+S42G+S57W+S59V+V76L+Q109R+S116D+T127V+S144P+A147H+G175D;

[1026] T1I+S13Y+T22P+S25P+S42G+S57W+S59V+V76L+Q109R+S116D+T127V+S144P+S167L+G175D;

[1027] T1I+S13Y+T22P+S25P+S42G+S57W+S59V+V76L+Q109R+S116D+T127V+A147H+S167L+G175D;

[1028] T1I+S13Y+T22P+S25P+S42G+S57W+S59V+V76L+Q109R+S116D+S144P+A147H+S167L+G175D;

[1029] T1I+S13Y+T22P+S25P+S42G+S57W+S59V+V76L+Q109R+T127V+S144P+A147H+S167L+G175D;

[1030] T1I+S13Y+T22P+S25P+S42G+S57W+S59V+V76L+S116D+T127V+S144P+A147H+S167L+G175D;

[1031] T1I+S13Y+T22P+S25P+S42G+S57W+S59V+Q109R+S116D+T127V+S144P+A147H+S167L+G175D;

[1032] T1I+S13Y+T22P+S25P+S42G+S57W+V76L+Q109R+S116D+T127V+S144P+A147H+S167L+G175D;

[1033] T1I+S13Y+T22P+S25P+S42G+S59V+V76L+Q109R+S116D+T127V+S144P+A147H+S167L+G175D;

[1034] T1I+S13Y+T22P+S25P+S57W+S59V+V76L+Q109R+S116D+T127V+S144P+A147H+S167L+G175D;

[1035] T1I+S13Y+T22P+S27L+S39P+S42G+S57W+S59V+V76L+Q109R+S116D+T127V+S144P+A147H;

[1036] T1I+S13Y+T22P+S27L+S39P+S42G+S57W+S59V+V76L+Q109R+S116D+T127V+S144P+S167L;

[1037] T1I+S13Y+T22P+S27L+S39P+S42G+S57W+S59V+V76L+Q109R+S116D+T127V+S144P+G175D;

[1038] T1I+S13Y+T22P+S27L+S39P+S42G+S57W+S59V+V76L+Q109R+S116D+T127V+A147H+S167L;

[1039] T1I+S13Y+T22P+S27L+S39P+S42G+S57W+S59V+V76L+Q109R+S116D+T127V+A147H+G175D;

[1040] T1I+S13Y+T22P+S27L+S39P+S42G+S57W+S59V+V76L+Q109R+S116D+T127V+S167L+G175D;

[1041] T1I+S13Y+T22P+S27L+S39P+S42G+S57W+S59V+V76L+Q109R+S116D+S144P+A147H+S167L;

[1042] T1I+S13Y+T22P+S27L+S39P+S42G+S57W+S59V+V76L+Q109R+S116D+S144P+A147H+G175D;

[1043] T1I+S13Y+T22P+S27L+S39P+S42G+S57W+S59V+V76L+Q109R+S116D+S144P+S167L+G175D;

[1044] T1I+S13Y+T22P+S27L+S39P+S42G+S57W+S59V+V76L+Q109R+S116D+A147H+S167L+G175D;

[1045] T1I+S13Y+T22P+S27L+S39P+S42G+S57W+S59V+V76L+Q109R+T127V+S144P+A147H+S167L;

[1046] T1I+S13Y+T22P+S27L+S39P+S42G+S57W+S59V+V76L+Q109R+T127V+S144P+A147H+G175D;

[1047] T1I+S13Y+T22P+S27L+S39P+S42G+S57W+S59V+V76L+Q109R+T127V+S144P+S167L+G175D;

[1048] T1I+S13Y+T22P+S27L+S39P+S42G+S57W+S59V+V76L+Q109R+T127V+A147H+S167L+G175D;

[1049] T1I+S13Y+T22P+S27L+S39P+S42G+S57W+S59V+V76L+Q109R+S144P+A147H+S167L+G175D;

[1050] T1I+S13Y+T22P+S27L+S39P+S42G+S57W+S59V+V76L+S116D+T127V+S144P+A147H+S167L;

[1051] T1I+S13Y+T22P+S27L+S39P+S42G+S57W+S59V+V76L+S116D+T127V+S144P+A147H+G175D;

[1052] T1I+S13Y+T22P+S27L+S39P+S42G+S57W+S59V+V76L+S116D+T127V+S144P+S167L+G175D;

[1053] T1I+S13Y+T22P+S27L+S39P+S42G+S57W+S59V+V76L+S116D+T127V+A147H+S167L+G175D;

[1054] T1I+S13Y+T22P+S27L+S39P+S42G+S57W+S59V+V76L+S116D+S144P+A147H+S167L+G175D;

[1055] T1I+S13Y+T22P+S27L+S39P+S42G+S57W+S59V+V76L+T127V+S144P+A147H+S167L+G175D;

[1056] T1I+S13Y+T22P+S27L+S39P+S42G+S57W+S59V+Q109R+S116D+T127V+S144P+A147H+S167L;

[1057] T1I+S13Y+T22P+S27L+S39P+S42G+S57W+S59V+Q109R+S116D+T127V+S144P+A147H+G175D;

[1058] T1I+S13Y+T22P+S27L+S39P+S42G+S57W+S59V+Q109R+S116D+T127V+S144P+S167L+G175D;

[1059] T1I+S13Y+T22P+S27L+S39P+S42G+S57W+S59V+Q109R+S116D+T127V+A147H+S167L+G175D;

[1060] T1I+S13Y+T22P+S27L+S39P+S42G+S57W+S59V+Q109R+S116D+S144P+A147H+S167L+G175D;

[1061] T1I+S13Y+T22P+S27L+S39P+S42G+S57W+S59V+Q109R+T127V+S144P+A147H+S167L+G175D;

[1062] T1I+S13Y+T22P+S27L+S39P+S42G+S57W+S59V+S116D+T127V+S144P+A147H+S167L+G175D;

[1063] T1I+S13Y+T22P+S27L+S39P+S42G+S57W+V76L+Q109R+S116D+T127V+S144P+A147H+S167L;

[1064] T1I+S13Y+T22P+S27L+S39P+S42G+S57W+V76L+Q109R+S116D+T127V+S144P+A147H+G175D;

[1065] T1I+S13Y+T22P+S27L+S39P+S42G+S57W+V76L+Q109R+S116D+T127V+S144P+S167L+G175D;

[1066] T1I+S13Y+T22P+S27L+S39P+S42G+S57W+V76L+Q109R+S116D+T127V+A147H+S167L+G175D;

[1067] T1I+S13Y+T22P+S27L+S39P+S42G+S57W+V76L+Q109R+S116D+S144P+A147H+S167L+G175D;

[1068] T1I+S13Y+T22P+S27L+S39P+S42G+S57W+V76L+Q109R+T127V+S144P+A147H+S167L+G175D;

[1069] T1I+S13Y+T22P+S27L+S39P+S42G+S57W+V76L+S116D+T127V+S144P+A147H+S167L+G175D;

[1070] T1I+S13Y+T22P+S27L+S39P+S42G+S57W+Q109R+S116D+T127V+S144P+A147H+S167L+G175D;

[1071] T1I+S13Y+T22P+S27L+S39P+S42G+S59V+V76L+Q109R+S116D+T127V+S144P+A147H+S167L;

[1072] T1I+S13Y+T22P+S27L+S39P+S42G+S59V+V76L+Q109R+S116D+T127V+S144P+A147H+G175D;

[1073] T1I+S13Y+T22P+S27L+S39P+S42G+S59V+V76L+Q109R+S116D+T127V+S144P+S167L+G175D;

[1074] T1I+S13Y+T22P+S27L+S39P+S42G+S59V+V76L+Q109R+S116D+T127V+A147H+S167L+G175D;

[1075] T1I+S13Y+T22P+S27L+S39P+S42G+S59V+V76L+Q109R+S116D+S144P+A147H+S167L+G175D;

[1076] T1I+S13Y+T22P+S27L+S39P+S42G+S59V+V76L+Q109R+T127V+S144P+A147H+S167L+G175D;

[1077] T1I+S13Y+T22P+S27L+S39P+S42G+S59V+V76L+S116D+T127V+S144P+A147H+S167L+G175D;

[1078] T1I+S13Y+T22P+S27L+S39P+S42G+S59V+Q109R+S116D+T127V+S144P+A147H+S167L+G175D;

[1079] T1I+S13Y+T22P+S27L+S39P+S42G+V76L+Q109R+S116D+T127V+S144P+A147H+S167L+G175D;

[1080] T1I+S13Y+T22P+S27L+S39P+S57W+S59V+V76L+Q109R+S116D+T127V+S144P+A147H+S167L;

[1081] T1I+S13Y+T22P+S27L+S39P+S57W+S59V+V76L+Q109R+S116D+T127V+S144P+A147H+G175D;

[1082] T1I+S13Y+T22P+S27L+S39P+S57W+S59V+V76L+Q109R+S116D+T127V+S144P+S167L+G175D;

[1083] T1I+S13Y+T22P+S27L+S39P+S57W+S59V+V76L+Q109R+S116D+T127V+A147H+S167L+G175D;

[1084] T1I+S13Y+T22P+S27L+S39P+S57W+S59V+V76L+Q109R+S116D+S144P+A147H+S167L+G175D;

[1085] T1I+S13Y+T22P+S27L+S39P+S57W+S59V+V76L+Q109R+T127V+S144P+A147H+S167L+G175D;

[1086] T1I+S13Y+T22P+S27L+S39P+S57W+S59V+V76L+S116D+T127V+S144P+A147H+S167L+G175D;

[1087] T1I+S13Y+T22P+S27L+S39P+S57W+S59V+Q109R+S116D+T127V+S144P+A147H+S167L+G175D;

[1088] T1I+S13Y+T22P+S27L+S39P+S57W+V76L+Q109R+S116D+T127V+S144P+A147H+S167L+G175D;

[1089] T1I+S13Y+T22P+S27L+S39P+S59V+V76L+Q109R+S116D+T127V+S144P+A147H+S167L+G175D;

[1090] T1I+S13Y+T22P+S27L+S42G+S57W+S59V+V76L+Q109R+S116D+T127V+S144P+A147H+S167L;

[1091] T1I+S13Y+T22P+S27L+S42G+S57W+S59V+V76L+Q109R+S116D+T127V+S144P+A147H+G175D;

[1092] T1I+S13Y+T22P+S27L+S42G+S57W+S59V+V76L+Q109R+S116D+T127V+S144P+S167L+G175D;

[1093] T1I+S13Y+T22P+S27L+S42G+S57W+S59V+V76L+Q109R+S116D+T127V+A147H+S167L+G175D;

[1094] T1I+S13Y+T22P+S27L+S42G+S57W+S59V+V76L+Q109R+S116D+S144P+A147H+S167L+G175D;

[1095] T1I+S13Y+T22P+S27L+S42G+S57W+S59V+V76L+Q109R+T127V+S144P+A147H+S167L+G175D;

[1096] T1I+S13Y+T22P+S27L+S42G+S57W+S59V+V76L+S116D+T127V+S144P+A147H+S167L+G175D;

[1097] T1I+S13Y+T22P+S27L+S42G+S57W+S59V+Q109R+S116D+T127V+S144P+A147H+S167L+G175D;

[1098] T1I+S13Y+T22P+S27L+S42G+S57W+V76L+Q109R+S116D+T127V+S144P+A147H+S167L+G175D;

[1099] T1I+S13Y+T22P+S27L+S42G+S59V+V76L+Q109R+S116D+T127V+S144P+A147H+S167L+G175D;

[1100] T1I+S13Y+T22P+S27L+S57W+S59V+V76L+Q109R+S116D+T127V+S144P+A147H+S167L+G175D;

[1101] T1I+S13Y+T22P+S39P+S42G+S57W+S59V+V76L+Q109R+S116D+T127V+S144P+A147H+S167L;

[1102] T1I+S13Y+T22P+S39P+S42G+S57W+S59V+V76L+Q109R+S116D+T127V+S144P+A147H+G175D;

[1103] T1I+S13Y+T22P+S39P+S42G+S57W+S59V+V76L+Q109R+S116D+T127V+S144P+S167L+G175D;

[1104] T1I+S13Y+T22P+S39P+S42G+S57W+S59V+V76L+Q109R+S116D+T127V+A147H+S167L+G175D;

[1105] T1I+S13Y+T22P+S39P+S42G+S57W+S59V+V76L+Q109R+S116D+S144P+A147H+S167L+G175D;

[1106] T1I+S13Y+T22P+S39P+S42G+S57W+S59V+V76L+Q109R+T127V+S144P+A147H+S167L+G175D;

[1107] T1I+S13Y+T22P+S39P+S42G+S57W+S59V+V76L+S116D+T127V+S144P+A147H+S167L+G175D;

[1108] T1I+S13Y+T22P+S39P+S42G+S57W+S59V+Q109R+S116D+T127V+S144P+A147H+S167L+G175D;

[1109] T1I+S13Y+T22P+S39P+S42G+S57W+V76L+Q109R+S116D+T127V+S144P+A147H+S167L+G175D;

[1110] T1I+S13Y+T22P+S39P+S42G+S59V+V76L+Q109R+S116D+T127V+S144P+A147H+S167L+G175D;

[1111] T1I+S13Y+T22P+S39P+S57W+S59V+V76L+Q109R+S116D+T127V+S144P+A147H+S167L+G175D;

[1112] T1I+S13Y+T22P+S42G+S57W+S59V+V76L+Q109R+S116D+T127V+S144P+A147H+S167L+G175D;

[1113] T1I+S13Y+S25P+S27L+S39P+S42G+S57W+S59V+V76L+Q109R+S116D+T127V+S144P+A147H;

[1114] T1I+S13Y+S25P+S27L+S39P+S42G+S57W+S59V+V76L+Q109R+S116D+T127V+S144P+S167L;

[1115] T1I+S13Y+S25P+S27L+S39P+S42G+S57W+S59V+V76L+Q109R+S116D+T127V+S144P+G175D;

[1116] T1I+S13Y+S25P+S27L+S39P+S42G+S57W+S59V+V76L+Q109R+S116D+T127V+A147H+S167L

[1117] T1I+S13Y+S25P+S27L+S39P+S42G+S57W+S59V+V76L+Q109R+S116D+T127V+A147H+G175D;

[1118] T1I+S13Y+S25P+S27L+S39P+S42G+S57W+S59V+V76L+Q109R+S116D+T127V+S167L+G175D;

[1119] T1I+S13Y+S25P+S27L+S39P+S42G+S57W+S59V+V76L+Q109R+S116D+S144P+A147H+S167L;

[1120] T1I+S13Y+S25P+S27L+S39P+S42G+S57W+S59V+V76L+Q109R+S116D+S144P+A147H+G175D;

[1121] T1I+S13Y+S25P+S27L+S39P+S42G+S57W+S59V+V76L+Q109R+S116D+S144P+S167L+G175D;

[1122] T1I+S13Y+S25P+S27L+S39P+S42G+S57W+S59V+V76L+Q109R+S116D+A147H+S167L+G175D;

[1123] T1I+S13Y+S25P+S27L+S39P+S42G+S57W+S59V+V76L+Q109R+T127V+S144P+A147H+S167L

[1124] T1I+S13Y+S25P+S27L+S39P+S42G+S57W+S59V+V76L+Q109R+T127V+S144P+A147H+G175D;

[1125] T1I+S13Y+S25P+S27L+S39P+S42G+S57W+S59V+V76L+Q109R+T127V+S144P+S167L+G175D;

[1126] T1I+S13Y+S25P+S27L+S39P+S42G+S57W+S59V+V76L+Q109R+T127V+A147H+S167L+G175D;

[1127] T1I+S13Y+S25P+S27L+S39P+S42G+S57W+S59V+V76L+Q109R+S144P+A147H+S167L+G175D;

[1128] T1I+S13Y+S25P+S27L+S39P+S42G+S57W+S59V+V76L+S116D+T127V+S144P+A147H+S167L;

[1129] T1I+S13Y+S25P+S27L+S39P+S42G+S57W+S59V+V76L+S116D+T127V+S144P+A147H+G175D;

[1130] T1I+S13Y+S25P+S27L+S39P+S42G+S57W+S59V+V76L+S116D+T127V+S144P+S167L+G175D;

[1131] T1I+S13Y+S25P+S27L+S39P+S42G+S57W+S59V+V76L+S116D+T127V+A147H+S167L+G175D;

[1132] T1I+S13Y+S25P+S27L+S39P+S42G+S57W+S59V+V76L+S116D+S144P+A147H+S167L+G175D;

[1133] T1I+S13Y+S25P+S27L+S39P+S42G+S57W+S59V+V76L+T127V+S144P+A147H+S167L+G175D;

[1134] T1I+S13Y+S25P+S27L+S39P+S42G+S57W+S59V+Q109R+S116D+T127V+S144P+A147H+S167L;

[1135] T1I+S13Y+S25P+S27L+S39P+S42G+S57W+S59V+Q109R+S116D+T127V+S144P+A147H+G175D;

[1136] T1I+S13Y+S25P+S27L+S39P+S42G+S57W+S59V+Q109R+S116D+T127V+S144P+S167L+G175D;

[1137] T1I+S13Y+S25P+S27L+S39P+S42G+S57W+S59V+Q109R+S116D+T127V+A147H+S167L+G175D;

[1138] T1I+S13Y+S25P+S27L+S39P+S42G+S57W+S59V+Q109R+S116D+S144P+A147H+S167L+G175D;

[1139] T1I+S13Y+S25P+S27L+S39P+S42G+S57W+S59V+Q109R+T127V+S144P+A147H+S167L+G175D;

[1140] T1I+S13Y+S25P+S27L+S39P+S42G+S57W+S59V+S116D+T127V+S144P+A147H+S167L+G175D;

[1141] T1I+S13Y+S25P+S27L+S39P+S42G+S57W+V76L+Q109R+S116D+T127V+S144P+A147H+S167L;

[1142] T1I+S13Y+S25P+S27L+S39P+S42G+S57W+V76L+Q109R+S116D+T127V+S144P+A147H+G175D;

[1143] T1I+S13Y+S25P+S27L+S39P+S42G+S57W+V76L+Q109R+S116D+T127V+S144P+S167L+G175D;

[1144] T1I+S13Y+S25P+S27L+S39P+S42G+S57W+V76L+Q109R+S116D+T127V+A147H+S167L+G175D;

[1145] T1I+S13Y+S25P+S27L+S39P+S42G+S57W+V76L+Q109R+S116D+S144P+A147H+S167L+G175D;

[1146] T1I+S13Y+S25P+S27L+S39P+S42G+S57W+V76L+Q109R+T127V+S144P+A147H+S167L+G175D;

[1147] T1I+S13Y+S25P+S27L+S39P+S42G+S57W+V76L+S116D+T127V+S144P+A147H+S167L+G175D;

[1148] T1I+S13Y+S25P+S27L+S39P+S42G+S57W+Q109R+S116D+T127V+S144P+A147H+S167L+G175D;

[1149] T1I+S13Y+S25P+S27L+S39P+S42G+S59V+V76L+Q109R+S116D+T127V+S144P+A147H+S167L;

[1150] T1I+S13Y+S25P+S27L+S39P+S42G+S59V+V76L+Q109R+S116D+T127V+S144P+A147H+G175D;

[1151] T1I+S13Y+S25P+S27L+S39P+S42G+S59V+V76L+Q109R+S116D+T127V+S144P+S167L+G175D;

[1152] T1I+S13Y+S25P+S27L+S39P+S42G+S59V+V76L+Q109R+S116D+T127V+A147H+S167L+G175D;

[1153] T1I+S13Y+S25P+S27L+S39P+S42G+S59V+V76L+Q109R+S116D+S144P+A147H+S167L+G175D;

[1154] T1I+S13Y+S25P+S27L+S39P+S42G+S59V+V76L+Q109R+T127V+S144P+A147H+S167L+G175D;

[1155] T1I+S13Y+S25P+S27L+S39P+S42G+S59V+V76L+S116D+T127V+S144P+A147H+S167L+G175D;

[1156] T1I+S13Y+S25P+S27L+S39P+S42G+S59V+Q109R+S116D+T127V+S144P+A147H+S167L+G175D;

[1157] T1I+S13Y+S25P+S27L+S39P+S42G+V76L+Q109R+S116D+T127V+S144P+A147H+S167L+G175D;

[1158] T1I+S13Y+S25P+S27L+S39P+S57W+S59V+V76L+Q109R+S116D+T127V+S144P+A147H+S167L;

[1159] T1I+S13Y+S25P+S27L+S39P+S57W+S59V+V76L+Q109R+S116D+T127V+S144P+A147H+G175D;

[1160] T1I+S13Y+S25P+S27L+S39P+S57W+S59V+V76L+Q109R+S116D+T127V+S144P+S167L+G175D;

[1161] T1I+S13Y+S25P+S27L+S39P+S57W+S59V+V76L+Q109R+S116D+T127V+A147H+S167L+G175D;

[1162] T1I+S13Y+S25P+S27L+S39P+S57W+S59V+V76L+Q109R+S116D+S144P+A147H+S167L+G175D;

[1163] T1I+S13Y+S25P+S27L+S39P+S57W+S59V+V76L+Q109R+T127V+S144P+A147H+S167L+G175D;

[1164] T1I+S13Y+S25P+S27L+S39P+S57W+S59V+V76L+S116D+T127V+S144P+A147H+S167L+G175D;

[1165] T1I+S13Y+S25P+S27L+S39P+S57W+S59V+Q109R+S116D+T127V+S144P+A147H+S167L+G175D;

[1166] T1I+S13Y+S25P+S27L+S39P+S57W+V76L+Q109R+S116D+T127V+S144P+A147H+S167L+G175D;

[1167] T1I+S13Y+S25P+S27L+S39P+S59V+V76L+Q109R+S116D+T127V+S144P+A147H+S167L+G175D;

[1168] T1I+S13Y+S25P+S27L+S42G+S57W+S59V+V76L+Q109R+S116D+T127V+S144P+A147H+S167L;

[1169] T1I+S13Y+S25P+S27L+S42G+S57W+S59V+V76L+Q109R+S116D+T127V+S144P+A147H+G175D;

[1170] T1I+S13Y+S25P+S27L+S42G+S57W+S59V+V76L+Q109R+S116D+T127V+S144P+S167L+G175D;

[1171] T1I+S13Y+S25P+S27L+S42G+S57W+S59V+V76L+Q109R+S116D+T127V+A147H+S167L+G175D;

[1172] T1I+S13Y+S25P+S27L+S42G+S57W+S59V+V76L+Q109R+S116D+S144P+A147H+S167L+G175D;

[1173] T1I+S13Y+S25P+S27L+S42G+S57W+S59V+V76L+Q109R+T127V+S144P+A147H+S167L+G175D;

[1174] T1I+S13Y+S25P+S27L+S42G+S57W+S59V+V76L+S116D+T127V+S144P+A147H+S167L+G175D;

[1175] T1I+S13Y+S25P+S27L+S42G+S57W+S59V+Q109R+S116D+T127V+S144P+A147H+S167L+G175D;

[1176] T1I+S13Y+S25P+S27L+S42G+S57W+V76L+Q109R+S116D+T127V+S144P+A147H+S167L+G175D;

[1177] T1I+S13Y+S25P+S27L+S42G+S59V+V76L+Q109R+S116D+T127V+S144P+A147H+S167L+G175D;

[1178] T1I+S13Y+S25P+S27L+S57W+S59V+V76L+Q109R+S116D+T127V+S144P+A147H+S167L+G175D;

[1179] T1I+S13Y+S25P+S39P+S42G+S57W+S59V+V76L+Q109R+S116D+T127V+S144P+A147H+S167L;

[1180] T1I+S13Y+S25P+S39P+S42G+S57W+S59V+V76L+Q109R+S116D+T127V+S144P+A147H+G175D;

[1181] T1I+S13Y+S25P+S39P+S42G+S57W+S59V+V76L+Q109R+S116D+T127V+S144P+S167L+G175D;

[1182] T1I+S13Y+S25P+S39P+S42G+S57W+S59V+V76L+Q109R+S116D+T127V+A147H+S167L+G175D;

[1183] T1I+S13Y+S25P+S39P+S42G+S57W+S59V+V76L+Q109R+S116D+S144P+A147H+S167L+G175D;

[1184] T1I+S13Y+S25P+S39P+S42G+S57W+S59V+V76L+Q109R+T127V+S144P+A147H+S167L+G175D;

[1185] T1I+S13Y+S25P+S39P+S42G+S57W+S59V+V76L+S116D+T127V+S144P+A147H+S167L+G175D;

[1186] T1I+S13Y+S25P+S39P+S42G+S57W+S59V+Q109R+S116D+T127V+S144P+A147H+S167L+G175D;

[1187] T1I+S13Y+S25P+S39P+S42G+S57W+V76L+Q109R+S116D+T127V+S144P+A147H+S167L+G175D;

[1188] T1I+S13Y+S25P+S39P+S42G+S59V+V76L+Q109R+S116D+T127V+S144P+A147H+S167L+G175D;

[1189] T1I+S13Y+S25P+S39P+S57W+S59V+V76L+Q109R+S116D+T127V+S144P+A147H+S167L+G175D;

[1190] T1I+S13Y+S25P+S42G+S57W+S59V+V76L+Q109R+S116D+T127V+S144P+A147H+S167L+G175D;

[1191] T1I+S13Y+S27L+S39P+S42G+S57W+S59V+V76L+Q109R+S116D+T127V+S144P+A147H+S167L;

[1192] T1I+S13Y+S27L+S39P+S42G+S57W+S59V+V76L+Q109R+S116D+T127V+S144P+A147H+G175D;

[1193] T1I+S13Y+S27L+S39P+S42G+S57W+S59V+V76L+Q109R+S116D+T127V+S144P+S167L+G175D;

[1194] T1I+S13Y+S27L+S39P+S42G+S57W+S59V+V76L+Q109R+S116D+T127V+A147H+S167L+G175D;

[1195] T1I+S13Y+S27L+S39P+S42G+S57W+S59V+V76L+Q109R+S116D+S144P+A147H+S167L+G175D;

[1196] T1I+S13Y+S27L+S39P+S42G+S57W+S59V+V76L+Q109R+T127V+S144P+A147H+S167L+G175D;

[1197] T1I+S13Y+S27L+S39P+S42G+S57W+S59V+V76L+S116D+T127V+S144P+A147H+S167L+G175D;

[1198] T1I+S13Y+S27L+S39P+S42G+S57W+S59V+Q109R+S116D+T127V+S144P+A147H+S167L+G175D;

[1199] T1I+S13Y+S27L+S39P+S42G+S57W+V76L+Q109R+S116D+T127V+S144P+A147H+S167L+G175D;

[1200] T1I+S13Y+S27L+S39P+S42G+S59V+V76L+Q109R+S116D+T127V+S144P+A147H+S167L+G175D;

[1201] T1I+S13Y+S27L+S39P+S57W+S59V+V76L+Q109R+S116D+T127V+S144P+A147H+S167L+G175D;

[1202] T1I+S13Y+S27L+S42G+S57W+S59V+V76L+Q109R+S116D+T127V+S144P+A147H+S167L+G175D;

[1203] T1I+S13Y+S39P+S42G+S57W+S59V+V76L+Q109R+S116D+T127V+S144P+A147H+S167L+G175D;

[1204] T1I+T22P+S25P+S27L+S39P+S42G+S57W+S59V+V76L+Q109R+S116D+T127V+S144P+A147H;

[1205] T1I+T22P+S25P+S27L+S39P+S42G+S57W+S59V+V76L+Q109R+S116D+T127V+S144P+S167L;

[1206] T1I+T22P+S25P+S27L+S39P+S42G+S57W+S59V+V76L+Q109R+S116D+T127V+S144P+G175D;

[1207] T1I+T22P+S25P+S27L+S39P+S42G+S57W+S59V+V76L+Q109R+S116D+T127V+A147H+S167L;

[1208] T1I+T22P+S25P+S27L+S39P+S42G+S57W+S59V+V76L+Q109R+S116D+T127V+A147H+G175D;

[1209] T1I+T22P+S25P+S27L+S39P+S42G+S57W+S59V+V76L+Q109R+S116D+T127V+S167L+G175D;

[1210] T1I+T22P+S25P+S27L+S39P+S42G+S57W+S59V+V76L+Q109R+S116D+S144P+A147H+S167L;

[1211] T1I+T22P+S25P+S27L+S39P+S42G+S57W+S59V+V76L+Q109R+S116D+S144P+A147H+G175D;

[1212] T1I+T22P+S25P+S27L+S39P+S42G+S57W+S59V+V76L+Q109R+S116D+S144P+S167L+G175D;

[1213] T1I+T22P+S25P+S27L+S39P+S42G+S57W+S59V+V76L+Q109R+S116D+A147H+S167L+G175D;

[1214] T1I+T22P+S25P+S27L+S39P+S42G+S57W+S59V+V76L+Q109R+T127V+S144P+A147H+S167L;

[1215] T1I+T22P+S25P+S27L+S39P+S42G+S57W+S59V+V76L+Q109R+T127V+S144P+A147H+G175D;

[1216] T1I+T22P+S25P+S27L+S39P+S42G+S57W+S59V+V76L+Q109R+T127V+S144P+S167L+G175D;

[1217] T1I+T22P+S25P+S27L+S39P+S42G+S57W+S59V+V76L+Q109R+T127V+A147H+S167L+G175D;

[1218] T1I+T22P+S25P+S27L+S39P+S42G+S57W+S59V+V76L+Q109R+S144P+A147H+S167L+G175D;

[1219] T1I+T22P+S25P+S27L+S39P+S42G+S57W+S59V+V76L+S116D+T127V+S144P+A147H+S167L;

[1220] T1I+T22P+S25P+S27L+S39P+S42G+S57W+S59V+V76L+S116D+T127V+S144P+A147H+G175D;

[1221] T1I+T22P+S25P+S27L+S39P+S42G+S57W+S59V+V76L+S116D+T127V+S144P+S167L+G175D;

[1222] T1I+T22P+S25P+S27L+S39P+S42G+S57W+S59V+V76L+S116D+T127V+A147H+S167L+G175D;

[1223] T1I+T22P+S25P+S27L+S39P+S42G+S57W+S59V+V76L+S116D+S144P+A147H+S167L+G175D;

[1224] T1I+T22P+S25P+S27L+S39P+S42G+S57W+S59V+V76L+T127V+S144P+A147H+S167L+G175D;

[1225] T1I+T22P+S25P+S27L+S39P+S42G+S57W+S59V+Q109R+S116D+T127V+S144P+A147H+S167L;

[1226] T1I+T22P+S25P+S27L+S39P+S42G+S57W+S59V+Q109R+S116D+T127V+S144P+A147H+G175D;

[1227] T1I+T22P+S25P+S27L+S39P+S42G+S57W+S59V+Q109R+S116D+T127V+S144P+S167L+G175D;

[1228] T1I+T22P+S25P+S27L+S39P+S42G+S57W+S59V+Q109R+S116D+T127V+A147H+S167L+G175D;

[1229] T1I+T22P+S25P+S27L+S39P+S42G+S57W+S59V+Q109R+S116D+S144P+A147H+S167L+G175D;

[1230] T1I+T22P+S25P+S27L+S39P+S42G+S57W+S59V+Q109R+T127V+S144P+A147H+S167L+G175D;

[1231] T1I+T22P+S25P+S27L+S39P+S42G+S57W+S59V+S116D+T127V+S144P+A147H+S167L+G175D;

[1232] T1I+T22P+S25P+S27L+S39P+S42G+S57W+V76L+Q109R+S116D+T127V+S144P+A147H+S167L;

[1233] T1I+T22P+S25P+S27L+S39P+S42G+S57W+V76L+Q109R+S116D+T127V+S144P+A147H+G175D;

[1234] T1I+T22P+S25P+S27L+S39P+S42G+S57W+V76L+Q109R+S116D+T127V+S144P+S167L+G175D;

[1235] T1I+T22P+S25P+S27L+S39P+S42G+S57W+V76L+Q109R+S116D+T127V+A147H+S167L+G175D;

[1236] T1I+T22P+S25P+S27L+S39P+S42G+S57W+V76L+Q109R+S116D+S144P+A147H+S167L+G175D;

[1237] T1I+T22P+S25P+S27L+S39P+S42G+S57W+V76L+Q109R+T127V+S144P+A147H+S167L+G175D;

[1238] T1I+T22P+S25P+S27L+S39P+S42G+S57W+V76L+S116D+T127V+S144P+A147H+S167L+G175D;

[1239] T1I+T22P+S25P+S27L+S39P+S42G+S57W+Q109R+S116D+T127V+S144P+A147H+S167L+G175D;

[1240] T1I+T22P+S25P+S27L+S39P+S42G+S59V+V76L+Q109R+S116D+T127V+S144P+A147H+S167L;

[1241] T1I+T22P+S25P+S27L+S39P+S42G+S59V+V76L+Q109R+S116D+T127V+S144P+A147H+G175D;

[1242] T1I+T22P+S25P+S27L+S39P+S42G+S59V+V76L+Q109R+S116D+T127V+S144P+S167L+G175D;

[1243] T1I+T22P+S25P+S27L+S39P+S42G+S59V+V76L+Q109R+S116D+T127V+A147H+S167L+G175D;

[1244] T1I+T22P+S25P+S27L+S39P+S42G+S59V+V76L+Q109R+S116D+S144P+A147H+S167L+G175D;

[1245] T1I+T22P+S25P+S27L+S39P+S42G+S59V+V76L+Q109R+T127V+S144P+A147H+S167L+G175D;

[1246] T1I+T22P+S25P+S27L+S39P+S42G+S59V+V76L+S116D+T127V+S144P+A147H+S167L+G175D;

[1247] T1I+T22P+S25P+S27L+S39P+S42G+S59V+Q109R+S116D+T127V+S144P+A147H+S167L+G175D;

[1248] T1I+T22P+S25P+S27L+S39P+S42G+V76L+Q109R+S116D+T127V+S144P+A147H+S167L+G175D;

[1249] T1I+T22P+S25P+S27L+S39P+S57W+S59V+V76L+Q109R+S116D+T127V+S144P+A147H+S167L;

[1250] T1I+T22P+S25P+S27L+S39P+S57W+S59V+V76L+Q109R+S116D+T127V+S144P+A147H+G175D;

[1251] T1I+T22P+S25P+S27L+S39P+S57W+S59V+V76L+Q109R+S...

Claims

1. A DNA enzyme variant, wherein the substitution of the DNA enzyme variant relative to SEQ ID NO: 1 is selected from the group consisting of: i.T1I+S13Y+T22P+S27L+S42G+S57W+S59V+V76L+Q109R+S116D+T127V+S144P+A147H+S167L+G175D; ii.T1I+S13Y+T22P+S25P+S27L+S39P+S42G+S57W+S59V+V76L+Q109R+S116D+T127V+S144P+A147H+S167L+G175D; iii.T1I+S13Y+T22P+S27L+S39P+S42G+D56I+S57W+S59V+V76L+Q109R+S116D+T127V+S144P+A147H+S167L+G175D; iv.T1I+S13Y+T22P+S25P+S27L+S39P+S42G+D56I+S57W+S59V+V76L+Q109R+S116D+T127V+S144P+A147H+S167L+G175D; v.T1I+S13Y+T22P+S25P+S27L+S39P+S42G+D56I+S57W+S59V+V76L+Q109R+S116D+T127V+S144P+A147H+Q166D+S167L+G175D; vi.T1I+S13Y+T22P+S27R+S39P+S42G+D56I+S57W+S59V+V76L+Q109R+S116D+T127V+S144P+A147H+S167L+G175D; vii.T1I+S13Y+T22P+S27L+S39P+S42G+D56I+S57W+S59V+T65L+V76L+Q109R+S116D+T127V+S144P+A147H+S167L+G175D; viii.T1I+S13Y+T22P+S27L+L33K+S39P+S42G+D56I+S57W+S59V+T65V+V76L+Q109R+S116D+T127V+S144P+A147H+S167L+G175D; ix.T1I+S13Y+T22P+S25P+S27R+S39P+S42G+S57W+S59V+S66W+V76L+T77Y+Q109R+S116D+T127V+S144P+A147H+S167L+G175D; x.T1I + S13Y + T22P + S25P + S27L + L33K + S39P + S42G + S57W + S59V + T65V + V76L + T77Y + Q109R + S116D + T127V + S144P + A147H + S167L + G175D; xi.T1I + S13Y + T22P + S25P + S27L + L33K + S39P + S42G + S57W + S59V + S66Y + V76L + T77Y + Q109R + S116D + T127V + S144P + A147H + S167L + G175D; xii.T1I + S13Y + T22P + S25P + S27L + S39P + S42G + S57W + S59V + T65V + S66Y + V76L + T77Y + Q109R + S116D + T127V + S144P + A147H + S167L + G175D; xiii.T1I + S13Y + T22P + S27L + S39P + S42G + D56L + S57W + S59V + T65V + V76L + Q109R + S116D + T127V + S144P + A147H + G162D + S167L + G175D; xiv.T1I + S13Y + T22P + S27R + S39P + S42G + D56L + S57W + S59V + T65V + V76L + Q109R + S116D + T127V + S144P + A147H + S167L + G175D; xv.T1I + S13Y + T22P + S27R + S39P + S42G + D56L + S57W + S59V + T65V + V76L + Q109R + S116D + T127V + S144P + A147H + G162D + S167L + G175D; xvi.T1I + S13Y + T22P + S27K + S39P + S42G + D56I + S57W + S59V + T65V + V76L + S106L + Q109R + S116D + T127V + S144P + A147H + S167L + G175D; xvii.T1I + S13Y + T22P + S27K + S39P + S42G + D56I + S57W + S59V + T65V + V76L + Q109R + S116D + T127V + S130A + S144P + A147H + S167L + G175D; xviii.T1I + S13Y + T22P + S27L + S39P + S42G + D56L + S57W + S59V + T65L + V76L + Q109R + S116D + T127V + S130A + S144P + A147H + S167L + G175D; and xix.T1I+S13Y+T22P+S27L+S39P+S42G+D56I+S57W+S59V+T65V+V76L+Q109R+S116D+T127V+S144P+A147H+G162D+S167L+G175D.

2. DNA enzyme variant according to claim 1, wherein compared with the DNA enzyme with the peptide sequence shown in SEQ ID NO:1, described DNA enzyme variant has improved stability, and the stability measurement of this improvement is to have> improvement factor of 1.0, wherein said improvement factor is residual activity ratio.

3. DNA enzyme variants according to claim 1 or 2, wherein compared with the DNA enzyme with the peptide sequence shown in SEQ ID NO:28, described DNA enzyme variants has improved stability, and the stability measurement of this improvement is with> improvement factor of 1.0, wherein said improvement factor is residual activity ratio.

4. according to the DNA enzyme variant described in any one of claims 1-3, wherein said DNA enzyme variant has the improved stability as measured by the half-life improvement factor, wherein the half-life improvement factor compared to SEQ ID NO:1 is at least 1.

2.

5. according to the DNA enzyme variant described in any one in claim 1-3, wherein said DNA enzyme variant has the improved stability as measured by the half-life improvement factor, and wherein the half-life improvement factor compared to SEQ ID NO:28 is at least 1.

2.

6. The DNA enzyme variant according to any one of claims 1-5, wherein the combination of substitutions provides a synergistic effect.

7. A method for obtaining a DNA enzyme variant, the method comprising: a) introducing the substitution defined in claim 1 into the parent DNA enzyme of SEQ ID NO: 1, b) and recovering the variant.

8. Use of the DNA enzyme variant according to any one of claims 1 to 6 for deep cleaning of an item, wherein the item is a textile or a hard surface.

9. An isolated polynucleotide encoding the DNase variant according to any one of claims 1 to 6.

10. A nucleic acid construct or an expression vector capable of expressing the polynucleotide according to claim 9. A host cell comprising the polynucleotide according to claim 9 .

12. A method for producing a DNA enzyme variant, the method comprising: i) culturing the host cell according to claim 11 under conditions suitable for expressing the DNase variant; as well as ii) recovering the DNase variant.

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