RU2783315C2 - Porcine trypsin options - Google Patents

Abstract

FIELD: biotechnology.

SUBSTANCE: invention relates to biotechnology, in particular to polypeptide options of porcine trypsin, nucleic acid molecules encoding these options, and host cells containing such nucleic acid molecules. It also relates to the use of these options in methods for the production of insulin.

EFFECT: invention allows for obtainment of an effective option of trypsin and obtainment of insulin with a high degree of efficiency.

16 cl, 4 dwg, 3 tbl, 8 ex

Description

The field of technology to which the invention belongs

The present invention relates to polypeptide variants of porcine trypsin, nucleic acid molecules encoding these variants, and host cells containing such nucleic acid molecules. It also relates to the use of these variants in methods for the production of insulin. The present invention further relates to the use of these variants as drugs, as food ingredients, or as feed ingredients, and to the use of these variants in a method for making a food ingredient or feed ingredient.

Background of the invention

Trypsin, which is an endopeptidase, is used to make human insulin, insulin analogs, and insulin derivatives. Human insulin, insulin analog or insulin derivative is prepared from preproinsulin (PPI) by enzymatic hydrolysis using trypsin, wherein the presequence and C-peptide are cleaved to obtain the respective products (see FIG. 1). Trypsin is specific for C-terminal cleavage of Arg and Lys residues in the polypeptide chain. In human preproinsulin and the preproinsulin glargine polypeptide, several Arg and Lys residues are present, including B29Lys, B31Arg and B32Arg at the junction of the B chain and C peptide. Therefore, human preproinsulin and preproinsulin glargine are trypsin-cleaved at the C-terminal side of Arg and Lys residues, including the C-terminal side of B29Lys and B31Arg and B32Arg. For the manufacture of insulin glargine, trypsin cleavage after B32Arg provides the final insulin glargine. Trypsin digestion after B29Lys and B31Arg results in the by-products desB30-Thr ("des-Thr")-insulin glargine and desB32Arg-insulin glargine (also referred to as "des-Arg"). Therefore, it is necessary to remove by-products during the manufacture of insulin glargine.

For the manufacture of human insulin, trypsin digestion after B31Arg results in human B31Arg-insulin (also referred to as "mono-Arg"), and after B32Arg to human B31Arg-B32Arg-insulin (also referred to as "di-Arg"). Both are intermediates in said manufacturing process. In the subsequent manufacturing process, both of these two intermediates are converted to the final human insulin. Trypsin digestion after B29Lys results in the by-product human desB30-Thr ("des-Thr")-insulin. Therefore, it is necessary to remove the by-product during the manufacture of human insulin.

In the case of human insulin and insulin glargine, trypsin digestion after B32Arg results in the final insulin glargine, and in the case of human insulin, it results in an intermediate that is converted to final human insulin in subsequent process steps.

Previous development to eliminate the undesirable formation of the desB30-Thr intermediate resulted in the porcine trypsin variant S172A, which reduces the likelihood of this miscleavage by increasing selectivity for the C-terminal side of Arg over Lys. The porcine trypsin S172A variant is described in WO 2007/031187 A1, which is incorporated herein by reference in its entirety.

At present, in the course of a thorough study by the present inventors of the kinetics of the S172A trypsin-catalyzed reaction, it has been found that the formation of incorrectly cleaved by-products will occur not only due to the primary incorrect cleavage of PPI by trypsin acting as an endoprotease, as described in the literature, but also due to the side activity inherent in porcine trypsin, as well as its S172A variant, which acts as an exopeptidase. Exopeptidase activity leads to the destruction of insulin glargine with the formation of also des-Arg and des-Thr.

Technical problems underlying the present invention

Thus, in the prior art, there has been a need to provide new trypsin variants with reduced side activities, especially with reduced exopeptidase activity. These novel trypsin variants will reduce the byproducts of PPI digestion and result in higher yields of insulin and insulin derivatives in the production of insulin (or insulin derivatives) from PPI.

The authors of the present invention have received new variants of porcine trypsin, which have reduced exopeptidase activity and/or show reduced by-product formation in the PPI cleavage reaction.

The above overview does not necessarily describe all the problems solved by the present invention.

Brief description of the invention

In a first aspect, the present invention provides a variant of porcine trypsin comprising or consisting of an amino acid sequence that shares at least 90% sequence identity with SEQ ID NO: 1, wherein said amino acid sequence differs from SEQ ID NO: 1 by at least one or multiple amino acid substitutions at one or more positions corresponding to F24, S44, D56, G78, Y131, S172 and W193 of native porcine trypsin according to SEQ ID NO: 1,

provided that the specified amino acid sequence is not a native porcine trypsin in accordance with SEQ ID NO: 1; and

provided that said amino acid sequence is not the porcine trypsin S172A variant according to SEQ ID NO: 2.

In a second aspect, the present invention relates to a nucleic acid molecule encoding a porcine trypsin variant according to the first aspect.

In a third aspect, the present invention relates to a host cell comprising a nucleic acid molecule according to the second aspect.

In a fourth aspect, the present invention relates to a method for producing a porcine trypsin variant according to the first aspect, comprising the step of

culturing the host cell according to the third aspect; and isolating the porcine trypsin variant from the culture medium or from the host cell.

In a fifth aspect, the present invention provides a porcine trypsin variant according to the first aspect, or a nucleic acid molecule according to the second aspect, or a host cell according to the third aspect for use as a drug; for use as a food ingredient; for use as a feed ingredient or for use in a process for the manufacture of a food ingredient or feed ingredient.

In a sixth aspect, the present invention relates to the use of the porcine trypsin variant according to the first aspect in a process for the preparation of human insulin, an insulin analogue or an insulin derivative.

In a seventh aspect, the present invention relates to the use of the porcine trypsin variant according to the first aspect to cleave a protein or peptide with the general formula A-Lys-Thr-Arg-Arg-B,

where A is an amino acid sequence consisting of one or more amino acids; and

where B is an amino acid sequence consisting of one or more amino acids.

In an eighth aspect, the present invention provides a composition comprising a porcine trypsin variant according to the first aspect, or a nucleic acid molecule according to the second aspect, or a host cell according to the third aspect.

Description of graphic materials

In FIG. 1 shows a schematic of the main tryptic cleavage sites for human insulin proproinsulins and insulin glargine. Solid triangles indicate cleavage sites to form product(s), open triangles indicate cleavage sites to form by-products. Disulfide bonds of prepreinsulins are not shown.

In FIG. 2 shows cleavage of preproinsulin glargine by trypsin variant S172A (SEQ ID NO: 2). In FIG. the time-dependent course of insulin glargine formation is shown, as well as the formation of the most prominent cleavage by-products, which are B31Arg-insulin glargine (cleavage after position 31 in the insulin glargine B chain, abbreviated "des-Arg" in the figure) and desB30Thr-insulin glargine (cleavage after position 29 of the insulin glargine B chain, abbreviated "des-Thr" in FIG.).

In FIG. 3 shows the cleavage of insulin glargine by the S172A trypsin variant (SEQ ID NO: 2) as an example of the exopeptidase activity of the S172A trypsin variant. In FIG. shows a time-dependent decrease in insulin glargine levels, as well as the formation of the main product, which is B32-desArg-insulin glargine (cleavage after position 31 in the B chain of insulin glargine, abbreviated "des-Arg" in Fig.) and small amounts of by-product , which is B30-desThr-insulin glargine (cleavage after position 29 of the insulin glargine B chain, abbreviated "des-Thr" in FIG.).

In FIG. 4 shows cleavage of preproinsulin glargine by trypsin variant #105 (SEQ ID NO: 105) as an example of cleavage by an optimized trypsin variant. In FIG. the time-dependent course of insulin glargine formation is shown, as well as the reduction in the formation of the most prominent cleavage by-products, which are B31Arg-insulin glargine (cleavage after position 31 in the B chain of insulin glargine, abbreviated "des-Arg" in Fig.) and desB30Thr- insulin glargine (cleavage after position 29 of the insulin glargine B chain, abbreviated "des-Thr" in FIG.).

Detailed description of the invention

Definitions

Although the present invention is described in detail below, it is to be understood that the present invention is not limited to the specific methods, protocols, and reagents described herein, as these may vary. It is also to be understood that the terminology used herein is for the purpose of describing specific embodiments only and is not intended to limit the scope of the present invention, which will be limited only by the appended claims. Unless otherwise defined, all technical and scientific terms used in this document have the same meaning as generally understood by a person skilled in the art to which the present invention belongs.

In the practice of the present invention will be used, unless otherwise indicated, the conventional methods of chemistry, biochemistry, cell biology, immunology and techniques of recombinant DNA, which are set forth in the literature in the art.

Throughout this description and in the following claims, unless the context otherwise implies, the word "comprise" and variations such as "comprises" and "comprising" will be intended to include the specified representative, an integer, or a stage or group of representatives, integers numbers or stages, but not the exclusion of any other representative, integer or stage or group of representatives, integers or stages, although in some embodiments, another such representative, integer or stage or group of representatives, integers or stages may be excluded, i.e. the subject matter is to include the specified representative, integer or stage or group of representatives, integers or stages. The terms singular and like reference used in the context of the description of the present invention (especially in the context of the claims) are intended to include the singular and plural unless otherwise specified herein or clearly contradicted by the context. The quoting of ranges of values in this document is only for the purposes of a shorthand way of referring to each individual value that is within the range. Unless otherwise specified in this document, each individual value is included in the description as if it were cited individually in this document. All methods described herein can be performed in any suitable order, unless otherwise indicated herein or otherwise contrary to the context. The use of any and each of the examples or illustrative language (eg, "such as") provided herein is only intended to more effectively describe the present invention and does not limit the scope of the present invention, unless otherwise indicated. Nothing in this specification should be construed as indicating any unclaimed element necessary for the practice of the present invention.

Several documents (for example: patents, patent applications, scientific publications, manufacturer's specifications, instructions, presented sequences under the GenBank accession number, etc.) are cited throughout the text of this description. Nothing in this document should be construed as an admission that the present invention has no reason to oppose such disclosure, based on an earlier invention. Some of the documents cited in this document are described as "incorporated by reference" or "incorporated by reference in their entirety". In the event of a conflict between the definitions or teachings of such incorporated references and the definitions or teachings set forth in this specification, the text of this specification shall prevail.

Sequences: All sequences mentioned herein are disclosed in the accompanying sequence listing, which, in its entirety and disclosure, forms part of this specification.

The term "naturally occurring" when used in connection with biological materials such as nucleic acid molecules, (poly)peptides, host cells, and the like, refers to materials that occur naturally and have not been exposed to human exposure.

As used herein, the term "amino acid" or "amino acid residue" refers to naturally occurring amino acids, non-naturally occurring amino acids, amino acid analogs, and amino acid mimetics that function similarly to naturally occurring amino acids, all of which are in the form D- and L-stereoisomers, if their structure allows the existence of such stereoisomeric forms. Amino acids are designated in this document by their names, by their commonly known three-letter designations, or by the one-letter designations recommended by the Commission on Biochemical Nomenclature of the IUPAC-IUB.

When referring to amino acids, the term "naturally occurring" refers to the 20 common amino acids (i.e., alanine (A), cysteine (C), aspartic acid (D), glutamic acid (E), phenylalanine (F), glycine (G ), histidine (H), isoleucine (I), lysine (K), leucine (L), methionine (M), asparagine (N), proline (P), glutamine (Q), arginine (R), serine (S ), threonine (T), valine (V), tryptophan (W) and tyrosine (Y)), as well as selenocysteine, pyrrolysine (PYL) and pyrrolinecarboxylysine (PCL). The terms "naturally occurring amino acids" and "encoded amino acids" are used interchangeably herein.

As used herein, the term "non-naturally occurring amino acid" is intended to refer to amino acids that are not naturally encoded or found in the genetic code of any organism.

As used herein, the term "amino acid analog" refers to compounds that have the same basic chemical structure as a naturally occurring amino acid.

As used herein, the term "amino acid mimetics" refers to chemical compounds with a structure that differs from the general chemical structure of an amino acid, but functions in a manner similar to a naturally occurring amino acid.

As used herein, the term "peptide" refers to a polymeric form of amino acids of any length, for example, containing two or more, or 3 or more, or 4 or more, or 6 or more, or 8 or more, or 9 or more, or 10 or more, or 13 or more, or 16 or more, or 21 or more amino acids covalently linked by peptide bonds. The term "polypeptide" refers to large peptides, in particular peptides with more than 100 amino acid residues. The terms "polypeptide" and "protein" are used interchangeably herein. This application provides information about the amino acid sequences of several peptides and polypeptides disclosed in this document. Amino acid sequences are presented herein from the amino terminus (left) to the carboxy terminus (right) of the peptide or polypeptide.

As used herein, the term "variant" of a (poly)peptide refers to a (poly)peptide that differs from a corresponding reference (poly)peptide by one or more amino acid modifications in its amino acid sequence. In one embodiment, a "variant" exhibits one or more biological activities of a reference (poly)peptide. The reference (poly)peptide may be a wild-type (poly)peptide or a (poly)peptide that does not occur naturally. The terms "variant" and "mutant" are used interchangeably herein.

As used herein, the term "variant porcine trypsin" refers to a polypeptide that differs from the wild-type porcine trypsin set forth in SEQ ID NO: 1 by one or more amino acid modifications in its amino acid sequence. The term "variant porcine trypsin" specifically refers to a polypeptide that differs from the wild-type porcine trypsin set forth under SEQ ID NO: 1 by one or more amino acid substitutions. In accordance with the present invention, a "variant porcine trypsin" exhibits an endoproteolytic activity similar to that of a reference trypsin (e.g. wild-type porcine trypsin according to SEQ ID NO: 1 or trypsin variant S172A according to SEQ ID NO: 2), but endoproteolytic the activities of "variant porcine trypsin" and reference trypsin may differ in their selectivity for certain cleavage sites.

The term "amino acid modifications" encompasses amino acid substitutions, amino acid deletions, and amino acid additions. The terms "amino acid substitution" and "amino acid substitution" are used interchangeably herein. The term "amino acid deletions" encompasses N-terminal truncations, internal deletions, and C-terminal truncations. The term "amino acid additions" encompasses N-terminal additions, amino acid insertions, and C-terminal additions.

The expression "Xaa24" (and similar expressions such as Xaa44, Xaa56, Xaa78, etc.) refers to a variable amino acid at position 24 (or 44, 56, 78, etc., respectively) of a given amino acid sequence. The sequence listing attached to the present application contains one protein sequence with variable amino acid positions, namely SEQ ID NO: 3. Variable amino acid positions in the protein sequence according to SEQ ID NO: 3 are indicated by the amino acid Xaa (in three letter code format). The number 24 in this example indicates that this variable position is the 24th amino acid of the protein sequence according to SEQ ID NO: 3. Similarly, the expression Xaa44 indicates that the 44th position of SEQ ID NO: 3 is a variable position of an amino acid, etc.

As used herein, the term "human insulin" refers to a human hormone whose structure and properties are well known. Human insulin contains two peptide chains (chains A and B) that are connected by disulfide bridges between cysteine residues, namely the A chain and the B chain. The A chain is a 21 amino acid peptide and the B chain is a 30 amino acid peptide, with the two chains connected by three disulfide bridges: one between the cysteines at positions 6 and 11 of the A chain; the second between the cysteine at position 7 of the A chain and the cysteine at position 7 of the B chain; and a third between a cysteine at position 20 of the A chain and a cysteine at position 19 of the B chain.

As used herein, the terms "insulin derivative" and "insulin derivative" refer to a peptide that has a molecular structure that can be formally derived from the structure of naturally occurring insulin, such as that of human insulin, in which one or more organic substituents (for example , fatty acid) bind to one or more amino acids. Optionally, one or more amino acids that are present in the naturally occurring insulin may be removed/or replaced with other amino acids, including non-coding amino acids, or amino acids, including non-coding ones, may be added to the naturally occurring insulin. Examples of insulin derivatives include, without limitation, the following:

(i). 'Insulin detemir', which differs from human insulin in that the C-terminal threonine at position B30 is removed and a fatty acid residue (myristic acid residue) is attached to the lysine epsilon amino functional group at position B29.

(ii). 'Insulin degludec', which differs from human insulin in that the last amino acid is removed from the B chain and a glutamyl bond is added between B29Lys and hexadecanedioic acid.

As used herein, the terms "insulin analog" and "insulin analog" refer to a peptide that has a molecular structure that can be formally derived from the structure of naturally occurring insulin, such as that of human insulin, by deletion and/or substitution of at least one amino acid residue found in naturally occurring insulin, and/or adding at least one amino acid residue. The added and/or substituted amino acid residue may be either an encoded amino acid residue, another naturally occurring residue, or a fully synthetic amino acid residue. Examples of insulin analogs include, without limitation, the following:

(i). 'Insulin aspart' is created by recombinant DNA technology such that the B28 amino acid in human insulin (i.e. amino acid number 28 in the human insulin B chain), which is proline, is replaced with aspartic acid;

(ii). 'Insulin lispro' is created by recombinant DNA technology such that the penultimate lysine and proline residues in the C-terminus of the B chain of human insulin are reversed (human insulin: B28Pro-B29Lys; insulin lispro: B28Lys-B29Pro);

(iii). 'Insulin glulisine' differs from human insulin in that the amino acid asparagine at position B3 is substituted with lysine and the lysine at position B29 is substituted with glutamic acid;

(iv). 'Insulin glargine' differs from human insulin in that the asparagine at position A21 is replaced by glycine and the B-chain is extended at the carboxy-terminus by two arginines.

As used herein, the term "yield" refers to the ratio of product (i.e., insulin or insulin derivative or insulin analog) obtained after trypsin digestion to preproinsulin (or preproinsulin derivative or preproinsulin analog) before trypsin digestion. The yield depends on the selectivity of the cleavage of the trypsin variant at the desired cleavage site and on its exonuclease activity. The higher the selectivity of the trypsin variant to cleave only at the desired cleavage site and the lower its exonuclease activity, the higher the yield.

For example, "yield of insulin glargine" or "yield of human B31Arg-B32Arg-insulin" refers to the ratio of produced insulin glargine or human B31Arg-B32Arg-insulin obtained after trypsin digestion and preproinsulin glargine or human preproinsulin, respectively, before trypsin digestion. The "yield of insulin glargine" and the "yield of human B31Arg-B32Arg-insulin" depend on the selectivity of cleavage of the trypsin variant only after B32Arg and on its exonuclease activity. The higher the selectivity of trypsin variant cleavage only after B32Arg and the lower its exonuclease activity, the higher the yield.

As used herein, the term "increased selectivity" refers to the property of a trypsin variant to cleave less of an unwanted cleavage site and more of a desired cleavage site compared to a reference trypsin. Increased selectivity leads to a reduction in the formation of undesirable by-products. The reference trypsin may be, for example, wild-type porcine trypsin according to SEQ ID NO: 1 or the S172A trypsin variant according to SEQ ID NO: 2.

"Sequence identity" of two amino acid sequences indicates the percentage of amino acids that are identical in the sequences. Optimal sequence alignment for comparison can be performed, in addition to being performed manually, using the local homology search algorithm of Smith and Waterman, 1981, Adv. app. Math. 2, 482-489 (incorporated herein by reference), using the local homology search algorithm of Needleman and Wunsch, 1970, J. Mol. Biol. 48, 443-453 (incorporated herein by reference), using the similarity search method of Pearson and Lipman, 1988, Proc. Natl Acad. sci. USA 85, 2444-2448 (incorporated herein by reference) or by computer programs that use these algorithms (GAP, BESTFIT, FASTA, BLAST P, BLAST N, and TFASTA in the Wisconsin Genetics, Genetics Computer Group, 575 Science Drive, Madison, Wisconsin).

In the context of the present invention, the degree of sequence identity between a modified sequence and the sequence from which it is derived is usually calculated relative to the total length of the unmodified sequence, unless explicitly stated otherwise. In embodiments where none of the sequences is a modified sequence, the degree of sequence identity is calculated relative to the total length of the sequence defined as the reference sequence, unless otherwise indicated. In embodiments where neither sequence can be considered a reference sequence, the degree of sequence identity is calculated relative to the longer of the two sequences, unless otherwise indicated.

A "nucleic acid molecule" according to some embodiments of the present invention is a deoxyribonucleic acid (DNA) or a ribonucleic acid (RNA). The nucleic acid molecule according to the present invention may be in the form of a molecule that is single-stranded or double-stranded; and may be linear or covalently closed to form a ring.

The term "DNA" refers to a molecule that contains deoxyribonucleotide residues and which may consist wholly or in large part of deoxyribonucleotide residues. "Deoxyribonucleotide" refers to a nucleotide that does not contain a hydroxyl group at the 2' position of the beta-D-ribofuranosyl group. The term "DNA" includes isolated DNA, such as partially or completely purified DNA, substantially pure DNA, synthetic DNA, and recombinantly generated DNA, and includes modified DNA that differs from naturally occurring DNA by the addition, deletion, substitution, and/or alteration of one or several nucleotides. Such changes may include the addition of non-nucleotide material, such as addition to the end(s) of the DNA or within the strand, for example, to one or more nucleotides of the DNA. Nucleotides in DNA molecules can also include non-standard nucleotides such as non-naturally occurring nucleotides or chemically synthesized nucleotides. These altered DNAs may be referred to as analogues or analogues of naturally occurring DNA. When used in connection with nucleotides, the term "naturally occurring" refers to the bases that are adenine (A), cytosine (C), guanine (G), thymine (T), and uracil (U).

The term "RNA" refers to a molecule that contains ribonucleotide residues and which may be wholly or largely composed of ribonucleotide residues. "Ribonucleotide" refers to a nucleotide with a hydroxyl group at the 2' position of the beta-D-ribofuranosyl group. The term "RNA" includes isolated RNA, such as partially or completely purified RNA, essentially pure RNA, synthetic RNA, and recombinantly generated RNA, and includes modified RNA that differs from naturally occurring RNA by the addition, deletion, substitution, and/or alteration of one or several nucleotides. Such changes may include the addition of non-nucleotide material, such as addition to the end(s) of the RNA or intrastrand, for example, to one or more nucleotides of the RNA. Nucleotides in RNA molecules may also include non-standard nucleotides such as non-naturally occurring nucleotides or chemically synthesized nucleotides or deoxynucleotides. These altered RNAs may be referred to as analogs or analogs of naturally occurring RNAs. In accordance with the present invention, "RNA" refers to single-stranded RNA or double-stranded RNA. In one embodiment, the RNA is mRNA, such as in vitro transcribed RNA (IVT RNA) or synthetic RNA. The RNA may also be modified, eg, with one or more modifications to increase the stability (eg, half-life) of the RNA. Such modifications are known to the person skilled in the art and include, for example, 5' caps or 5' cap analogs.

The nucleic acid molecule according to the present invention may be in/be included in the vector. As used herein, the term "vector" includes all vectors known to those skilled in the art, including plasmid vectors, cosmid vectors, phage vectors such as lambda phage vectors, viral vectors such as adenovirus or baculovirus vectors, or vectors based on artificial chromosomes such as bacterial artificial chromosomes (BAC), yeast artificial chromosomes (YAC) or P1 artificial chromosomes (PAC). These vectors include expression vectors as well as cloning vectors. Expression vectors contain plasmids as well as viral vectors, and typically contain the desired coding sequence and the corresponding DNA sequences necessary to express the operably linked coding sequence in a particular host organism (e.g., bacteria, yeast, plant, insect, or mammal) or in vitro expression systems. Typically, cloning vectors are used to design and amplify a particular desired DNA fragment and may lack the functional sequences required to express the desired DNA fragments.

Alternatively, the nucleic acid molecule in accordance with the present invention may be integrated into the genome, for example, the genome of the host cell. Means and methods for integrating a particular nucleic acid molecule into the genome are known to those skilled in the art.

The term "cell" or "host cell" generally refers to an intact cell, ie, a cell with an intact membrane that has not released its normal intracellular components such as enzymes, organelles, or genetic material. An intact cell is usually a viable cell, ie a living cell capable of performing its normal metabolic functions. As used herein, this term refers to any cell that can be transfected or transformed with exogenous nucleic acid. In some embodiments, the cell, when transfected or transformed with an exogenous nucleic acid and transferred to a recipient, is capable of expressing the nucleic acid in the recipient. The term "cell" includes prokaryotic cells such as bacterial cells and eukaryotic cells such as yeast cells, fungal cells or mammalian cells. Bacterial cells include cells from Gram-negative bacterial strains such as Escherichia coli, Proteus and Pseudomonas strains and Gram-positive bacterial strains such as Bacillus, Streptomyces, Staphylococcus and Lactococcus strains. Fungal cells include cells from Trichoderma, Neurospora and Aspergillus species. Yeast cells include cells from Saccharomyces species (eg Saccharomyces cerevisiae), Schizosaccharomyces (eg Schizosaccharomyces pombe), Pichia (eg Pichia pastoris and Pichia methanolica), Komagataella (eg Komagataella pastoris and Komagataella phaffii) and Hansenula. Mammalian cells include, for example, CHO cells, BHK cells, HeLa cells, COS cells, HEK293 cells, and the like. Amphibian cells, insect cells, plant cells, and any other cells used in the art to express heterologous proteins can also be used. Mammalian cells are particularly suitable for adoptive transfer, for example cells from humans, mice, hamsters, pigs, goats and primates. Cells can be obtained from a wide variety of tissue types and include primary cells and cell lines such as cells of the immune system, in particular antigen presenting cells such as dendritic cells and T cells, stem cells such as hematopoietic stem cells and mesenchymal stem cells. , and other types of cells. An antigen presenting cell is a cell that presents an antigen in combination with a major histocompatibility complex on its surface. T cells are able to recognize this complex using their T cell receptor (TCR). A "cell" or "host cell" may be isolated or be part of a tissue or organism, in particular a "non-human organism".

As used herein, the term "non-human organism" is intended to include non-human primates or other animals, in particular mammals such as cows, horses, pigs, sheep, goats, dogs, cats, rabbits, or rodents such as like mice, rats, guinea pigs and hamsters.

Used in this document, the term "drug" refers to a substance/composition used in therapy, for example, to treat a disease or disorder.

In accordance with the present invention, the term "disease or disorder" refers to any pathological or unhealthy condition.

By "treatment" is meant the administration of a compound or composition, or a combination of compounds or compositions, to a subject for the purpose of preventing or eliminating a disease or disorder; stopping or slowing down the course of the disease or disorder in the subject; suppressing or slowing down the development of a new disease or disorder in a subject; reducing the frequency or severity of symptoms and/or relapses in a subject who currently suffers from or previously suffered from a disease or disorder; and/or renewals, i.e. increase, life expectancy of the subject.

In particular, the term "treatment/treatment of a disease or disorder" includes curing, shortening, alleviating, preventing, slowing or suppressing the progression or aggravation, or preventing or slowing the onset of a disease or disorder or symptoms thereof.

The term "subject" in accordance with the present invention means a subject to be treated, in particular a subject suffering from a disease (also referred to as a "patient"), including humans, non-human primates or other animals, in particular mammals such as cows, horses, pigs, sheep, goats, dogs, cats, rabbits or rodents such as mice, rats, guinea pigs and hamsters. In one embodiment, the subject/patient is a human.

A "composition" according to the present invention contains an effective amount of the porcine trypsin variant of the present invention (also referred to herein as the "active agent"). The composition according to the present invention can be used (a) in a manufacturing process, in particular in a manufacturing process for making peptides, in particular in a manufacturing process for making human insulin, insulin derivatives and/or insulin analogues; (b) as a food ingredient; (c) as a feed ingredient (d) or for use in a process for the manufacture of a food ingredient or feed ingredient. The composition according to the present invention can also be used to treat a disease or disorder and thus can be a pharmaceutical composition.

A "pharmaceutical composition" according to the present invention contains a therapeutically effective amount of the porcine trypsin variant, nucleic acid molecule or host cell of the present invention (also referred to herein as "active agents") to produce the desired response or desired effect. The pharmaceutical composition according to the present invention may further comprise at least one other active agent(s).

In one embodiment, the compositions are provided in the form of a uniform dosage form and can be obtained using a known method per se. The composition may, for example, be in the form of a solution or suspension.

The composition or pharmaceutical composition may further contain one or more excipients. In one embodiment of the pharmaceutical composition, one or more of the excipients are all pharmaceutically acceptable. Used in this document, the expression "pharmaceutically acceptable" refers to physiologically well tolerated by mammals or humans. In particular, it means approved by a regulatory agency of the federal or state government or listed in the USP or other generally accepted pharmacopoeia for use in animals and, more specifically, in humans.

The composition or pharmaceutical composition according to the present invention may be present in the form of a composition wherein various active agents and excipients (e.g. diluents and/or carriers) are mixed with each other, or may take the form of a combined preparation wherein the active agents are present in part or completely different form. An example of such a combination or combined preparation is a set of parts.

Unless otherwise indicated in the context, the terms "active agent" and "active ingredient" refer to the porcine trypsin variants of the present invention, the nucleic acid molecule of the present invention, and the host cell of the present invention. The terms "active agent" and "active ingredient" are used interchangeably herein.

The term "carrier" refers to an organic or inorganic component of natural or synthetic origin with which the active agent is combined for the purpose of facilitating, improving or enabling use. In accordance with the present invention, the term "carrier" also includes one or more solid or liquid fillers, diluents, adjuvants, excipients, carriers or encapsulating substances. Such carriers may be sterile liquids such as saline solutions in water and oils, including petroleum, animal, vegetable or synthetic oils such as peanut oil, soybean oil, mineral oil, sesame oil, and the like. Saline solutions and aqueous dextrose solution as well as glycerol solutions can also be used as liquid carriers. Suitable excipients include starch, glucose, lactose, sucrose, gelatin, malt, rice, flour, chalk, silica gel, sodium stearate, glycerol monostearate, talc, sodium chloride, skimmed milk powder, glycerin, propylene, glycol, water, ethanol, etc. The composition or pharmaceutical composition, if necessary, may also contain minor amounts of wetting or emulsifying agents, or buffering agents for adjusting the pH. These compositions may take the form of solutions, suspensions, emulsions, tablets, pills, capsules, powders, sustained release formulations, and the like. The composition or pharmaceutical composition may be formulated as a suppository with conventional binders and carriers such as triglycerides. The compounds of the present invention may be formulated in neutral or salt forms. Pharmaceutically acceptable salts include salts formed with free amino groups, such as those derived from hydrochloric, phosphoric, acetic, oxalic, tartaric acids, etc., and salts formed with free carboxyl groups, such as those derived from sodium, potassium hydroxides. , ammonium, calcium, iron, isopropylamine, triethylamine, 2-ethylaminoethanol, histidine, procaine, etc. Such compositions will contain an effective amount of the compound, in some embodiments in purified form, together with a suitable amount of carrier. The composition must correspond to the route of administration.

The term "excipient" as used herein is intended to include all substances that may be present in a composition or pharmaceutical composition that are not active agents, such as salts, binders (e.g. lactose, dextrose, sucrose, trehalose , sorbitol, mannitol), carriers, lubricants, thickeners, surfactants, preservatives (e.g. antioxidants, citric acid, sodium citrate, benzalkonium chloride, chlorobutanol, cysteine, methionine, parabens, thimerosal), emulsifiers, buffers, fragrances or dyes.

Salts are included in the present invention and may also be pharmaceutically acceptable salts. Salts or pharmaceutically acceptable salts of this type include, without limitation, salts derived from the following acids: hydrochloric, hydrobromic, sulfuric, nitric, phosphoric, maleic, acetic, salicylic, citric, formic, malonic, succinic acids, etc. Salts and pharmaceutically acceptable salts can also be obtained in the form of alkali metal salts or alkaline earth metal salts such as sodium salts, potassium salts or calcium salts. Salts can be added to adjust ionic strength or tonicity.

Suitable buffering agents for use in a composition or pharmaceutical composition according to the present invention include acetic acid in salt, citric acid in salt, boric acid in salt and phosphoric acid in salt.

The composition or pharmaceutical composition may also be formulated as a stable lyophilized product which is reconstituted with a suitable diluent which optionally contains one or more excipients as defined above.

The active agents and pharmaceutical compositions described herein may be administered in therapeutically effective amounts. "Therapeutically effective amount" refers to the amount at which the desired therapeutic response or the desired therapeutic effect is achieved alone or together with additional doses, in one embodiment, without causing unacceptable side effects. In the case of treating a particular disease or a particular condition, the desired response may relate to the suppression of the course of the disease. This includes slowing down the progression of the disease, and in particular arresting or reversing the progression of the disease. The desired response in the treatment of a disease or condition may also be to delay the onset or prevent the onset of said disease or said condition. The effective amount of the agent or composition described herein will depend on the condition being treated, the severity of the disease, the individual parameters of the subject (including age, physiological condition, size and weight), the duration of treatment, the type of concomitant therapy (if any), the specific route introduction and similar factors. Accordingly, the administered doses of the agents described herein may depend on several such parameters. In the event that a subject's response is insufficient at the initial dose, higher doses may be used (or effectively higher doses achieved via a different, more localized route of administration).

As used herein, the term "set of parts (short set)" refers to an article containing one or more containers and optionally a storage medium. Said one or more containers may be filled with one or more of the aforementioned agents or reagents. Additional containers may be included in the kit, which contain, for example, diluents, buffers, and additional reagents. Said storage medium may be a non-electronic storage medium, such as a graphical storage medium such as an information leaflet, information leaflet, barcode or access code, or an electronic storage medium such as a compact disc (CD), digital versatile disc (DVD) , microchip or other electronic storage medium based on semiconductors. The access code may provide access to a database, such as an Internet database, centralized or decentralized databases. Said data carrier may contain instructions for the use of the active agents of the present invention. For example, the data carrier may contain instructions for using the active agents (a) in a manufacturing method, in particular in a manufacturing method for preparing peptides, in particular in a manufacturing method for preparing human insulin, insulin derivatives and/or insulin analogs; (b) as a food ingredient; (c) as a feed ingredient (d) or for use in a process for the manufacture of a food ingredient or feed ingredient.

As used herein, the term "preproinsulin" or the abbreviation "PPI" refers to a single chain insulin precursor starting at the N-terminus with a presequence followed by a B chain, a C peptide and an A chain (see FIG. 1).

Embodiments of the present invention

In the following paragraphs, certain elements of the present invention will be described. These elements may be listed in certain embodiments, however, it should be understood that they can be combined in any manner and in any amount to obtain additional embodiments. The variously described examples and embodiments should not limit the present invention to the expressly described embodiments. This description is to be understood as supporting and covering embodiments that combine the expressly described embodiments with any number of disclosed elements. In addition, any permutations and combinations of all described elements in this application should be considered as disclosed by the description of this application, unless the context indicates otherwise.

In a first aspect, the present invention is directed to a variant of porcine trypsin comprising or consisting of an amino acid sequence that shares at least 90% sequence identity with SEQ ID NO: 1, wherein said amino acid sequence differs from SEQ ID NO: 1 by at least one or multiple amino acid substitutions at one or more positions corresponding to F24, S44, D56, G78, Y131, S172 and W193 of native porcine trypsin according to SEQ ID NO: 1,

provided that the specified amino acid sequence is not a native porcine trypsin in accordance with SEQ ID NO: 1; and

provided that said amino acid sequence is not the porcine trypsin S172A variant according to SEQ ID NO: 2.

In one embodiment of the first aspect

the amino acid at the position corresponding to F24 is not replaced by Asp, Glu or Gly;

the amino acid at the position corresponding to S44 is not replaced by Tyr;

the amino acid at the position corresponding to Y131 is not replaced by Lys or Pro; and/or

the amino acid at the position corresponding to W193 is not replaced by Asn or Cys.

In one embodiment of the first aspect

the amino acid at the position corresponding to F24 is replaced by an amino acid selected from the group consisting of Ala, Asn, Arg, Gln, Ile, Leu, Lys, Met, Ser, Thr and Val;

the amino acid at the position corresponding to S44 is replaced by an amino acid selected from the group consisting of Leu and Pro;

the amino acid at the position corresponding to D56 is replaced by an amino acid selected from the group consisting of Ala, Asn, His and Trp;

the amino acid at the position corresponding to G78 is replaced by an amino acid selected from the group consisting of Ala, Glu; Pro, Ser and Tyr;

the amino acid at the position corresponding to Y131 is replaced by an amino acid selected from the group consisting of Ala, Asn, Asp, Cys, Gln; Glu, Gly, His, Ile, Leu, Met, Ser, Thr, Trp and Val;

the amino acid at the position corresponding to S172 is replaced by an amino acid selected from the group consisting of Ala, Cys and Thr; and/or

the amino acid at the position corresponding to W193 is replaced by an amino acid selected from the group consisting of Phe, Ser, Thr and Tyr.

In a further embodiment of the first aspect

the amino acid at the position corresponding to F24 is replaced by Arg;

the amino acid at the position corresponding to S44 is replaced by Pro;

the amino acid at the position corresponding to D56 is replaced by His;

the amino acid at the position corresponding to G78 is replaced by Pro;

the amino acid at the position corresponding to Y131 is replaced by Met;

the amino acid at the position corresponding to S172 is replaced by Ala; and/or

the amino acid at the position corresponding to W193 is replaced by Ser.

In one embodiment of the first aspect, said amino acid sequence further differs from SEQ ID NO: 1 by at least one or more amino acid substitutions at one or more positions corresponding to R99, R107, K125 and K170 of native porcine trypsin according to SEQ ID NO: 1 .

In one embodiment of the first aspect

the amino acid at the position corresponding to R107 is not replaced by Pro; and/or

the amino acid at the position corresponding to K170 is not replaced by Ile or Phe.

In a further embodiment of the first aspect

the amino acid at the position corresponding to R99 is replaced by an amino acid selected from the group consisting of Ala, Asn, Asp, Glu, Gly, His, Leu, Phe, Thr, Trp and Tyr;

the amino acid at the position corresponding to R107 is replaced by an amino acid selected from the group consisting of Asp, Gly, Pro, Ser and Thr;

the amino acid at the position corresponding to K125 is replaced by an amino acid selected from the group consisting of Ala, Cys, Gln, Glu, Gly, His, Leu, Ser, and Tyr; and/or

the amino acid at the position corresponding to K170 is replaced by an amino acid selected from the group consisting of Ala, Asn, Gly and Tyr.

In a further embodiment of the first aspect

the amino acid at the position corresponding to R99 is replaced by an amino acid selected from the group consisting of Ala, His and Asn;

the amino acid at the position corresponding to R107 is replaced by Thr;

the amino acid at the position corresponding to K125 is replaced by an amino acid selected from the group consisting of Ala, Cys and Ser; and/or

the amino acid at the position corresponding to K170 is replaced by Ala.

In one embodiment of the first aspect, said amino acid sequence is characterized by at least 91%, or at least 92%, or at least 93%, or at least 94%, or at least 95%, or at least 96% , or at least 97%, or at least 98%, or at least 99% sequence identity with wild-type porcine trypsin according to SEQ ID NO: 1.

In one embodiment of the first aspect, said amino acid sequence differs from wild-type porcine trypsin according to SEQ ID NO: 1 by 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, or 23 amino acid substitutions.

In one embodiment of the first aspect, said amino acid sequence is SEQ ID NO: 3, where

Xaa24 is an amino acid selected from the group consisting of Ala, Asn, Arg, Gln, Ile, Leu, Lys, Met, Phe, Ser, Thr and Val;

Xaa44 is an amino acid selected from the group consisting of Leu, Pro and Ser;

Xaa56 is an amino acid selected from the group consisting of Ala, Asn, Asp, His and Trp;

Xaa78 is an amino acid selected from the group consisting of Ala, Glu, Gly, Pro, Ser and Tyr;

Xaa99 is an amino acid selected from the group consisting of Ala, Arg, Asn, Asp, Glu, Gly, His, Leu, Phe, Thr, Trp and Tyr;

Xaa107 is an amino acid selected from the group consisting of Arg, Asp, Gly, Pro, Ser and Thr;

Xaa125 is an amino acid selected from the group consisting of Ala, Cys, Gln, Glu, Gly, His, Leu, Lys, Ser, and Tyr;

Xaa131 is an amino acid selected from the group consisting of Ala, Asn, Asp, Cys, Gln; Glu, Gly, His, Ile, Leu, Met, Ser, Thr, Trp, Tyr and Val;

Xaa170 is an amino acid selected from the group consisting of Ala, Asn, Gly, Lys and Tyr;

Xaa172 is an amino acid selected from the group consisting of Ala, Cys, Ser and Thr; and/or

Xaa193 is an amino acid selected from the group consisting of Phe, Ser, Thr, Trp and Tyr;

provided that SEQ ID NO: 3 is not wild-type porcine trypsin according to SEQ ID NO: 1; and

provided that SEQ ID NO: 3 is not the S172A porcine trypsin variant of SEQ ID NO: 2.

In a further embodiment of the first aspect, said amino acid sequence is SEQ ID NO: 3, where

Xaa24 is an amino acid selected from the group consisting of Phe and Arg;

Xaa44 is an amino acid selected from the group consisting of Ser and Pro;

Xaa56 is an amino acid selected from the group consisting of Asp and His;

Xaa78 is an amino acid selected from the group consisting of Gly and Pro;

Xaa99 is an amino acid selected from the group consisting of Ala, Arg, His and Asn;

Xaa107 is an amino acid selected from the group consisting of Arg and Thr;

Xaa125 is an amino acid selected from the group consisting of Ala, Cys, Lys and Ser;

Xaa131 is an amino acid selected from the group consisting of Met and Tyr;

Xaa170 is an amino acid selected from the group consisting of Ala and Lys;

Xaa172 is an amino acid selected from the group consisting of Ala and Ser; and/or

Xaa193 is an amino acid selected from the group consisting of Ser and Trp;

provided that SEQ ID NO: 3 is not wild-type porcine trypsin according to SEQ ID NO: 1; and

provided that SEQ ID NO: 3 is not the S172A porcine trypsin variant of SEQ ID NO: 2.

In one embodiment of the first aspect, said amino acid sequence is selected from the group consisting of SEQ ID NOs: 4-119. In one embodiment, said amino acid sequence is selected from the group consisting of SEQ ID NO: 89, SEQ ID NO: 90, SEQ ID NO: 91, SEQ ID NO: 92, SEQ ID NO: 93, SEQ ID NO: 94, SEQ ID NO: 95, SEQ ID NO: 96, SEQ ID NO: 97 and SEQ ID NO: 98.

In one embodiment of the first aspect, said variant is capable of cleaving a peptide with the general formula A-Lys-Thr-Arg-Arg-B to produce a cleavage product with the general formula A-Lys-Thr-Arg-Arg in a yield of at least 80 % (for example, at least 81%, at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88 %, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98% or at least 99%),

where A is an amino acid sequence consisting of one or more amino acids; and

where B is an amino acid sequence consisting of one or more amino acids.

In additional embodiments of the first aspect, A is an amino acid sequence consisting of 1-105 amino acids (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15 , 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 100 , or 105 amino acids), and B is an amino acid sequence consisting of 1-105 amino acids (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 100, or 105 amino acids).

In one embodiment of the first aspect, the variant exhibits increased selectivity for cleavage of human preproinsulin, preproinsulin glargine, preproinsulin lispro, preproinsulin aspart, or preproinsulin glulisine at the C-terminal position relative to the B32-Arg position compared to the porcine trypsin variant S172A according to SEQ ID NO : 2.

In some embodiments of the first aspect of the present invention, the porcine trypsin variant is lyophilized.

In a second aspect, the present invention is directed to a nucleic acid molecule encoding

- porcine trypsin variant as defined in the first aspect of the present invention; or

a trypsinogen precursor molecule of the porcine trypsin variant as defined in the first aspect of the present invention.

In one embodiment of the second aspect, said nucleic acid molecule is contained in a vector or integrated into the genome.

In a third aspect, the present invention is directed to a host cell comprising the nucleic acid molecule as defined in the second aspect. In one embodiment, the host cell is a recombinant host cell.

In a fourth aspect, the present invention is directed to a method for producing the porcine trypsin variant of the present invention, comprising the steps of:

culturing the host cell according to the third aspect; and

isolating the trypsinogen variant porcine trypsin precursor molecule from the culture medium or from the host cell.

In one embodiment of the fourth aspect, the method includes the additional step of:

activation of the precursor molecule, which is trypsinogen, thus obtaining the specified variant of porcine trypsin.

The activation of the trypsinogen precursor molecule can be carried out chemically or enzymatically. For example, the pH can be adjusted in such a way that auto-activation occurs. In another example, a protease (eg, enterokinase) can be added to enzymatically activate trypsinogen.

In a fifth aspect, the present invention is directed to a porcine trypsin variant of the present invention, or a nucleic acid molecule of the present invention, or a host cell of the present invention for use as a drug; for use as a food ingredient; for use as a feed ingredient or for use in a process for the manufacture of a food ingredient or feed ingredient.

One embodiment of the fifth aspect is directed to the porcine trypsin variant of the present invention for use as a food ingredient; for use as a feed ingredient or for use in a process for the manufacture of a food ingredient or feed ingredient.

In one embodiment of the fifth aspect, the drug is for use in the treatment of edema (especially trauma-induced edema), inflammation (especially trauma-induced inflammation), thrombophlebitis, activated arthrosis, digestive enzyme deficiency (eg, caused by pancreatic insufficiency), and/or digestive disorders.

In other words, the present invention is directed to a porcine trypsin variant of the present invention, or a nucleic acid molecule of the present invention, or a host cell of the present invention for use in the treatment of edema (especially trauma-induced edema), inflammation (especially trauma-induced inflammation) , thrombophlebitis, activated arthrosis, deficiency of digestive enzymes (for example, caused by pancreatic insufficiency) and / or digestive disorders.

With regard to use as a food ingredient, as a feed ingredient, or in a manufacturing process, the fifth aspect can alternatively be stated as follows:

the use of the porcine trypsin variant of the present invention as a food ingredient.

Use of the porcine trypsin variant of the present invention as a feed ingredient.

The use of the variant porcine trypsin of the present invention in a process for the manufacture of a food ingredient or feed ingredient.

In a sixth aspect, the present invention is directed to the use of the porcine trypsin variant of the present invention in a process for the preparation of human insulin, an insulin analog, or an insulin derivative.

A sixth aspect encompasses the use of the porcine trypsin variants of the present invention in both in vitro and in vivo methods for the production of human insulin, insulin analog or insulin derivative.

With regard to the use of methods in vivo, the sixth aspect can be alternatively formulated as follows:

A variant of porcine trypsin according to the first aspect for use in in vivo production of human insulin, insulin analogue or insulin derivative.

In one embodiment of the sixth aspect, the insulin analog is selected from the group consisting of insulin aspart, insulin lispro, insulin glulisine, and insulin glargine.

In a seventh aspect, the present invention is directed to the use of the porcine trypsin variant of the present invention to cleave a protein or peptide with the general formula A-Lys-Thr-Arg-Arg-B,

where A is an amino acid sequence consisting of one or more amino acids; and

where B is an amino acid sequence consisting of one or more amino acids.

In additional embodiments of the seventh aspect, A is an amino acid sequence consisting of 1-105 amino acids (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15 , 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 100 , or 105 amino acids), and B is an amino acid sequence consisting of 1-105 amino acids (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 100, or 105 amino acids).

In one embodiment of the seventh aspect, cleavage results in a cleavage product of A-Lys-Thr-Arg-Arg. In further embodiments of the seventh aspect, cleavage results in a cleavage product of A-Lys-Thr-Arg-Arg in at least 80% yield (e.g., at least 81%, at least 82%, at least 83% , at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%).

In an eighth aspect, the present invention is directed to a composition comprising a porcine trypsin variant of the present invention, or a nucleic acid molecule of the present invention, or a host cell of the present invention.

In one embodiment of the eighth aspect, the composition further comprises an excipient and/or at least one other active agent.

In some embodiments of the eighth aspect, the composition is a pharmaceutical composition.

In some embodiments of the eighth aspect, the composition or pharmaceutical composition is lyophilized.

The present invention also contemplates the combination of a porcine trypsin variant of the present invention (or a nucleic acid molecule of the present invention or a host cell of the present invention) with at least one other active pharmaceutical ingredient. In one embodiment, the combination of the porcine trypsin variant of the present invention and at least one other active pharmaceutical ingredient can be administered either by separately administering the active pharmaceutical ingredient to a patient or as a product combination in which multiple active pharmaceutical ingredients are present in a single pharmaceutical composition. When administered separately, administration may occur simultaneously or sequentially in any order. The amount of the active agent of the present invention and the other active pharmaceutical ingredient(s) and the relative timing of administration will be chosen to achieve the desired combined therapeutic effect. The administration of the combination can be carried out simultaneously in: (1) one pharmaceutical composition containing all active pharmaceutical ingredients; or (2) separate pharmaceutical compositions, each containing at least one of the active pharmaceutical ingredients. Alternatively, the combination may be administered separately in a sequential manner, with one treatment agent administered first and the other administered second, or vice versa. Such sequential administration can be carried out at close time intervals or at remote time intervals.

Examples

The following examples are set forth to provide those skilled in the art with a complete disclosure and description of how to make and use the compounds, compositions and methods of the present invention and are not intended to limit the scope of the present invention in any way as set forth in the appended invention formula. Efforts have been made to ensure accuracy with respect to the numbers used (eg amounts, temperatures, etc.), but some experimental errors and deviations must be taken into account. Unless otherwise indicated, parts are parts by weight, molecular weight is average molecular weight, temperature is in degrees Celsius, and pressure is atmospheric or near atmospheric.

Example 1: Mutagenesis and cloning of a synthetic nucleotide sequence encoding porcine trypsin

Enzyme improvement can be achieved through enzyme engineering. This technique involves the development of variants of the original enzyme with improved properties. To provide enzyme variants, mutations at the gene level were first introduced into the synthetic nucleotide sequence encoding porcine pancreatic trypsin using conventional molecular biological methods. The enzyme variant genes were cloned into the pLE1A17 expression vector (pRSF-1b derivative, Novagen). The resulting plasmids were used to transform E. coli BL21 (DE3) cells.

Example 2: Expression of a Porcine Trypsinogen Variant

Recombinant expression of porcine trypsinogen in E. coli was performed by autoinduction according to F.W. Studier (F.W. Studier (2005) Prot. Exp. Pur. 41:207-234) using ZYM-5052 medium supplemented with kanamycin (50 mg/l) at 30°C. Once the glucose in the culture is metabolized, the presence of lactose induces expression. Glycerol was used as the C-source. Expression was carried out in a microplate, as well as in a shake flask with a scale.

For shake flask experiments, a preculture in 20 ml of ZYM-505 medium supplemented with kanamycin (50 mg/l) with 50 μl of glycerol (36%) was incubated for 24 h at 30°C and 200 rpm. An OD600 value of 12.5-14 was achieved. For the main culture, 500 μl of preculture was used to inoculate 100 ml of ZYM-5052 medium supplemented with kanamycin (50 mg/l). After incubation for 24 hours at 30° C. and 200 rpm, an OD600 value in the range of 6.5-7.5 was reached. After centrifugation, the cell pellets were stored at -20°C.

Example 3 Cell Destruction and Trypsinogen Activation

Periplasmic trypsinogen was released by osmotic shock. 3.6 g of cells (obtained from Example 2) were resuspended in 10 ml of lysis buffer (10 mM Tris-HCl, pH=8, 25 mM MgCl ₂ , 200 mM NaCl) until a homogeneous suspension was obtained. The suspension was stirred for 24 hours at 5° C. and 180 rpm. After centrifugation, the supernatant was subjected to activation.

Enterokinase from porcine intestine (Sigma, item no. E0885; 40 units; definition of units: one unit of enterokinase will produce 1.0 nanomoles of trypsin from trypsinogen per minute at pH 5.6 at 25° C.) was used to activate trypsinogen.

Enterokinase was added to the cell-free extract to a final concentration of 17 mU/ml. The reaction mixture was incubated for 16-24 h at 30°C. The resulting trypsin solution was stored at -20°C.

Example 4 Tryptic Activity Assay of a Purified Variant of Recombinant Porcine Trypsin

Trypsin activity was determined using TRY chromozyme (Roche Diagnostics GmbH) in 100 mM Tris, pH 8.0, 20 mM CaCl ₂ at 25°C. Chromozyme TRY is a substrate (carbobenzoxy-L-valyl-L-glycyl-L-arginine-4-nitranilide acetate) for reliable photometric determination of the activity of proteases that hydrolyze peptides on the carboxyl side of arginine (trypsin, Arg-C endoproteinase, and others). Photometric measurement was carried out at 405 nm.

Example 5: Screening results with human preproinsulin.

Initial screening of all trypsin variants was performed with human preproinsulin (PPI) as a model system. PPI was dissolved in a buffer solution (pH=8.3) to a concentration of 0.75 g/l. A trypsin variant solution was prepared according to Example 3 and added to the reaction mixture without further processing. Samples were periodically taken, acidified with 1 N. HCl and analyzed by HPLC (example 7).

Two aspects were considered: (1) whether the trypsin variant exhibits any PPI cleavage activity (results are shown in Table 1, column A); (2) whether the trypsin variant exhibits PPI cleavage activity: if trypsin cleavage selectivity is improved compared to SEQ ID NO: 2 (results are shown in Table 1, column B). To evaluate the selectivity of di-Arg trypsin variants, human B31Arg-B32Arg-insulin production was monitored by HPLC (see Example 7) and compared with SEQ ID NO: 2. Selectivity was calculated according to

where

[diArg] = concentration of human B31Arg-B32Arg-insulin

[monoArg] = concentration of human desB32Arg-insulin

[desThr] = concentration of human desB30Thr-insulin

[UI] = concentration of unknown cleavage intermediate

Table 1 - Screening results with human preproinsulin

Mutant A B SEQID NO: PPI cleavage activity Selectivity 2 Yes 62.1% four Yes >90% 5 Yes >90% 6 Yes >90% 7 Yes >90% eight Yes >90% 9 Yes >90% ten Yes >90% eleven Yes >90% 12 Yes >90% 13 Yes >90% fourteen Yes >90% fifteen Yes >90% 16 Yes >90% 17 Yes >80% eighteen Yes >90% 19 Yes >90% twenty Yes >90% 21 Yes >90% 22 Yes >90% 23 Yes >90% 24 Yes >90% 25 Yes >90% 26 Yes >80% 27 Yes >80% 28 Yes >90% 29 Yes >90% thirty Yes >70% 31 Yes >62.1% 32 Yes >90% 33 Yes >90% 34 Yes >90% 35 Yes >90% 36 Yes >90% 37 Yes >90% 38 Yes >80% 39 Yes >90% 40 Yes >80% 41 Yes >90% 42 Yes >90% 43 Yes >80% 44 Yes >80% 45 Yes >65% 46 Yes >70% 47 Yes >90% 48 Yes >80% 49 Yes >80% fifty Yes >65% 51 Yes >70% 52 Yes >90% 53 Yes >90% 54 Yes >90% 55 Yes >90% 56 Yes >90% 57 Yes >90% 58 Yes >90% 59 Yes >90% 60 Yes >90% 61 Yes >80% 62 Yes >90% 63 Yes >90% 64 Yes >90% 65 Yes >80% 66 Yes >90% 67 Yes >90% 68 Yes >70% 69 Yes >70% 70 Yes >70% 71 Yes >62.1% 72 Yes >90% 73 Yes >90% 74 Yes >90% 75 Yes >90% 76 Yes >90% 77 Yes >90% 78 Yes >80% 79 Yes >90% 80 Yes >90% 81 Yes >90% 82 Yes >90% 83 Yes >90% 84 Yes >90% 85 Yes >90% 86 Yes >90% 87 Yes >90% 88 Yes >90% 89 Yes >90% 90 Yes >90% 91 Yes >90% 92 Yes >90% 93 Yes >90% 94 Yes >90% 95 Yes >90% 96 Yes >90% 97 Yes >90% 98 Yes >90% 99 Yes >80% 100 Yes >90% 101 Yes >90% 102 Yes >90% 103 Yes >90% 104 Yes >90% 105 Yes >90% 106 Yes >90% 107 Yes >90% 108 Yes >80% 109 Yes >90% 110 Yes >90% 111 Yes >80% 112 Yes >90% 113 Yes >90% 114 Yes >90% 115 Yes >90% 116 Yes >90% 117 Yes >90% 118 Yes >90% 119 Yes >90% 120 Yes 1.4% 121 Yes 4.6% 122 Yes 59.4% 123 Yes 60.7% 124 Yes 27.6% 125 Yes 12.2% 126 Yes 7.2% 127 Yes 20.5% 128 Yes 57.1% 129 Yes 44.8% 130 Yes 33.8%

Example 6 Cleavage of preproinsulin glargine using recombinant SEQ ID NO: 2 (trypsin variant S172A) and optimized variants thereof

These experiments were carried out at 20°C and a pH value of 8.3 (buffer solution), and the volume was adjusted to 1000 ml on the scale.

The solution of preproinsulin glargine was filled into an appropriate thermostated reaction vessel and the reaction was started by adding the variant porcine trypsin solution. Samples were taken at certain time intervals; the enzymatic reaction was immediately stopped by acidifying the sample solution using 1N. or 2 n. HCl solution. Samples were analyzed by HPLC (see example 7).

Figure 2 shows the reaction kinetics of SEQ ID NO: 2. The figure shows the time-dependent course of formation of insulin glargine, as well as the formation of the most prominent by-products of the B31Arg-insulin glargine cleavage (cleavage after position 31 in the B chain of insulin glargine, called "des -Arg") and desB30Thr-insulin glargine (cleavage after position 29 of the B chain of insulin glargine, called "des-Thr").

Preproinsulin glargine was cleaved with the selected options. The results are shown in Table 2. The contents of insulin glargine, desB32Arg-insulin glargine and desB29Thr-insulin glargine are shown as the peak area on the analytical HPLC chromatogram at the end of digestion and represent the ratio of insulin glargine to by-products. In addition, the molar yield of cleavage is presented according to

where

n(insulin glargine) = number of moles of insulin glargine produced

n(preproinsulin glargine) = number of moles of preproinsulin glargine used

Figure 4 shows the reaction kinetics of porcine trypsin variant No. 105 (SEQ ID NO: 105) as an example of an optimized trypsin variant.

Table 2 - Comparison of splitting results with preproinsulin glargine

Mutant Purity Exit SEQID NO: insulin glargine
% area deArg
% area deThr
% area % 2 53.6 19.1 5.2 65 four 86.4 <0.5 2.9 >90 9 85.2 <0.5 3.6 >90 ten 82.7 <0.5 2.6 >90 eleven 84.1 <0.5 2.2 >90 fourteen 84.5 <0.5 2.4 >90 eighteen 82.6 <0.5 1.9 >80 19 81.0 <0.5 3.3 >80 24 82.3 <0.5 2.8 >80 25 80.4 <0.5 2.8 >80 36 87.6 <0.5 1.5 >90 41 82.8 <0.5 1.9 >90 53 81.3 <0.5 2.1 >80 56 85.3 <0.5 2.0 >90 57 87.5 <0.5 2.3 >90 73 83.0 <0.5 2.2 >90 74 82.4 <0.5 2.1 >80 77 88.2 0.5 1.6 >90 78 85.1 0.5 1.6 >90 79 87.8 0.7 0.3 >90 80 90.1 0.9 0.5 >90 81 89.7 1.2 0.5 >90 82 83.8 0.6 0.3 >80 83 89.0 <0.5 1.2 >90 84 85.4 0.6 1.6 >90 85 85.4 1.1 0.2 >90 86 79.3 0.8 0.1 >80 87 83.8 0.6 0.3 >80 88 84.3 0.7 1.9 >90 89 88.1 0.5 1.2 >90 90 89.5 0.9 0.2 >90 91 86.8 1.0 0.3 >90 92 88.1 0.9 0.6 >90 93 71.3 0.7 0.3 >75 94 85.2 0.8 0.3 >90 95 90.0 1.0 0.6 >90 96 78.7 0.7 0.3 >80 97 89.4 <0.5 1.2 >90 98 89.4 0.9 2.7 >90 99 89.2 0.9 1.0 >90 100 80.0 0.7 0.3 >80 101 89.2 1.0 0.4 >90 102 88.7 0.4 1.2 >90 103 90.7 0.9 1.4 >90 104 89.3 1.1 1.0 >90 105 88.9 1.0 0.9 >90 106 86.8 0.7 1.0 >90 107 88.1 0.4 1.3 >90 108 88.8 0.8 0.7 >90 109 85.9 0.7 0.5 >80 110 90.6 0.9 0.6 >90 111 85.9 0.7 2.6 >90 113 86.1 0.5 1.7 >90 115 83.5 0.8 0.6 >90 116 82.7 0.5 0.6 >90 117 85.0 0.6 0.8 >90 118 87.6 0.6 1.0 >90 119 85.2 0.6 1.0 >90

As clearly seen in Table 2, the optimized trypsin variants exhibit increased cleavage selectivity for preproinsulin glargine, resulting in a significant increase in yield accompanied by a decrease in impurity profile when considering B31Arg-insulin glargine (desArg) and desB30Thr-insulin glargine (desThr). Comparison of tables 1 and 2 shows a very high comparability of the splitting results obtained with human preproinsulin and glargine preproinsulin.

Example 7 HPLC Method

Method High performance liquid chromatography

Column Manufacturer: Macherey & Nagel

Trade name: Nucleosil 120-5 C18

Size: 250mm x 4mm

Mobile phase A: 45 mM sodium phosphate buffer (pH 2.5). 315 mM NaCl. 25% (v/v) acetonitrile

Mobile phase B 45 mM sodium phosphate buffer (pH 2.5). 55 mM NaCl. 65% (v/v) acetonitrile

Gradient time/min Mobile phase A Mobile phase B

0 min 94% 6%

30 min linear 90% 10%

Injection volume: 5 µl

Detection: UV (215nm)

Example 8 Exopeptidase Activity

These experiments were carried out at 8°C and a pH value of 8.3 (buffer solution).

A solution of insulin glargine (0.5 g/l) was filled into an appropriate thermostated reaction vessel and the reaction started by adding a solution of SEQ ID NO: 2 (100 U/g _insulin ). Samples were taken at regular intervals and the enzymatic reaction was immediately stopped by acidifying the sample solution using 1N sodium hydroxide. HCl solution. Samples were analyzed by HPLC (see example 7). The reaction kinetics is shown in Fig. 3.

As clearly seen in FIG. 3, the concentration of insulin glargine decreases over time. In addition to the formation of a small amount of B30-desThr-insulin glargine, the main product formed in this reaction is B32-desArg-insulin glargine, indicating that the trypsin of SEQ ID NO: 2 is able to cleave a single arginine at position B32 through exopeptidase activity. .

Table 3: List of trypsin variants

SEQID NO: F24 S44 D56 G78 R99 R107 K125 Y131 K170 S172 W193 one F S D G R R K Y K S W 2 F S D G R R K Y K A W 3 X X X X X X X X X X X four R S D G A R K Y K A S 5 R S D G L R K Y K A S 6 R S D G F R K Y K A S 7 R S D G W R K Y K A S eight R S D G G R K Y K A S 9 R S D G Y R K Y K A S ten R S D G T R K Y K A S eleven R S D G N R K Y K A S 12 R S D G D R K Y K A S 13 R S D G E R K Y K A S fourteen R S D G H R K Y K A S fifteen R S D G R R A Y K A S 16 R S D G R R G Y K A S 17 R S D G R R Y Y K A S eighteen R S D G R R S Y K A S 19 R S D G R R C Y K A S twenty R S D G R R E Y K A S 21 R S D G R R H Y K A S 22 R S D G R R L Y K A S 23 R S D G R R Q Y K A S 24 R S D G R R K Y Y A S 25 R S D G R R K Y A A S 26 R S D G R R K Y G A S 27 R S D G R R K Y N A S 28 V S D G R R K Y K A S 29 L S D G R R K Y K A S thirty S S D G R R K Y K A S 31 Q S D G R R K Y K A S 32 R S D G R R K Y K A S 33 R S D G R T K Y K A S 34 R S D G R R K A K A S 35 R S D G R R K L K A S 36 R S D G R R K M K A S 37 R S D G R R K T K A S 38 R S D G R R K C K A S 39 R S D G R R K N K A S 40 R S D G R R K E K A S 41 R S D G R R K H K A S 42 R S D G R R K Y K A S 43 R S D G R R K Y K T S 44 R S D G R R K Y K S S 45 R S D G R R K Y K C S 46 A S D G R R K Y K A S 47 I S D G R R K Y K A S 48 M S D G R R K Y K A S 49 T S D G R R K Y K A S fifty N S D G R R K Y K A S 51 K S D G R R K Y K A S 52 R L D G R R K Y K A S 53 R P D G R R K Y K A S 54 R S A G R R K Y K A S 55 R S W G R R K Y K A S 56 R S N G R R K Y K A S 57 R S H G R R K Y K A S 58 R S D G R P K Y K A S 59 R S D G R G K Y K A S 60 R S D G R S K Y K A S 61 R S D G R D K Y K A S 62 R S D G R R K V K A S 63 R S D G R R K I K A S 64 R S D G R R K W K A S 65 R S D G R R K G K A S 66 R S D G R R K S K A S 67 R S D G R R K Q K A S 68 R S D G R R K D K A S 69 R S D G R R K Y K A F 70 R S D G R R K Y K A Y 71 R S D G R R K Y K A T 72 R S D S R R K Y K A S 73 R S D A R R K Y K A S 74 R S D P R R K Y K A S 75 R S D Y R R K Y K A S 76 R S D E R R K Y K A S 77 R P H G H T C M K A S 78 R S H P H T C Y K A S 79 R P H P H R A M K A S 80 R S H P H T S M K A S 81 R S H P H T A M K A S 82 R P H P H R C M K A S 83 R P H P N T S M K A S 84 R P H P N T S M K S S 85 R P H P H T C M K A S 86 R P D P A T S M K A S 87 R P H P H T S M K A S 88 R P H P H T S M K S S 89 R P H P H T A M K A S 90 R P H P H T A M K S S 91 R P H P A T S M K A S 92 R P H P A T C M K A S 93 R S D P N T C M A A S 94 R P H G N R C M A A S 95 R P H P H R S M A A S 96 R P D P H R S M A A S 97 R P H P A T A M A A S 98 R P H P A T A M A S S 99 R S H P N T S Y A A S 100 R P D P H T C M A A S 101 R S H P N T S M A A S 102 R P H P H T S M A A S 103 R P H P H T S M A S S 104 R S H P N T C M A A S 105 R S H P N T A Y A A S 106 R S H P A T C M A A S 107 R P H P H T C M A A S 108 R P D P A T C M A A S 109 R S H G N T C M A A S 110 R S H P H R C M A A S 111 R P H P H T S Y A A S 112 R P H P H T S Y A S S 113 R P H G A T A Y A A S 114 R P H G A T A Y A S S 115 R P H G N T S M K A S 116 R P H G N T S M A A S 117 R P H G N T A M K A S 118 R P H G H T A M A A S 119 R P H G A T A M K A S 120 R S D G R R P Y K A S 121 R S D G R R K Y I A S 122 R S D G R R K Y F A S 123 G S D G R R K Y K A S 124 R S D G R R K K K A S 125 D S D G R R K Y K A S 126 E S D G R R K Y K A S 127 R Y D G R R K Y K A S 128 R S D G R R K P K A S 129 R S D G R R K Y K A N 130 R S D G R R K Y K A C

Full-length wild-type porcine trypsin sequences and selected trypsin variants:

Wild-type porcine trypsin, SEQ ID NO: 1:

IVGGYTCAAN SIPYQVSLNS GSHFCGGSLI NSQWVVSAAH CYKSRIQVRL GEHNIDVLEG NEQFINAAKI ITHPNFNGNT LDNDIMLIKL SSPATLNSRV ATVSLPRSCA AAGTECLISG WGNTKSSGSS YPSLLQCLKA PVLSDSSCKS SYPGQITGNM ICVGFLEGGK DSCQGDSGGP VVCNGQLQGI VSWGYGCAQK NKPGVYTKVC NYVNWIQQTI AAN

Variant S172A, SEQ ID NO: 2:

IVGGYTCAAN SIPYQVSLNS GSHFCGGSLI NSQWVVSAAH CYKSRIQVRL GEHNIDVLEG NEQFINAAKI ITHPNFNGNT LDNDIMLIKL SSPATLNSRV ATVSLPRSCA AAGTECLISG WGNTKSSGSS YPSLLQCLKA PVLSDSSCKS SYPGQITGNM ICVGFLEGGK DACQGDSGGP VVCNGQLQGI VSWGYGCAQK NKPGVYTKVC NYVNWIQQTI AAN

General sequence of porcine trypsin with variable positions at amino acid positions 24, 44, 56, 78, 99, 107, 125, 131, 170, 172 and 193; SEQ ID NO: 3:

IVGGYTCAAN SIPYQVSLNS GSHXCGGSLI NSQWVVSAAH CYKXRIQVRL GEHNIXVLEG NEQFINAAKI ITHPNFNXNT LDNDIMLIKL SSPATLNSXV ATVSLPXSCA AAGTECLISG WGNTXSSGSS XPSLLQCLKA PVLSDSSCKS SYPGQITGNM ICVGFLEGGX DXCQGDSGGP VVCNGQLQGI VSXGYGCAQK NKPGVYTKVC NYVNWIQQTI AAN

SEQ ID NO: 4:

IVGGYTCAAN SIPYQVSLNS GSHRCGGSLI NSQWVVSAAH CYKSRIQVRL GEHNIDVLEG NEQFINAAKI ITHPNFNGNT LDNDIMLIKL SSPATLNSAV ATVSLPRSCA AAGTECLISG WGNTKSSGSS YPSLLQCLKA PVLSDSSCKS SYPGQITGNM ICVGFLEGGK DACQGDSGGP VVCNGQLQGI VSSGYGCAQK NKPGVYTKVC NYVNWIQQTI AAN

SEQ ID NO: 5:

IVGGYTCAAN SIPYQVSLNS GSHRCGGSLI NSQWVVSAAH CYKSRIQVRL GEHNIDVLEG NEQFINAAKI ITHPNFNGNT LDNDIMLIKL SSPATLNSLV ATVSLPRSCA AAGTECLISG WGNTKSSGSS YPSLLQCLKA PVLSDSSCKS SYPGQITGNM ICVGFLEGGK DACQGDSGGP VVCNGQLQGI VSSGYGCAQK NKPGVYTKVC NYVNWIQQTI AAN

SEQ ID NO: 6:

IVGGYTCAAN SIPYQVSLNS GSHRCGGSLI NSQWVVSAAH CYKSRIQVRL GEHNIDVLEG NEQFINAAKI ITHPNFNGNT LDNDIMLIKL SSPATLNSFV ATVSLPRSCA AAGTECLISG WGNTKSSGSS YPSLLQCLKA PVLSDSSCKS SYPGQITGNM ICVGFLEGGK DACQGDSGGP VVCNGQLQGI VSSGYGCAQK NKPGVYTKVC NYVNWIQQTI AAN

SEQ ID NO: 7:

IVGGYTCAAN SIPYQVSLNS GSHRCGGSLI NSQWVVSAAH CYKSRIQVRL GEHNIDVLEG NEQFINAAKI ITHPNFNGNT LDNDIMLIKL SSPATLNSWV ATVSLPRSCA AAGTECLISG WGNTKSSGSS YPSLLQCLKA PVLSDSSCKS SYPGQITGNM ICVGFLEGGK DACQGDSGGP VVCNGQLQGI VSSGYGCAQK NKPGVYTKVC NYVNWIQQTI AAN

SEQ ID NO: 8:

IVGGYTCAAN SIPYQVSLNS GSHRCGGSLI NSQWVVSAAH CYKSRIQVRL GEHNIDVLEG NEQFINAAKI ITHPNFNGNT LDNDIMLIKL SSPATLNSGV ATVSLPRSCA AAGTECLISG WGNTKSSGSS YPSLLQCLKA PVLSDSSCKS SYPGQITGNM ICVGFLEGGK DACQGDSGGP VVCNGQLQGI VSSGYGCAQK NKPGVYTKVC NYVNWIQQTI AAN

SEQ ID NO: 9:

IVGGYTCAAN SIPYQVSLNS GSHRCGGSLI NSQWVVSAAH CYKSRIQVRL GEHNIDVLEG NEQFINAAKI ITHPNFNGNT LDNDIMLIKL SSPATLNSYV ATVSLPRSCA AAGTECLISG WGNTKSSGSS YPSLLQCLKA PVLSDSSCKS SYPGQITGNM ICVGFLEGGK DACQGDSGGP VVCNGQLQGI VSSGYGCAQK NKPGVYTKVC NYVNWIQQTI AAN

SEQ ID NO: 10:

IVGGYTCAAN SIPYQVSLNS GSHRCGGSLI NSQWVVSAAH CYKSRIQVRL GEHNIDVLEG NEQFINAAKI ITHPNFNGNT LDNDIMLIKL SSPATLNSTV ATVSLPRSCA AAGTECLISG WGNTKSSGSS YPSLLQCLKA PVLSDSSCKS SYPGQITGNM ICVGFLEGGK DACQGDSGGP VVCNGQLQGI VSSGYGCAQK NKPGVYTKVC NYVNWIQQTI AAN

SEQ ID NO: 11:

IVGGYTCAAN SIPYQVSLNS GSHRCGGSLI NSQWVVSAAH CYKSRIQVRL GEHNIDVLEG NEQFINAAKI ITHPNFNGNT LDNDIMLIKL SSPATLNSNV ATVSLPRSCA AAGTECLISG WGNTKSSGSS YPSLLQCLKA PVLSDSSCKS SYPGQITGNM ICVGFLEGGK DACQGDSGGP VVCNGQLQGI VSSGYGCAQK NKPGVYTKVC NYVNWIQQTI AAN

SEQ ID NO: 12:

IVGGYTCAAN SIPYQVSLNS GSHRCGGSLI NSQWVVSAAH CYKSRIQVRL GEHNIDVLEG NEQFINAAKI ITHPNFNGNT LDNDIMLIKL SSPATLNSDV ATVSLPRSCA AAGTECLISG WGNTKSSGSS YPSLLQCLKA PVLSDSSCKS SYPGQITGNM ICVGFLEGGK DACQGDSGGP VVCNGQLQGI VSSGYGCAQK NKPGVYTKVC NYVNWIQQTI AAN

SEQ ID NO: 13:

IVGGYTCAAN SIPYQVSLNS GSHRCGGSLI NSQWVVSAAH CYKSRIQVRL GEHNIDVLEG NEQFINAAKI ITHPNFNGNT LDNDIMLIKL SSPATLNSEV ATVSLPRSCA AAGTECLISG WGNTKSSGSS YPSLLQCLKA PVLSDSSCKS SYPGQITGNM ICVGFLEGGK DACQGDSGGP VVCNGQLQGI VSSGYGCAQK NKPGVYTKVC NYVNWIQQTI AAN

SEQ ID NO: 14:

IVGGYTCAAN SIPYQVSLNS GSHRCGGSLI NSQWVVSAAH CYKSRIQVRL GEHNIDVLEG NEQFINAAKI ITHPNFNGNT LDNDIMLIKL SSPATLNSHV ATVSLPRSCA AAGTECLISG WGNTKSSGSS YPSLLQCLKA PVLSDSSCKS SYPGQITGNM ICVGFLEGGK DACQGDSGGP VVCNGQLQGI VSSGYGCAQK NKPGVYTKVC NYVNWIQQTI AAN

SEQ ID NO: 15:

IVGGYTCAAN SIPYQVSLNS GSHRCGGSLI NSQWVVSAAH CYKSRIQVRL GEHNIDVLEG NEQFINAAKI ITHPNFNGNT LDNDIMLIKL SSPATLNSRV ATVSLPRSCA AAGTECLISG WGNTASSGSS YPSLLQCLKA PVLSDSSCKS SYPGQITGNM ICVGFLEGGK DACQGDSGGP VVCNGQLQGI VSSGYGCAQK NKPGVYTKVC NYVNWIQQTI AAN

SEQ ID NO: 16:

IVGGYTCAAN SIPYQVSLNS GSHRCGGSLI NSQWVVSAAH CYKSRIQVRL GEHNIDVLEG NEQFINAAKI ITHPNFNGNT LDNDIMLIKL SSPATLNSRV ATVSLPRSCA AAGTECLISG WGNTGSSGSS YPSLLQCLKA PVLSDSSCKS SYPGQITGNM ICVGFLEGGK DACQGDSGGP VVCNGQLQGI VSSGYGCAQK NKPGVYTKVC NYVNWIQQTI AAN

SEQ ID NO: 17:

IVGGYTCAAN SIPYQVSLNS GSHRCGGSLI NSQWVVSAAH CYKSRIQVRL GEHNIDVLEG NEQFINAAKI ITHPNFNGNT LDNDIMLIKL SSPATLNSRV ATVSLPRSCA AAGTECLISG WGNTYSSGSS YPSLLQCLKA PVLSDSSCKS SYPGQITGNM ICVGFLEGGK DACQGDSGGP VVCNGQLQGI VSSGYGCAQK NKPGVYTKVC NYVNWIQQTI AAN

SEQ ID NO: 18:

IVGGYTCAAN SIPYQVSLNS GSHRCGGSLI NSQWVVSAAH CYKSRIQVRL GEHNIDVLEG NEQFINAAKI ITHPNFNGNT LDNDIMLIKL SSPATLNSRV ATVSLPRSCA AAGTECLISG WGNTSSSGSS YPSLLQCLKA PVLSDSSCKS SYPGQITGNM ICVGFLEGGK DACQGDSGGP VVCNGQLQGI VSSGYGCAQK NKPGVYTKVC NYVNWIQQTI AAN

SEQ ID NO: 19:

IVGGYTCAAN SIPYQVSLNS GSHRCGGSLI NSQWVVSAAH CYKSRIQVRL GEHNIDVLEG NEQFINAAKI ITHPNFNGNT LDNDIMLIKL SSPATLNSRV ATVSLPRSCA AAGTECLISG WGNTCSSGSS YPSLLQCLKA PVLSDSSCKS SYPGQITGNM ICVGFLEGGK DACQGDSGGP VVCNGQLQGI VSSGYGCAQK NKPGVYTKVC NYVNWIQQTI AAN

SEQ ID NO: 20:

IVGGYTCAAN SIPYQVSLNS GSHRCGGSLI NSQWVVSAAH CYKSRIQVRL GEHNIDVLEG NEQFINAAKI ITHPNFNGNT LDNDIMLIKL SSPATLNSRV ATVSLPRSCA AAGTECLISG WGNTESSGSS YPSLLQCLKA PVLSDSSCKS SYPGQITGNM ICVGFLEGGK DACQGDSGGP VVCNGQLQGI VSSGYGCAQK NKPGVYTKVC NYVNWIQQTI AAN

SEQ ID NO: 21:

IVGGYTCAAN SIPYQVSLNS GSHRCGGSLI NSQWVVSAAH CYKSRIQVRL GEHNIDVLEG NEQFINAAKI ITHPNFNGNT LDNDIMLIKL SSPATLNSRV ATVSLPRSCA AAGTECLISG WGNTHSSGSS YPSLLQCLKA PVLSDSSCKS SYPGQITGNM ICVGFLEGGK DACQGDSGGP VVCNGQLQGI VSSGYGCAQK NKPGVYTKVC NYVNWIQQTI AAN

SEQ ID NO: 22:

IVGGYTCAAN SIPYQVSLNS GSHRCGGSLI NSQWVVSAAH CYKSRIQVRL GEHNIDVLEG NEQFINAAKI ITHPNFNGNT LDNDIMLIKL SSPATLNSRV ATVSLPRSCA AAGTECLISG WGNTLSSGSS YPSLLQCLKA PVLSDSSCKS SYPGQITGNM ICVGFLEGGK DACQGDSGGP VVCNGQLQGI VSSGYGCAQK NKPGVYTKVC NYVNWIQQTI AAN

SEQ ID NO: 23:

IVGGYTCAAN SIPYQVSLNS GSHRCGGSLI NSQWVVSAAH CYKSRIQVRL GEHNIDVLEG NEQFINAAKI ITHPNFNGNT LDNDIMLIKL SSPATLNSRV ATVSLPRSCA AAGTECLISG WGNTQSSGSS YPSLLQCLKA PVLSDSSCKS SYPGQITGNM ICVGFLEGGK DACQGDSGGP VVCNGQLQGI VSSGYGCAQK NKPGVYTKVC NYVNWIQQTI AAN

SEQ ID NO: 24:

IVGGYTCAAN SIPYQVSLNS GSHRCGGSLI NSQWVVSAAH CYKSRIQVRL GEHNIDVLEG NEQFINAAKI ITHPNFNGNT LDNDIMLIKL SSPATLNSRV ATVSLPRSCA AAGTECLISG WGNTKSSGSS YPSLLQCLKA PVLSDSSCKS SYPGQITGNM ICVGFLEGGY DACQGDSGGP VVCNGQLQGI VSSGYGCAQK NKPGVYTKVC NYVNWIQQTI AAN

SEQ ID NO: 25:

IVGGYTCAAN SIPYQVSLNS GSHRCGGSLI NSQWVVSAAH CYKSRIQVRL GEHNIDVLEG NEQFINAAKI ITHPNFNGNT LDNDIMLIKL SSPATLNSRV ATVSLPRSCA AAGTECLISG WGNTKSSGSS YPSLLQCLKA PVLSDSSCKS SYPGQITGNM ICVGFLEGGA DACQGDSGGP VVCNGQLQGI VSSGYGCAQK NKPGVYTKVC NYVNWIQQTI AAN

SEQ ID NO: 26:

IVGGYTCAAN SIPYQVSLNS GSHRCGGSLI NSQWVVSAAH CYKSRIQVRL GEHNIDVLEG NEQFINAAKI ITHPNFNGNT LDNDIMLIKL SSPATLNSRV ATVSLPRSCA AAGTECLISG WGNTKSSGSS YPSLLQCLKA PVLSDSSCKS SYPGQITGNM ICVGFLEGGG DACQGDSGGP VVCNGQLQGI VSSGYGCAQK NKPGVYTKVC NYVNWIQQTI AAN

SEQ ID NO: 27:

IVGGYTCAAN SIPYQVSLNS GSHRCGGSLI NSQWVVSAAH CYKSRIQVRL GEHNIDVLEG NEQFINAAKI ITHPNFNGNT LDNDIMLIKL SSPATLNSRV ATVSLPRSCA AAGTECLISG WGNTKSSGSS YPSLLQCLKA PVLSDSSCKS SYPGQITGNM ICVGFLEGGN DACQGDSGGP VVCNGQLQGI VSSGYGCAQK NKPGVYTKVC NYVNWIQQTI AAN

SEQ ID NO: 28:

IVGGYTCAAN SIPYQVSLNS GSHVCGGSLI NSQWVVSAAH CYKSRIQVRL GEHNIDVLEG NEQFINAAKI ITHPNFNGNT LDNDIMLIKL SSPATLNSRV ATVSLPRSCA AAGTECLISG WGNTKSSGSS YPSLLQCLKA PVLSDSSCKS SYPGQITGNM ICVGFLEGGK DACQGDSGGP VVCNGQLQGI VSSGYGCAQK NKPGVYTKVC NYVNWIQQTI AAN

SEQ ID NO: 29:

IVGGYTCAAN SIPYQVSLNS GSHLCGGSLI NSQWVVSAAH CYKSRIQVRL GEHNIDVLEG NEQFINAAKI ITHPNFNGNT LDNDIMLIKL SSPATLNSRV ATVSLPRSCA AAGTECLISG WGNTKSSGSS YPSLLQCLKA PVLSDSSCKS SYPGQITGNM ICVGFLEGGK DACQGDSGGP VVCNGQLQGI VSSGYGCAQK NKPGVYTKVC NYVNWIQQTI AAN

SEQ ID NO: 30:

IVGGYTCAAN SIPYQVSLNS GSHSCGGSLI NSQWVVSAAH CYKSRIQVRL GEHNIDVLEG NEQFINAAKI ITHPNFNGNT LDNDIMLIKL SSPATLNSRV ATVSLPRSCA AAGTECLISG WGNTKSSGSS YPSLLQCLKA PVLSDSSCKS SYPGQITGNM ICVGFLEGGK DACQGDSGGP VVCNGQLQGI VSSGYGCAQK NKPGVYTKVC NYVNWIQQTI AAN

SEQ ID NO: 31:

IVGGYTCAAN SIPYQVSLNS GSHQCGGSLI NSQWVVSAAH CYKSRIQVRL GEHNIDVLEG NEQFINAAKI ITHPNFNGNT LDNDIMLIKL SSPATLNSRV ATVSLPRSCA AAGTECLISG WGNTKSSGSS YPSLLQCLKA PVLSDSSCKS SYPGQITGNM ICVGFLEGGK DACQGDSGGP VVCNGQLQGI VSSGYGCAQK NKPGVYTKVC NYVNWIQQTI AAN

SEQ ID NO: 32:

IVGGYTCAAN SIPYQVSLNS GSHRCGGSLI NSQWVVSAAH CYKSRIQVRL GEHNIDVLEG NEQFINAAKI ITHPNFNGNT LDNDIMLIKL SSPATLNSRV ATVSLPRSCA AAGTECLISG WGNTKSSGSS YPSLLQCLKA PVLSDSSCKS SYPGQITGNM ICVGFLEGGK DACQGDSGGP VVCNGQLQGI VSSGYGCAQK NKPGVYTKVC NYVNWIQQTI AAN

SEQ ID NO: 33:

IVGGYTCAAN SIPYQVSLNS GSHRCGGSLI NSQWVVSAAH CYKSRIQVRL GEHNIDVLEG NEQFINAAKI ITHPNFNGNT LDNDIMLIKL SSPATLNSRV ATVSLPTSCA AAGTECLISG WGNTKSSGSS YPSLLQCLKA PVLSDSSCKS SYPGQITGNM ICVGFLEGGK DACQGDSGGP VVCNGQLQGI VSSGYGCAQK NKPGVYTKVC NYVNWIQQTI AAN

SEQ ID NO: 34:

IVGGYTCAAN SIPYQVSLNS GSHRCGGSLI NSQWVVSAAH CYKSRIQVRL GEHNIDVLEG NEQFINAAKI ITHPNFNGNT LDNDIMLIKL SSPATLNSRV ATVSLPRSCA AAGTECLISG WGNTKSSGSS APSLLQCLKA PVLSDSSCKS SYPGQITGNM ICVGFLEGGK DACQGDSGGP VVCNGQLQGI VSSGYGCAQK NKPGVYTKVC NYVNWIQQTI AAN

SEQ ID NO: 35:

IVGGYTCAAN SIPYQVSLNS GSHRCGGSLI NSQWVVSAAH CYKSRIQVRL GEHNIDVLEG NEQFINAAKI ITHPNFNGNT LDNDIMLIKL SSPATLNSRV ATVSLPRSCA AAGTECLISG WGNTKSSGSS LPSLLQCLKA PVLSDSSCKS SYPGQITGNM ICVGFLEGGK DACQGDSGGP VVCNGQLQGI VSSGYGCAQK NKPGVYTKVC NYVNWIQQTI AAN

SEQ ID NO: 36:

IVGGYTCAAN SIPYQVSLNS GSHRCGGSLI NSQWVVSAAH CYKSRIQVRL GEHNIDVLEG NEQFINAAKI ITHPNFNGNT LDNDIMLIKL SSPATLNSRV ATVSLPRSCA AAGTECLISG WGNTKSSGSS MPSLLQCLKA PVLSDSSCKS SYPGQITGNM ICVGFLEGGK DACQGDSGGP VVCNGQLQGI VSSGYGCAQK NKPGVYTKVC NYVNWIQQTI AAN

SEQ ID NO: 37:

IVGGYTCAAN SIPYQVSLNS GSHRCGGSLI NSQWVVSAAH CYKSRIQVRL GEHNIDVLEG NEQFINAAKI ITHPNFNGNT LDNDIMLIKL SSPATLNSRV ATVSLPRSCA AAGTECLISG WGNTKSSGSS TPSLLQCLKA PVLSDSSCKS SYPGQITGNM ICVGFLEGGK DACQGDSGGP VVCNGQLQGI VSSGYGCAQK NKPGVYTKVC NYVNWIQQTI AAN

SEQ ID NO: 38:

IVGGYTCAAN SIPYQVSLNS GSHRCGGSLI NSQWVVSAAH CYKSRIQVRL GEHNIDVLEG NEQFINAAKI ITHPNFNGNT LDNDIMLIKL SSPATLNSRV ATVSLPRSCA AAGTECLISG WGNTKSSGSS CPSLLQCLKA PVLSDSSCKS SYPGQITGNM ICVGFLEGGK DACQGDSGGP VVCNGQLQGI VSSGYGCAQK NKPGVYTKVC NYVNWIQQTI AAN

SEQ ID NO: 39:

IVGGYTCAAN SIPYQVSLNS GSHRCGGSLI NSQWVVSAAH CYKSRIQVRL GEHNIDVLEG NEQFINAAKI ITHPNFNGNT LDNDIMLIKL SSPATLNSRV ATVSLPRSCA AAGTECLISG WGNTKSSGSS NPSLLQCLKA PVLSDSSCKS SYPGQITGNM ICVGFLEGGK DACQGDSGGP VVCNGQLQGI VSSGYGCAQK NKPGVYTKVC NYVNWIQQTI AAN

SEQ ID NO: 40:

IVGGYTCAAN SIPYQVSLNS GSHRCGGSLI NSQWVVSAAH CYKSRIQVRL GEHNIDVLEG NEQFINAAKI ITHPNFNGNT LDNDIMLIKL SSPATLNSRV ATVSLPRSCA AAGTECLISG WGNTKSSGSS EPSLLQCLKA PVLSDSSCKS SYPGQITGNM ICVGFLEGGK DACQGDSGGP VVCNGQLQGI VSSGYGCAQK NKPGVYTKVC NYVNWIQQTI AAN

SEQ ID NO: 41:

IVGGYTCAAN SIPYQVSLNS GSHRCGGSLI NSQWVVSAAH CYKSRIQVRL GEHNIDVLEG NEQFINAAKI ITHPNFNGNT LDNDIMLIKL SSPATLNSRV ATVSLPRSCA AAGTECLISG WGNTKSSGSS HPSLLQCLKA PVLSDSSCKS SYPGQITGNM ICVGFLEGGK DACQGDSGGP VVCNGQLQGI VSSGYGCAQK NKPGVYTKVC NYVNWIQQTI AAN

SEQ ID NO: 42:

IVGGYTCAAN SIPYQVSLNS GSHRCGGSLI NSQWVVSAAH CYKSRIQVRL GEHNIDVLEG NEQFINAAKI ITHPNFNGNT LDNDIMLIKL SSPATLNSRV ATVSLPRSCA AAGTECLISG WGNTKSSGSS YPSLLQCLKA PVLSDSSCKS SYPGQITGNM ICVGFLEGGK DACQGDSGGP VVCNGQLQGI VSSGYGCAQK NKPGVYTKVC NYVNWIQQTI AAN

SEQ ID NO: 43:

IVGGYTCAAN SIPYQVSLNS GSHRCGGSLI NSQWVVSAAH CYKSRIQVRL GEHNIDVLEG NEQFINAAKI ITHPNFNGNT LDNDIMLIKL SSPATLNSRV ATVSLPRSCA AAGTECLISG WGNTKSSGSS YPSLLQCLKA PVLSDSSCKS SYPGQITGNM ICVGFLEGGK DTCQGDSGGP VVCNGQLQGI VSSGYGCAQK NKPGVYTKVC NYVNWIQQTI AAN

SEQ ID NO: 44:

IVGGYTCAAN SIPYQVSLNS GSHRCGGSLI NSQWVVSAAH CYKSRIQVRL GEHNIDVLEG NEQFINAAKI ITHPNFNGNT LDNDIMLIKL SSPATLNSRV ATVSLPRSCA AAGTECLISG WGNTKSSGSS YPSLLQCLKA PVLSDSSCKS SYPGQITGNM ICVGFLEGGK DSCQGDSGGP VVCNGQLQGI VSSGYGCAQK NKPGVYTKVC NYVNWIQQTI AAN

SEQ ID NO: 45:

IVGGYTCAAN SIPYQVSLNS GSHRCGGSLI NSQWVVSAAH CYKSRIQVRL GEHNIDVLEG NEQFINAAKI ITHPNFNGNT LDNDIMLIKL SSPATLNSRV ATVSLPRSCA AAGTECLISG WGNTKSSGSS YPSLLQCLKA PVLSDSSCKS SYPGQITGNM ICVGFLEGGK DCCQGDSGGP VVCNGQLQGI VSSGYGCAQK NKPGVYTKVC NYVNWIQQTI AAN

SEQ ID NO: 46:

IVGGYTCAAN SIPYQVSLNS GSHACGGSLI NSQWVVSAAH CYKSRIQVRL GEHNIDVLEG NEQFINAAKI ITHPNFNGNT LDNDIMLIKL SSPATLNSRV ATVSLPRSCA AAGTECLISG WGNTKSSGSS YPSLLQCLKA PVLSDSSCKS SYPGQITGNM ICVGFLEGGK DACQGDSGGP VVCNGQLQGI VSSGYGCAQK NKPGVYTKVC NYVNWIQQTI AAN

SEQ ID NO: 47:

IVGGYTCAAN SIPYQVSLNS GSHICGGSLI NSQWVVSAAH CYKSRIQVRL GEHNIDVLEG NEQFINAAKI ITHPNFNGNT LDNDIMLIKL SSPATLNSRV ATVSLPRSCA AAGTECLISG WGNTKSSGSS YPSLLQCLKA PVLSDSSCKS SYPGQITGNM ICVGFLEGGK DACQGDSGGP VVCNGQLQGI VSSGYGCAQK NKPGVYTKVC NYVNWIQQTI AAN

SEQ ID NO: 48:

IVGGYTCAAN SIPYQVSLNS GSHMCGGSLI NSQWVVSAAH CYKSRIQVRL GEHNIDVLEG NEQFINAAKI ITHPNFNGNT LDNDIMLIKL SSPATLNSRV ATVSLPRSCA AAGTECLISG WGNTKSSGSS YPSLLQCLKA PVLSDSSCKS SYPGQITGNM ICVGFLEGGK DACQGDSGGP VVCNGQLQGI VSSGYGCAQK NKPGVYTKVC NYVNWIQQTI AAN

SEQ ID NO: 49:

IVGGYTCAAN SIPYQVSLNS GSHTCGGSLI NSQWVVSAAH CYKSRIQVRL GEHNIDVLEG NEQFINAAKI ITHPNFNGNT LDNDIMLIKL SSPATLNSRV ATVSLPRSCA AAGTECLISG WGNTKSSGSS YPSLLQCLKA PVLSDSSCKS SYPGQITGNM ICVGFLEGGK DACQGDSGGP VVCNGQLQGI VSSGYGCAQK NKPGVYTKVC NYVNWIQQTI AAN

SEQ ID NO: 50:

IVGGYTCAAN SIPYQVSLNS GSHNCGGSLI NSQWVVSAAH CYKSRIQVRL GEHNIDVLEG NEQFINAAKI ITHPNFNGNT LDNDIMLIKL SSPATLNSRV ATVSLPRSCA AAGTECLISG WGNTKSSGSS YPSLLQCLKA PVLSDSSCKS SYPGQITGNM ICVGFLEGGK DACQGDSGGP VVCNGQLQGI VSSGYGCAQK NKPGVYTKVC NYVNWIQQTI AAN

SEQ ID NO: 51:

IVGGYTCAAN SIPYQVSLNS GSHKCGGSLI NSQWVVSAAH CYKSRIQVRL GEHNIDVLEG NEQFINAAKI ITHPNFNGNT LDNDIMLIKL SSPATLNSRV ATVSLPRSCA AAGTECLISG WGNTKSSGSS YPSLLQCLKA PVLSDSSCKS SYPGQITGNM ICVGFLEGGK DACQGDSGGP VVCNGQLQGI VSSGYGCAQK NKPGVYTKVC NYVNWIQQTI AAN

SEQ ID NO: 52:

IVGGYTCAAN SIPYQVSLNS GSHRCGGSLI NSQWVVSAAH CYKLRIQVRL GEHNIDVLEG NEQFINAAKI ITHPNFNGNT LDNDIMLIKL SSPATLNSRV ATVSLPRSCA AAGTECLISG WGNTKSSGSS YPSLLQCLKA PVLSDSSCKS SYPGQITGNM ICVGFLEGGK DACQGDSGGP VVCNGQLQGI VSSGYGCAQK NKPGVYTKVC NYVNWIQQTI AAN

SEQ ID NO: 53:

IVGGYTCAAN SIPYQVSLNS GSHRCGGSLI NSQWVVSAAH CYKPRIQVRL GEHNIDVLEG NEQFINAAKI ITHPNFNGNT LDNDIMLIKL SSPATLNSRV ATVSLPRSCA AAGTECLISG WGNTKSSGSS YPSLLQCLKA PVLSDSSCKS SYPGQITGNM ICVGFLEGGK DACQGDSGGP VVCNGQLQGI VSSGYGCAQK NKPGVYTKVC NYVNWIQQTI AAN

SEQ ID NO: 54:

IVGGYTCAAN SIPYQVSLNS GSHRCGGSLI NSQWVVSAAH CYKSRIQVRL GEHNIAVLEG NEQFINAAKI ITHPNFNGNT LDNDIMLIKL SSPATLNSRV ATVSLPRSCA AAGTECLISG WGNTKSSGSS YPSLLQCLKA PVLSDSSCKS SYPGQITGNM ICVGFLEGGK DACQGDSGGP VVCNGQLQGI VSSGYGCAQK NKPGVYTKVC NYVNWIQQTI AAN

SEQ ID NO: 55:

IVGGYTCAAN SIPYQVSLNS GSHRCGGSLI NSQWVVSAAH CYKSRIQVRL GEHNIWVLEG NEQFINAAKI ITHPNFNGNT LDNDIMLIKL SSPATLNSRV ATVSLPRSCA AAGTECLISG WGNTKSSGSS YPSLLQCLKA PVLSDSSCKS SYPGQITGNM ICVGFLEGGK DACQGDSGGP VVCNGQLQGI VSSGYGCAQK NKPGVYTKVC NYVNWIQQTI AAN

SEQ ID NO: 56:

IVGGYTCAAN SIPYQVSLNS GSHRCGGSLI NSQWVVSAAH CYKSRIQVRL GEHNINVLEG NEQFINAAKI ITHPNFNGNT LDNDIMLIKL SSPATLNSRV ATVSLPRSCA AAGTECLISG WGNTKSSGSS YPSLLQCLKA PVLSDSSCKS SYPGQITGNM ICVGFLEGGK DACQGDSGGP VVCNGQLQGI VSSGYGCAQK NKPGVYTKVC NYVNWIQQTI AAN

SEQ ID NO: 57:

IVGGYTCAAN SIPYQVSLNS GSHRCGGSLI NSQWVVSAAH CYKSRIQVRL GEHNIHVLEG NEQFINAAKI ITHPNFNGNT LDNDIMLIKL SSPATLNSRV ATVSLPRSCA AAGTECLISG WGNTKSSGSS YPSLLQCLKA PVLSDSSCKS SYPGQITGNM ICVGFLEGGK DACQGDSGGP VVCNGQLQGI VSSGYGCAQK NKPGVYTKVC NYVNWIQQTI AAN

SEQ ID NO: 58:

IVGGYTCAAN SIPYQVSLNS GSHRCGGSLI NSQWVVSAAH CYKSRIQVRL GEHNIDVLEG NEQFINAAKI ITHPNFNGNT LDNDIMLIKL SSPATLNSRV ATVSLPPSCA AAGTECLISG WGNTKSSGSS YPSLLQCLKA PVLSDSSCKS SYPGQITGNM ICVGFLEGGK DACQGDSGGP VVCNGQLQGI VSSGYGCAQK NKPGVYTKVC NYVNWIQQTI AAN

SEQ ID NO: 59:

IVGGYTCAAN SIPYQVSLNS GSHRCGGSLI NSQWVVSAAH CYKSRIQVRL GEHNIDVLEG NEQFINAAKI ITHPNFNGNT LDNDIMLIKL SSPATLNSRV ATVSLPGSCA AAGTECLISG WGNTKSSGSS YPSLLQCLKA PVLSDSSCKS SYPGQITGNM ICVGFLEGGK DACQGDSGGP VVCNGQLQGI VSSGYGCAQK NKPGVYTKVC NYVNWIQQTI AAN

SEQ ID NO: 60:

IVGGYTCAAN SIPYQVSLNS GSHRCGGSLI NSQWVVSAAH CYKSRIQVRL GEHNIDVLEG NEQFINAAKI ITHPNFNGNT LDNDIMLIKL SSPATLNSRV ATVSLPSSCA AAGTECLISG WGNTKSSGSS YPSLLQCLKA PVLSDSSCKS SYPGQITGNM ICVGFLEGGK DACQGDSGGP VVCNGQLQGI VSSGYGCAQK NKPGVYTKVC NYVNWIQQTI AAN

SEQ ID NO: 61:

IVGGYTCAAN SIPYQVSLNS GSHRCGGSLI NSQWVVSAAH CYKSRIQVRL GEHNIDVLEG NEQFINAAKI ITHPNFNGNT LDNDIMLIKL SSPATLNSRV ATVSLPDSCA AAGTECLISG WGNTKSSGSS YPSLLQCLKA PVLSDSSCKS SYPGQITGNM ICVGFLEGGK DACQGDSGGP VVCNGQLQGI VSSGYGCAQK NKPGVYTKVC NYVNWIQQTI AAN

SEQ ID NO: 62:

IVGGYTCAAN SIPYQVSLNS GSHRCGGSLI NSQWVVSAAH CYKSRIQVRL GEHNIDVLEG NEQFINAAKI ITHPNFNGNT LDNDIMLIKL SSPATLNSRV ATVSLPRSCA AAGTECLISG WGNTKSSGSS VPSLLQCLKA PVLSDSSCKS SYPGQITGNM ICVGFLEGGK DACQGDSGGP VVCNGQLQGI VSSGYGCAQK NKPGVYTKVC NYVNWIQQTI AAN

SEQ ID NO: 63:

IVGGYTCAAN SIPYQVSLNS GSHRCGGSLI NSQWVVSAAH CYKSRIQVRL GEHNIDVLEG NEQFINAAKI ITHPNFNGNT LDNDIMLIKL SSPATLNSRV ATVSLPRSCA AAGTECLISG WGNTKSSGSS IPSLLQCLKA PVLSDSSCKS SYPGQITGNM ICVGFLEGGK DACQGDSGGP VVCNGQLQGI VSSGYGCAQK NKPGVYTKVC NYVNWIQQTI AAN

SEQ ID NO: 64:

IVGGYTCAAN SIPYQVSLNS GSHRCGGSLI NSQWVVSAAH CYKSRIQVRL GEHNIDVLEG NEQFINAAKI ITHPNFNGNT LDNDIMLIKL SSPATLNSRV ATVSLPRSCA AAGTECLISG WGNTKSSGSS WPSLLQCLKA PVLSDSSCKS SYPGQITGNM ICVGFLEGGK DACQGDSGGP VVCNGQLQGI VSSGYGCAQK NKPGVYTKVC NYVNWIQQTI AAN

SEQ ID NO: 65:

IVGGYTCAAN SIPYQVSLNS GSHRCGGSLI NSQWVVSAAH CYKSRIQVRL GEHNIDVLEG NEQFINAAKI ITHPNFNGNT LDNDIMLIKL SSPATLNSRV ATVSLPRSCA AAGTECLISG WGNTKSSGSS GPSLLQCLKA PVLSDSSCKS SYPGQITGNM ICVGFLEGGK DACQGDSGGP VVCNGQLQGI VSSGYGCAQK NKPGVYTKVC NYVNWIQQTI AAN

SEQ ID NO: 66:

IVGGYTCAAN SIPYQVSLNS GSHRCGGSLI NSQWVVSAAH CYKSRIQVRL GEHNIDVLEG NEQFINAAKI ITHPNFNGNT LDNDIMLIKL SSPATLNSRV ATVSLPRSCA AAGTECLISG WGNTKSSGSS SPSLLQCLKA PVLSDSSCKS SYPGQITGNM ICVGFLEGGK DACQGDSGGP VVCNGQLQGI VSSGYGCAQK NKPGVYTKVC NYVNWIQQTI AAN

SEQ ID NO: 67:

IVGGYTCAAN SIPYQVSLNS GSHRCGGSLI NSQWVVSAAH CYKSRIQVRL GEHNIDVLEG NEQFINAAKI ITHPNFNGNT LDNDIMLIKL SSPATLNSRV ATVSLPRSCA AAGTECLISG WGNTKSSGSS QPSLLQCLKA PVLSDSSCKS SYPGQITGNM ICVGFLEGGK DACQGDSGGP VVCNGQLQGI VSSGYGCAQK NKPGVYTKVC NYVNWIQQTI AAN

SEQ ID NO: 68:

IVGGYTCAAN SIPYQVSLNS GSHRCGGSLI NSQWVVSAAH CYKSRIQVRL GEHNIDVLEG NEQFINAAKI ITHPNFNGNT LDNDIMLIKL SSPATLNSRV ATVSLPRSCA AAGTECLISG WGNTKSSGSS DPSLLQCLKA PVLSDSSCKS SYPGQITGNM ICVGFLEGGK DACQGDSGGP VVCNGQLQGI VSSGYGCAQK NKPGVYTKVC NYVNWIQQTI AAN

SEQ ID NO: 69:

IVGGYTCAAN SIPYQVSLNS GSHRCGGSLI NSQWVVSAAH CYKSRIQVRL GEHNIDVLEG NEQFINAAKI ITHPNFNGNT LDNDIMLIKL SSPATLNSRV ATVSLPRSCA AAGTECLISG WGNTKSSGSS YPSLLQCLKA PVLSDSSCKS SYPGQITGNM ICVGFLEGGK DACQGDSGGP VVCNGQLQGI VSFGYGCAQK NKPGVYTKVC NYVNWIQQTI AAN

SEQ ID NO: 70:

IVGGYTCAAN SIPYQVSLNS GSHRCGGSLI NSQWVVSAAH CYKSRIQVRL GEHNIDVLEG NEQFINAAKI ITHPNFNGNT LDNDIMLIKL SSPATLNSRV ATVSLPRSCA AAGTECLISG WGNTKSSGSS YPSLLQCLKA PVLSDSSCKS SYPGQITGNM ICVGFLEGGK DACQGDSGGP VVCNGQLQGI VSYGYGCAQK NKPGVYTKVC NYVNWIQQTI AAN

SEQ ID NO: 71:

IVGGYTCAAN SIPYQVSLNS GSHRCGGSLI NSQWVVSAAH CYKSRIQVRL GEHNIDVLEG NEQFINAAKI ITHPNFNGNT LDNDIMLIKL SSPATLNSRV ATVSLPRSCA AAGTECLISG WGNTKSSGSS YPSLLQCLKA PVLSDSSCKS SYPGQITGNM ICVGFLEGGK DACQGDSGGP VVCNGQLQGI VSTGYGCAQK NKPGVYTKVC NYVNWIQQTI AAN

SEQ ID NO: 72:

IVGGYTCAAN SIPYQVSLNS GSHRCGGSLI NSQWVVSAAH CYKSRIQVRL GEHNIDVLEG NEQFINAAKI ITHPNFNSNT LDNDIMLIKL SSPATLNSRV ATVSLPRSCA AAGTECLISG WGNTKSSGSS YPSLLQCLKA PVLSDSSCKS SYPGQITGNM ICVGFLEGGK DACQGDSGGP VVCNGQLQGI VSSGYGCAQK NKPGVYTKVC NYVNWIQQTI AAN

SEQ ID NO: 73:

IVGGYTCAAN SIPYQVSLNS GSHRCGGSLI NSQWVVSAAH CYKSRIQVRL GEHNIDVLEG NEQFINAAKI ITHPNFNANT LDNDIMLIKL SSPATLNSRV ATVSLPRSCA AAGTECLISG WGNTKSSGSS YPSLLQCLKA PVLSDSSCKS SYPGQITGNM ICVGFLEGGK DACQGDSGGP VVCNGQLQGI VSSGYGCAQK NKPGVYTKVC NYVNWIQQTI AAN

SEQ ID NO: 74:

IVGGYTCAAN SIPYQVSLNS GSHRCGGSLI NSQWVVSAAH CYKSRIQVRL GEHNIDVLEG NEQFINAAKI ITHPNFNPNT LDNDIMLIKL SSPATLNSRV ATVSLPRSCA AAGTECLISG WGNTKSSGSS YPSLLQCLKA PVLSDSSCKS SYPGQITGNM ICVGFLEGGK DACQGDSGGP VVCNGQLQGI VSSGYGCAQK NKPGVYTKVC NYVNWIQQTI AAN

SEQ ID NO: 75:

IVGGYTCAAN SIPYQVSLNS GSHRCGGSLI NSQWVVSAAH CYKSRIQVRL GEHNIDVLEG NEQFINAAKI ITHPNFNYNT LDNDIMLIKL SSPATLNSRV ATVSLPRSCA AAGTECLISG WGNTKSSGSS YPSLLQCLKA PVLSDSSCKS SYPGQITGNM ICVGFLEGGK DACQGDSGGP VVCNGQLQGI VSSGYGCAQK NKPGVYTKVC NYVNWIQQTI AAN

SEQ ID NO: 76:

IVGGYTCAAN SIPYQVSLNS GSHRCGGSLI NSQWVVSAAH CYKSRIQVRL GEHNIDVLEG NEQFINAAKI ITHPNFNENT LDNDIMLIKL SSPATLNSRV ATVSLPRSCA AAGTECLISG WGNTKSSGSS YPSLLQCLKA PVLSDSSCKS SYPGQITGNM ICVGFLEGGK DACQGDSGGP VVCNGQLQGI VSSGYGCAQK NKPGVYTKVC NYVNWIQQTI AAN

SEQ ID NO: 77:

IVGGYTCAAN SIPYQVSLNS GSHRCGGSLI NSQWVVSAAH CYKPRIQVRL GEHNIHVLEG NEQFINAAKI ITHPNFNGNT LDNDIMLIKL SSPATLNSHV ATVSLPTSCA AAGTECLISG WGNTCSSGSS MPSLLQCLKA PVLSDSSCKS SYPGQITGNM ICVGFLEGGK DACQGDSGGP VVCNGQLQGI VSSGYGCAQK NKPGVYTKVC NYVNWIQQTI AAN

SEQ ID NO: 78:

IVGGYTCAAN SIPYQVSLNS GSHRCGGSLI NSQWVVSAAH CYKSRIQVRL GEHNIHVLEG NEQFINAAKI ITHPNFNPNT LDNDIMLIKL SSPATLNSHV ATVSLPTSCA AAGTECLISG WGNTCSSGSS YPSLLQCLKA PVLSDSSCKS SYPGQITGNM ICVGFLEGGK DACQGDSGGP VVCNGQLQGI VSSGYGCAQK NKPGVYTKVC NYVNWIQQTI AAN

SEQ ID NO: 79:

IVGGYTCAAN SIPYQVSLNS GSHRCGGSLI NSQWVVSAAH CYKPRIQVRL GEHNIHVLEG NEQFINAAKI ITHPNFNPNT LDNDIMLIKL SSPATLNSHV ATVSLPRSCA AAGTECLISG WGNTASSGSS MPSLLQCLKA PVLSDSSCKS SYPGQITGNM ICVGFLEGGK DACQGDSGGP VVCNGQLQGI VSSGYGCAQK NKPGVYTKVC NYVNWIQQTI AAN

SEQ ID NO: 80:

IVGGYTCAAN SIPYQVSLNS GSHRCGGSLI NSQWVVSAAH CYKSRIQVRL GEHNIHVLEG NEQFINAAKI ITHPNFNPNT LDNDIMLIKL SSPATLNSHV ATVSLPTSCA AAGTECLISG WGNTSSSGSS MPSLLQCLKA PVLSDSSCKS SYPGQITGNM ICVGFLEGGK DACQGDSGGP VVCNGQLQGI VSSGYGCAQK NKPGVYTKVC NYVNWIQQTI AAN

SEQ ID NO: 81:

IVGGYTCAAN SIPYQVSLNS GSHRCGGSLI NSQWVVSAAH CYKSRIQVRL GEHNIHVLEG NEQFINAAKI ITHPNFNPNT LDNDIMLIKL SSPATLNSHV ATVSLPTSCA AAGTECLISG WGNTASSGSS MPSLLQCLKA PVLSDSSCKS SYPGQITGNM ICVGFLEGGK DACQGDSGGP VVCNGQLQGI VSSGYGCAQK NKPGVYTKVC NYVNWIQQTI AAN

SEQ ID NO: 82:

IVGGYTCAAN SIPYQVSLNS GSHRCGGSLI NSQWVVSAAH CYKPRIQVRL GEHNIHVLEG NEQFINAAKI ITHPNFNPNT LDNDIMLIKL SSPATLNSHV ATVSLPRSCA AAGTECLISG WGNTCSSGSS MPSLLQCLKA PVLSDSSCKS SYPGQITGNM ICVGFLEGGK DACQGDSGGP VVCNGQLQGI VSSGYGCAQK NKPGVYTKVC NYVNWIQQTI AAN

SEQ ID NO: 83:

IVGGYTCAAN SIPYQVSLNS GSHRCGGSLI NSQWVVSAAH CYKPRIQVRL GEHNIHVLEG NEQFINAAKI ITHPNFNPNT LDNDIMLIKL SSPATLNSNV ATVSLPTSCA AAGTECLISG WGNTSSSGSS MPSLLQCLKA PVLSDSSCKS SYPGQITGNM ICVGFLEGGK DACQGDSGGP VVCNGQLQGI VSSGYGCAQK NKPGVYTKVC NYVNWIQQTI AAN

SEQ ID NO: 84:

IVGGYTCAAN SIPYQVSLNS GSHRCGGSLI NSQWVVSAAH CYKPRIQVRL GEHNIHVLEG NEQFINAAKI ITHPNFNPNT LDNDIMLIKL SSPATLNSNV ATVSLPTSCA AAGTECLISG WGNTSSSGSS MPSLLQCLKA PVLSDSSCKS SYPGQITGNM ICVGFLEGGK DSCQGDSGGP VVCNGQLQGI VSSGYGCAQK NKPGVYTKVC NYVNWIQQTI AAN

SEQ ID NO: 85:

IVGGYTCAAN SIPYQVSLNS GSHRCGGSLI NSQWVVSAAH CYKPRIQVRL GEHNIHVLEG NEQFINAAKI ITHPNFNPNT LDNDIMLIKL SSPATLNSHV ATVSLPTSCA AAGTECLISG WGNTCSSGSS MPSLLQCLKA PVLSDSSCKS SYPGQITGNM ICVGFLEGGK DACQGDSGGP VVCNGQLQGI VSSGYGCAQK NKPGVYTKVC NYVNWIQQTI AAN

SEQ ID NO: 86:

IVGGYTCAAN SIPYQVSLNS GSHRCGGSLI NSQWVVSAAH CYKPRIQVRL GEHNIDVLEG NEQFINAAKI ITHPNFNPNT LDNDIMLIKL SSPATLNSAV ATVSLPTSCA AAGTECLISG WGNTSSSGSS MPSLLQCLKA PVLSDSSCKS SYPGQITGNM ICVGFLEGGK DACQGDSGGP VVCNGQLQGI VSSGYGCAQK NKPGVYTKVC NYVNWIQQTI AAN

SEQ ID NO: 87:

IVGGYTCAAN SIPYQVSLNS GSHRCGGSLI NSQWVVSAAH CYKPRIQVRL GEHNIHVLEG NEQFINAAKI ITHPNFNPNT LDNDIMLIKL SSPATLNSHV ATVSLPTSCA AAGTECLISG WGNTSSSGSS MPSLLQCLKA PVLSDSSCKS SYPGQITGNM ICVGFLEGGK DACQGDSGGP VVCNGQLQGI VSSGYGCAQK NKPGVYTKVC NYVNWIQQTI AAN

SEQ ID NO: 88:

IVGGYTCAAN SIPYQVSLNS GSHRCGGSLI NSQWVVSAAH CYKPRIQVRL GEHNIHVLEG NEQFINAAKI ITHPNFNPNT LDNDIMLIKL SSPATLNSHV ATVSLPTSCA AAGTECLISG WGNTSSSGSS MPSLLQCLKA PVLSDSSCKS SYPGQITGNM ICVGFLEGGK DSCQGDSGGP VVCNGQLQGI VSSGYGCAQK NKPGVYTKVC NYVNWIQQTI AAN

SEQ ID NO: 89:

IVGGYTCAAN SIPYQVSLNS GSHRCGGSLI NSQWVVSAAH CYKPRIQVRL GEHNIHVLEG NEQFINAAKI ITHPNFNPNT LDNDIMLIKL SSPATLNSHV ATVSLPTSCA AAGTECLISG WGNTASSGSS MPSLLQCLKA PVLSDSSCKS SYPGQITGNM ICVGFLEGGK DACQGDSGGP VVCNGQLQGI VSSGYGCAQK NKPGVYTKVC NYVNWIQQTI AAN

SEQ ID NO: 90:

IVGGYTCAAN SIPYQVSLNS GSHRCGGSLI NSQWVVSAAH CYKPRIQVRL GEHNIHVLEG NEQFINAAKI ITHPNFNPNT LDNDIMLIKL SSPATLNSHV ATVSLPTSCA AAGTECLISG WGNTASSGSS MPSLLQCLKA PVLSDSSCKS SYPGQITGNM ICVGFLEGGK DSCQGDSGGP VVCNGQLQGI VSSGYGCAQK NKPGVYTKVC NYVNWIQQTI AAN

SEQ ID NO: 91:

IVGGYTCAAN SIPYQVSLNS GSHRCGGSLI NSQWVVSAAH CYKPRIQVRL GEHNIHVLEG NEQFINAAKI ITHPNFNPNT LDNDIMLIKL SSPATLNSAV ATVSLPTSCA AAGTECLISG WGNTSSSGSS MPSLLQCLKA PVLSDSSCKS SYPGQITGNM ICVGFLEGGK DACQGDSGGP VVCNGQLQGI VSSGYGCAQK NKPGVYTKVC NYVNWIQQTI AAN

SEQ ID NO: 92:

IVGGYTCAAN SIPYQVSLNS GSHRCGGSLI NSQWVVSAAH CYKPRIQVRL GEHNIHVLEG NEQFINAAKI ITHPNFNPNT LDNDIMLIKL SSPATLNSAV ATVSLPTSCA AAGTECLISG WGNTCSSGSS MPSLLQCLKA PVLSDSSCKS SYPGQITGNM ICVGFLEGGK DACQGDSGGP VVCNGQLQGI VSSGYGCAQK NKPGVYTKVC NYVNWIQQTI AAN

SEQ ID NO: 93:

IVGGYTCAAN SIPYQVSLNS GSHRCGGSLI NSQWVVSAAH CYKSRIQVRL GEHNIDVLEG NEQFINAAKI ITHPNFNPNT LDNDIMLIKL SSPATLNSNV ATVSLPTSCA AAGTECLISG WGNTCSSGSS MPSLLQCLKA PVLSDSSCKS SYPGQITGNM ICVGFLEGGA DACQGDSGGP VVCNGQLQGI VSSGYGCAQK NKPGVYTKVC NYVNWIQQTI AAN

SEQ ID NO: 94:

IVGGYTCAAN SIPYQVSLNS GSHRCGGSLI NSQWVVSAAH CYKPRIQVRL GEHNIHVLEG NEQFINAAKI ITHPNFNGNT LDNDIMLIKL SSPATLNSNV ATVSLPRSCA AAGTECLISG WGNTCSSGSS MPSLLQCLKA PVLSDSSCKS SYPGQITGNM ICVGFLEGGA DACQGDSGGP VVCNGQLQGI VSSGYGCAQK NKPGVYTKVC NYVNWIQQTI AAN

SEQ ID NO: 95:

IVGGYTCAAN SIPYQVSLNS GSHRCGGSLI NSQWVVSAAH CYKPRIQVRL GEHNIHVLEG NEQFINAAKI ITHPNFNPNT LDNDIMLIKL SSPATLNSHV ATVSLPRSCA AAGTECLISG WGNTSSSGSS MPSLLQCLKA PVLSDSSCKS SYPGQITGNM ICVGFLEGGA DACQGDSGGP VVCNGQLQGI VSSGYGCAQK NKPGVYTKVC NYVNWIQQTI AAN

SEQ ID NO: 96:

IVGGYTCAAN SIPYQVSLNS GSHRCGGSLI NSQWVVSAAH CYKPRIQVRL GEHNIDVLEG NEQFINAAKI ITHPNFNPNT LDNDIMLIKL SSPATLNSHV ATVSLPRSCA AAGTECLISG WGNTSSSGSS MPSLLQCLKA PVLSDSSCKS SYPGQITGNM ICVGFLEGGA DACQGDSGGP VVCNGQLQGI VSSGYGCAQK NKPGVYTKVC NYVNWIQQTI AAN

SEQ ID NO: 97:

IVGGYTCAAN SIPYQVSLNS GSHRCGGSLI NSQWVVSAAH CYKPRIQVRL GEHNIHVLEG NEQFINAAKI ITHPNFNPNT LDNDIMLIKL SSPATLNSAV ATVSLPTSCA AAGTECLISG WGNTASSGSS MPSLLQCLKA PVLSDSSCKS SYPGQITGNM ICVGFLEGGA DACQGDSGGP VVCNGQLQGI VSSGYGCAQK NKPGVYTKVC NYVNWIQQTI AAN

SEQ ID NO: 98:

IVGGYTCAAN SIPYQVSLNS GSHRCGGSLI NSQWVVSAAH CYKPRIQVRL GEHNIHVLEG NEQFINAAKI ITHPNFNPNT LDNDIMLIKL SSPATLNSAV ATVSLPTSCA AAGTECLISG WGNTASSGSS MPSLLQCLKA PVLSDSSCKS SYPGQITGNM ICVGFLEGGA DSCQGDSGGP VVCNGQLQGI VSSGYGCAQK NKPGVYTKVC NYVNWIQQTI AAN

SEQ ID NO: 99:

IVGGYTCAAN SIPYQVSLNS GSHRCGGSLI NSQWVVSAAH CYKSRIQVRL GEHNIHVLEG NEQFINAAKI ITHPNFNPNT LDNDIMLIKL SSPATLNSNV ATVSLPTSCA AAGTECLISG WGNTSSSGSS YPSLLQCLKA PVLSDSSCKS SYPGQITGNM ICVGFLEGGA DACQGDSGGP VVCNGQLQGI VSSGYGCAQK NKPGVYTKVC NYVNWIQQTI AAN

SEQ ID NO: 100:

IVGGYTCAAN SIPYQVSLNS GSHRCGGSLI NSQWVVSAAH CYKPRIQVRL GEHNIDVLEG NEQFINAAKI ITHPNFNPNT LDNDIMLIKL SSPATLNSHV ATVSLPTSCA AAGTECLISG WGNTCSSGSS MPSLLQCLKA PVLSDSSCKS SYPGQITGNM ICVGFLEGGA DACQGDSGGP VVCNGQLQGI VSSGYGCAQK NKPGVYTKVC NYVNWIQQTI AAN

SEQ ID NO: 101:

IVGGYTCAAN SIPYQVSLNS GSHRCGGSLI NSQWVVSAAH CYKSRIQVRL GEHNIHVLEG NEQFINAAKI ITHPNFNPNT LDNDIMLIKL SSPATLNSNV ATVSLPTSCA AAGTECLISG WGNTSSSGSS MPSLLQCLKA PVLSDSSCKS SYPGQITGNM ICVGFLEGGA DACQGDSGGP VVCNGQLQGI VSSGYGCAQK NKPGVYTKVC NYVNWIQQTI AAN

SEQ ID NO: 102:

IVGGYTCAAN SIPYQVSLNS GSHRCGGSLI NSQWVVSAAH CYKPRIQVRL GEHNIHVLEG NEQFINAAKI ITHPNFNPNT LDNDIMLIKL SSPATLNSHV ATVSLPTSCA AAGTECLISG WGNTSSSGSS MPSLLQCLKA PVLSDSSCKS SYPGQITGNM ICVGFLEGGA DACQGDSGGP VVCNGQLQGI VSSGYGCAQK NKPGVYTKVC NYVNWIQQTI AAN

SEQ ID NO: 103:

IVGGYTCAAN SIPYQVSLNS GSHRCGGSLI NSQWVVSAAH CYKPRIQVRL GEHNIHVLEG NEQFINAAKI ITHPNFNPNT LDNDIMLIKL SSPATLNSHV ATVSLPTSCA AAGTECLISG WGNTSSSGSS MPSLLQCLKA PVLSDSSCKS SYPGQITGNM ICVGFLEGGA DSCQGDSGGP VVCNGQLQGI VSSGYGCAQK NKPGVYTKVC NYVNWIQQTI AAN

SEQ ID NO: 104:

IVGGYTCAAN SIPYQVSLNS GSHRCGGSLI NSQWVVSAAH CYKSRIQVRL GEHNIHVLEG NEQFINAAKI ITHPNFNPNT LDNDIMLIKL SSPATLNSNV ATVSLPTSCA AAGTECLISG WGNTCSSGSS MPSLLQCLKA PVLSDSSCKS SYPGQITGNM ICVGFLEGGA DACQGDSGGP VVCNGQLQGI VSSGYGCAQK NKPGVYTKVC NYVNWIQQTI AAN

SEQ ID NO: 105:

IVGGYTCAAN SIPYQVSLNS GSHRCGGSLI NSQWVVSAAH CYKSRIQVRL GEHNIHVLEG NEQFINAAKI ITHPNFNPNT LDNDIMLIKL SSPATLNSNV ATVSLPTSCA AAGTECLISG WGNTASSGSS YPSLLQCLKA PVLSDSSCKS SYPGQITGNM ICVGFLEGGA DACQGDSGGP VVCNGQLQGI VSSGYGCAQK NKPGVYTKVC NYVNWIQQTI AAN

SEQ ID NO: 106:

IVGGYTCAAN SIPYQVSLNS GSHRCGGSLI NSQWVVSAAH CYKSRIQVRL GEHNIHVLEG NEQFINAAKI ITHPNFNPNT LDNDIMLIKL SSPATLNSAV ATVSLPTSCA AAGTECLISG WGNTCSSGSS MPSLLQCLKA PVLSDSSCKS SYPGQITGNM ICVGFLEGGA DACQGDSGGP VVCNGQLQGI VSSGYGCAQK NKPGVYTKVC NYVNWIQQTI AAN

SEQ ID NO: 107:

IVGGYTCAAN SIPYQVSLNS GSHRCGGSLI NSQWVVSAAH CYKPRIQVRL GEHNIHVLEG NEQFINAAKI ITHPNFNPNT LDNDIMLIKL SSPATLNSHV ATVSLPTSCA AAGTECLISG WGNTCSSGSS MPSLLQCLKA PVLSDSSCKS SYPGQITGNM ICVGFLEGGA DACQGDSGGP VVCNGQLQGI VSSGYGCAQK NKPGVYTKVC NYVNWIQQTI AAN

SEQ ID NO: 108:

IVGGYTCAAN SIPYQVSLNS GSHRCGGSLI NSQWVVSAAH CYKPRIQVRL GEHNIDVLEG NEQFINAAKI ITHPNFNPNT LDNDIMLIKL SSPATLNSAV ATVSLPTSCA AAGTECLISG WGNTCSSGSS MPSLLQCLKA PVLSDSSCKS SYPGQITGNM ICVGFLEGGA DACQGDSGGP VVCNGQLQGI VSSGYGCAQK NKPGVYTKVC NYVNWIQQTI AAN

SEQ ID NO: 109:

IVGGYTCAAN SIPYQVSLNS GSHRCGGSLI NSQWVVSAAH CYKSRIQVRL GEHNIHVLEG NEQFINAAKI ITHPNFNGNT LDNDIMLIKL SSPATLNSNV ATVSLPTSCA AAGTECLISG WGNTCSSGSS MPSLLQCLKA PVLSDSSCKS SYPGQITGNM ICVGFLEGGA DACQGDSGGP VVCNGQLQGI VSSGYGCAQK NKPGVYTKVC NYVNWIQQTI AAN

SEQ ID NO: 110:

IVGGYTCAAN SIPYQVSLNS GSHRCGGSLI NSQWVVSAAH CYKSRIQVRL GEHNIHVLEG NEQFINAAKI ITHPNFNPNT LDNDIMLIKL SSPATLNSHV ATVSLPRSCA AAGTECLISG WGNTCSSGSS MPSLLQCLKA PVLSDSSCKS SYPGQITGNM ICVGFLEGGA DACQGDSGGP VVCNGQLQGI VSSGYGCAQK NKPGVYTKVC NYVNWIQQTI AAN

SEQ ID NO: 111:

IVGGYTCAAN SIPYQVSLNS GSHRCGGSLI NSQWVVSAAH CYKPRIQVRL GEHNIHVLEG NEQFINAAKI ITHPNFNPNT LDNDIMLIKL SSPATLNSHV ATVSLPTSCA AAGTECLISG WGNTSSSGSS YPSLLQCLKA PVLSDSSCKS SYPGQITGNM ICVGFLEGGA DACQGDSGGP VVCNGQLQGI VSSGYGCAQK NKPGVYTKVC NYVNWIQQTI AAN

SEQ ID NO: 112:

IVGGYTCAAN SIPYQVSLNS GSHRCGGSLI NSQWVVSAAH CYKPRIQVRL GEHNIHVLEG NEQFINAAKI ITHPNFNPNT LDNDIMLIKL SSPATLNSHV ATVSLPTSCA AAGTECLISG WGNTSSSGSS YPSLLQCLKA PVLSDSSCKS SYPGQITGNM ICVGFLEGGA DSCQGDSGGP VVCNGQLQGI VSSGYGCAQK NKPGVYTKVC NYVNWIQQTI AAN

SEQ ID NO: 113:

IVGGYTCAAN SIPYQVSLNS GSHRCGGSLI NSQWVVSAAH CYKPRIQVRL GEHNIHVLEG NEQFINAAKI ITHPNFNGNT LDNDIMLIKL SSPATLNSAV ATVSLPTSCA AAGTECLISG WGNTASSGSS YPSLLQCLKA PVLSDSSCKS SYPGQITGNM ICVGFLEGGA DACQGDSGGP VVCNGQLQGI VSSGYGCAQK NKPGVYTKVC NYVNWIQQTI AAN

SEQ ID NO: 114:

IVGGYTCAAN SIPYQVSLNS GSHRCGGSLI NSQWVVSAAH CYKPRIQVRL GEHNIHVLEG NEQFINAAKI ITHPNFNGNT LDNDIMLIKL SSPATLNSAV ATVSLPTSCA AAGTECLISG WGNTASSGSS YPSLLQCLKA PVLSDSSCKS SYPGQITGNM ICVGFLEGGA DSCQGDSGGP VVCNGQLQGI VSSGYGCAQK NKPGVYTKVC NYVNWIQQTI AAN

SEQ ID NO: 115:

IVGGYTCAAN SIPYQVSLNS GSHRCGGSLI NSQWVVSAAH CYKPRIQVRL GEHNIHVLEG NEQFINAAKI ITHPNFNGNT LDNDIMLIKL SSPATLNSNV ATVSLPTSCA AAGTECLISG WGNTSSSGSS MPSLLQCLKA PVLSDSSCKS SYPGQITGNM ICVGFLEGGK DACQGDSGGP VVCNGQLQGI VSSGYGCAQK NKPGVYTKVC NYVNWIQQTI AAN

SEQ ID NO: 116:

IVGGYTCAAN SIPYQVSLNS GSHRCGGSLI NSQWVVSAAH CYKPRIQVRL GEHNIHVLEG NEQFINAAKI ITHPNFNGNT LDNDIMLIKL SSPATLNSNV ATVSLPTSCA AAGTECLISG WGNTSSSGSS MPSLLQCLKA PVLSDSSCKS SYPGQITGNM ICVGFLEGGA DACQGDSGGP VVCNGQLQGI VSSGYGCAQK NKPGVYTKVC NYVNWIQQTI AAN

SEQ ID NO: 117:

IVGGYTCAAN SIPYQVSLNS GSHRCGGSLI NSQWVVSAAH CYKPRIQVRL GEHNIHVLEG NEQFINAAKI ITHPNFNGNT LDNDIMLIKL SSPATLNSNV ATVSLPTSCA AAGTECLISG WGNTASSGSS MPSLLQCLKA PVLSDSSCKS SYPGQITGNM ICVGFLEGGK DACQGDSGGP VVCNGQLQGI VSSGYGCAQK NKPGVYTKVC NYVNWIQQTI AAN

SEQ ID NO: 118:

IVGGYTCAAN SIPYQVSLNS GSHRCGGSLI NSQWVVSAAH CYKPRIQVRL GEHNIHVLEG NEQFINAAKI ITHPNFNGNT LDNDIMLIKL SSPATLNSHV ATVSLPTSCA AAGTECLISG WGNTASSGSS MPSLLQCLKA PVLSDSSCKS SYPGQITGNM ICVGFLEGGA DACQGDSGGP VVCNGQLQGI VSSGYGCAQK NKPGVYTKVC NYVNWIQQTI AAN

SEQ ID NO: 119:

IVGGYTCAAN SIPYQVSLNS GSHRCGGSLI NSQWVVSAAH CYKPRIQVRL GEHNIHVLEG NEQFINAAKI ITHPNFNGNT LDNDIMLIKL SSPATLNSAV ATVSLPTSCA AAGTECLISG WGNTASSGSS MPSLLQCLKA PVLSDSSCKS SYPGQITGNM ICVGFLEGGK DACQGDSGGP VVCNGQLQGI VSSGYGCAQK NKPGVYTKVC NYVNWIQQTI AAN

--->

SEQUENCE LIST

<110> SANOFI-AVENTIS DEUTSCHLAND GMBH

<120> PORK TRIPSIN VARIANTS

<130> 589-276 PCT

<150>EP18158034.1

<151> 2018-02-22

<160> 130

<170> PatentIn version 3.5

<210> 1

<211> 223

<212> PROTEIN

<213> Sus scrofa

<400> 1

Ile Val Gly Gly Tyr Thr Cys Ala Ala Asn Ser Ile Pro Tyr Gln Val

1 5 10 15

Ser Leu Asn Ser Gly Ser His Phe Cys Gly Gly Ser Leu Ile Asn Ser

20 25 30

Gln Trp Val Val Ser Ala Ala His Cys Tyr Lys Ser Arg Ile Gln Val

35 40 45

Arg Leu Gly Glu His Asn Ile Asp Val Leu Glu Gly Asn Glu Gln Phe

50 55 60

Ile Asn Ala Ala Lys Ile Ile Thr His Pro Asn Phe Asn Gly Asn Thr

65 70 75 80

Leu Asp Asn Asp Ile Met Leu Ile Lys Leu Ser Ser Pro Ala Thr Leu

85 90 95

Asn Ser Arg Val Ala Thr Val Ser Leu Pro Arg Ser Cys Ala Ala Ala

100 105 110

Gly Thr Glu Cys Leu Ile Ser Gly Trp Gly Asn Thr Lys Ser Ser Gly

115 120 125

Ser Ser Tyr Pro Ser Leu Leu Gln Cys Leu Lys Ala Pro Val Leu Ser

130 135 140

Asp Ser Ser Cys Lys Ser Ser Tyr Pro Gly Gln Ile Thr Gly Asn Met

145 150 155 160

Ile Cys Val Gly Phe Leu Glu Gly Gly Lys Asp Ser Cys Gln Gly Asp

165 170 175

Ser Gly Gly Pro Val Val Cys Asn Gly Gln Leu Gln Gly Ile Val Ser

180 185 190

Trp Gly Tyr Gly Cys Ala Gln Lys Asn Lys Pro Gly Val Tyr Thr Lys

195 200 205

Val Cys Asn Tyr Val Asn Trp Ile Gln Gln Thr Ile Ala Ala Asn

210 215 220

<210> 2

<211> 223

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Porcine trypsin variant S172A

<400> 2

Ile Val Gly Gly Tyr Thr Cys Ala Ala Asn Ser Ile Pro Tyr Gln Val

1 5 10 15

Ser Leu Asn Ser Gly Ser His Phe Cys Gly Gly Ser Leu Ile Asn Ser

20 25 30

Gln Trp Val Val Ser Ala Ala His Cys Tyr Lys Ser Arg Ile Gln Val

35 40 45

Arg Leu Gly Glu His Asn Ile Asp Val Leu Glu Gly Asn Glu Gln Phe

50 55 60

Ile Asn Ala Ala Lys Ile Ile Thr His Pro Asn Phe Asn Gly Asn Thr

65 70 75 80

Leu Asp Asn Asp Ile Met Leu Ile Lys Leu Ser Ser Pro Ala Thr Leu

85 90 95

Asn Ser Arg Val Ala Thr Val Ser Leu Pro Arg Ser Cys Ala Ala Ala

100 105 110

Gly Thr Glu Cys Leu Ile Ser Gly Trp Gly Asn Thr Lys Ser Ser Gly

115 120 125

Ser Ser Tyr Pro Ser Leu Leu Gln Cys Leu Lys Ala Pro Val Leu Ser

130 135 140

Asp Ser Ser Cys Lys Ser Ser Tyr Pro Gly Gln Ile Thr Gly Asn Met

145 150 155 160

Ile Cys Val Gly Phe Leu Glu Gly Gly Lys Asp Ala Cys Gln Gly Asp

165 170 175

Ser Gly Gly Pro Val Val Cys Asn Gly Gln Leu Gln Gly Ile Val Ser

180 185 190

Trp Gly Tyr Gly Cys Ala Gln Lys Asn Lys Pro Gly Val Tyr Thr Lys

195 200 205

Val Cys Asn Tyr Val Asn Trp Ile Gln Gln Thr Ile Ala Ala Asn

210 215 220

<210> 3

<211> 223

<212> PROTEIN

<213> Artificial sequence

<220>

<223> General porcine trypsin sequence with variable positions in

amino acid positions 24, 44, 56, 78, 99, 107, 125, 131, 170, 172 and 193

<220>

<221> MISC_FEATURE

<222> (24)..(24)

<223> Xaa is selected from the group consisting of Ala, Asn, Arg, Gln, Ile, Leu, Lys, Met,

Phe, Ser, Thr and Val

<220>

<221> MISC_FEATURE

<222> (44)..(44)

<223> Xaa is selected from the group consisting of Leu, Pro and Ser

<220>

<221> MISC_FEATURE

<222> (56)..(56)

<223> Xaa is selected from the group consisting of Ala, Asn, Asp, His and Trp

<220>

<221> MISC_FEATURE

<222> (78)..(78)

<223> Xaa is selected from the group consisting of Ala, Glu, Gly, Pro, Ser and Tyr

<220>

<221> MISC_FEATURE

<222> (99)..(99)

<223> Xaa is selected from the group consisting of Ala, Arg, Asn, Asp, Glu, Gly, His, Leu,

Phe, Thr, Trp and Tyr

<220>

<221> MISC_FEATURE

<222> (107)..(107)

<223> Xaa is selected from the group consisting of Arg, Asp, Gly, Pro, Ser and Thr

<220>

<221> MISC_FEATURE

<222> (125)..(125)

<223> Xaa is selected from the group consisting of Ala, Cys, Gln, Glu, Gly, His, Leu, Lys,

Ser and Tyr

<220>

<221> MISC_FEATURE

<222> (131)..(131)

<223> Xaa is selected from the group consisting of Ala, Asn, Asp, Cys, Gln; Glu, Gly, His,

Ile, Leu, Met, Ser, Thr, Trp, Tyr and Val

<220>

<221> MISC_FEATURE

<222> (170)..(170)

<223> Xaa is selected from the group consisting of Ala, Asn, Gly, Lys and Tyr

<220>

<221> MISC_FEATURE

<222> (172)..(172)

<223> Xaa is selected from the group consisting of Ala, Cys, Ser and Thr

<220>

<221> MISC_FEATURE

<222> (193)..(193)

<223> Xaa is selected from the group consisting of Phe, Ser, Thr, Trp and Tyr

<400> 3

Ile Val Gly Gly Tyr Thr Cys Ala Ala Asn Ser Ile Pro Tyr Gln Val

1 5 10 15

Ser Leu Asn Ser Gly Ser His Xaa Cys Gly Gly Ser Leu Ile Asn Ser

20 25 30

Gln Trp Val Val Ser Ala Ala His Cys Tyr Lys Xaa Arg Ile Gln Val

35 40 45

Arg Leu Gly Glu His Asn Ile Xaa Val Leu Glu Gly Asn Glu Gln Phe

50 55 60

Ile Asn Ala Ala Lys Ile Ile Thr His Pro Asn Phe Asn Xaa Asn Thr

65 70 75 80

Leu Asp Asn Asp Ile Met Leu Ile Lys Leu Ser Ser Pro Ala Thr Leu

85 90 95

Asn Ser Xaa Val Ala Thr Val Ser Leu Pro Xaa Ser Cys Ala Ala Ala

100 105 110

Gly Thr Glu Cys Leu Ile Ser Gly Trp Gly Asn Thr Xaa Ser Ser Gly

115 120 125

Ser Ser Xaa Pro Ser Leu Leu Gln Cys Leu Lys Ala Pro Val Leu Ser

130 135 140

Asp Ser Ser Cys Lys Ser Ser Tyr Pro Gly Gln Ile Thr Gly Asn Met

145 150 155 160

Ile Cys Val Gly Phe Leu Glu Gly Gly Xaa Asp Xaa Cys Gln Gly Asp

165 170 175

Ser Gly Gly Pro Val Val Cys Asn Gly Gln Leu Gln Gly Ile Val Ser

180 185 190

Xaa Gly Tyr Gly Cys Ala Gln Lys Asn Lys Pro Gly Val Tyr Thr Lys

195 200 205

Val Cys Asn Tyr Val Asn Trp Ile Gln Gln Thr Ile Ala Ala Asn

210 215 220

<210> 4

<211> 223

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Porcine trypsin variant #4

<400> 4

Ile Val Gly Gly Tyr Thr Cys Ala Ala Asn Ser Ile Pro Tyr Gln Val

1 5 10 15

Ser Leu Asn Ser Gly Ser His Arg Cys Gly Gly Ser Leu Ile Asn Ser

20 25 30

Gln Trp Val Val Ser Ala Ala His Cys Tyr Lys Ser Arg Ile Gln Val

35 40 45

Arg Leu Gly Glu His Asn Ile Asp Val Leu Glu Gly Asn Glu Gln Phe

50 55 60

Ile Asn Ala Ala Lys Ile Ile Thr His Pro Asn Phe Asn Gly Asn Thr

65 70 75 80

Leu Asp Asn Asp Ile Met Leu Ile Lys Leu Ser Ser Pro Ala Thr Leu

85 90 95

Asn Ser Ala Val Ala Thr Val Ser Leu Pro Arg Ser Cys Ala Ala Ala

100 105 110

Gly Thr Glu Cys Leu Ile Ser Gly Trp Gly Asn Thr Lys Ser Ser Gly

115 120 125

Ser Ser Tyr Pro Ser Leu Leu Gln Cys Leu Lys Ala Pro Val Leu Ser

130 135 140

Asp Ser Ser Cys Lys Ser Ser Tyr Pro Gly Gln Ile Thr Gly Asn Met

145 150 155 160

Ile Cys Val Gly Phe Leu Glu Gly Gly Lys Asp Ala Cys Gln Gly Asp

165 170 175

Ser Gly Gly Pro Val Val Cys Asn Gly Gln Leu Gln Gly Ile Val Ser

180 185 190

Ser Gly Tyr Gly Cys Ala Gln Lys Asn Lys Pro Gly Val Tyr Thr Lys

195 200 205

Val Cys Asn Tyr Val Asn Trp Ile Gln Gln Thr Ile Ala Ala Asn

210 215 220

<210> 5

<211> 223

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Porcine trypsin variant #5

<400> 5

Ile Val Gly Gly Tyr Thr Cys Ala Ala Asn Ser Ile Pro Tyr Gln Val

1 5 10 15

Ser Leu Asn Ser Gly Ser His Arg Cys Gly Gly Ser Leu Ile Asn Ser

20 25 30

Gln Trp Val Val Ser Ala Ala His Cys Tyr Lys Ser Arg Ile Gln Val

35 40 45

Arg Leu Gly Glu His Asn Ile Asp Val Leu Glu Gly Asn Glu Gln Phe

50 55 60

Ile Asn Ala Ala Lys Ile Ile Thr His Pro Asn Phe Asn Gly Asn Thr

65 70 75 80

Leu Asp Asn Asp Ile Met Leu Ile Lys Leu Ser Ser Pro Ala Thr Leu

85 90 95

Asn Ser Leu Val Ala Thr Val Ser Leu Pro Arg Ser Cys Ala Ala Ala

100 105 110

Gly Thr Glu Cys Leu Ile Ser Gly Trp Gly Asn Thr Lys Ser Ser Gly

115 120 125

Ser Ser Tyr Pro Ser Leu Leu Gln Cys Leu Lys Ala Pro Val Leu Ser

130 135 140

Asp Ser Ser Cys Lys Ser Ser Tyr Pro Gly Gln Ile Thr Gly Asn Met

145 150 155 160

Ile Cys Val Gly Phe Leu Glu Gly Gly Lys Asp Ala Cys Gln Gly Asp

165 170 175

Ser Gly Gly Pro Val Val Cys Asn Gly Gln Leu Gln Gly Ile Val Ser

180 185 190

Ser Gly Tyr Gly Cys Ala Gln Lys Asn Lys Pro Gly Val Tyr Thr Lys

195 200 205

Val Cys Asn Tyr Val Asn Trp Ile Gln Gln Thr Ile Ala Ala Asn

210 215 220

<210> 6

<211> 223

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Porcine trypsin variant #6

<400> 6

Ile Val Gly Gly Tyr Thr Cys Ala Ala Asn Ser Ile Pro Tyr Gln Val

1 5 10 15

Ser Leu Asn Ser Gly Ser His Arg Cys Gly Gly Ser Leu Ile Asn Ser

20 25 30

Gln Trp Val Val Ser Ala Ala His Cys Tyr Lys Ser Arg Ile Gln Val

35 40 45

Arg Leu Gly Glu His Asn Ile Asp Val Leu Glu Gly Asn Glu Gln Phe

50 55 60

Ile Asn Ala Ala Lys Ile Ile Thr His Pro Asn Phe Asn Gly Asn Thr

65 70 75 80

Leu Asp Asn Asp Ile Met Leu Ile Lys Leu Ser Ser Pro Ala Thr Leu

85 90 95

Asn Ser Phe Val Ala Thr Val Ser Leu Pro Arg Ser Cys Ala Ala Ala

100 105 110

Gly Thr Glu Cys Leu Ile Ser Gly Trp Gly Asn Thr Lys Ser Ser Gly

115 120 125

Ser Ser Tyr Pro Ser Leu Leu Gln Cys Leu Lys Ala Pro Val Leu Ser

130 135 140

Asp Ser Ser Cys Lys Ser Ser Tyr Pro Gly Gln Ile Thr Gly Asn Met

145 150 155 160

Ile Cys Val Gly Phe Leu Glu Gly Gly Lys Asp Ala Cys Gln Gly Asp

165 170 175

Ser Gly Gly Pro Val Val Cys Asn Gly Gln Leu Gln Gly Ile Val Ser

180 185 190

Ser Gly Tyr Gly Cys Ala Gln Lys Asn Lys Pro Gly Val Tyr Thr Lys

195 200 205

Val Cys Asn Tyr Val Asn Trp Ile Gln Gln Thr Ile Ala Ala Asn

210 215 220

<210> 7

<211> 223

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Porcine trypsin variant #7

<400> 7

Ile Val Gly Gly Tyr Thr Cys Ala Ala Asn Ser Ile Pro Tyr Gln Val

1 5 10 15

Ser Leu Asn Ser Gly Ser His Arg Cys Gly Gly Ser Leu Ile Asn Ser

20 25 30

Gln Trp Val Val Ser Ala Ala His Cys Tyr Lys Ser Arg Ile Gln Val

35 40 45

Arg Leu Gly Glu His Asn Ile Asp Val Leu Glu Gly Asn Glu Gln Phe

50 55 60

Ile Asn Ala Ala Lys Ile Ile Thr His Pro Asn Phe Asn Gly Asn Thr

65 70 75 80

Leu Asp Asn Asp Ile Met Leu Ile Lys Leu Ser Ser Pro Ala Thr Leu

85 90 95

Asn Ser Trp Val Ala Thr Val Ser Leu Pro Arg Ser Cys Ala Ala Ala

100 105 110

Gly Thr Glu Cys Leu Ile Ser Gly Trp Gly Asn Thr Lys Ser Ser Gly

115 120 125

Ser Ser Tyr Pro Ser Leu Leu Gln Cys Leu Lys Ala Pro Val Leu Ser

130 135 140

Asp Ser Ser Cys Lys Ser Ser Tyr Pro Gly Gln Ile Thr Gly Asn Met

145 150 155 160

Ile Cys Val Gly Phe Leu Glu Gly Gly Lys Asp Ala Cys Gln Gly Asp

165 170 175

Ser Gly Gly Pro Val Val Cys Asn Gly Gln Leu Gln Gly Ile Val Ser

180 185 190

Ser Gly Tyr Gly Cys Ala Gln Lys Asn Lys Pro Gly Val Tyr Thr Lys

195 200 205

Val Cys Asn Tyr Val Asn Trp Ile Gln Gln Thr Ile Ala Ala Asn

210 215 220

<210> 8

<211> 223

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Porcine trypsin variant #8

<400> 8

Ile Val Gly Gly Tyr Thr Cys Ala Ala Asn Ser Ile Pro Tyr Gln Val

1 5 10 15

Ser Leu Asn Ser Gly Ser His Arg Cys Gly Gly Ser Leu Ile Asn Ser

20 25 30

Gln Trp Val Val Ser Ala Ala His Cys Tyr Lys Ser Arg Ile Gln Val

35 40 45

Arg Leu Gly Glu His Asn Ile Asp Val Leu Glu Gly Asn Glu Gln Phe

50 55 60

Ile Asn Ala Ala Lys Ile Ile Thr His Pro Asn Phe Asn Gly Asn Thr

65 70 75 80

Leu Asp Asn Asp Ile Met Leu Ile Lys Leu Ser Ser Pro Ala Thr Leu

85 90 95

Asn Ser Gly Val Ala Thr Val Ser Leu Pro Arg Ser Cys Ala Ala Ala

100 105 110

Gly Thr Glu Cys Leu Ile Ser Gly Trp Gly Asn Thr Lys Ser Ser Gly

115 120 125

Ser Ser Tyr Pro Ser Leu Leu Gln Cys Leu Lys Ala Pro Val Leu Ser

130 135 140

Asp Ser Ser Cys Lys Ser Ser Tyr Pro Gly Gln Ile Thr Gly Asn Met

145 150 155 160

Ile Cys Val Gly Phe Leu Glu Gly Gly Lys Asp Ala Cys Gln Gly Asp

165 170 175

Ser Gly Gly Pro Val Val Cys Asn Gly Gln Leu Gln Gly Ile Val Ser

180 185 190

Ser Gly Tyr Gly Cys Ala Gln Lys Asn Lys Pro Gly Val Tyr Thr Lys

195 200 205

Val Cys Asn Tyr Val Asn Trp Ile Gln Gln Thr Ile Ala Ala Asn

210 215 220

<210> 9

<211> 223

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Porcine trypsin variant #9

<400> 9

Ile Val Gly Gly Tyr Thr Cys Ala Ala Asn Ser Ile Pro Tyr Gln Val

1 5 10 15

Ser Leu Asn Ser Gly Ser His Arg Cys Gly Gly Ser Leu Ile Asn Ser

20 25 30

Gln Trp Val Val Ser Ala Ala His Cys Tyr Lys Ser Arg Ile Gln Val

35 40 45

Arg Leu Gly Glu His Asn Ile Asp Val Leu Glu Gly Asn Glu Gln Phe

50 55 60

Ile Asn Ala Ala Lys Ile Ile Thr His Pro Asn Phe Asn Gly Asn Thr

65 70 75 80

Leu Asp Asn Asp Ile Met Leu Ile Lys Leu Ser Ser Pro Ala Thr Leu

85 90 95

Asn Ser Tyr Val Ala Thr Val Ser Leu Pro Arg Ser Cys Ala Ala Ala

100 105 110

Gly Thr Glu Cys Leu Ile Ser Gly Trp Gly Asn Thr Lys Ser Ser Gly

115 120 125

Ser Ser Tyr Pro Ser Leu Leu Gln Cys Leu Lys Ala Pro Val Leu Ser

130 135 140

Asp Ser Ser Cys Lys Ser Ser Tyr Pro Gly Gln Ile Thr Gly Asn Met

145 150 155 160

Ile Cys Val Gly Phe Leu Glu Gly Gly Lys Asp Ala Cys Gln Gly Asp

165 170 175

Ser Gly Gly Pro Val Val Cys Asn Gly Gln Leu Gln Gly Ile Val Ser

180 185 190

Ser Gly Tyr Gly Cys Ala Gln Lys Asn Lys Pro Gly Val Tyr Thr Lys

195 200 205

Val Cys Asn Tyr Val Asn Trp Ile Gln Gln Thr Ile Ala Ala Asn

210 215 220

<210> 10

<211> 223

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Porcine trypsin variant #10

<400> 10

Ile Val Gly Gly Tyr Thr Cys Ala Ala Asn Ser Ile Pro Tyr Gln Val

1 5 10 15

Ser Leu Asn Ser Gly Ser His Arg Cys Gly Gly Ser Leu Ile Asn Ser

20 25 30

Gln Trp Val Val Ser Ala Ala His Cys Tyr Lys Ser Arg Ile Gln Val

35 40 45

Arg Leu Gly Glu His Asn Ile Asp Val Leu Glu Gly Asn Glu Gln Phe

50 55 60

Ile Asn Ala Ala Lys Ile Ile Thr His Pro Asn Phe Asn Gly Asn Thr

65 70 75 80

Leu Asp Asn Asp Ile Met Leu Ile Lys Leu Ser Ser Pro Ala Thr Leu

85 90 95

Asn Ser Thr Val Ala Thr Val Ser Leu Pro Arg Ser Cys Ala Ala Ala

100 105 110

Gly Thr Glu Cys Leu Ile Ser Gly Trp Gly Asn Thr Lys Ser Ser Gly

115 120 125

Ser Ser Tyr Pro Ser Leu Leu Gln Cys Leu Lys Ala Pro Val Leu Ser

130 135 140

Asp Ser Ser Cys Lys Ser Ser Tyr Pro Gly Gln Ile Thr Gly Asn Met

145 150 155 160

Ile Cys Val Gly Phe Leu Glu Gly Gly Lys Asp Ala Cys Gln Gly Asp

165 170 175

Ser Gly Gly Pro Val Val Cys Asn Gly Gln Leu Gln Gly Ile Val Ser

180 185 190

Ser Gly Tyr Gly Cys Ala Gln Lys Asn Lys Pro Gly Val Tyr Thr Lys

195 200 205

Val Cys Asn Tyr Val Asn Trp Ile Gln Gln Thr Ile Ala Ala Asn

210 215 220

<210> 11

<211> 223

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Porcine trypsin variant #11

<400> 11

Ile Val Gly Gly Tyr Thr Cys Ala Ala Asn Ser Ile Pro Tyr Gln Val

1 5 10 15

Ser Leu Asn Ser Gly Ser His Arg Cys Gly Gly Ser Leu Ile Asn Ser

20 25 30

Gln Trp Val Val Ser Ala Ala His Cys Tyr Lys Ser Arg Ile Gln Val

35 40 45

Arg Leu Gly Glu His Asn Ile Asp Val Leu Glu Gly Asn Glu Gln Phe

50 55 60

Ile Asn Ala Ala Lys Ile Ile Thr His Pro Asn Phe Asn Gly Asn Thr

65 70 75 80

Leu Asp Asn Asp Ile Met Leu Ile Lys Leu Ser Ser Pro Ala Thr Leu

85 90 95

Asn Ser Asn Val Ala Thr Val Ser Leu Pro Arg Ser Cys Ala Ala Ala

100 105 110

Gly Thr Glu Cys Leu Ile Ser Gly Trp Gly Asn Thr Lys Ser Ser Gly

115 120 125

Ser Ser Tyr Pro Ser Leu Leu Gln Cys Leu Lys Ala Pro Val Leu Ser

130 135 140

Asp Ser Ser Cys Lys Ser Ser Tyr Pro Gly Gln Ile Thr Gly Asn Met

145 150 155 160

Ile Cys Val Gly Phe Leu Glu Gly Gly Lys Asp Ala Cys Gln Gly Asp

165 170 175

Ser Gly Gly Pro Val Val Cys Asn Gly Gln Leu Gln Gly Ile Val Ser

180 185 190

Ser Gly Tyr Gly Cys Ala Gln Lys Asn Lys Pro Gly Val Tyr Thr Lys

195 200 205

Val Cys Asn Tyr Val Asn Trp Ile Gln Gln Thr Ile Ala Ala Asn

210 215 220

<210> 12

<211> 223

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Porcine trypsin variant #12

<400> 12

Ile Val Gly Gly Tyr Thr Cys Ala Ala Asn Ser Ile Pro Tyr Gln Val

1 5 10 15

Ser Leu Asn Ser Gly Ser His Arg Cys Gly Gly Ser Leu Ile Asn Ser

20 25 30

Gln Trp Val Val Ser Ala Ala His Cys Tyr Lys Ser Arg Ile Gln Val

35 40 45

Arg Leu Gly Glu His Asn Ile Asp Val Leu Glu Gly Asn Glu Gln Phe

50 55 60

Ile Asn Ala Ala Lys Ile Ile Thr His Pro Asn Phe Asn Gly Asn Thr

65 70 75 80

Leu Asp Asn Asp Ile Met Leu Ile Lys Leu Ser Ser Pro Ala Thr Leu

85 90 95

Asn Ser Asp Val Ala Thr Val Ser Leu Pro Arg Ser Cys Ala Ala Ala

100 105 110

Gly Thr Glu Cys Leu Ile Ser Gly Trp Gly Asn Thr Lys Ser Ser Gly

115 120 125

Ser Ser Tyr Pro Ser Leu Leu Gln Cys Leu Lys Ala Pro Val Leu Ser

130 135 140

Asp Ser Ser Cys Lys Ser Ser Tyr Pro Gly Gln Ile Thr Gly Asn Met

145 150 155 160

Ile Cys Val Gly Phe Leu Glu Gly Gly Lys Asp Ala Cys Gln Gly Asp

165 170 175

Ser Gly Gly Pro Val Val Cys Asn Gly Gln Leu Gln Gly Ile Val Ser

180 185 190

Ser Gly Tyr Gly Cys Ala Gln Lys Asn Lys Pro Gly Val Tyr Thr Lys

195 200 205

Val Cys Asn Tyr Val Asn Trp Ile Gln Gln Thr Ile Ala Ala Asn

210 215 220

<210> 13

<211> 223

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Porcine trypsin variant #13

<400> 13

Ile Val Gly Gly Tyr Thr Cys Ala Ala Asn Ser Ile Pro Tyr Gln Val

1 5 10 15

Ser Leu Asn Ser Gly Ser His Arg Cys Gly Gly Ser Leu Ile Asn Ser

20 25 30

Gln Trp Val Val Ser Ala Ala His Cys Tyr Lys Ser Arg Ile Gln Val

35 40 45

Arg Leu Gly Glu His Asn Ile Asp Val Leu Glu Gly Asn Glu Gln Phe

50 55 60

Ile Asn Ala Ala Lys Ile Ile Thr His Pro Asn Phe Asn Gly Asn Thr

65 70 75 80

Leu Asp Asn Asp Ile Met Leu Ile Lys Leu Ser Ser Pro Ala Thr Leu

85 90 95

Asn Ser Glu Val Ala Thr Val Ser Leu Pro Arg Ser Cys Ala Ala Ala

100 105 110

Gly Thr Glu Cys Leu Ile Ser Gly Trp Gly Asn Thr Lys Ser Ser Gly

115 120 125

Ser Ser Tyr Pro Ser Leu Leu Gln Cys Leu Lys Ala Pro Val Leu Ser

130 135 140

Asp Ser Ser Cys Lys Ser Ser Tyr Pro Gly Gln Ile Thr Gly Asn Met

145 150 155 160

Ile Cys Val Gly Phe Leu Glu Gly Gly Lys Asp Ala Cys Gln Gly Asp

165 170 175

Ser Gly Gly Pro Val Val Cys Asn Gly Gln Leu Gln Gly Ile Val Ser

180 185 190

Ser Gly Tyr Gly Cys Ala Gln Lys Asn Lys Pro Gly Val Tyr Thr Lys

195 200 205

Val Cys Asn Tyr Val Asn Trp Ile Gln Gln Thr Ile Ala Ala Asn

210 215 220

<210> 14

<211> 223

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Porcine trypsin variant #14

<400> 14

Ile Val Gly Gly Tyr Thr Cys Ala Ala Asn Ser Ile Pro Tyr Gln Val

1 5 10 15

Ser Leu Asn Ser Gly Ser His Arg Cys Gly Gly Ser Leu Ile Asn Ser

20 25 30

Gln Trp Val Val Ser Ala Ala His Cys Tyr Lys Ser Arg Ile Gln Val

35 40 45

Arg Leu Gly Glu His Asn Ile Asp Val Leu Glu Gly Asn Glu Gln Phe

50 55 60

Ile Asn Ala Ala Lys Ile Ile Thr His Pro Asn Phe Asn Gly Asn Thr

65 70 75 80

Leu Asp Asn Asp Ile Met Leu Ile Lys Leu Ser Ser Pro Ala Thr Leu

85 90 95

Asn Ser His Val Ala Thr Val Ser Leu Pro Arg Ser Cys Ala Ala Ala

100 105 110

Gly Thr Glu Cys Leu Ile Ser Gly Trp Gly Asn Thr Lys Ser Ser Gly

115 120 125

Ser Ser Tyr Pro Ser Leu Leu Gln Cys Leu Lys Ala Pro Val Leu Ser

130 135 140

Asp Ser Ser Cys Lys Ser Ser Tyr Pro Gly Gln Ile Thr Gly Asn Met

145 150 155 160

Ile Cys Val Gly Phe Leu Glu Gly Gly Lys Asp Ala Cys Gln Gly Asp

165 170 175

Ser Gly Gly Pro Val Val Cys Asn Gly Gln Leu Gln Gly Ile Val Ser

180 185 190

Ser Gly Tyr Gly Cys Ala Gln Lys Asn Lys Pro Gly Val Tyr Thr Lys

195 200 205

Val Cys Asn Tyr Val Asn Trp Ile Gln Gln Thr Ile Ala Ala Asn

210 215 220

<210> 15

<211> 223

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Porcine trypsin variant #15

<400> 15

Ile Val Gly Gly Tyr Thr Cys Ala Ala Asn Ser Ile Pro Tyr Gln Val

1 5 10 15

Ser Leu Asn Ser Gly Ser His Arg Cys Gly Gly Ser Leu Ile Asn Ser

20 25 30

Gln Trp Val Val Ser Ala Ala His Cys Tyr Lys Ser Arg Ile Gln Val

35 40 45

Arg Leu Gly Glu His Asn Ile Asp Val Leu Glu Gly Asn Glu Gln Phe

50 55 60

Ile Asn Ala Ala Lys Ile Ile Thr His Pro Asn Phe Asn Gly Asn Thr

65 70 75 80

Leu Asp Asn Asp Ile Met Leu Ile Lys Leu Ser Ser Pro Ala Thr Leu

85 90 95

Asn Ser Arg Val Ala Thr Val Ser Leu Pro Arg Ser Cys Ala Ala Ala

100 105 110

Gly Thr Glu Cys Leu Ile Ser Gly Trp Gly Asn Thr Ala Ser Ser Gly

115 120 125

Ser Ser Tyr Pro Ser Leu Leu Gln Cys Leu Lys Ala Pro Val Leu Ser

130 135 140

Asp Ser Ser Cys Lys Ser Ser Tyr Pro Gly Gln Ile Thr Gly Asn Met

145 150 155 160

Ile Cys Val Gly Phe Leu Glu Gly Gly Lys Asp Ala Cys Gln Gly Asp

165 170 175

Ser Gly Gly Pro Val Val Cys Asn Gly Gln Leu Gln Gly Ile Val Ser

180 185 190

Ser Gly Tyr Gly Cys Ala Gln Lys Asn Lys Pro Gly Val Tyr Thr Lys

195 200 205

Val Cys Asn Tyr Val Asn Trp Ile Gln Gln Thr Ile Ala Ala Asn

210 215 220

<210> 16

<211> 223

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Porcine trypsin variant #16

<400> 16

Ile Val Gly Gly Tyr Thr Cys Ala Ala Asn Ser Ile Pro Tyr Gln Val

1 5 10 15

Ser Leu Asn Ser Gly Ser His Arg Cys Gly Gly Ser Leu Ile Asn Ser

20 25 30

Gln Trp Val Val Ser Ala Ala His Cys Tyr Lys Ser Arg Ile Gln Val

35 40 45

Arg Leu Gly Glu His Asn Ile Asp Val Leu Glu Gly Asn Glu Gln Phe

50 55 60

Ile Asn Ala Ala Lys Ile Ile Thr His Pro Asn Phe Asn Gly Asn Thr

65 70 75 80

Leu Asp Asn Asp Ile Met Leu Ile Lys Leu Ser Ser Pro Ala Thr Leu

85 90 95

Asn Ser Arg Val Ala Thr Val Ser Leu Pro Arg Ser Cys Ala Ala Ala

100 105 110

Gly Thr Glu Cys Leu Ile Ser Gly Trp Gly Asn Thr Gly Ser Ser Gly

115 120 125

Ser Ser Tyr Pro Ser Leu Leu Gln Cys Leu Lys Ala Pro Val Leu Ser

130 135 140

Asp Ser Ser Cys Lys Ser Ser Tyr Pro Gly Gln Ile Thr Gly Asn Met

145 150 155 160

Ile Cys Val Gly Phe Leu Glu Gly Gly Lys Asp Ala Cys Gln Gly Asp

165 170 175

Ser Gly Gly Pro Val Val Cys Asn Gly Gln Leu Gln Gly Ile Val Ser

180 185 190

Ser Gly Tyr Gly Cys Ala Gln Lys Asn Lys Pro Gly Val Tyr Thr Lys

195 200 205

Val Cys Asn Tyr Val Asn Trp Ile Gln Gln Thr Ile Ala Ala Asn

210 215 220

<210> 17

<211> 223

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Porcine trypsin variant #17

<400> 17

Ile Val Gly Gly Tyr Thr Cys Ala Ala Asn Ser Ile Pro Tyr Gln Val

1 5 10 15

Ser Leu Asn Ser Gly Ser His Arg Cys Gly Gly Ser Leu Ile Asn Ser

20 25 30

Gln Trp Val Val Ser Ala Ala His Cys Tyr Lys Ser Arg Ile Gln Val

35 40 45

Arg Leu Gly Glu His Asn Ile Asp Val Leu Glu Gly Asn Glu Gln Phe

50 55 60

Ile Asn Ala Ala Lys Ile Ile Thr His Pro Asn Phe Asn Gly Asn Thr

65 70 75 80

Leu Asp Asn Asp Ile Met Leu Ile Lys Leu Ser Ser Pro Ala Thr Leu

85 90 95

Asn Ser Arg Val Ala Thr Val Ser Leu Pro Arg Ser Cys Ala Ala Ala

100 105 110

Gly Thr Glu Cys Leu Ile Ser Gly Trp Gly Asn Thr Tyr Ser Ser Gly

115 120 125

Ser Ser Tyr Pro Ser Leu Leu Gln Cys Leu Lys Ala Pro Val Leu Ser

130 135 140

Asp Ser Ser Cys Lys Ser Ser Tyr Pro Gly Gln Ile Thr Gly Asn Met

145 150 155 160

Ile Cys Val Gly Phe Leu Glu Gly Gly Lys Asp Ala Cys Gln Gly Asp

165 170 175

Ser Gly Gly Pro Val Val Cys Asn Gly Gln Leu Gln Gly Ile Val Ser

180 185 190

Ser Gly Tyr Gly Cys Ala Gln Lys Asn Lys Pro Gly Val Tyr Thr Lys

195 200 205

Val Cys Asn Tyr Val Asn Trp Ile Gln Gln Thr Ile Ala Ala Asn

210 215 220

<210> 18

<211> 223

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Porcine trypsin variant #18

<400> 18

Ile Val Gly Gly Tyr Thr Cys Ala Ala Asn Ser Ile Pro Tyr Gln Val

1 5 10 15

Ser Leu Asn Ser Gly Ser His Arg Cys Gly Gly Ser Leu Ile Asn Ser

20 25 30

Gln Trp Val Val Ser Ala Ala His Cys Tyr Lys Ser Arg Ile Gln Val

35 40 45

Arg Leu Gly Glu His Asn Ile Asp Val Leu Glu Gly Asn Glu Gln Phe

50 55 60

Ile Asn Ala Ala Lys Ile Ile Thr His Pro Asn Phe Asn Gly Asn Thr

65 70 75 80

Leu Asp Asn Asp Ile Met Leu Ile Lys Leu Ser Ser Pro Ala Thr Leu

85 90 95

Asn Ser Arg Val Ala Thr Val Ser Leu Pro Arg Ser Cys Ala Ala Ala

100 105 110

Gly Thr Glu Cys Leu Ile Ser Gly Trp Gly Asn Thr Ser Ser Ser Gly

115 120 125

Ser Ser Tyr Pro Ser Leu Leu Gln Cys Leu Lys Ala Pro Val Leu Ser

130 135 140

Asp Ser Ser Cys Lys Ser Ser Tyr Pro Gly Gln Ile Thr Gly Asn Met

145 150 155 160

Ile Cys Val Gly Phe Leu Glu Gly Gly Lys Asp Ala Cys Gln Gly Asp

165 170 175

Ser Gly Gly Pro Val Val Cys Asn Gly Gln Leu Gln Gly Ile Val Ser

180 185 190

Ser Gly Tyr Gly Cys Ala Gln Lys Asn Lys Pro Gly Val Tyr Thr Lys

195 200 205

Val Cys Asn Tyr Val Asn Trp Ile Gln Gln Thr Ile Ala Ala Asn

210 215 220

<210> 19

<211> 223

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Porcine trypsin variant #19

<400> 19

Ile Val Gly Gly Tyr Thr Cys Ala Ala Asn Ser Ile Pro Tyr Gln Val

1 5 10 15

Ser Leu Asn Ser Gly Ser His Arg Cys Gly Gly Ser Leu Ile Asn Ser

20 25 30

Gln Trp Val Val Ser Ala Ala His Cys Tyr Lys Ser Arg Ile Gln Val

35 40 45

Arg Leu Gly Glu His Asn Ile Asp Val Leu Glu Gly Asn Glu Gln Phe

50 55 60

Ile Asn Ala Ala Lys Ile Ile Thr His Pro Asn Phe Asn Gly Asn Thr

65 70 75 80

Leu Asp Asn Asp Ile Met Leu Ile Lys Leu Ser Ser Pro Ala Thr Leu

85 90 95

Asn Ser Arg Val Ala Thr Val Ser Leu Pro Arg Ser Cys Ala Ala Ala

100 105 110

Gly Thr Glu Cys Leu Ile Ser Gly Trp Gly Asn Thr Cys Ser Ser Gly

115 120 125

Ser Ser Tyr Pro Ser Leu Leu Gln Cys Leu Lys Ala Pro Val Leu Ser

130 135 140

Asp Ser Ser Cys Lys Ser Ser Tyr Pro Gly Gln Ile Thr Gly Asn Met

145 150 155 160

Ile Cys Val Gly Phe Leu Glu Gly Gly Lys Asp Ala Cys Gln Gly Asp

165 170 175

Ser Gly Gly Pro Val Val Cys Asn Gly Gln Leu Gln Gly Ile Val Ser

180 185 190

Ser Gly Tyr Gly Cys Ala Gln Lys Asn Lys Pro Gly Val Tyr Thr Lys

195 200 205

Val Cys Asn Tyr Val Asn Trp Ile Gln Gln Thr Ile Ala Ala Asn

210 215 220

<210> 20

<211> 223

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Porcine trypsin variant #20

<400> 20

Ile Val Gly Gly Tyr Thr Cys Ala Ala Asn Ser Ile Pro Tyr Gln Val

1 5 10 15

Ser Leu Asn Ser Gly Ser His Arg Cys Gly Gly Ser Leu Ile Asn Ser

20 25 30

Gln Trp Val Val Ser Ala Ala His Cys Tyr Lys Ser Arg Ile Gln Val

35 40 45

Arg Leu Gly Glu His Asn Ile Asp Val Leu Glu Gly Asn Glu Gln Phe

50 55 60

Ile Asn Ala Ala Lys Ile Ile Thr His Pro Asn Phe Asn Gly Asn Thr

65 70 75 80

Leu Asp Asn Asp Ile Met Leu Ile Lys Leu Ser Ser Pro Ala Thr Leu

85 90 95

Asn Ser Arg Val Ala Thr Val Ser Leu Pro Arg Ser Cys Ala Ala Ala

100 105 110

Gly Thr Glu Cys Leu Ile Ser Gly Trp Gly Asn Thr Glu Ser Ser Gly

115 120 125

Ser Ser Tyr Pro Ser Leu Leu Gln Cys Leu Lys Ala Pro Val Leu Ser

130 135 140

Asp Ser Ser Cys Lys Ser Ser Tyr Pro Gly Gln Ile Thr Gly Asn Met

145 150 155 160

Ile Cys Val Gly Phe Leu Glu Gly Gly Lys Asp Ala Cys Gln Gly Asp

165 170 175

Ser Gly Gly Pro Val Val Cys Asn Gly Gln Leu Gln Gly Ile Val Ser

180 185 190

Ser Gly Tyr Gly Cys Ala Gln Lys Asn Lys Pro Gly Val Tyr Thr Lys

195 200 205

Val Cys Asn Tyr Val Asn Trp Ile Gln Gln Thr Ile Ala Ala Asn

210 215 220

<210> 21

<211> 223

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Porcine trypsin variant #21

<400> 21

Ile Val Gly Gly Tyr Thr Cys Ala Ala Asn Ser Ile Pro Tyr Gln Val

1 5 10 15

Ser Leu Asn Ser Gly Ser His Arg Cys Gly Gly Ser Leu Ile Asn Ser

20 25 30

Gln Trp Val Val Ser Ala Ala His Cys Tyr Lys Ser Arg Ile Gln Val

35 40 45

Arg Leu Gly Glu His Asn Ile Asp Val Leu Glu Gly Asn Glu Gln Phe

50 55 60

Ile Asn Ala Ala Lys Ile Ile Thr His Pro Asn Phe Asn Gly Asn Thr

65 70 75 80

Leu Asp Asn Asp Ile Met Leu Ile Lys Leu Ser Ser Pro Ala Thr Leu

85 90 95

Asn Ser Arg Val Ala Thr Val Ser Leu Pro Arg Ser Cys Ala Ala Ala

100 105 110

Gly Thr Glu Cys Leu Ile Ser Gly Trp Gly Asn Thr His Ser Ser Gly

115 120 125

Ser Ser Tyr Pro Ser Leu Leu Gln Cys Leu Lys Ala Pro Val Leu Ser

130 135 140

Asp Ser Ser Cys Lys Ser Ser Tyr Pro Gly Gln Ile Thr Gly Asn Met

145 150 155 160

Ile Cys Val Gly Phe Leu Glu Gly Gly Lys Asp Ala Cys Gln Gly Asp

165 170 175

Ser Gly Gly Pro Val Val Cys Asn Gly Gln Leu Gln Gly Ile Val Ser

180 185 190

Ser Gly Tyr Gly Cys Ala Gln Lys Asn Lys Pro Gly Val Tyr Thr Lys

195 200 205

Val Cys Asn Tyr Val Asn Trp Ile Gln Gln Thr Ile Ala Ala Asn

210 215 220

<210> 22

<211> 223

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Porcine trypsin variant #22

<400> 22

Ile Val Gly Gly Tyr Thr Cys Ala Ala Asn Ser Ile Pro Tyr Gln Val

1 5 10 15

Ser Leu Asn Ser Gly Ser His Arg Cys Gly Gly Ser Leu Ile Asn Ser

20 25 30

Gln Trp Val Val Ser Ala Ala His Cys Tyr Lys Ser Arg Ile Gln Val

35 40 45

Arg Leu Gly Glu His Asn Ile Asp Val Leu Glu Gly Asn Glu Gln Phe

50 55 60

Ile Asn Ala Ala Lys Ile Ile Thr His Pro Asn Phe Asn Gly Asn Thr

65 70 75 80

Leu Asp Asn Asp Ile Met Leu Ile Lys Leu Ser Ser Pro Ala Thr Leu

85 90 95

Asn Ser Arg Val Ala Thr Val Ser Leu Pro Arg Ser Cys Ala Ala Ala

100 105 110

Gly Thr Glu Cys Leu Ile Ser Gly Trp Gly Asn Thr Leu Ser Ser Gly

115 120 125

Ser Ser Tyr Pro Ser Leu Leu Gln Cys Leu Lys Ala Pro Val Leu Ser

130 135 140

Asp Ser Ser Cys Lys Ser Ser Tyr Pro Gly Gln Ile Thr Gly Asn Met

145 150 155 160

Ile Cys Val Gly Phe Leu Glu Gly Gly Lys Asp Ala Cys Gln Gly Asp

165 170 175

Ser Gly Gly Pro Val Val Cys Asn Gly Gln Leu Gln Gly Ile Val Ser

180 185 190

Ser Gly Tyr Gly Cys Ala Gln Lys Asn Lys Pro Gly Val Tyr Thr Lys

195 200 205

Val Cys Asn Tyr Val Asn Trp Ile Gln Gln Thr Ile Ala Ala Asn

210 215 220

<210> 23

<211> 223

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Porcine trypsin variant #23

<400> 23

Ile Val Gly Gly Tyr Thr Cys Ala Ala Asn Ser Ile Pro Tyr Gln Val

1 5 10 15

Ser Leu Asn Ser Gly Ser His Arg Cys Gly Gly Ser Leu Ile Asn Ser

20 25 30

Gln Trp Val Val Ser Ala Ala His Cys Tyr Lys Ser Arg Ile Gln Val

35 40 45

Arg Leu Gly Glu His Asn Ile Asp Val Leu Glu Gly Asn Glu Gln Phe

50 55 60

Ile Asn Ala Ala Lys Ile Ile Thr His Pro Asn Phe Asn Gly Asn Thr

65 70 75 80

Leu Asp Asn Asp Ile Met Leu Ile Lys Leu Ser Ser Pro Ala Thr Leu

85 90 95

Asn Ser Arg Val Ala Thr Val Ser Leu Pro Arg Ser Cys Ala Ala Ala

100 105 110

Gly Thr Glu Cys Leu Ile Ser Gly Trp Gly Asn Thr Gln Ser Ser Gly

115 120 125

Ser Ser Tyr Pro Ser Leu Leu Gln Cys Leu Lys Ala Pro Val Leu Ser

130 135 140

Asp Ser Ser Cys Lys Ser Ser Tyr Pro Gly Gln Ile Thr Gly Asn Met

145 150 155 160

Ile Cys Val Gly Phe Leu Glu Gly Gly Lys Asp Ala Cys Gln Gly Asp

165 170 175

Ser Gly Gly Pro Val Val Cys Asn Gly Gln Leu Gln Gly Ile Val Ser

180 185 190

Ser Gly Tyr Gly Cys Ala Gln Lys Asn Lys Pro Gly Val Tyr Thr Lys

195 200 205

Val Cys Asn Tyr Val Asn Trp Ile Gln Gln Thr Ile Ala Ala Asn

210 215 220

<210> 24

<211> 223

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Porcine trypsin variant #24

<400> 24

Ile Val Gly Gly Tyr Thr Cys Ala Ala Asn Ser Ile Pro Tyr Gln Val

1 5 10 15

Ser Leu Asn Ser Gly Ser His Arg Cys Gly Gly Ser Leu Ile Asn Ser

20 25 30

Gln Trp Val Val Ser Ala Ala His Cys Tyr Lys Ser Arg Ile Gln Val

35 40 45

Arg Leu Gly Glu His Asn Ile Asp Val Leu Glu Gly Asn Glu Gln Phe

50 55 60

Ile Asn Ala Ala Lys Ile Ile Thr His Pro Asn Phe Asn Gly Asn Thr

65 70 75 80

Leu Asp Asn Asp Ile Met Leu Ile Lys Leu Ser Ser Pro Ala Thr Leu

85 90 95

Asn Ser Arg Val Ala Thr Val Ser Leu Pro Arg Ser Cys Ala Ala Ala

100 105 110

Gly Thr Glu Cys Leu Ile Ser Gly Trp Gly Asn Thr Lys Ser Ser Gly

115 120 125

Ser Ser Tyr Pro Ser Leu Leu Gln Cys Leu Lys Ala Pro Val Leu Ser

130 135 140

Asp Ser Ser Cys Lys Ser Ser Tyr Pro Gly Gln Ile Thr Gly Asn Met

145 150 155 160

Ile Cys Val Gly Phe Leu Glu Gly Gly Tyr Asp Ala Cys Gln Gly Asp

165 170 175

Ser Gly Gly Pro Val Val Cys Asn Gly Gln Leu Gln Gly Ile Val Ser

180 185 190

Ser Gly Tyr Gly Cys Ala Gln Lys Asn Lys Pro Gly Val Tyr Thr Lys

195 200 205

Val Cys Asn Tyr Val Asn Trp Ile Gln Gln Thr Ile Ala Ala Asn

210 215 220

<210> 25

<211> 223

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Porcine trypsin variant #25

<400> 25

Ile Val Gly Gly Tyr Thr Cys Ala Ala Asn Ser Ile Pro Tyr Gln Val

1 5 10 15

Ser Leu Asn Ser Gly Ser His Arg Cys Gly Gly Ser Leu Ile Asn Ser

20 25 30

Gln Trp Val Val Ser Ala Ala His Cys Tyr Lys Ser Arg Ile Gln Val

35 40 45

Arg Leu Gly Glu His Asn Ile Asp Val Leu Glu Gly Asn Glu Gln Phe

50 55 60

Ile Asn Ala Ala Lys Ile Ile Thr His Pro Asn Phe Asn Gly Asn Thr

65 70 75 80

Leu Asp Asn Asp Ile Met Leu Ile Lys Leu Ser Ser Pro Ala Thr Leu

85 90 95

Asn Ser Arg Val Ala Thr Val Ser Leu Pro Arg Ser Cys Ala Ala Ala

100 105 110

Gly Thr Glu Cys Leu Ile Ser Gly Trp Gly Asn Thr Lys Ser Ser Gly

115 120 125

Ser Ser Tyr Pro Ser Leu Leu Gln Cys Leu Lys Ala Pro Val Leu Ser

130 135 140

Asp Ser Ser Cys Lys Ser Ser Tyr Pro Gly Gln Ile Thr Gly Asn Met

145 150 155 160

Ile Cys Val Gly Phe Leu Glu Gly Gly Ala Asp Ala Cys Gln Gly Asp

165 170 175

Ser Gly Gly Pro Val Val Cys Asn Gly Gln Leu Gln Gly Ile Val Ser

180 185 190

Ser Gly Tyr Gly Cys Ala Gln Lys Asn Lys Pro Gly Val Tyr Thr Lys

195 200 205

Val Cys Asn Tyr Val Asn Trp Ile Gln Gln Thr Ile Ala Ala Asn

210 215 220

<210> 26

<211> 223

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Porcine trypsin variant #26

<400> 26

Ile Val Gly Gly Tyr Thr Cys Ala Ala Asn Ser Ile Pro Tyr Gln Val

1 5 10 15

Ser Leu Asn Ser Gly Ser His Arg Cys Gly Gly Ser Leu Ile Asn Ser

20 25 30

Gln Trp Val Val Ser Ala Ala His Cys Tyr Lys Ser Arg Ile Gln Val

35 40 45

Arg Leu Gly Glu His Asn Ile Asp Val Leu Glu Gly Asn Glu Gln Phe

50 55 60

Ile Asn Ala Ala Lys Ile Ile Thr His Pro Asn Phe Asn Gly Asn Thr

65 70 75 80

Leu Asp Asn Asp Ile Met Leu Ile Lys Leu Ser Ser Pro Ala Thr Leu

85 90 95

Asn Ser Arg Val Ala Thr Val Ser Leu Pro Arg Ser Cys Ala Ala Ala

100 105 110

Gly Thr Glu Cys Leu Ile Ser Gly Trp Gly Asn Thr Lys Ser Ser Gly

115 120 125

Ser Ser Tyr Pro Ser Leu Leu Gln Cys Leu Lys Ala Pro Val Leu Ser

130 135 140

Asp Ser Ser Cys Lys Ser Ser Tyr Pro Gly Gln Ile Thr Gly Asn Met

145 150 155 160

Ile Cys Val Gly Phe Leu Glu Gly Gly Gly Asp Ala Cys Gln Gly Asp

165 170 175

Ser Gly Gly Pro Val Val Cys Asn Gly Gln Leu Gln Gly Ile Val Ser

180 185 190

Ser Gly Tyr Gly Cys Ala Gln Lys Asn Lys Pro Gly Val Tyr Thr Lys

195 200 205

Val Cys Asn Tyr Val Asn Trp Ile Gln Gln Thr Ile Ala Ala Asn

210 215 220

<210> 27

<211> 223

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Porcine trypsin variant #27

<400> 27

Ile Val Gly Gly Tyr Thr Cys Ala Ala Asn Ser Ile Pro Tyr Gln Val

1 5 10 15

Ser Leu Asn Ser Gly Ser His Arg Cys Gly Gly Ser Leu Ile Asn Ser

20 25 30

Gln Trp Val Val Ser Ala Ala His Cys Tyr Lys Ser Arg Ile Gln Val

35 40 45

Arg Leu Gly Glu His Asn Ile Asp Val Leu Glu Gly Asn Glu Gln Phe

50 55 60

Ile Asn Ala Ala Lys Ile Ile Thr His Pro Asn Phe Asn Gly Asn Thr

65 70 75 80

Leu Asp Asn Asp Ile Met Leu Ile Lys Leu Ser Ser Pro Ala Thr Leu

85 90 95

Asn Ser Arg Val Ala Thr Val Ser Leu Pro Arg Ser Cys Ala Ala Ala

100 105 110

Gly Thr Glu Cys Leu Ile Ser Gly Trp Gly Asn Thr Lys Ser Ser Gly

115 120 125

Ser Ser Tyr Pro Ser Leu Leu Gln Cys Leu Lys Ala Pro Val Leu Ser

130 135 140

Asp Ser Ser Cys Lys Ser Ser Tyr Pro Gly Gln Ile Thr Gly Asn Met

145 150 155 160

Ile Cys Val Gly Phe Leu Glu Gly Gly Asn Asp Ala Cys Gln Gly Asp

165 170 175

Ser Gly Gly Pro Val Val Cys Asn Gly Gln Leu Gln Gly Ile Val Ser

180 185 190

Ser Gly Tyr Gly Cys Ala Gln Lys Asn Lys Pro Gly Val Tyr Thr Lys

195 200 205

Val Cys Asn Tyr Val Asn Trp Ile Gln Gln Thr Ile Ala Ala Asn

210 215 220

<210> 28

<211> 223

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Porcine trypsin variant #28

<400> 28

Ile Val Gly Gly Tyr Thr Cys Ala Ala Asn Ser Ile Pro Tyr Gln Val

1 5 10 15

Ser Leu Asn Ser Gly Ser His Val Cys Gly Gly Ser Leu Ile Asn Ser

20 25 30

Gln Trp Val Val Ser Ala Ala His Cys Tyr Lys Ser Arg Ile Gln Val

35 40 45

Arg Leu Gly Glu His Asn Ile Asp Val Leu Glu Gly Asn Glu Gln Phe

50 55 60

Ile Asn Ala Ala Lys Ile Ile Thr His Pro Asn Phe Asn Gly Asn Thr

65 70 75 80

Leu Asp Asn Asp Ile Met Leu Ile Lys Leu Ser Ser Pro Ala Thr Leu

85 90 95

Asn Ser Arg Val Ala Thr Val Ser Leu Pro Arg Ser Cys Ala Ala Ala

100 105 110

Gly Thr Glu Cys Leu Ile Ser Gly Trp Gly Asn Thr Lys Ser Ser Gly

115 120 125

Ser Ser Tyr Pro Ser Leu Leu Gln Cys Leu Lys Ala Pro Val Leu Ser

130 135 140

Asp Ser Ser Cys Lys Ser Ser Tyr Pro Gly Gln Ile Thr Gly Asn Met

145 150 155 160

Ile Cys Val Gly Phe Leu Glu Gly Gly Lys Asp Ala Cys Gln Gly Asp

165 170 175

Ser Gly Gly Pro Val Val Cys Asn Gly Gln Leu Gln Gly Ile Val Ser

180 185 190

Ser Gly Tyr Gly Cys Ala Gln Lys Asn Lys Pro Gly Val Tyr Thr Lys

195 200 205

Val Cys Asn Tyr Val Asn Trp Ile Gln Gln Thr Ile Ala Ala Asn

210 215 220

<210> 29

<211> 223

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Porcine trypsin variant #29

<400> 29

Ile Val Gly Gly Tyr Thr Cys Ala Ala Asn Ser Ile Pro Tyr Gln Val

1 5 10 15

Ser Leu Asn Ser Gly Ser His Leu Cys Gly Gly Ser Leu Ile Asn Ser

20 25 30

Gln Trp Val Val Ser Ala Ala His Cys Tyr Lys Ser Arg Ile Gln Val

35 40 45

Arg Leu Gly Glu His Asn Ile Asp Val Leu Glu Gly Asn Glu Gln Phe

50 55 60

Ile Asn Ala Ala Lys Ile Ile Thr His Pro Asn Phe Asn Gly Asn Thr

65 70 75 80

Leu Asp Asn Asp Ile Met Leu Ile Lys Leu Ser Ser Pro Ala Thr Leu

85 90 95

Asn Ser Arg Val Ala Thr Val Ser Leu Pro Arg Ser Cys Ala Ala Ala

100 105 110

Gly Thr Glu Cys Leu Ile Ser Gly Trp Gly Asn Thr Lys Ser Ser Gly

115 120 125

Ser Ser Tyr Pro Ser Leu Leu Gln Cys Leu Lys Ala Pro Val Leu Ser

130 135 140

Asp Ser Ser Cys Lys Ser Ser Tyr Pro Gly Gln Ile Thr Gly Asn Met

145 150 155 160

Ile Cys Val Gly Phe Leu Glu Gly Gly Lys Asp Ala Cys Gln Gly Asp

165 170 175

Ser Gly Gly Pro Val Val Cys Asn Gly Gln Leu Gln Gly Ile Val Ser

180 185 190

Ser Gly Tyr Gly Cys Ala Gln Lys Asn Lys Pro Gly Val Tyr Thr Lys

195 200 205

Val Cys Asn Tyr Val Asn Trp Ile Gln Gln Thr Ile Ala Ala Asn

210 215 220

<210> 30

<211> 223

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Porcine trypsin variant #30

<400> 30

Ile Val Gly Gly Tyr Thr Cys Ala Ala Asn Ser Ile Pro Tyr Gln Val

1 5 10 15

Ser Leu Asn Ser Gly Ser His Ser Cys Gly Gly Ser Leu Ile Asn Ser

20 25 30

Gln Trp Val Val Ser Ala Ala His Cys Tyr Lys Ser Arg Ile Gln Val

35 40 45

Arg Leu Gly Glu His Asn Ile Asp Val Leu Glu Gly Asn Glu Gln Phe

50 55 60

Ile Asn Ala Ala Lys Ile Ile Thr His Pro Asn Phe Asn Gly Asn Thr

65 70 75 80

Leu Asp Asn Asp Ile Met Leu Ile Lys Leu Ser Ser Pro Ala Thr Leu

85 90 95

Asn Ser Arg Val Ala Thr Val Ser Leu Pro Arg Ser Cys Ala Ala Ala

100 105 110

Gly Thr Glu Cys Leu Ile Ser Gly Trp Gly Asn Thr Lys Ser Ser Gly

115 120 125

Ser Ser Tyr Pro Ser Leu Leu Gln Cys Leu Lys Ala Pro Val Leu Ser

130 135 140

Asp Ser Ser Cys Lys Ser Ser Tyr Pro Gly Gln Ile Thr Gly Asn Met

145 150 155 160

Ile Cys Val Gly Phe Leu Glu Gly Gly Lys Asp Ala Cys Gln Gly Asp

165 170 175

Ser Gly Gly Pro Val Val Cys Asn Gly Gln Leu Gln Gly Ile Val Ser

180 185 190

Ser Gly Tyr Gly Cys Ala Gln Lys Asn Lys Pro Gly Val Tyr Thr Lys

195 200 205

Val Cys Asn Tyr Val Asn Trp Ile Gln Gln Thr Ile Ala Ala Asn

210 215 220

<210> 31

<211> 223

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Porcine trypsin variant #31

<400> 31

Ile Val Gly Gly Tyr Thr Cys Ala Ala Asn Ser Ile Pro Tyr Gln Val

1 5 10 15

Ser Leu Asn Ser Gly Ser His Gln Cys Gly Gly Ser Leu Ile Asn Ser

20 25 30

Gln Trp Val Val Ser Ala Ala His Cys Tyr Lys Ser Arg Ile Gln Val

35 40 45

Arg Leu Gly Glu His Asn Ile Asp Val Leu Glu Gly Asn Glu Gln Phe

50 55 60

Ile Asn Ala Ala Lys Ile Ile Thr His Pro Asn Phe Asn Gly Asn Thr

65 70 75 80

Leu Asp Asn Asp Ile Met Leu Ile Lys Leu Ser Ser Pro Ala Thr Leu

85 90 95

Asn Ser Arg Val Ala Thr Val Ser Leu Pro Arg Ser Cys Ala Ala Ala

100 105 110

Gly Thr Glu Cys Leu Ile Ser Gly Trp Gly Asn Thr Lys Ser Ser Gly

115 120 125

Ser Ser Tyr Pro Ser Leu Leu Gln Cys Leu Lys Ala Pro Val Leu Ser

130 135 140

Asp Ser Ser Cys Lys Ser Ser Tyr Pro Gly Gln Ile Thr Gly Asn Met

145 150 155 160

Ile Cys Val Gly Phe Leu Glu Gly Gly Lys Asp Ala Cys Gln Gly Asp

165 170 175

Ser Gly Gly Pro Val Val Cys Asn Gly Gln Leu Gln Gly Ile Val Ser

180 185 190

Ser Gly Tyr Gly Cys Ala Gln Lys Asn Lys Pro Gly Val Tyr Thr Lys

195 200 205

Val Cys Asn Tyr Val Asn Trp Ile Gln Gln Thr Ile Ala Ala Asn

210 215 220

<210> 32

<211> 223

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Porcine trypsin variant #32

<400> 32

Ile Val Gly Gly Tyr Thr Cys Ala Ala Asn Ser Ile Pro Tyr Gln Val

1 5 10 15

Ser Leu Asn Ser Gly Ser His Arg Cys Gly Gly Ser Leu Ile Asn Ser

20 25 30

Gln Trp Val Val Ser Ala Ala His Cys Tyr Lys Ser Arg Ile Gln Val

35 40 45

Arg Leu Gly Glu His Asn Ile Asp Val Leu Glu Gly Asn Glu Gln Phe

50 55 60

Ile Asn Ala Ala Lys Ile Ile Thr His Pro Asn Phe Asn Gly Asn Thr

65 70 75 80

Leu Asp Asn Asp Ile Met Leu Ile Lys Leu Ser Ser Pro Ala Thr Leu

85 90 95

Asn Ser Arg Val Ala Thr Val Ser Leu Pro Arg Ser Cys Ala Ala Ala

100 105 110

Gly Thr Glu Cys Leu Ile Ser Gly Trp Gly Asn Thr Lys Ser Ser Gly

115 120 125

Ser Ser Tyr Pro Ser Leu Leu Gln Cys Leu Lys Ala Pro Val Leu Ser

130 135 140

Asp Ser Ser Cys Lys Ser Ser Tyr Pro Gly Gln Ile Thr Gly Asn Met

145 150 155 160

Ile Cys Val Gly Phe Leu Glu Gly Gly Lys Asp Ala Cys Gln Gly Asp

165 170 175

Ser Gly Gly Pro Val Val Cys Asn Gly Gln Leu Gln Gly Ile Val Ser

180 185 190

Ser Gly Tyr Gly Cys Ala Gln Lys Asn Lys Pro Gly Val Tyr Thr Lys

195 200 205

Val Cys Asn Tyr Val Asn Trp Ile Gln Gln Thr Ile Ala Ala Asn

210 215 220

<210> 33

<211> 223

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Porcine trypsin variant #33

<400> 33

Ile Val Gly Gly Tyr Thr Cys Ala Ala Asn Ser Ile Pro Tyr Gln Val

1 5 10 15

Ser Leu Asn Ser Gly Ser His Arg Cys Gly Gly Ser Leu Ile Asn Ser

20 25 30

Gln Trp Val Val Ser Ala Ala His Cys Tyr Lys Ser Arg Ile Gln Val

35 40 45

Arg Leu Gly Glu His Asn Ile Asp Val Leu Glu Gly Asn Glu Gln Phe

50 55 60

Ile Asn Ala Ala Lys Ile Ile Thr His Pro Asn Phe Asn Gly Asn Thr

65 70 75 80

Leu Asp Asn Asp Ile Met Leu Ile Lys Leu Ser Ser Pro Ala Thr Leu

85 90 95

Asn Ser Arg Val Ala Thr Val Ser Leu Pro Thr Ser Cys Ala Ala Ala

100 105 110

Gly Thr Glu Cys Leu Ile Ser Gly Trp Gly Asn Thr Lys Ser Ser Gly

115 120 125

Ser Ser Tyr Pro Ser Leu Leu Gln Cys Leu Lys Ala Pro Val Leu Ser

130 135 140

Asp Ser Ser Cys Lys Ser Ser Tyr Pro Gly Gln Ile Thr Gly Asn Met

145 150 155 160

Ile Cys Val Gly Phe Leu Glu Gly Gly Lys Asp Ala Cys Gln Gly Asp

165 170 175

Ser Gly Gly Pro Val Val Cys Asn Gly Gln Leu Gln Gly Ile Val Ser

180 185 190

Ser Gly Tyr Gly Cys Ala Gln Lys Asn Lys Pro Gly Val Tyr Thr Lys

195 200 205

Val Cys Asn Tyr Val Asn Trp Ile Gln Gln Thr Ile Ala Ala Asn

210 215 220

<210> 34

<211> 223

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Porcine trypsin variant #34

<400> 34

Ile Val Gly Gly Tyr Thr Cys Ala Ala Asn Ser Ile Pro Tyr Gln Val

1 5 10 15

Ser Leu Asn Ser Gly Ser His Arg Cys Gly Gly Ser Leu Ile Asn Ser

20 25 30

Gln Trp Val Val Ser Ala Ala His Cys Tyr Lys Ser Arg Ile Gln Val

35 40 45

Arg Leu Gly Glu His Asn Ile Asp Val Leu Glu Gly Asn Glu Gln Phe

50 55 60

Ile Asn Ala Ala Lys Ile Ile Thr His Pro Asn Phe Asn Gly Asn Thr

65 70 75 80

Leu Asp Asn Asp Ile Met Leu Ile Lys Leu Ser Ser Pro Ala Thr Leu

85 90 95

Asn Ser Arg Val Ala Thr Val Ser Leu Pro Arg Ser Cys Ala Ala Ala

100 105 110

Gly Thr Glu Cys Leu Ile Ser Gly Trp Gly Asn Thr Lys Ser Ser Gly

115 120 125

Ser Ser Ala Pro Ser Leu Leu Gln Cys Leu Lys Ala Pro Val Leu Ser

130 135 140

Asp Ser Ser Cys Lys Ser Ser Tyr Pro Gly Gln Ile Thr Gly Asn Met

145 150 155 160

Ile Cys Val Gly Phe Leu Glu Gly Gly Lys Asp Ala Cys Gln Gly Asp

165 170 175

Ser Gly Gly Pro Val Val Cys Asn Gly Gln Leu Gln Gly Ile Val Ser

180 185 190

Ser Gly Tyr Gly Cys Ala Gln Lys Asn Lys Pro Gly Val Tyr Thr Lys

195 200 205

Val Cys Asn Tyr Val Asn Trp Ile Gln Gln Thr Ile Ala Ala Asn

210 215 220

<210> 35

<211> 223

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Porcine trypsin variant #35

<400> 35

Ile Val Gly Gly Tyr Thr Cys Ala Ala Asn Ser Ile Pro Tyr Gln Val

1 5 10 15

Ser Leu Asn Ser Gly Ser His Arg Cys Gly Gly Ser Leu Ile Asn Ser

20 25 30

Gln Trp Val Val Ser Ala Ala His Cys Tyr Lys Ser Arg Ile Gln Val

35 40 45

Arg Leu Gly Glu His Asn Ile Asp Val Leu Glu Gly Asn Glu Gln Phe

50 55 60

Ile Asn Ala Ala Lys Ile Ile Thr His Pro Asn Phe Asn Gly Asn Thr

65 70 75 80

Leu Asp Asn Asp Ile Met Leu Ile Lys Leu Ser Ser Pro Ala Thr Leu

85 90 95

Asn Ser Arg Val Ala Thr Val Ser Leu Pro Arg Ser Cys Ala Ala Ala

100 105 110

Gly Thr Glu Cys Leu Ile Ser Gly Trp Gly Asn Thr Lys Ser Ser Gly

115 120 125

Ser Ser Leu Pro Ser Leu Leu Gln Cys Leu Lys Ala Pro Val Leu Ser

130 135 140

Asp Ser Ser Cys Lys Ser Ser Tyr Pro Gly Gln Ile Thr Gly Asn Met

145 150 155 160

Ile Cys Val Gly Phe Leu Glu Gly Gly Lys Asp Ala Cys Gln Gly Asp

165 170 175

Ser Gly Gly Pro Val Val Cys Asn Gly Gln Leu Gln Gly Ile Val Ser

180 185 190

Ser Gly Tyr Gly Cys Ala Gln Lys Asn Lys Pro Gly Val Tyr Thr Lys

195 200 205

Val Cys Asn Tyr Val Asn Trp Ile Gln Gln Thr Ile Ala Ala Asn

210 215 220

<210> 36

<211> 223

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Porcine trypsin variant #36

<400> 36

Ile Val Gly Gly Tyr Thr Cys Ala Ala Asn Ser Ile Pro Tyr Gln Val

1 5 10 15

Ser Leu Asn Ser Gly Ser His Arg Cys Gly Gly Ser Leu Ile Asn Ser

20 25 30

Gln Trp Val Val Ser Ala Ala His Cys Tyr Lys Ser Arg Ile Gln Val

35 40 45

Arg Leu Gly Glu His Asn Ile Asp Val Leu Glu Gly Asn Glu Gln Phe

50 55 60

Ile Asn Ala Ala Lys Ile Ile Thr His Pro Asn Phe Asn Gly Asn Thr

65 70 75 80

Leu Asp Asn Asp Ile Met Leu Ile Lys Leu Ser Ser Pro Ala Thr Leu

85 90 95

Asn Ser Arg Val Ala Thr Val Ser Leu Pro Arg Ser Cys Ala Ala Ala

100 105 110

Gly Thr Glu Cys Leu Ile Ser Gly Trp Gly Asn Thr Lys Ser Ser Gly

115 120 125

Ser Ser Met Pro Ser Leu Leu Gln Cys Leu Lys Ala Pro Val Leu Ser

130 135 140

Asp Ser Ser Cys Lys Ser Ser Tyr Pro Gly Gln Ile Thr Gly Asn Met

145 150 155 160

Ile Cys Val Gly Phe Leu Glu Gly Gly Lys Asp Ala Cys Gln Gly Asp

165 170 175

Ser Gly Gly Pro Val Val Cys Asn Gly Gln Leu Gln Gly Ile Val Ser

180 185 190

Ser Gly Tyr Gly Cys Ala Gln Lys Asn Lys Pro Gly Val Tyr Thr Lys

195 200 205

Val Cys Asn Tyr Val Asn Trp Ile Gln Gln Thr Ile Ala Ala Asn

210 215 220

<210> 37

<211> 223

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Porcine trypsin variant #37

<400> 37

Ile Val Gly Gly Tyr Thr Cys Ala Ala Asn Ser Ile Pro Tyr Gln Val

1 5 10 15

Ser Leu Asn Ser Gly Ser His Arg Cys Gly Gly Ser Leu Ile Asn Ser

20 25 30

Gln Trp Val Val Ser Ala Ala His Cys Tyr Lys Ser Arg Ile Gln Val

35 40 45

Arg Leu Gly Glu His Asn Ile Asp Val Leu Glu Gly Asn Glu Gln Phe

50 55 60

Ile Asn Ala Ala Lys Ile Ile Thr His Pro Asn Phe Asn Gly Asn Thr

65 70 75 80

Leu Asp Asn Asp Ile Met Leu Ile Lys Leu Ser Ser Pro Ala Thr Leu

85 90 95

Asn Ser Arg Val Ala Thr Val Ser Leu Pro Arg Ser Cys Ala Ala Ala

100 105 110

Gly Thr Glu Cys Leu Ile Ser Gly Trp Gly Asn Thr Lys Ser Ser Gly

115 120 125

Ser Ser Thr Pro Ser Leu Leu Gln Cys Leu Lys Ala Pro Val Leu Ser

130 135 140

Asp Ser Ser Cys Lys Ser Ser Tyr Pro Gly Gln Ile Thr Gly Asn Met

145 150 155 160

Ile Cys Val Gly Phe Leu Glu Gly Gly Lys Asp Ala Cys Gln Gly Asp

165 170 175

Ser Gly Gly Pro Val Val Cys Asn Gly Gln Leu Gln Gly Ile Val Ser

180 185 190

Ser Gly Tyr Gly Cys Ala Gln Lys Asn Lys Pro Gly Val Tyr Thr Lys

195 200 205

Val Cys Asn Tyr Val Asn Trp Ile Gln Gln Thr Ile Ala Ala Asn

210 215 220

<210> 38

<211> 223

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Porcine trypsin variant #38

<400> 38

Ile Val Gly Gly Tyr Thr Cys Ala Ala Asn Ser Ile Pro Tyr Gln Val

1 5 10 15

Ser Leu Asn Ser Gly Ser His Arg Cys Gly Gly Ser Leu Ile Asn Ser

20 25 30

Gln Trp Val Val Ser Ala Ala His Cys Tyr Lys Ser Arg Ile Gln Val

35 40 45

Arg Leu Gly Glu His Asn Ile Asp Val Leu Glu Gly Asn Glu Gln Phe

50 55 60

Ile Asn Ala Ala Lys Ile Ile Thr His Pro Asn Phe Asn Gly Asn Thr

65 70 75 80

Leu Asp Asn Asp Ile Met Leu Ile Lys Leu Ser Ser Pro Ala Thr Leu

85 90 95

Asn Ser Arg Val Ala Thr Val Ser Leu Pro Arg Ser Cys Ala Ala Ala

100 105 110

Gly Thr Glu Cys Leu Ile Ser Gly Trp Gly Asn Thr Lys Ser Ser Gly

115 120 125

Ser Ser Cys Pro Ser Leu Leu Gln Cys Leu Lys Ala Pro Val Leu Ser

130 135 140

Asp Ser Ser Cys Lys Ser Ser Tyr Pro Gly Gln Ile Thr Gly Asn Met

145 150 155 160

Ile Cys Val Gly Phe Leu Glu Gly Gly Lys Asp Ala Cys Gln Gly Asp

165 170 175

Ser Gly Gly Pro Val Val Cys Asn Gly Gln Leu Gln Gly Ile Val Ser

180 185 190

Ser Gly Tyr Gly Cys Ala Gln Lys Asn Lys Pro Gly Val Tyr Thr Lys

195 200 205

Val Cys Asn Tyr Val Asn Trp Ile Gln Gln Thr Ile Ala Ala Asn

210 215 220

<210> 39

<211> 223

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Porcine trypsin variant #39

<400> 39

Ile Val Gly Gly Tyr Thr Cys Ala Ala Asn Ser Ile Pro Tyr Gln Val

1 5 10 15

Ser Leu Asn Ser Gly Ser His Arg Cys Gly Gly Ser Leu Ile Asn Ser

20 25 30

Gln Trp Val Val Ser Ala Ala His Cys Tyr Lys Ser Arg Ile Gln Val

35 40 45

Arg Leu Gly Glu His Asn Ile Asp Val Leu Glu Gly Asn Glu Gln Phe

50 55 60

Ile Asn Ala Ala Lys Ile Ile Thr His Pro Asn Phe Asn Gly Asn Thr

65 70 75 80

Leu Asp Asn Asp Ile Met Leu Ile Lys Leu Ser Ser Pro Ala Thr Leu

85 90 95

Asn Ser Arg Val Ala Thr Val Ser Leu Pro Arg Ser Cys Ala Ala Ala

100 105 110

Gly Thr Glu Cys Leu Ile Ser Gly Trp Gly Asn Thr Lys Ser Ser Gly

115 120 125

Ser Ser Asn Pro Ser Leu Leu Gln Cys Leu Lys Ala Pro Val Leu Ser

130 135 140

Asp Ser Ser Cys Lys Ser Ser Tyr Pro Gly Gln Ile Thr Gly Asn Met

145 150 155 160

Ile Cys Val Gly Phe Leu Glu Gly Gly Lys Asp Ala Cys Gln Gly Asp

165 170 175

Ser Gly Gly Pro Val Val Cys Asn Gly Gln Leu Gln Gly Ile Val Ser

180 185 190

Ser Gly Tyr Gly Cys Ala Gln Lys Asn Lys Pro Gly Val Tyr Thr Lys

195 200 205

Val Cys Asn Tyr Val Asn Trp Ile Gln Gln Thr Ile Ala Ala Asn

210 215 220

<210> 40

<211> 223

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Porcine trypsin variant #40

<400> 40

Ile Val Gly Gly Tyr Thr Cys Ala Ala Asn Ser Ile Pro Tyr Gln Val

1 5 10 15

Ser Leu Asn Ser Gly Ser His Arg Cys Gly Gly Ser Leu Ile Asn Ser

20 25 30

Gln Trp Val Val Ser Ala Ala His Cys Tyr Lys Ser Arg Ile Gln Val

35 40 45

Arg Leu Gly Glu His Asn Ile Asp Val Leu Glu Gly Asn Glu Gln Phe

50 55 60

Ile Asn Ala Ala Lys Ile Ile Thr His Pro Asn Phe Asn Gly Asn Thr

65 70 75 80

Leu Asp Asn Asp Ile Met Leu Ile Lys Leu Ser Ser Pro Ala Thr Leu

85 90 95

Asn Ser Arg Val Ala Thr Val Ser Leu Pro Arg Ser Cys Ala Ala Ala

100 105 110

Gly Thr Glu Cys Leu Ile Ser Gly Trp Gly Asn Thr Lys Ser Ser Gly

115 120 125

Ser Ser Glu Pro Ser Leu Leu Gln Cys Leu Lys Ala Pro Val Leu Ser

130 135 140

Asp Ser Ser Cys Lys Ser Ser Tyr Pro Gly Gln Ile Thr Gly Asn Met

145 150 155 160

Ile Cys Val Gly Phe Leu Glu Gly Gly Lys Asp Ala Cys Gln Gly Asp

165 170 175

Ser Gly Gly Pro Val Val Cys Asn Gly Gln Leu Gln Gly Ile Val Ser

180 185 190

Ser Gly Tyr Gly Cys Ala Gln Lys Asn Lys Pro Gly Val Tyr Thr Lys

195 200 205

Val Cys Asn Tyr Val Asn Trp Ile Gln Gln Thr Ile Ala Ala Asn

210 215 220

<210> 41

<211> 223

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Porcine trypsin variant #41

<400> 41

Ile Val Gly Gly Tyr Thr Cys Ala Ala Asn Ser Ile Pro Tyr Gln Val

1 5 10 15

Ser Leu Asn Ser Gly Ser His Arg Cys Gly Gly Ser Leu Ile Asn Ser

20 25 30

Gln Trp Val Val Ser Ala Ala His Cys Tyr Lys Ser Arg Ile Gln Val

35 40 45

Arg Leu Gly Glu His Asn Ile Asp Val Leu Glu Gly Asn Glu Gln Phe

50 55 60

Ile Asn Ala Ala Lys Ile Ile Thr His Pro Asn Phe Asn Gly Asn Thr

65 70 75 80

Leu Asp Asn Asp Ile Met Leu Ile Lys Leu Ser Ser Pro Ala Thr Leu

85 90 95

Asn Ser Arg Val Ala Thr Val Ser Leu Pro Arg Ser Cys Ala Ala Ala

100 105 110

Gly Thr Glu Cys Leu Ile Ser Gly Trp Gly Asn Thr Lys Ser Ser Gly

115 120 125

Ser Ser His Pro Ser Leu Leu Gln Cys Leu Lys Ala Pro Val Leu Ser

130 135 140

Asp Ser Ser Cys Lys Ser Ser Tyr Pro Gly Gln Ile Thr Gly Asn Met

145 150 155 160

Ile Cys Val Gly Phe Leu Glu Gly Gly Lys Asp Ala Cys Gln Gly Asp

165 170 175

Ser Gly Gly Pro Val Val Cys Asn Gly Gln Leu Gln Gly Ile Val Ser

180 185 190

Ser Gly Tyr Gly Cys Ala Gln Lys Asn Lys Pro Gly Val Tyr Thr Lys

195 200 205

Val Cys Asn Tyr Val Asn Trp Ile Gln Gln Thr Ile Ala Ala Asn

210 215 220

<210> 42

<211> 223

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Porcine trypsin variant #42

<400> 42

Ile Val Gly Gly Tyr Thr Cys Ala Ala Asn Ser Ile Pro Tyr Gln Val

1 5 10 15

Ser Leu Asn Ser Gly Ser His Arg Cys Gly Gly Ser Leu Ile Asn Ser

20 25 30

Gln Trp Val Val Ser Ala Ala His Cys Tyr Lys Ser Arg Ile Gln Val

35 40 45

Arg Leu Gly Glu His Asn Ile Asp Val Leu Glu Gly Asn Glu Gln Phe

50 55 60

Ile Asn Ala Ala Lys Ile Ile Thr His Pro Asn Phe Asn Gly Asn Thr

65 70 75 80

Leu Asp Asn Asp Ile Met Leu Ile Lys Leu Ser Ser Pro Ala Thr Leu

85 90 95

Asn Ser Arg Val Ala Thr Val Ser Leu Pro Arg Ser Cys Ala Ala Ala

100 105 110

Gly Thr Glu Cys Leu Ile Ser Gly Trp Gly Asn Thr Lys Ser Ser Gly

115 120 125

Ser Ser Tyr Pro Ser Leu Leu Gln Cys Leu Lys Ala Pro Val Leu Ser

130 135 140

Asp Ser Ser Cys Lys Ser Ser Tyr Pro Gly Gln Ile Thr Gly Asn Met

145 150 155 160

Ile Cys Val Gly Phe Leu Glu Gly Gly Lys Asp Ala Cys Gln Gly Asp

165 170 175

Ser Gly Gly Pro Val Val Cys Asn Gly Gln Leu Gln Gly Ile Val Ser

180 185 190

Ser Gly Tyr Gly Cys Ala Gln Lys Asn Lys Pro Gly Val Tyr Thr Lys

195 200 205

Val Cys Asn Tyr Val Asn Trp Ile Gln Gln Thr Ile Ala Ala Asn

210 215 220

<210> 43

<211> 223

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Porcine trypsin variant #43

<400> 43

Ile Val Gly Gly Tyr Thr Cys Ala Ala Asn Ser Ile Pro Tyr Gln Val

1 5 10 15

Ser Leu Asn Ser Gly Ser His Arg Cys Gly Gly Ser Leu Ile Asn Ser

20 25 30

Gln Trp Val Val Ser Ala Ala His Cys Tyr Lys Ser Arg Ile Gln Val

35 40 45

Arg Leu Gly Glu His Asn Ile Asp Val Leu Glu Gly Asn Glu Gln Phe

50 55 60

Ile Asn Ala Ala Lys Ile Ile Thr His Pro Asn Phe Asn Gly Asn Thr

65 70 75 80

Leu Asp Asn Asp Ile Met Leu Ile Lys Leu Ser Ser Pro Ala Thr Leu

85 90 95

Asn Ser Arg Val Ala Thr Val Ser Leu Pro Arg Ser Cys Ala Ala Ala

100 105 110

Gly Thr Glu Cys Leu Ile Ser Gly Trp Gly Asn Thr Lys Ser Ser Gly

115 120 125

Ser Ser Tyr Pro Ser Leu Leu Gln Cys Leu Lys Ala Pro Val Leu Ser

130 135 140

Asp Ser Ser Cys Lys Ser Ser Tyr Pro Gly Gln Ile Thr Gly Asn Met

145 150 155 160

Ile Cys Val Gly Phe Leu Glu Gly Gly Lys Asp Thr Cys Gln Gly Asp

165 170 175

Ser Gly Gly Pro Val Val Cys Asn Gly Gln Leu Gln Gly Ile Val Ser

180 185 190

Ser Gly Tyr Gly Cys Ala Gln Lys Asn Lys Pro Gly Val Tyr Thr Lys

195 200 205

Val Cys Asn Tyr Val Asn Trp Ile Gln Gln Thr Ile Ala Ala Asn

210 215 220

<210> 44

<211> 223

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Porcine trypsin variant #44

<400> 44

Ile Val Gly Gly Tyr Thr Cys Ala Ala Asn Ser Ile Pro Tyr Gln Val

1 5 10 15

Ser Leu Asn Ser Gly Ser His Arg Cys Gly Gly Ser Leu Ile Asn Ser

20 25 30

Gln Trp Val Val Ser Ala Ala His Cys Tyr Lys Ser Arg Ile Gln Val

35 40 45

Arg Leu Gly Glu His Asn Ile Asp Val Leu Glu Gly Asn Glu Gln Phe

50 55 60

Ile Asn Ala Ala Lys Ile Ile Thr His Pro Asn Phe Asn Gly Asn Thr

65 70 75 80

Leu Asp Asn Asp Ile Met Leu Ile Lys Leu Ser Ser Pro Ala Thr Leu

85 90 95

Asn Ser Arg Val Ala Thr Val Ser Leu Pro Arg Ser Cys Ala Ala Ala

100 105 110

Gly Thr Glu Cys Leu Ile Ser Gly Trp Gly Asn Thr Lys Ser Ser Gly

115 120 125

Ser Ser Tyr Pro Ser Leu Leu Gln Cys Leu Lys Ala Pro Val Leu Ser

130 135 140

Asp Ser Ser Cys Lys Ser Ser Tyr Pro Gly Gln Ile Thr Gly Asn Met

145 150 155 160

Ile Cys Val Gly Phe Leu Glu Gly Gly Lys Asp Ser Cys Gln Gly Asp

165 170 175

Ser Gly Gly Pro Val Val Cys Asn Gly Gln Leu Gln Gly Ile Val Ser

180 185 190

Ser Gly Tyr Gly Cys Ala Gln Lys Asn Lys Pro Gly Val Tyr Thr Lys

195 200 205

Val Cys Asn Tyr Val Asn Trp Ile Gln Gln Thr Ile Ala Ala Asn

210 215 220

<210> 45

<211> 223

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Porcine trypsin variant #45

<400> 45

Ile Val Gly Gly Tyr Thr Cys Ala Ala Asn Ser Ile Pro Tyr Gln Val

1 5 10 15

Ser Leu Asn Ser Gly Ser His Arg Cys Gly Gly Ser Leu Ile Asn Ser

20 25 30

Gln Trp Val Val Ser Ala Ala His Cys Tyr Lys Ser Arg Ile Gln Val

35 40 45

Arg Leu Gly Glu His Asn Ile Asp Val Leu Glu Gly Asn Glu Gln Phe

50 55 60

Ile Asn Ala Ala Lys Ile Ile Thr His Pro Asn Phe Asn Gly Asn Thr

65 70 75 80

Leu Asp Asn Asp Ile Met Leu Ile Lys Leu Ser Ser Pro Ala Thr Leu

85 90 95

Asn Ser Arg Val Ala Thr Val Ser Leu Pro Arg Ser Cys Ala Ala Ala

100 105 110

Gly Thr Glu Cys Leu Ile Ser Gly Trp Gly Asn Thr Lys Ser Ser Gly

115 120 125

Ser Ser Tyr Pro Ser Leu Leu Gln Cys Leu Lys Ala Pro Val Leu Ser

130 135 140

Asp Ser Ser Cys Lys Ser Ser Tyr Pro Gly Gln Ile Thr Gly Asn Met

145 150 155 160

Ile Cys Val Gly Phe Leu Glu Gly Gly Lys Asp Cys Cys Gln Gly Asp

165 170 175

Ser Gly Gly Pro Val Val Cys Asn Gly Gln Leu Gln Gly Ile Val Ser

180 185 190

Ser Gly Tyr Gly Cys Ala Gln Lys Asn Lys Pro Gly Val Tyr Thr Lys

195 200 205

Val Cys Asn Tyr Val Asn Trp Ile Gln Gln Thr Ile Ala Ala Asn

210 215 220

<210> 46

<211> 223

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Porcine trypsin variant #46

<400> 46

Ile Val Gly Gly Tyr Thr Cys Ala Ala Asn Ser Ile Pro Tyr Gln Val

1 5 10 15

Ser Leu Asn Ser Gly Ser His Ala Cys Gly Gly Ser Leu Ile Asn Ser

20 25 30

Gln Trp Val Val Ser Ala Ala His Cys Tyr Lys Ser Arg Ile Gln Val

35 40 45

Arg Leu Gly Glu His Asn Ile Asp Val Leu Glu Gly Asn Glu Gln Phe

50 55 60

Ile Asn Ala Ala Lys Ile Ile Thr His Pro Asn Phe Asn Gly Asn Thr

65 70 75 80

Leu Asp Asn Asp Ile Met Leu Ile Lys Leu Ser Ser Pro Ala Thr Leu

85 90 95

Asn Ser Arg Val Ala Thr Val Ser Leu Pro Arg Ser Cys Ala Ala Ala

100 105 110

Gly Thr Glu Cys Leu Ile Ser Gly Trp Gly Asn Thr Lys Ser Ser Gly

115 120 125

Ser Ser Tyr Pro Ser Leu Leu Gln Cys Leu Lys Ala Pro Val Leu Ser

130 135 140

Asp Ser Ser Cys Lys Ser Ser Tyr Pro Gly Gln Ile Thr Gly Asn Met

145 150 155 160

Ile Cys Val Gly Phe Leu Glu Gly Gly Lys Asp Ala Cys Gln Gly Asp

165 170 175

Ser Gly Gly Pro Val Val Cys Asn Gly Gln Leu Gln Gly Ile Val Ser

180 185 190

Ser Gly Tyr Gly Cys Ala Gln Lys Asn Lys Pro Gly Val Tyr Thr Lys

195 200 205

Val Cys Asn Tyr Val Asn Trp Ile Gln Gln Thr Ile Ala Ala Asn

210 215 220

<210> 47

<211> 223

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Porcine trypsin variant #47

<400> 47

Ile Val Gly Gly Tyr Thr Cys Ala Ala Asn Ser Ile Pro Tyr Gln Val

1 5 10 15

Ser Leu Asn Ser Gly Ser His Ile Cys Gly Gly Ser Leu Ile Asn Ser

20 25 30

Gln Trp Val Val Ser Ala Ala His Cys Tyr Lys Ser Arg Ile Gln Val

35 40 45

Arg Leu Gly Glu His Asn Ile Asp Val Leu Glu Gly Asn Glu Gln Phe

50 55 60

Ile Asn Ala Ala Lys Ile Ile Thr His Pro Asn Phe Asn Gly Asn Thr

65 70 75 80

Leu Asp Asn Asp Ile Met Leu Ile Lys Leu Ser Ser Pro Ala Thr Leu

85 90 95

Asn Ser Arg Val Ala Thr Val Ser Leu Pro Arg Ser Cys Ala Ala Ala

100 105 110

Gly Thr Glu Cys Leu Ile Ser Gly Trp Gly Asn Thr Lys Ser Ser Gly

115 120 125

Ser Ser Tyr Pro Ser Leu Leu Gln Cys Leu Lys Ala Pro Val Leu Ser

130 135 140

Asp Ser Ser Cys Lys Ser Ser Tyr Pro Gly Gln Ile Thr Gly Asn Met

145 150 155 160

Ile Cys Val Gly Phe Leu Glu Gly Gly Lys Asp Ala Cys Gln Gly Asp

165 170 175

Ser Gly Gly Pro Val Val Cys Asn Gly Gln Leu Gln Gly Ile Val Ser

180 185 190

Ser Gly Tyr Gly Cys Ala Gln Lys Asn Lys Pro Gly Val Tyr Thr Lys

195 200 205

Val Cys Asn Tyr Val Asn Trp Ile Gln Gln Thr Ile Ala Ala Asn

210 215 220

<210> 48

<211> 223

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Porcine trypsin variant #48

<400> 48

Ile Val Gly Gly Tyr Thr Cys Ala Ala Asn Ser Ile Pro Tyr Gln Val

1 5 10 15

Ser Leu Asn Ser Gly Ser His Met Cys Gly Gly Ser Leu Ile Asn Ser

20 25 30

Gln Trp Val Val Ser Ala Ala His Cys Tyr Lys Ser Arg Ile Gln Val

35 40 45

Arg Leu Gly Glu His Asn Ile Asp Val Leu Glu Gly Asn Glu Gln Phe

50 55 60

Ile Asn Ala Ala Lys Ile Ile Thr His Pro Asn Phe Asn Gly Asn Thr

65 70 75 80

Leu Asp Asn Asp Ile Met Leu Ile Lys Leu Ser Ser Pro Ala Thr Leu

85 90 95

Asn Ser Arg Val Ala Thr Val Ser Leu Pro Arg Ser Cys Ala Ala Ala

100 105 110

Gly Thr Glu Cys Leu Ile Ser Gly Trp Gly Asn Thr Lys Ser Ser Gly

115 120 125

Ser Ser Tyr Pro Ser Leu Leu Gln Cys Leu Lys Ala Pro Val Leu Ser

130 135 140

Asp Ser Ser Cys Lys Ser Ser Tyr Pro Gly Gln Ile Thr Gly Asn Met

145 150 155 160

Ile Cys Val Gly Phe Leu Glu Gly Gly Lys Asp Ala Cys Gln Gly Asp

165 170 175

Ser Gly Gly Pro Val Val Cys Asn Gly Gln Leu Gln Gly Ile Val Ser

180 185 190

Ser Gly Tyr Gly Cys Ala Gln Lys Asn Lys Pro Gly Val Tyr Thr Lys

195 200 205

Val Cys Asn Tyr Val Asn Trp Ile Gln Gln Thr Ile Ala Ala Asn

210 215 220

<210> 49

<211> 223

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Porcine trypsin variant #49

<400> 49

Ile Val Gly Gly Tyr Thr Cys Ala Ala Asn Ser Ile Pro Tyr Gln Val

1 5 10 15

Ser Leu Asn Ser Gly Ser His Thr Cys Gly Gly Ser Leu Ile Asn Ser

20 25 30

Gln Trp Val Val Ser Ala Ala His Cys Tyr Lys Ser Arg Ile Gln Val

35 40 45

Arg Leu Gly Glu His Asn Ile Asp Val Leu Glu Gly Asn Glu Gln Phe

50 55 60

Ile Asn Ala Ala Lys Ile Ile Thr His Pro Asn Phe Asn Gly Asn Thr

65 70 75 80

Leu Asp Asn Asp Ile Met Leu Ile Lys Leu Ser Ser Pro Ala Thr Leu

85 90 95

Asn Ser Arg Val Ala Thr Val Ser Leu Pro Arg Ser Cys Ala Ala Ala

100 105 110

Gly Thr Glu Cys Leu Ile Ser Gly Trp Gly Asn Thr Lys Ser Ser Gly

115 120 125

Ser Ser Tyr Pro Ser Leu Leu Gln Cys Leu Lys Ala Pro Val Leu Ser

130 135 140

Asp Ser Ser Cys Lys Ser Ser Tyr Pro Gly Gln Ile Thr Gly Asn Met

145 150 155 160

Ile Cys Val Gly Phe Leu Glu Gly Gly Lys Asp Ala Cys Gln Gly Asp

165 170 175

Ser Gly Gly Pro Val Val Cys Asn Gly Gln Leu Gln Gly Ile Val Ser

180 185 190

Ser Gly Tyr Gly Cys Ala Gln Lys Asn Lys Pro Gly Val Tyr Thr Lys

195 200 205

Val Cys Asn Tyr Val Asn Trp Ile Gln Gln Thr Ile Ala Ala Asn

210 215 220

<210> 50

<211> 223

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Porcine trypsin variant #50

<400> 50

Ile Val Gly Gly Tyr Thr Cys Ala Ala Asn Ser Ile Pro Tyr Gln Val

1 5 10 15

Ser Leu Asn Ser Gly Ser His Asn Cys Gly Gly Ser Leu Ile Asn Ser

20 25 30

Gln Trp Val Val Ser Ala Ala His Cys Tyr Lys Ser Arg Ile Gln Val

35 40 45

Arg Leu Gly Glu His Asn Ile Asp Val Leu Glu Gly Asn Glu Gln Phe

50 55 60

Ile Asn Ala Ala Lys Ile Ile Thr His Pro Asn Phe Asn Gly Asn Thr

65 70 75 80

Leu Asp Asn Asp Ile Met Leu Ile Lys Leu Ser Ser Pro Ala Thr Leu

85 90 95

Asn Ser Arg Val Ala Thr Val Ser Leu Pro Arg Ser Cys Ala Ala Ala

100 105 110

Gly Thr Glu Cys Leu Ile Ser Gly Trp Gly Asn Thr Lys Ser Ser Gly

115 120 125

Ser Ser Tyr Pro Ser Leu Leu Gln Cys Leu Lys Ala Pro Val Leu Ser

130 135 140

Asp Ser Ser Cys Lys Ser Ser Tyr Pro Gly Gln Ile Thr Gly Asn Met

145 150 155 160

Ile Cys Val Gly Phe Leu Glu Gly Gly Lys Asp Ala Cys Gln Gly Asp

165 170 175

Ser Gly Gly Pro Val Val Cys Asn Gly Gln Leu Gln Gly Ile Val Ser

180 185 190

Ser Gly Tyr Gly Cys Ala Gln Lys Asn Lys Pro Gly Val Tyr Thr Lys

195 200 205

Val Cys Asn Tyr Val Asn Trp Ile Gln Gln Thr Ile Ala Ala Asn

210 215 220

<210> 51

<211> 223

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Porcine trypsin variant #51

<400> 51

Ile Val Gly Gly Tyr Thr Cys Ala Ala Asn Ser Ile Pro Tyr Gln Val

1 5 10 15

Ser Leu Asn Ser Gly Ser His Lys Cys Gly Gly Ser Leu Ile Asn Ser

20 25 30

Gln Trp Val Val Ser Ala Ala His Cys Tyr Lys Ser Arg Ile Gln Val

35 40 45

Arg Leu Gly Glu His Asn Ile Asp Val Leu Glu Gly Asn Glu Gln Phe

50 55 60

Ile Asn Ala Ala Lys Ile Ile Thr His Pro Asn Phe Asn Gly Asn Thr

65 70 75 80

Leu Asp Asn Asp Ile Met Leu Ile Lys Leu Ser Ser Pro Ala Thr Leu

85 90 95

Asn Ser Arg Val Ala Thr Val Ser Leu Pro Arg Ser Cys Ala Ala Ala

100 105 110

Gly Thr Glu Cys Leu Ile Ser Gly Trp Gly Asn Thr Lys Ser Ser Gly

115 120 125

Ser Ser Tyr Pro Ser Leu Leu Gln Cys Leu Lys Ala Pro Val Leu Ser

130 135 140

Asp Ser Ser Cys Lys Ser Ser Tyr Pro Gly Gln Ile Thr Gly Asn Met

145 150 155 160

Ile Cys Val Gly Phe Leu Glu Gly Gly Lys Asp Ala Cys Gln Gly Asp

165 170 175

Ser Gly Gly Pro Val Val Cys Asn Gly Gln Leu Gln Gly Ile Val Ser

180 185 190

Ser Gly Tyr Gly Cys Ala Gln Lys Asn Lys Pro Gly Val Tyr Thr Lys

195 200 205

Val Cys Asn Tyr Val Asn Trp Ile Gln Gln Thr Ile Ala Ala Asn

210 215 220

<210> 52

<211> 223

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Porcine trypsin variant #52

<400> 52

Ile Val Gly Gly Tyr Thr Cys Ala Ala Asn Ser Ile Pro Tyr Gln Val

1 5 10 15

Ser Leu Asn Ser Gly Ser His Arg Cys Gly Gly Ser Leu Ile Asn Ser

20 25 30

Gln Trp Val Val Ser Ala Ala His Cys Tyr Lys Leu Arg Ile Gln Val

35 40 45

Arg Leu Gly Glu His Asn Ile Asp Val Leu Glu Gly Asn Glu Gln Phe

50 55 60

Ile Asn Ala Ala Lys Ile Ile Thr His Pro Asn Phe Asn Gly Asn Thr

65 70 75 80

Leu Asp Asn Asp Ile Met Leu Ile Lys Leu Ser Ser Pro Ala Thr Leu

85 90 95

Asn Ser Arg Val Ala Thr Val Ser Leu Pro Arg Ser Cys Ala Ala Ala

100 105 110

Gly Thr Glu Cys Leu Ile Ser Gly Trp Gly Asn Thr Lys Ser Ser Gly

115 120 125

Ser Ser Tyr Pro Ser Leu Leu Gln Cys Leu Lys Ala Pro Val Leu Ser

130 135 140

Asp Ser Ser Cys Lys Ser Ser Tyr Pro Gly Gln Ile Thr Gly Asn Met

145 150 155 160

Ile Cys Val Gly Phe Leu Glu Gly Gly Lys Asp Ala Cys Gln Gly Asp

165 170 175

Ser Gly Gly Pro Val Val Cys Asn Gly Gln Leu Gln Gly Ile Val Ser

180 185 190

Ser Gly Tyr Gly Cys Ala Gln Lys Asn Lys Pro Gly Val Tyr Thr Lys

195 200 205

Val Cys Asn Tyr Val Asn Trp Ile Gln Gln Thr Ile Ala Ala Asn

210 215 220

<210> 53

<211> 223

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Porcine trypsin variant #53

<400> 53

Ile Val Gly Gly Tyr Thr Cys Ala Ala Asn Ser Ile Pro Tyr Gln Val

1 5 10 15

Ser Leu Asn Ser Gly Ser His Arg Cys Gly Gly Ser Leu Ile Asn Ser

20 25 30

Gln Trp Val Val Ser Ala Ala His Cys Tyr Lys Pro Arg Ile Gln Val

35 40 45

Arg Leu Gly Glu His Asn Ile Asp Val Leu Glu Gly Asn Glu Gln Phe

50 55 60

Ile Asn Ala Ala Lys Ile Ile Thr His Pro Asn Phe Asn Gly Asn Thr

65 70 75 80

Leu Asp Asn Asp Ile Met Leu Ile Lys Leu Ser Ser Pro Ala Thr Leu

85 90 95

Asn Ser Arg Val Ala Thr Val Ser Leu Pro Arg Ser Cys Ala Ala Ala

100 105 110

Gly Thr Glu Cys Leu Ile Ser Gly Trp Gly Asn Thr Lys Ser Ser Gly

115 120 125

Ser Ser Tyr Pro Ser Leu Leu Gln Cys Leu Lys Ala Pro Val Leu Ser

130 135 140

Asp Ser Ser Cys Lys Ser Ser Tyr Pro Gly Gln Ile Thr Gly Asn Met

145 150 155 160

Ile Cys Val Gly Phe Leu Glu Gly Gly Lys Asp Ala Cys Gln Gly Asp

165 170 175

Ser Gly Gly Pro Val Val Cys Asn Gly Gln Leu Gln Gly Ile Val Ser

180 185 190

Ser Gly Tyr Gly Cys Ala Gln Lys Asn Lys Pro Gly Val Tyr Thr Lys

195 200 205

Val Cys Asn Tyr Val Asn Trp Ile Gln Gln Thr Ile Ala Ala Asn

210 215 220

<210> 54

<211> 223

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Porcine trypsin variant #54

<400> 54

Ile Val Gly Gly Tyr Thr Cys Ala Ala Asn Ser Ile Pro Tyr Gln Val

1 5 10 15

Ser Leu Asn Ser Gly Ser His Arg Cys Gly Gly Ser Leu Ile Asn Ser

20 25 30

Gln Trp Val Val Ser Ala Ala His Cys Tyr Lys Ser Arg Ile Gln Val

35 40 45

Arg Leu Gly Glu His Asn Ile Ala Val Leu Glu Gly Asn Glu Gln Phe

50 55 60

Ile Asn Ala Ala Lys Ile Ile Thr His Pro Asn Phe Asn Gly Asn Thr

65 70 75 80

Leu Asp Asn Asp Ile Met Leu Ile Lys Leu Ser Ser Pro Ala Thr Leu

85 90 95

Asn Ser Arg Val Ala Thr Val Ser Leu Pro Arg Ser Cys Ala Ala Ala

100 105 110

Gly Thr Glu Cys Leu Ile Ser Gly Trp Gly Asn Thr Lys Ser Ser Gly

115 120 125

Ser Ser Tyr Pro Ser Leu Leu Gln Cys Leu Lys Ala Pro Val Leu Ser

130 135 140

Asp Ser Ser Cys Lys Ser Ser Tyr Pro Gly Gln Ile Thr Gly Asn Met

145 150 155 160

Ile Cys Val Gly Phe Leu Glu Gly Gly Lys Asp Ala Cys Gln Gly Asp

165 170 175

Ser Gly Gly Pro Val Val Cys Asn Gly Gln Leu Gln Gly Ile Val Ser

180 185 190

Ser Gly Tyr Gly Cys Ala Gln Lys Asn Lys Pro Gly Val Tyr Thr Lys

195 200 205

Val Cys Asn Tyr Val Asn Trp Ile Gln Gln Thr Ile Ala Ala Asn

210 215 220

<210> 55

<211> 223

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Porcine trypsin variant #55

<400> 55

Ile Val Gly Gly Tyr Thr Cys Ala Ala Asn Ser Ile Pro Tyr Gln Val

1 5 10 15

Ser Leu Asn Ser Gly Ser His Arg Cys Gly Gly Ser Leu Ile Asn Ser

20 25 30

Gln Trp Val Val Ser Ala Ala His Cys Tyr Lys Ser Arg Ile Gln Val

35 40 45

Arg Leu Gly Glu His Asn Ile Trp Val Leu Glu Gly Asn Glu Gln Phe

50 55 60

Ile Asn Ala Ala Lys Ile Ile Thr His Pro Asn Phe Asn Gly Asn Thr

65 70 75 80

Leu Asp Asn Asp Ile Met Leu Ile Lys Leu Ser Ser Pro Ala Thr Leu

85 90 95

Asn Ser Arg Val Ala Thr Val Ser Leu Pro Arg Ser Cys Ala Ala Ala

100 105 110

Gly Thr Glu Cys Leu Ile Ser Gly Trp Gly Asn Thr Lys Ser Ser Gly

115 120 125

Ser Ser Tyr Pro Ser Leu Leu Gln Cys Leu Lys Ala Pro Val Leu Ser

130 135 140

Asp Ser Ser Cys Lys Ser Ser Tyr Pro Gly Gln Ile Thr Gly Asn Met

145 150 155 160

Ile Cys Val Gly Phe Leu Glu Gly Gly Lys Asp Ala Cys Gln Gly Asp

165 170 175

Ser Gly Gly Pro Val Val Cys Asn Gly Gln Leu Gln Gly Ile Val Ser

180 185 190

Ser Gly Tyr Gly Cys Ala Gln Lys Asn Lys Pro Gly Val Tyr Thr Lys

195 200 205

Val Cys Asn Tyr Val Asn Trp Ile Gln Gln Thr Ile Ala Ala Asn

210 215 220

<210> 56

<211> 223

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Porcine trypsin variant #56

<400> 56

Ile Val Gly Gly Tyr Thr Cys Ala Ala Asn Ser Ile Pro Tyr Gln Val

1 5 10 15

Ser Leu Asn Ser Gly Ser His Arg Cys Gly Gly Ser Leu Ile Asn Ser

20 25 30

Gln Trp Val Val Ser Ala Ala His Cys Tyr Lys Ser Arg Ile Gln Val

35 40 45

Arg Leu Gly Glu His Asn Ile Asn Val Leu Glu Gly Asn Glu Gln Phe

50 55 60

Ile Asn Ala Ala Lys Ile Ile Thr His Pro Asn Phe Asn Gly Asn Thr

65 70 75 80

Leu Asp Asn Asp Ile Met Leu Ile Lys Leu Ser Ser Pro Ala Thr Leu

85 90 95

Asn Ser Arg Val Ala Thr Val Ser Leu Pro Arg Ser Cys Ala Ala Ala

100 105 110

Gly Thr Glu Cys Leu Ile Ser Gly Trp Gly Asn Thr Lys Ser Ser Gly

115 120 125

Ser Ser Tyr Pro Ser Leu Leu Gln Cys Leu Lys Ala Pro Val Leu Ser

130 135 140

Asp Ser Ser Cys Lys Ser Ser Tyr Pro Gly Gln Ile Thr Gly Asn Met

145 150 155 160

Ile Cys Val Gly Phe Leu Glu Gly Gly Lys Asp Ala Cys Gln Gly Asp

165 170 175

Ser Gly Gly Pro Val Val Cys Asn Gly Gln Leu Gln Gly Ile Val Ser

180 185 190

Ser Gly Tyr Gly Cys Ala Gln Lys Asn Lys Pro Gly Val Tyr Thr Lys

195 200 205

Val Cys Asn Tyr Val Asn Trp Ile Gln Gln Thr Ile Ala Ala Asn

210 215 220

<210> 57

<211> 223

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Porcine trypsin variant #57

<400> 57

Ile Val Gly Gly Tyr Thr Cys Ala Ala Asn Ser Ile Pro Tyr Gln Val

1 5 10 15

Ser Leu Asn Ser Gly Ser His Arg Cys Gly Gly Ser Leu Ile Asn Ser

20 25 30

Gln Trp Val Val Ser Ala Ala His Cys Tyr Lys Ser Arg Ile Gln Val

35 40 45

Arg Leu Gly Glu His Asn Ile His Val Leu Glu Gly Asn Glu Gln Phe

50 55 60

Ile Asn Ala Ala Lys Ile Ile Thr His Pro Asn Phe Asn Gly Asn Thr

65 70 75 80

Leu Asp Asn Asp Ile Met Leu Ile Lys Leu Ser Ser Pro Ala Thr Leu

85 90 95

Asn Ser Arg Val Ala Thr Val Ser Leu Pro Arg Ser Cys Ala Ala Ala

100 105 110

Gly Thr Glu Cys Leu Ile Ser Gly Trp Gly Asn Thr Lys Ser Ser Gly

115 120 125

Ser Ser Tyr Pro Ser Leu Leu Gln Cys Leu Lys Ala Pro Val Leu Ser

130 135 140

Asp Ser Ser Cys Lys Ser Ser Tyr Pro Gly Gln Ile Thr Gly Asn Met

145 150 155 160

Ile Cys Val Gly Phe Leu Glu Gly Gly Lys Asp Ala Cys Gln Gly Asp

165 170 175

Ser Gly Gly Pro Val Val Cys Asn Gly Gln Leu Gln Gly Ile Val Ser

180 185 190

Ser Gly Tyr Gly Cys Ala Gln Lys Asn Lys Pro Gly Val Tyr Thr Lys

195 200 205

Val Cys Asn Tyr Val Asn Trp Ile Gln Gln Thr Ile Ala Ala Asn

210 215 220

<210> 58

<211> 223

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Porcine trypsin variant #58

<400> 58

Ile Val Gly Gly Tyr Thr Cys Ala Ala Asn Ser Ile Pro Tyr Gln Val

1 5 10 15

Ser Leu Asn Ser Gly Ser His Arg Cys Gly Gly Ser Leu Ile Asn Ser

20 25 30

Gln Trp Val Val Ser Ala Ala His Cys Tyr Lys Ser Arg Ile Gln Val

35 40 45

Arg Leu Gly Glu His Asn Ile Asp Val Leu Glu Gly Asn Glu Gln Phe

50 55 60

Ile Asn Ala Ala Lys Ile Ile Thr His Pro Asn Phe Asn Gly Asn Thr

65 70 75 80

Leu Asp Asn Asp Ile Met Leu Ile Lys Leu Ser Ser Pro Ala Thr Leu

85 90 95

Asn Ser Arg Val Ala Thr Val Ser Leu Pro Pro Ser Cys Ala Ala Ala

100 105 110

Gly Thr Glu Cys Leu Ile Ser Gly Trp Gly Asn Thr Lys Ser Ser Gly

115 120 125

Ser Ser Tyr Pro Ser Leu Leu Gln Cys Leu Lys Ala Pro Val Leu Ser

130 135 140

Asp Ser Ser Cys Lys Ser Ser Tyr Pro Gly Gln Ile Thr Gly Asn Met

145 150 155 160

Ile Cys Val Gly Phe Leu Glu Gly Gly Lys Asp Ala Cys Gln Gly Asp

165 170 175

Ser Gly Gly Pro Val Val Cys Asn Gly Gln Leu Gln Gly Ile Val Ser

180 185 190

Ser Gly Tyr Gly Cys Ala Gln Lys Asn Lys Pro Gly Val Tyr Thr Lys

195 200 205

Val Cys Asn Tyr Val Asn Trp Ile Gln Gln Thr Ile Ala Ala Asn

210 215 220

<210> 59

<211> 223

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Porcine trypsin variant #59

<400> 59

Ile Val Gly Gly Tyr Thr Cys Ala Ala Asn Ser Ile Pro Tyr Gln Val

1 5 10 15

Ser Leu Asn Ser Gly Ser His Arg Cys Gly Gly Ser Leu Ile Asn Ser

20 25 30

Gln Trp Val Val Ser Ala Ala His Cys Tyr Lys Ser Arg Ile Gln Val

35 40 45

Arg Leu Gly Glu His Asn Ile Asp Val Leu Glu Gly Asn Glu Gln Phe

50 55 60

Ile Asn Ala Ala Lys Ile Ile Thr His Pro Asn Phe Asn Gly Asn Thr

65 70 75 80

Leu Asp Asn Asp Ile Met Leu Ile Lys Leu Ser Ser Pro Ala Thr Leu

85 90 95

Asn Ser Arg Val Ala Thr Val Ser Leu Pro Gly Ser Cys Ala Ala Ala

100 105 110

Gly Thr Glu Cys Leu Ile Ser Gly Trp Gly Asn Thr Lys Ser Ser Gly

115 120 125

Ser Ser Tyr Pro Ser Leu Leu Gln Cys Leu Lys Ala Pro Val Leu Ser

130 135 140

Asp Ser Ser Cys Lys Ser Ser Tyr Pro Gly Gln Ile Thr Gly Asn Met

145 150 155 160

Ile Cys Val Gly Phe Leu Glu Gly Gly Lys Asp Ala Cys Gln Gly Asp

165 170 175

Ser Gly Gly Pro Val Val Cys Asn Gly Gln Leu Gln Gly Ile Val Ser

180 185 190

Ser Gly Tyr Gly Cys Ala Gln Lys Asn Lys Pro Gly Val Tyr Thr Lys

195 200 205

Val Cys Asn Tyr Val Asn Trp Ile Gln Gln Thr Ile Ala Ala Asn

210 215 220

<210> 60

<211> 223

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Porcine trypsin variant #60

<400> 60

Ile Val Gly Gly Tyr Thr Cys Ala Ala Asn Ser Ile Pro Tyr Gln Val

1 5 10 15

Ser Leu Asn Ser Gly Ser His Arg Cys Gly Gly Ser Leu Ile Asn Ser

20 25 30

Gln Trp Val Val Ser Ala Ala His Cys Tyr Lys Ser Arg Ile Gln Val

35 40 45

Arg Leu Gly Glu His Asn Ile Asp Val Leu Glu Gly Asn Glu Gln Phe

50 55 60

Ile Asn Ala Ala Lys Ile Ile Thr His Pro Asn Phe Asn Gly Asn Thr

65 70 75 80

Leu Asp Asn Asp Ile Met Leu Ile Lys Leu Ser Ser Pro Ala Thr Leu

85 90 95

Asn Ser Arg Val Ala Thr Val Ser Leu Pro Ser Ser Cys Ala Ala Ala

100 105 110

Gly Thr Glu Cys Leu Ile Ser Gly Trp Gly Asn Thr Lys Ser Ser Gly

115 120 125

Ser Ser Tyr Pro Ser Leu Leu Gln Cys Leu Lys Ala Pro Val Leu Ser

130 135 140

Asp Ser Ser Cys Lys Ser Ser Tyr Pro Gly Gln Ile Thr Gly Asn Met

145 150 155 160

Ile Cys Val Gly Phe Leu Glu Gly Gly Lys Asp Ala Cys Gln Gly Asp

165 170 175

Ser Gly Gly Pro Val Val Cys Asn Gly Gln Leu Gln Gly Ile Val Ser

180 185 190

Ser Gly Tyr Gly Cys Ala Gln Lys Asn Lys Pro Gly Val Tyr Thr Lys

195 200 205

Val Cys Asn Tyr Val Asn Trp Ile Gln Gln Thr Ile Ala Ala Asn

210 215 220

<210> 61

<211> 223

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Porcine trypsin variant #61

<400> 61

Ile Val Gly Gly Tyr Thr Cys Ala Ala Asn Ser Ile Pro Tyr Gln Val

1 5 10 15

Ser Leu Asn Ser Gly Ser His Arg Cys Gly Gly Ser Leu Ile Asn Ser

20 25 30

Gln Trp Val Val Ser Ala Ala His Cys Tyr Lys Ser Arg Ile Gln Val

35 40 45

Arg Leu Gly Glu His Asn Ile Asp Val Leu Glu Gly Asn Glu Gln Phe

50 55 60

Ile Asn Ala Ala Lys Ile Ile Thr His Pro Asn Phe Asn Gly Asn Thr

65 70 75 80

Leu Asp Asn Asp Ile Met Leu Ile Lys Leu Ser Ser Pro Ala Thr Leu

85 90 95

Asn Ser Arg Val Ala Thr Val Ser Leu Pro Asp Ser Cys Ala Ala Ala

100 105 110

Gly Thr Glu Cys Leu Ile Ser Gly Trp Gly Asn Thr Lys Ser Ser Gly

115 120 125

Ser Ser Tyr Pro Ser Leu Leu Gln Cys Leu Lys Ala Pro Val Leu Ser

130 135 140

Asp Ser Ser Cys Lys Ser Ser Tyr Pro Gly Gln Ile Thr Gly Asn Met

145 150 155 160

Ile Cys Val Gly Phe Leu Glu Gly Gly Lys Asp Ala Cys Gln Gly Asp

165 170 175

Ser Gly Gly Pro Val Val Cys Asn Gly Gln Leu Gln Gly Ile Val Ser

180 185 190

Ser Gly Tyr Gly Cys Ala Gln Lys Asn Lys Pro Gly Val Tyr Thr Lys

195 200 205

Val Cys Asn Tyr Val Asn Trp Ile Gln Gln Thr Ile Ala Ala Asn

210 215 220

<210> 62

<211> 223

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Porcine trypsin variant #62

<400> 62

Ile Val Gly Gly Tyr Thr Cys Ala Ala Asn Ser Ile Pro Tyr Gln Val

1 5 10 15

Ser Leu Asn Ser Gly Ser His Arg Cys Gly Gly Ser Leu Ile Asn Ser

20 25 30

Gln Trp Val Val Ser Ala Ala His Cys Tyr Lys Ser Arg Ile Gln Val

35 40 45

Arg Leu Gly Glu His Asn Ile Asp Val Leu Glu Gly Asn Glu Gln Phe

50 55 60

Ile Asn Ala Ala Lys Ile Ile Thr His Pro Asn Phe Asn Gly Asn Thr

65 70 75 80

Leu Asp Asn Asp Ile Met Leu Ile Lys Leu Ser Ser Pro Ala Thr Leu

85 90 95

Asn Ser Arg Val Ala Thr Val Ser Leu Pro Arg Ser Cys Ala Ala Ala

100 105 110

Gly Thr Glu Cys Leu Ile Ser Gly Trp Gly Asn Thr Lys Ser Ser Gly

115 120 125

Ser Ser Val Pro Ser Leu Leu Gln Cys Leu Lys Ala Pro Val Leu Ser

130 135 140

Asp Ser Ser Cys Lys Ser Ser Tyr Pro Gly Gln Ile Thr Gly Asn Met

145 150 155 160

Ile Cys Val Gly Phe Leu Glu Gly Gly Lys Asp Ala Cys Gln Gly Asp

165 170 175

Ser Gly Gly Pro Val Val Cys Asn Gly Gln Leu Gln Gly Ile Val Ser

180 185 190

Ser Gly Tyr Gly Cys Ala Gln Lys Asn Lys Pro Gly Val Tyr Thr Lys

195 200 205

Val Cys Asn Tyr Val Asn Trp Ile Gln Gln Thr Ile Ala Ala Asn

210 215 220

<210> 63

<211> 223

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Porcine trypsin variant #63

<400> 63

Ile Val Gly Gly Tyr Thr Cys Ala Ala Asn Ser Ile Pro Tyr Gln Val

1 5 10 15

Ser Leu Asn Ser Gly Ser His Arg Cys Gly Gly Ser Leu Ile Asn Ser

20 25 30

Gln Trp Val Val Ser Ala Ala His Cys Tyr Lys Ser Arg Ile Gln Val

35 40 45

Arg Leu Gly Glu His Asn Ile Asp Val Leu Glu Gly Asn Glu Gln Phe

50 55 60

Ile Asn Ala Ala Lys Ile Ile Thr His Pro Asn Phe Asn Gly Asn Thr

65 70 75 80

Leu Asp Asn Asp Ile Met Leu Ile Lys Leu Ser Ser Pro Ala Thr Leu

85 90 95

Asn Ser Arg Val Ala Thr Val Ser Leu Pro Arg Ser Cys Ala Ala Ala

100 105 110

Gly Thr Glu Cys Leu Ile Ser Gly Trp Gly Asn Thr Lys Ser Ser Gly

115 120 125

Ser Ser Ile Pro Ser Leu Leu Gln Cys Leu Lys Ala Pro Val Leu Ser

130 135 140

Asp Ser Ser Cys Lys Ser Ser Tyr Pro Gly Gln Ile Thr Gly Asn Met

145 150 155 160

Ile Cys Val Gly Phe Leu Glu Gly Gly Lys Asp Ala Cys Gln Gly Asp

165 170 175

Ser Gly Gly Pro Val Val Cys Asn Gly Gln Leu Gln Gly Ile Val Ser

180 185 190

Ser Gly Tyr Gly Cys Ala Gln Lys Asn Lys Pro Gly Val Tyr Thr Lys

195 200 205

Val Cys Asn Tyr Val Asn Trp Ile Gln Gln Thr Ile Ala Ala Asn

210 215 220

<210> 64

<211> 223

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Porcine trypsin variant #64

<400> 64

Ile Val Gly Gly Tyr Thr Cys Ala Ala Asn Ser Ile Pro Tyr Gln Val

1 5 10 15

Ser Leu Asn Ser Gly Ser His Arg Cys Gly Gly Ser Leu Ile Asn Ser

20 25 30

Gln Trp Val Val Ser Ala Ala His Cys Tyr Lys Ser Arg Ile Gln Val

35 40 45

Arg Leu Gly Glu His Asn Ile Asp Val Leu Glu Gly Asn Glu Gln Phe

50 55 60

Ile Asn Ala Ala Lys Ile Ile Thr His Pro Asn Phe Asn Gly Asn Thr

65 70 75 80

Leu Asp Asn Asp Ile Met Leu Ile Lys Leu Ser Ser Pro Ala Thr Leu

85 90 95

Asn Ser Arg Val Ala Thr Val Ser Leu Pro Arg Ser Cys Ala Ala Ala

100 105 110

Gly Thr Glu Cys Leu Ile Ser Gly Trp Gly Asn Thr Lys Ser Ser Gly

115 120 125

Ser Ser Trp Pro Ser Leu Leu Gln Cys Leu Lys Ala Pro Val Leu Ser

130 135 140

Asp Ser Ser Cys Lys Ser Ser Tyr Pro Gly Gln Ile Thr Gly Asn Met

145 150 155 160

Ile Cys Val Gly Phe Leu Glu Gly Gly Lys Asp Ala Cys Gln Gly Asp

165 170 175

Ser Gly Gly Pro Val Val Cys Asn Gly Gln Leu Gln Gly Ile Val Ser

180 185 190

Ser Gly Tyr Gly Cys Ala Gln Lys Asn Lys Pro Gly Val Tyr Thr Lys

195 200 205

Val Cys Asn Tyr Val Asn Trp Ile Gln Gln Thr Ile Ala Ala Asn

210 215 220

<210> 65

<211> 223

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Porcine trypsin variant #65

<400> 65

Ile Val Gly Gly Tyr Thr Cys Ala Ala Asn Ser Ile Pro Tyr Gln Val

1 5 10 15

Ser Leu Asn Ser Gly Ser His Arg Cys Gly Gly Ser Leu Ile Asn Ser

20 25 30

Gln Trp Val Val Ser Ala Ala His Cys Tyr Lys Ser Arg Ile Gln Val

35 40 45

Arg Leu Gly Glu His Asn Ile Asp Val Leu Glu Gly Asn Glu Gln Phe

50 55 60

Ile Asn Ala Ala Lys Ile Ile Thr His Pro Asn Phe Asn Gly Asn Thr

65 70 75 80

Leu Asp Asn Asp Ile Met Leu Ile Lys Leu Ser Ser Pro Ala Thr Leu

85 90 95

Asn Ser Arg Val Ala Thr Val Ser Leu Pro Arg Ser Cys Ala Ala Ala

100 105 110

Gly Thr Glu Cys Leu Ile Ser Gly Trp Gly Asn Thr Lys Ser Ser Gly

115 120 125

Ser Ser Gly Pro Ser Leu Leu Gln Cys Leu Lys Ala Pro Val Leu Ser

130 135 140

Asp Ser Ser Cys Lys Ser Ser Tyr Pro Gly Gln Ile Thr Gly Asn Met

145 150 155 160

Ile Cys Val Gly Phe Leu Glu Gly Gly Lys Asp Ala Cys Gln Gly Asp

165 170 175

Ser Gly Gly Pro Val Val Cys Asn Gly Gln Leu Gln Gly Ile Val Ser

180 185 190

Ser Gly Tyr Gly Cys Ala Gln Lys Asn Lys Pro Gly Val Tyr Thr Lys

195 200 205

Val Cys Asn Tyr Val Asn Trp Ile Gln Gln Thr Ile Ala Ala Asn

210 215 220

<210> 66

<211> 223

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Porcine trypsin variant #66

<400> 66

Ile Val Gly Gly Tyr Thr Cys Ala Ala Asn Ser Ile Pro Tyr Gln Val

1 5 10 15

Ser Leu Asn Ser Gly Ser His Arg Cys Gly Gly Ser Leu Ile Asn Ser

20 25 30

Gln Trp Val Val Ser Ala Ala His Cys Tyr Lys Ser Arg Ile Gln Val

35 40 45

Arg Leu Gly Glu His Asn Ile Asp Val Leu Glu Gly Asn Glu Gln Phe

50 55 60

Ile Asn Ala Ala Lys Ile Ile Thr His Pro Asn Phe Asn Gly Asn Thr

65 70 75 80

Leu Asp Asn Asp Ile Met Leu Ile Lys Leu Ser Ser Pro Ala Thr Leu

85 90 95

Asn Ser Arg Val Ala Thr Val Ser Leu Pro Arg Ser Cys Ala Ala Ala

100 105 110

Gly Thr Glu Cys Leu Ile Ser Gly Trp Gly Asn Thr Lys Ser Ser Gly

115 120 125

Ser Ser Ser Pro Ser Leu Leu Gln Cys Leu Lys Ala Pro Val Leu Ser

130 135 140

Asp Ser Ser Cys Lys Ser Ser Tyr Pro Gly Gln Ile Thr Gly Asn Met

145 150 155 160

Ile Cys Val Gly Phe Leu Glu Gly Gly Lys Asp Ala Cys Gln Gly Asp

165 170 175

Ser Gly Gly Pro Val Val Cys Asn Gly Gln Leu Gln Gly Ile Val Ser

180 185 190

Ser Gly Tyr Gly Cys Ala Gln Lys Asn Lys Pro Gly Val Tyr Thr Lys

195 200 205

Val Cys Asn Tyr Val Asn Trp Ile Gln Gln Thr Ile Ala Ala Asn

210 215 220

<210> 67

<211> 223

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Porcine trypsin variant #67

<400> 67

Ile Val Gly Gly Tyr Thr Cys Ala Ala Asn Ser Ile Pro Tyr Gln Val

1 5 10 15

Ser Leu Asn Ser Gly Ser His Arg Cys Gly Gly Ser Leu Ile Asn Ser

20 25 30

Gln Trp Val Val Ser Ala Ala His Cys Tyr Lys Ser Arg Ile Gln Val

35 40 45

Arg Leu Gly Glu His Asn Ile Asp Val Leu Glu Gly Asn Glu Gln Phe

50 55 60

Ile Asn Ala Ala Lys Ile Ile Thr His Pro Asn Phe Asn Gly Asn Thr

65 70 75 80

Leu Asp Asn Asp Ile Met Leu Ile Lys Leu Ser Ser Pro Ala Thr Leu

85 90 95

Asn Ser Arg Val Ala Thr Val Ser Leu Pro Arg Ser Cys Ala Ala Ala

100 105 110

Gly Thr Glu Cys Leu Ile Ser Gly Trp Gly Asn Thr Lys Ser Ser Gly

115 120 125

Ser Ser Gln Pro Ser Leu Leu Gln Cys Leu Lys Ala Pro Val Leu Ser

130 135 140

Asp Ser Ser Cys Lys Ser Ser Tyr Pro Gly Gln Ile Thr Gly Asn Met

145 150 155 160

Ile Cys Val Gly Phe Leu Glu Gly Gly Lys Asp Ala Cys Gln Gly Asp

165 170 175

Ser Gly Gly Pro Val Val Cys Asn Gly Gln Leu Gln Gly Ile Val Ser

180 185 190

Ser Gly Tyr Gly Cys Ala Gln Lys Asn Lys Pro Gly Val Tyr Thr Lys

195 200 205

Val Cys Asn Tyr Val Asn Trp Ile Gln Gln Thr Ile Ala Ala Asn

210 215 220

<210> 68

<211> 223

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Porcine trypsin variant #68

<400> 68

Ile Val Gly Gly Tyr Thr Cys Ala Ala Asn Ser Ile Pro Tyr Gln Val

1 5 10 15

Ser Leu Asn Ser Gly Ser His Arg Cys Gly Gly Ser Leu Ile Asn Ser

20 25 30

Gln Trp Val Val Ser Ala Ala His Cys Tyr Lys Ser Arg Ile Gln Val

35 40 45

Arg Leu Gly Glu His Asn Ile Asp Val Leu Glu Gly Asn Glu Gln Phe

50 55 60

Ile Asn Ala Ala Lys Ile Ile Thr His Pro Asn Phe Asn Gly Asn Thr

65 70 75 80

Leu Asp Asn Asp Ile Met Leu Ile Lys Leu Ser Ser Pro Ala Thr Leu

85 90 95

Asn Ser Arg Val Ala Thr Val Ser Leu Pro Arg Ser Cys Ala Ala Ala

100 105 110

Gly Thr Glu Cys Leu Ile Ser Gly Trp Gly Asn Thr Lys Ser Ser Gly

115 120 125

Ser Ser Asp Pro Ser Leu Leu Gln Cys Leu Lys Ala Pro Val Leu Ser

130 135 140

Asp Ser Ser Cys Lys Ser Ser Tyr Pro Gly Gln Ile Thr Gly Asn Met

145 150 155 160

Ile Cys Val Gly Phe Leu Glu Gly Gly Lys Asp Ala Cys Gln Gly Asp

165 170 175

Ser Gly Gly Pro Val Val Cys Asn Gly Gln Leu Gln Gly Ile Val Ser

180 185 190

Ser Gly Tyr Gly Cys Ala Gln Lys Asn Lys Pro Gly Val Tyr Thr Lys

195 200 205

Val Cys Asn Tyr Val Asn Trp Ile Gln Gln Thr Ile Ala Ala Asn

210 215 220

<210> 69

<211> 223

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Porcine trypsin variant #69

<400> 69

Ile Val Gly Gly Tyr Thr Cys Ala Ala Asn Ser Ile Pro Tyr Gln Val

1 5 10 15

Ser Leu Asn Ser Gly Ser His Arg Cys Gly Gly Ser Leu Ile Asn Ser

20 25 30

Gln Trp Val Val Ser Ala Ala His Cys Tyr Lys Ser Arg Ile Gln Val

35 40 45

Arg Leu Gly Glu His Asn Ile Asp Val Leu Glu Gly Asn Glu Gln Phe

50 55 60

Ile Asn Ala Ala Lys Ile Ile Thr His Pro Asn Phe Asn Gly Asn Thr

65 70 75 80

Leu Asp Asn Asp Ile Met Leu Ile Lys Leu Ser Ser Pro Ala Thr Leu

85 90 95

Asn Ser Arg Val Ala Thr Val Ser Leu Pro Arg Ser Cys Ala Ala Ala

100 105 110

Gly Thr Glu Cys Leu Ile Ser Gly Trp Gly Asn Thr Lys Ser Ser Gly

115 120 125

Ser Ser Tyr Pro Ser Leu Leu Gln Cys Leu Lys Ala Pro Val Leu Ser

130 135 140

Asp Ser Ser Cys Lys Ser Ser Tyr Pro Gly Gln Ile Thr Gly Asn Met

145 150 155 160

Ile Cys Val Gly Phe Leu Glu Gly Gly Lys Asp Ala Cys Gln Gly Asp

165 170 175

Ser Gly Gly Pro Val Val Cys Asn Gly Gln Leu Gln Gly Ile Val Ser

180 185 190

Phe Gly Tyr Gly Cys Ala Gln Lys Asn Lys Pro Gly Val Tyr Thr Lys

195 200 205

Val Cys Asn Tyr Val Asn Trp Ile Gln Gln Thr Ile Ala Ala Asn

210 215 220

<210> 70

<211> 223

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Porcine trypsin variant #70

<400> 70

Ile Val Gly Gly Tyr Thr Cys Ala Ala Asn Ser Ile Pro Tyr Gln Val

1 5 10 15

Ser Leu Asn Ser Gly Ser His Arg Cys Gly Gly Ser Leu Ile Asn Ser

20 25 30

Gln Trp Val Val Ser Ala Ala His Cys Tyr Lys Ser Arg Ile Gln Val

35 40 45

Arg Leu Gly Glu His Asn Ile Asp Val Leu Glu Gly Asn Glu Gln Phe

50 55 60

Ile Asn Ala Ala Lys Ile Ile Thr His Pro Asn Phe Asn Gly Asn Thr

65 70 75 80

Leu Asp Asn Asp Ile Met Leu Ile Lys Leu Ser Ser Pro Ala Thr Leu

85 90 95

Asn Ser Arg Val Ala Thr Val Ser Leu Pro Arg Ser Cys Ala Ala Ala

100 105 110

Gly Thr Glu Cys Leu Ile Ser Gly Trp Gly Asn Thr Lys Ser Ser Gly

115 120 125

Ser Ser Tyr Pro Ser Leu Leu Gln Cys Leu Lys Ala Pro Val Leu Ser

130 135 140

Asp Ser Ser Cys Lys Ser Ser Tyr Pro Gly Gln Ile Thr Gly Asn Met

145 150 155 160

Ile Cys Val Gly Phe Leu Glu Gly Gly Lys Asp Ala Cys Gln Gly Asp

165 170 175

Ser Gly Gly Pro Val Val Cys Asn Gly Gln Leu Gln Gly Ile Val Ser

180 185 190

Tyr Gly Tyr Gly Cys Ala Gln Lys Asn Lys Pro Gly Val Tyr Thr Lys

195 200 205

Val Cys Asn Tyr Val Asn Trp Ile Gln Gln Thr Ile Ala Ala Asn

210 215 220

<210> 71

<211> 223

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Porcine trypsin variant #71

<400> 71

Ile Val Gly Gly Tyr Thr Cys Ala Ala Asn Ser Ile Pro Tyr Gln Val

1 5 10 15

Ser Leu Asn Ser Gly Ser His Arg Cys Gly Gly Ser Leu Ile Asn Ser

20 25 30

Gln Trp Val Val Ser Ala Ala His Cys Tyr Lys Ser Arg Ile Gln Val

35 40 45

Arg Leu Gly Glu His Asn Ile Asp Val Leu Glu Gly Asn Glu Gln Phe

50 55 60

Ile Asn Ala Ala Lys Ile Ile Thr His Pro Asn Phe Asn Gly Asn Thr

65 70 75 80

Leu Asp Asn Asp Ile Met Leu Ile Lys Leu Ser Ser Pro Ala Thr Leu

85 90 95

Asn Ser Arg Val Ala Thr Val Ser Leu Pro Arg Ser Cys Ala Ala Ala

100 105 110

Gly Thr Glu Cys Leu Ile Ser Gly Trp Gly Asn Thr Lys Ser Ser Gly

115 120 125

Ser Ser Tyr Pro Ser Leu Leu Gln Cys Leu Lys Ala Pro Val Leu Ser

130 135 140

Asp Ser Ser Cys Lys Ser Ser Tyr Pro Gly Gln Ile Thr Gly Asn Met

145 150 155 160

Ile Cys Val Gly Phe Leu Glu Gly Gly Lys Asp Ala Cys Gln Gly Asp

165 170 175

Ser Gly Gly Pro Val Val Cys Asn Gly Gln Leu Gln Gly Ile Val Ser

180 185 190

Thr Gly Tyr Gly Cys Ala Gln Lys Asn Lys Pro Gly Val Tyr Thr Lys

195 200 205

Val Cys Asn Tyr Val Asn Trp Ile Gln Gln Thr Ile Ala Ala Asn

210 215 220

<210> 72

<211> 223

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Porcine trypsin variant #72

<400> 72

Ile Val Gly Gly Tyr Thr Cys Ala Ala Asn Ser Ile Pro Tyr Gln Val

1 5 10 15

Ser Leu Asn Ser Gly Ser His Arg Cys Gly Gly Ser Leu Ile Asn Ser

20 25 30

Gln Trp Val Val Ser Ala Ala His Cys Tyr Lys Ser Arg Ile Gln Val

35 40 45

Arg Leu Gly Glu His Asn Ile Asp Val Leu Glu Gly Asn Glu Gln Phe

50 55 60

Ile Asn Ala Ala Lys Ile Ile Thr His Pro Asn Phe Asn Ser Asn Thr

65 70 75 80

Leu Asp Asn Asp Ile Met Leu Ile Lys Leu Ser Ser Pro Ala Thr Leu

85 90 95

Asn Ser Arg Val Ala Thr Val Ser Leu Pro Arg Ser Cys Ala Ala Ala

100 105 110

Gly Thr Glu Cys Leu Ile Ser Gly Trp Gly Asn Thr Lys Ser Ser Gly

115 120 125

Ser Ser Tyr Pro Ser Leu Leu Gln Cys Leu Lys Ala Pro Val Leu Ser

130 135 140

Asp Ser Ser Cys Lys Ser Ser Tyr Pro Gly Gln Ile Thr Gly Asn Met

145 150 155 160

Ile Cys Val Gly Phe Leu Glu Gly Gly Lys Asp Ala Cys Gln Gly Asp

165 170 175

Ser Gly Gly Pro Val Val Cys Asn Gly Gln Leu Gln Gly Ile Val Ser

180 185 190

Ser Gly Tyr Gly Cys Ala Gln Lys Asn Lys Pro Gly Val Tyr Thr Lys

195 200 205

Val Cys Asn Tyr Val Asn Trp Ile Gln Gln Thr Ile Ala Ala Asn

210 215 220

<210> 73

<211> 223

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Porcine trypsin variant #73

<400> 73

Ile Val Gly Gly Tyr Thr Cys Ala Ala Asn Ser Ile Pro Tyr Gln Val

1 5 10 15

Ser Leu Asn Ser Gly Ser His Arg Cys Gly Gly Ser Leu Ile Asn Ser

20 25 30

Gln Trp Val Val Ser Ala Ala His Cys Tyr Lys Ser Arg Ile Gln Val

35 40 45

Arg Leu Gly Glu His Asn Ile Asp Val Leu Glu Gly Asn Glu Gln Phe

50 55 60

Ile Asn Ala Ala Lys Ile Ile Thr His Pro Asn Phe Asn Ala Asn Thr

65 70 75 80

Leu Asp Asn Asp Ile Met Leu Ile Lys Leu Ser Ser Pro Ala Thr Leu

85 90 95

Asn Ser Arg Val Ala Thr Val Ser Leu Pro Arg Ser Cys Ala Ala Ala

100 105 110

Gly Thr Glu Cys Leu Ile Ser Gly Trp Gly Asn Thr Lys Ser Ser Gly

115 120 125

Ser Ser Tyr Pro Ser Leu Leu Gln Cys Leu Lys Ala Pro Val Leu Ser

130 135 140

Asp Ser Ser Cys Lys Ser Ser Tyr Pro Gly Gln Ile Thr Gly Asn Met

145 150 155 160

Ile Cys Val Gly Phe Leu Glu Gly Gly Lys Asp Ala Cys Gln Gly Asp

165 170 175

Ser Gly Gly Pro Val Val Cys Asn Gly Gln Leu Gln Gly Ile Val Ser

180 185 190

Ser Gly Tyr Gly Cys Ala Gln Lys Asn Lys Pro Gly Val Tyr Thr Lys

195 200 205

Val Cys Asn Tyr Val Asn Trp Ile Gln Gln Thr Ile Ala Ala Asn

210 215 220

<210> 74

<211> 223

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Porcine trypsin variant #74

<400> 74

Ile Val Gly Gly Tyr Thr Cys Ala Ala Asn Ser Ile Pro Tyr Gln Val

1 5 10 15

Ser Leu Asn Ser Gly Ser His Arg Cys Gly Gly Ser Leu Ile Asn Ser

20 25 30

Gln Trp Val Val Ser Ala Ala His Cys Tyr Lys Ser Arg Ile Gln Val

35 40 45

Arg Leu Gly Glu His Asn Ile Asp Val Leu Glu Gly Asn Glu Gln Phe

50 55 60

Ile Asn Ala Ala Lys Ile Ile Thr His Pro Asn Phe Asn Pro Asn Thr

65 70 75 80

Leu Asp Asn Asp Ile Met Leu Ile Lys Leu Ser Ser Pro Ala Thr Leu

85 90 95

Asn Ser Arg Val Ala Thr Val Ser Leu Pro Arg Ser Cys Ala Ala Ala

100 105 110

Gly Thr Glu Cys Leu Ile Ser Gly Trp Gly Asn Thr Lys Ser Ser Gly

115 120 125

Ser Ser Tyr Pro Ser Leu Leu Gln Cys Leu Lys Ala Pro Val Leu Ser

130 135 140

Asp Ser Ser Cys Lys Ser Ser Tyr Pro Gly Gln Ile Thr Gly Asn Met

145 150 155 160

Ile Cys Val Gly Phe Leu Glu Gly Gly Lys Asp Ala Cys Gln Gly Asp

165 170 175

Ser Gly Gly Pro Val Val Cys Asn Gly Gln Leu Gln Gly Ile Val Ser

180 185 190

Ser Gly Tyr Gly Cys Ala Gln Lys Asn Lys Pro Gly Val Tyr Thr Lys

195 200 205

Val Cys Asn Tyr Val Asn Trp Ile Gln Gln Thr Ile Ala Ala Asn

210 215 220

<210> 75

<211> 223

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Porcine trypsin variant #75

<400> 75

Ile Val Gly Gly Tyr Thr Cys Ala Ala Asn Ser Ile Pro Tyr Gln Val

1 5 10 15

Ser Leu Asn Ser Gly Ser His Arg Cys Gly Gly Ser Leu Ile Asn Ser

20 25 30

Gln Trp Val Val Ser Ala Ala His Cys Tyr Lys Ser Arg Ile Gln Val

35 40 45

Arg Leu Gly Glu His Asn Ile Asp Val Leu Glu Gly Asn Glu Gln Phe

50 55 60

Ile Asn Ala Ala Lys Ile Ile Thr His Pro Asn Phe Asn Tyr Asn Thr

65 70 75 80

Leu Asp Asn Asp Ile Met Leu Ile Lys Leu Ser Ser Pro Ala Thr Leu

85 90 95

Asn Ser Arg Val Ala Thr Val Ser Leu Pro Arg Ser Cys Ala Ala Ala

100 105 110

Gly Thr Glu Cys Leu Ile Ser Gly Trp Gly Asn Thr Lys Ser Ser Gly

115 120 125

Ser Ser Tyr Pro Ser Leu Leu Gln Cys Leu Lys Ala Pro Val Leu Ser

130 135 140

Asp Ser Ser Cys Lys Ser Ser Tyr Pro Gly Gln Ile Thr Gly Asn Met

145 150 155 160

Ile Cys Val Gly Phe Leu Glu Gly Gly Lys Asp Ala Cys Gln Gly Asp

165 170 175

Ser Gly Gly Pro Val Val Cys Asn Gly Gln Leu Gln Gly Ile Val Ser

180 185 190

Ser Gly Tyr Gly Cys Ala Gln Lys Asn Lys Pro Gly Val Tyr Thr Lys

195 200 205

Val Cys Asn Tyr Val Asn Trp Ile Gln Gln Thr Ile Ala Ala Asn

210 215 220

<210> 76

<211> 223

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Porcine trypsin variant #76

<400> 76

Ile Val Gly Gly Tyr Thr Cys Ala Ala Asn Ser Ile Pro Tyr Gln Val

1 5 10 15

Ser Leu Asn Ser Gly Ser His Arg Cys Gly Gly Ser Leu Ile Asn Ser

20 25 30

Gln Trp Val Val Ser Ala Ala His Cys Tyr Lys Ser Arg Ile Gln Val

35 40 45

Arg Leu Gly Glu His Asn Ile Asp Val Leu Glu Gly Asn Glu Gln Phe

50 55 60

Ile Asn Ala Ala Lys Ile Ile Thr His Pro Asn Phe Asn Glu Asn Thr

65 70 75 80

Leu Asp Asn Asp Ile Met Leu Ile Lys Leu Ser Ser Pro Ala Thr Leu

85 90 95

Asn Ser Arg Val Ala Thr Val Ser Leu Pro Arg Ser Cys Ala Ala Ala

100 105 110

Gly Thr Glu Cys Leu Ile Ser Gly Trp Gly Asn Thr Lys Ser Ser Gly

115 120 125

Ser Ser Tyr Pro Ser Leu Leu Gln Cys Leu Lys Ala Pro Val Leu Ser

130 135 140

Asp Ser Ser Cys Lys Ser Ser Tyr Pro Gly Gln Ile Thr Gly Asn Met

145 150 155 160

Ile Cys Val Gly Phe Leu Glu Gly Gly Lys Asp Ala Cys Gln Gly Asp

165 170 175

Ser Gly Gly Pro Val Val Cys Asn Gly Gln Leu Gln Gly Ile Val Ser

180 185 190

Ser Gly Tyr Gly Cys Ala Gln Lys Asn Lys Pro Gly Val Tyr Thr Lys

195 200 205

Val Cys Asn Tyr Val Asn Trp Ile Gln Gln Thr Ile Ala Ala Asn

210 215 220

<210> 77

<211> 223

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Porcine trypsin variant #77

<400> 77

Ile Val Gly Gly Tyr Thr Cys Ala Ala Asn Ser Ile Pro Tyr Gln Val

1 5 10 15

Ser Leu Asn Ser Gly Ser His Arg Cys Gly Gly Ser Leu Ile Asn Ser

20 25 30

Gln Trp Val Val Ser Ala Ala His Cys Tyr Lys Pro Arg Ile Gln Val

35 40 45

Arg Leu Gly Glu His Asn Ile His Val Leu Glu Gly Asn Glu Gln Phe

50 55 60

Ile Asn Ala Ala Lys Ile Ile Thr His Pro Asn Phe Asn Gly Asn Thr

65 70 75 80

Leu Asp Asn Asp Ile Met Leu Ile Lys Leu Ser Ser Pro Ala Thr Leu

85 90 95

Asn Ser His Val Ala Thr Val Ser Leu Pro Thr Ser Cys Ala Ala Ala

100 105 110

Gly Thr Glu Cys Leu Ile Ser Gly Trp Gly Asn Thr Cys Ser Ser Gly

115 120 125

Ser Ser Met Pro Ser Leu Leu Gln Cys Leu Lys Ala Pro Val Leu Ser

130 135 140

Asp Ser Ser Cys Lys Ser Ser Tyr Pro Gly Gln Ile Thr Gly Asn Met

145 150 155 160

Ile Cys Val Gly Phe Leu Glu Gly Gly Lys Asp Ala Cys Gln Gly Asp

165 170 175

Ser Gly Gly Pro Val Val Cys Asn Gly Gln Leu Gln Gly Ile Val Ser

180 185 190

Ser Gly Tyr Gly Cys Ala Gln Lys Asn Lys Pro Gly Val Tyr Thr Lys

195 200 205

Val Cys Asn Tyr Val Asn Trp Ile Gln Gln Thr Ile Ala Ala Asn

210 215 220

<210> 78

<211> 223

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Porcine trypsin variant #78

<400> 78

Ile Val Gly Gly Tyr Thr Cys Ala Ala Asn Ser Ile Pro Tyr Gln Val

1 5 10 15

Ser Leu Asn Ser Gly Ser His Arg Cys Gly Gly Ser Leu Ile Asn Ser

20 25 30

Gln Trp Val Val Ser Ala Ala His Cys Tyr Lys Ser Arg Ile Gln Val

35 40 45

Arg Leu Gly Glu His Asn Ile His Val Leu Glu Gly Asn Glu Gln Phe

50 55 60

Ile Asn Ala Ala Lys Ile Ile Thr His Pro Asn Phe Asn Pro Asn Thr

65 70 75 80

Leu Asp Asn Asp Ile Met Leu Ile Lys Leu Ser Ser Pro Ala Thr Leu

85 90 95

Asn Ser His Val Ala Thr Val Ser Leu Pro Thr Ser Cys Ala Ala Ala

100 105 110

Gly Thr Glu Cys Leu Ile Ser Gly Trp Gly Asn Thr Cys Ser Ser Gly

115 120 125

Ser Ser Tyr Pro Ser Leu Leu Gln Cys Leu Lys Ala Pro Val Leu Ser

130 135 140

Asp Ser Ser Cys Lys Ser Ser Tyr Pro Gly Gln Ile Thr Gly Asn Met

145 150 155 160

Ile Cys Val Gly Phe Leu Glu Gly Gly Lys Asp Ala Cys Gln Gly Asp

165 170 175

Ser Gly Gly Pro Val Val Cys Asn Gly Gln Leu Gln Gly Ile Val Ser

180 185 190

Ser Gly Tyr Gly Cys Ala Gln Lys Asn Lys Pro Gly Val Tyr Thr Lys

195 200 205

Val Cys Asn Tyr Val Asn Trp Ile Gln Gln Thr Ile Ala Ala Asn

210 215 220

<210> 79

<211> 223

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Porcine trypsin variant #79

<400> 79

Ile Val Gly Gly Tyr Thr Cys Ala Ala Asn Ser Ile Pro Tyr Gln Val

1 5 10 15

Ser Leu Asn Ser Gly Ser His Arg Cys Gly Gly Ser Leu Ile Asn Ser

20 25 30

Gln Trp Val Val Ser Ala Ala His Cys Tyr Lys Pro Arg Ile Gln Val

35 40 45

Arg Leu Gly Glu His Asn Ile His Val Leu Glu Gly Asn Glu Gln Phe

50 55 60

Ile Asn Ala Ala Lys Ile Ile Thr His Pro Asn Phe Asn Pro Asn Thr

65 70 75 80

Leu Asp Asn Asp Ile Met Leu Ile Lys Leu Ser Ser Pro Ala Thr Leu

85 90 95

Asn Ser His Val Ala Thr Val Ser Leu Pro Arg Ser Cys Ala Ala Ala

100 105 110

Gly Thr Glu Cys Leu Ile Ser Gly Trp Gly Asn Thr Ala Ser Ser Gly

115 120 125

Ser Ser Met Pro Ser Leu Leu Gln Cys Leu Lys Ala Pro Val Leu Ser

130 135 140

Asp Ser Ser Cys Lys Ser Ser Tyr Pro Gly Gln Ile Thr Gly Asn Met

145 150 155 160

Ile Cys Val Gly Phe Leu Glu Gly Gly Lys Asp Ala Cys Gln Gly Asp

165 170 175

Ser Gly Gly Pro Val Val Cys Asn Gly Gln Leu Gln Gly Ile Val Ser

180 185 190

Ser Gly Tyr Gly Cys Ala Gln Lys Asn Lys Pro Gly Val Tyr Thr Lys

195 200 205

Val Cys Asn Tyr Val Asn Trp Ile Gln Gln Thr Ile Ala Ala Asn

210 215 220

<210> 80

<211> 223

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Porcine trypsin variant #80

<400> 80

Ile Val Gly Gly Tyr Thr Cys Ala Ala Asn Ser Ile Pro Tyr Gln Val

1 5 10 15

Ser Leu Asn Ser Gly Ser His Arg Cys Gly Gly Ser Leu Ile Asn Ser

20 25 30

Gln Trp Val Val Ser Ala Ala His Cys Tyr Lys Ser Arg Ile Gln Val

35 40 45

Arg Leu Gly Glu His Asn Ile His Val Leu Glu Gly Asn Glu Gln Phe

50 55 60

Ile Asn Ala Ala Lys Ile Ile Thr His Pro Asn Phe Asn Pro Asn Thr

65 70 75 80

Leu Asp Asn Asp Ile Met Leu Ile Lys Leu Ser Ser Pro Ala Thr Leu

85 90 95

Asn Ser His Val Ala Thr Val Ser Leu Pro Thr Ser Cys Ala Ala Ala

100 105 110

Gly Thr Glu Cys Leu Ile Ser Gly Trp Gly Asn Thr Ser Ser Ser Gly

115 120 125

Ser Ser Met Pro Ser Leu Leu Gln Cys Leu Lys Ala Pro Val Leu Ser

130 135 140

Asp Ser Ser Cys Lys Ser Ser Tyr Pro Gly Gln Ile Thr Gly Asn Met

145 150 155 160

Ile Cys Val Gly Phe Leu Glu Gly Gly Lys Asp Ala Cys Gln Gly Asp

165 170 175

Ser Gly Gly Pro Val Val Cys Asn Gly Gln Leu Gln Gly Ile Val Ser

180 185 190

Ser Gly Tyr Gly Cys Ala Gln Lys Asn Lys Pro Gly Val Tyr Thr Lys

195 200 205

Val Cys Asn Tyr Val Asn Trp Ile Gln Gln Thr Ile Ala Ala Asn

210 215 220

<210> 81

<211> 223

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Porcine trypsin variant #81

<400> 81

Ile Val Gly Gly Tyr Thr Cys Ala Ala Asn Ser Ile Pro Tyr Gln Val

1 5 10 15

Ser Leu Asn Ser Gly Ser His Arg Cys Gly Gly Ser Leu Ile Asn Ser

20 25 30

Gln Trp Val Val Ser Ala Ala His Cys Tyr Lys Ser Arg Ile Gln Val

35 40 45

Arg Leu Gly Glu His Asn Ile His Val Leu Glu Gly Asn Glu Gln Phe

50 55 60

Ile Asn Ala Ala Lys Ile Ile Thr His Pro Asn Phe Asn Pro Asn Thr

65 70 75 80

Leu Asp Asn Asp Ile Met Leu Ile Lys Leu Ser Ser Pro Ala Thr Leu

85 90 95

Asn Ser His Val Ala Thr Val Ser Leu Pro Thr Ser Cys Ala Ala Ala

100 105 110

Gly Thr Glu Cys Leu Ile Ser Gly Trp Gly Asn Thr Ala Ser Ser Gly

115 120 125

Ser Ser Met Pro Ser Leu Leu Gln Cys Leu Lys Ala Pro Val Leu Ser

130 135 140

Asp Ser Ser Cys Lys Ser Ser Tyr Pro Gly Gln Ile Thr Gly Asn Met

145 150 155 160

Ile Cys Val Gly Phe Leu Glu Gly Gly Lys Asp Ala Cys Gln Gly Asp

165 170 175

Ser Gly Gly Pro Val Val Cys Asn Gly Gln Leu Gln Gly Ile Val Ser

180 185 190

Ser Gly Tyr Gly Cys Ala Gln Lys Asn Lys Pro Gly Val Tyr Thr Lys

195 200 205

Val Cys Asn Tyr Val Asn Trp Ile Gln Gln Thr Ile Ala Ala Asn

210 215 220

<210> 82

<211> 223

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Porcine trypsin variant #82

<400> 82

Ile Val Gly Gly Tyr Thr Cys Ala Ala Asn Ser Ile Pro Tyr Gln Val

1 5 10 15

Ser Leu Asn Ser Gly Ser His Arg Cys Gly Gly Ser Leu Ile Asn Ser

20 25 30

Gln Trp Val Val Ser Ala Ala His Cys Tyr Lys Pro Arg Ile Gln Val

35 40 45

Arg Leu Gly Glu His Asn Ile His Val Leu Glu Gly Asn Glu Gln Phe

50 55 60

Ile Asn Ala Ala Lys Ile Ile Thr His Pro Asn Phe Asn Pro Asn Thr

65 70 75 80

Leu Asp Asn Asp Ile Met Leu Ile Lys Leu Ser Ser Pro Ala Thr Leu

85 90 95

Asn Ser His Val Ala Thr Val Ser Leu Pro Arg Ser Cys Ala Ala Ala

100 105 110

Gly Thr Glu Cys Leu Ile Ser Gly Trp Gly Asn Thr Cys Ser Ser Gly

115 120 125

Ser Ser Met Pro Ser Leu Leu Gln Cys Leu Lys Ala Pro Val Leu Ser

130 135 140

Asp Ser Ser Cys Lys Ser Ser Tyr Pro Gly Gln Ile Thr Gly Asn Met

145 150 155 160

Ile Cys Val Gly Phe Leu Glu Gly Gly Lys Asp Ala Cys Gln Gly Asp

165 170 175

Ser Gly Gly Pro Val Val Cys Asn Gly Gln Leu Gln Gly Ile Val Ser

180 185 190

Ser Gly Tyr Gly Cys Ala Gln Lys Asn Lys Pro Gly Val Tyr Thr Lys

195 200 205

Val Cys Asn Tyr Val Asn Trp Ile Gln Gln Thr Ile Ala Ala Asn

210 215 220

<210> 83

<211> 223

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Porcine trypsin variant #83

<400> 83

Ile Val Gly Gly Tyr Thr Cys Ala Ala Asn Ser Ile Pro Tyr Gln Val

1 5 10 15

Ser Leu Asn Ser Gly Ser His Arg Cys Gly Gly Ser Leu Ile Asn Ser

20 25 30

Gln Trp Val Val Ser Ala Ala His Cys Tyr Lys Pro Arg Ile Gln Val

35 40 45

Arg Leu Gly Glu His Asn Ile His Val Leu Glu Gly Asn Glu Gln Phe

50 55 60

Ile Asn Ala Ala Lys Ile Ile Thr His Pro Asn Phe Asn Pro Asn Thr

65 70 75 80

Leu Asp Asn Asp Ile Met Leu Ile Lys Leu Ser Ser Pro Ala Thr Leu

85 90 95

Asn Ser Asn Val Ala Thr Val Ser Leu Pro Thr Ser Cys Ala Ala Ala

100 105 110

Gly Thr Glu Cys Leu Ile Ser Gly Trp Gly Asn Thr Ser Ser Ser Gly

115 120 125

Ser Ser Met Pro Ser Leu Leu Gln Cys Leu Lys Ala Pro Val Leu Ser

130 135 140

Asp Ser Ser Cys Lys Ser Ser Tyr Pro Gly Gln Ile Thr Gly Asn Met

145 150 155 160

Ile Cys Val Gly Phe Leu Glu Gly Gly Lys Asp Ala Cys Gln Gly Asp

165 170 175

Ser Gly Gly Pro Val Val Cys Asn Gly Gln Leu Gln Gly Ile Val Ser

180 185 190

Ser Gly Tyr Gly Cys Ala Gln Lys Asn Lys Pro Gly Val Tyr Thr Lys

195 200 205

Val Cys Asn Tyr Val Asn Trp Ile Gln Gln Thr Ile Ala Ala Asn

210 215 220

<210> 84

<211> 223

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Porcine trypsin variant #84

<400> 84

Ile Val Gly Gly Tyr Thr Cys Ala Ala Asn Ser Ile Pro Tyr Gln Val

1 5 10 15

Ser Leu Asn Ser Gly Ser His Arg Cys Gly Gly Ser Leu Ile Asn Ser

20 25 30

Gln Trp Val Val Ser Ala Ala His Cys Tyr Lys Pro Arg Ile Gln Val

35 40 45

Arg Leu Gly Glu His Asn Ile His Val Leu Glu Gly Asn Glu Gln Phe

50 55 60

Ile Asn Ala Ala Lys Ile Ile Thr His Pro Asn Phe Asn Pro Asn Thr

65 70 75 80

Leu Asp Asn Asp Ile Met Leu Ile Lys Leu Ser Ser Pro Ala Thr Leu

85 90 95

Asn Ser Asn Val Ala Thr Val Ser Leu Pro Thr Ser Cys Ala Ala Ala

100 105 110

Gly Thr Glu Cys Leu Ile Ser Gly Trp Gly Asn Thr Ser Ser Ser Gly

115 120 125

Ser Ser Met Pro Ser Leu Leu Gln Cys Leu Lys Ala Pro Val Leu Ser

130 135 140

Asp Ser Ser Cys Lys Ser Ser Tyr Pro Gly Gln Ile Thr Gly Asn Met

145 150 155 160

Ile Cys Val Gly Phe Leu Glu Gly Gly Lys Asp Ser Cys Gln Gly Asp

165 170 175

Ser Gly Gly Pro Val Val Cys Asn Gly Gln Leu Gln Gly Ile Val Ser

180 185 190

Ser Gly Tyr Gly Cys Ala Gln Lys Asn Lys Pro Gly Val Tyr Thr Lys

195 200 205

Val Cys Asn Tyr Val Asn Trp Ile Gln Gln Thr Ile Ala Ala Asn

210 215 220

<210> 85

<211> 223

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Porcine trypsin variant #85

<400> 85

Ile Val Gly Gly Tyr Thr Cys Ala Ala Asn Ser Ile Pro Tyr Gln Val

1 5 10 15

Ser Leu Asn Ser Gly Ser His Arg Cys Gly Gly Ser Leu Ile Asn Ser

20 25 30

Gln Trp Val Val Ser Ala Ala His Cys Tyr Lys Pro Arg Ile Gln Val

35 40 45

Arg Leu Gly Glu His Asn Ile His Val Leu Glu Gly Asn Glu Gln Phe

50 55 60

Ile Asn Ala Ala Lys Ile Ile Thr His Pro Asn Phe Asn Pro Asn Thr

65 70 75 80

Leu Asp Asn Asp Ile Met Leu Ile Lys Leu Ser Ser Pro Ala Thr Leu

85 90 95

Asn Ser His Val Ala Thr Val Ser Leu Pro Thr Ser Cys Ala Ala Ala

100 105 110

Gly Thr Glu Cys Leu Ile Ser Gly Trp Gly Asn Thr Cys Ser Ser Gly

115 120 125

Ser Ser Met Pro Ser Leu Leu Gln Cys Leu Lys Ala Pro Val Leu Ser

130 135 140

Asp Ser Ser Cys Lys Ser Ser Tyr Pro Gly Gln Ile Thr Gly Asn Met

145 150 155 160

Ile Cys Val Gly Phe Leu Glu Gly Gly Lys Asp Ala Cys Gln Gly Asp

165 170 175

Ser Gly Gly Pro Val Val Cys Asn Gly Gln Leu Gln Gly Ile Val Ser

180 185 190

Ser Gly Tyr Gly Cys Ala Gln Lys Asn Lys Pro Gly Val Tyr Thr Lys

195 200 205

Val Cys Asn Tyr Val Asn Trp Ile Gln Gln Thr Ile Ala Ala Asn

210 215 220

<210> 86

<211> 223

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Porcine trypsin variant #86

<400> 86

Ile Val Gly Gly Tyr Thr Cys Ala Ala Asn Ser Ile Pro Tyr Gln Val

1 5 10 15

Ser Leu Asn Ser Gly Ser His Arg Cys Gly Gly Ser Leu Ile Asn Ser

20 25 30

Gln Trp Val Val Ser Ala Ala His Cys Tyr Lys Pro Arg Ile Gln Val

35 40 45

Arg Leu Gly Glu His Asn Ile Asp Val Leu Glu Gly Asn Glu Gln Phe

50 55 60

Ile Asn Ala Ala Lys Ile Ile Thr His Pro Asn Phe Asn Pro Asn Thr

65 70 75 80

Leu Asp Asn Asp Ile Met Leu Ile Lys Leu Ser Ser Pro Ala Thr Leu

85 90 95

Asn Ser Ala Val Ala Thr Val Ser Leu Pro Thr Ser Cys Ala Ala Ala

100 105 110

Gly Thr Glu Cys Leu Ile Ser Gly Trp Gly Asn Thr Ser Ser Ser Gly

115 120 125

Ser Ser Met Pro Ser Leu Leu Gln Cys Leu Lys Ala Pro Val Leu Ser

130 135 140

Asp Ser Ser Cys Lys Ser Ser Tyr Pro Gly Gln Ile Thr Gly Asn Met

145 150 155 160

Ile Cys Val Gly Phe Leu Glu Gly Gly Lys Asp Ala Cys Gln Gly Asp

165 170 175

Ser Gly Gly Pro Val Val Cys Asn Gly Gln Leu Gln Gly Ile Val Ser

180 185 190

Ser Gly Tyr Gly Cys Ala Gln Lys Asn Lys Pro Gly Val Tyr Thr Lys

195 200 205

Val Cys Asn Tyr Val Asn Trp Ile Gln Gln Thr Ile Ala Ala Asn

210 215 220

<210> 87

<211> 223

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Porcine trypsin variant #87

<400> 87

Ile Val Gly Gly Tyr Thr Cys Ala Ala Asn Ser Ile Pro Tyr Gln Val

1 5 10 15

Ser Leu Asn Ser Gly Ser His Arg Cys Gly Gly Ser Leu Ile Asn Ser

20 25 30

Gln Trp Val Val Ser Ala Ala His Cys Tyr Lys Pro Arg Ile Gln Val

35 40 45

Arg Leu Gly Glu His Asn Ile His Val Leu Glu Gly Asn Glu Gln Phe

50 55 60

Ile Asn Ala Ala Lys Ile Ile Thr His Pro Asn Phe Asn Pro Asn Thr

65 70 75 80

Leu Asp Asn Asp Ile Met Leu Ile Lys Leu Ser Ser Pro Ala Thr Leu

85 90 95

Asn Ser His Val Ala Thr Val Ser Leu Pro Thr Ser Cys Ala Ala Ala

100 105 110

Gly Thr Glu Cys Leu Ile Ser Gly Trp Gly Asn Thr Ser Ser Ser Gly

115 120 125

Ser Ser Met Pro Ser Leu Leu Gln Cys Leu Lys Ala Pro Val Leu Ser

130 135 140

Asp Ser Ser Cys Lys Ser Ser Tyr Pro Gly Gln Ile Thr Gly Asn Met

145 150 155 160

Ile Cys Val Gly Phe Leu Glu Gly Gly Lys Asp Ala Cys Gln Gly Asp

165 170 175

Ser Gly Gly Pro Val Val Cys Asn Gly Gln Leu Gln Gly Ile Val Ser

180 185 190

Ser Gly Tyr Gly Cys Ala Gln Lys Asn Lys Pro Gly Val Tyr Thr Lys

195 200 205

Val Cys Asn Tyr Val Asn Trp Ile Gln Gln Thr Ile Ala Ala Asn

210 215 220

<210> 88

<211> 223

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Porcine trypsin variant #88

<400> 88

Ile Val Gly Gly Tyr Thr Cys Ala Ala Asn Ser Ile Pro Tyr Gln Val

1 5 10 15

Ser Leu Asn Ser Gly Ser His Arg Cys Gly Gly Ser Leu Ile Asn Ser

20 25 30

Gln Trp Val Val Ser Ala Ala His Cys Tyr Lys Pro Arg Ile Gln Val

35 40 45

Arg Leu Gly Glu His Asn Ile His Val Leu Glu Gly Asn Glu Gln Phe

50 55 60

Ile Asn Ala Ala Lys Ile Ile Thr His Pro Asn Phe Asn Pro Asn Thr

65 70 75 80

Leu Asp Asn Asp Ile Met Leu Ile Lys Leu Ser Ser Pro Ala Thr Leu

85 90 95

Asn Ser His Val Ala Thr Val Ser Leu Pro Thr Ser Cys Ala Ala Ala

100 105 110

Gly Thr Glu Cys Leu Ile Ser Gly Trp Gly Asn Thr Ser Ser Ser Gly

115 120 125

Ser Ser Met Pro Ser Leu Leu Gln Cys Leu Lys Ala Pro Val Leu Ser

130 135 140

Asp Ser Ser Cys Lys Ser Ser Tyr Pro Gly Gln Ile Thr Gly Asn Met

145 150 155 160

Ile Cys Val Gly Phe Leu Glu Gly Gly Lys Asp Ser Cys Gln Gly Asp

165 170 175

Ser Gly Gly Pro Val Val Cys Asn Gly Gln Leu Gln Gly Ile Val Ser

180 185 190

Ser Gly Tyr Gly Cys Ala Gln Lys Asn Lys Pro Gly Val Tyr Thr Lys

195 200 205

Val Cys Asn Tyr Val Asn Trp Ile Gln Gln Thr Ile Ala Ala Asn

210 215 220

<210> 89

<211> 223

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Porcine trypsin variant #89

<400> 89

Ile Val Gly Gly Tyr Thr Cys Ala Ala Asn Ser Ile Pro Tyr Gln Val

1 5 10 15

Ser Leu Asn Ser Gly Ser His Arg Cys Gly Gly Ser Leu Ile Asn Ser

20 25 30

Gln Trp Val Val Ser Ala Ala His Cys Tyr Lys Pro Arg Ile Gln Val

35 40 45

Arg Leu Gly Glu His Asn Ile His Val Leu Glu Gly Asn Glu Gln Phe

50 55 60

Ile Asn Ala Ala Lys Ile Ile Thr His Pro Asn Phe Asn Pro Asn Thr

65 70 75 80

Leu Asp Asn Asp Ile Met Leu Ile Lys Leu Ser Ser Pro Ala Thr Leu

85 90 95

Asn Ser His Val Ala Thr Val Ser Leu Pro Thr Ser Cys Ala Ala Ala

100 105 110

Gly Thr Glu Cys Leu Ile Ser Gly Trp Gly Asn Thr Ala Ser Ser Gly

115 120 125

Ser Ser Met Pro Ser Leu Leu Gln Cys Leu Lys Ala Pro Val Leu Ser

130 135 140

Asp Ser Ser Cys Lys Ser Ser Tyr Pro Gly Gln Ile Thr Gly Asn Met

145 150 155 160

Ile Cys Val Gly Phe Leu Glu Gly Gly Lys Asp Ala Cys Gln Gly Asp

165 170 175

Ser Gly Gly Pro Val Val Cys Asn Gly Gln Leu Gln Gly Ile Val Ser

180 185 190

Ser Gly Tyr Gly Cys Ala Gln Lys Asn Lys Pro Gly Val Tyr Thr Lys

195 200 205

Val Cys Asn Tyr Val Asn Trp Ile Gln Gln Thr Ile Ala Ala Asn

210 215 220

<210> 90

<211> 223

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Porcine trypsin variant #90

<400> 90

Ile Val Gly Gly Tyr Thr Cys Ala Ala Asn Ser Ile Pro Tyr Gln Val

1 5 10 15

Ser Leu Asn Ser Gly Ser His Arg Cys Gly Gly Ser Leu Ile Asn Ser

20 25 30

Gln Trp Val Val Ser Ala Ala His Cys Tyr Lys Pro Arg Ile Gln Val

35 40 45

Arg Leu Gly Glu His Asn Ile His Val Leu Glu Gly Asn Glu Gln Phe

50 55 60

Ile Asn Ala Ala Lys Ile Ile Thr His Pro Asn Phe Asn Pro Asn Thr

65 70 75 80

Leu Asp Asn Asp Ile Met Leu Ile Lys Leu Ser Ser Pro Ala Thr Leu

85 90 95

Asn Ser His Val Ala Thr Val Ser Leu Pro Thr Ser Cys Ala Ala Ala

100 105 110

Gly Thr Glu Cys Leu Ile Ser Gly Trp Gly Asn Thr Ala Ser Ser Gly

115 120 125

Ser Ser Met Pro Ser Leu Leu Gln Cys Leu Lys Ala Pro Val Leu Ser

130 135 140

Asp Ser Ser Cys Lys Ser Ser Tyr Pro Gly Gln Ile Thr Gly Asn Met

145 150 155 160

Ile Cys Val Gly Phe Leu Glu Gly Gly Lys Asp Ser Cys Gln Gly Asp

165 170 175

Ser Gly Gly Pro Val Val Cys Asn Gly Gln Leu Gln Gly Ile Val Ser

180 185 190

Ser Gly Tyr Gly Cys Ala Gln Lys Asn Lys Pro Gly Val Tyr Thr Lys

195 200 205

Val Cys Asn Tyr Val Asn Trp Ile Gln Gln Thr Ile Ala Ala Asn

210 215 220

<210> 91

<211> 223

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Porcine trypsin variant #91

<400> 91

Ile Val Gly Gly Tyr Thr Cys Ala Ala Asn Ser Ile Pro Tyr Gln Val

1 5 10 15

Ser Leu Asn Ser Gly Ser His Arg Cys Gly Gly Ser Leu Ile Asn Ser

20 25 30

Gln Trp Val Val Ser Ala Ala His Cys Tyr Lys Pro Arg Ile Gln Val

35 40 45

Arg Leu Gly Glu His Asn Ile His Val Leu Glu Gly Asn Glu Gln Phe

50 55 60

Ile Asn Ala Ala Lys Ile Ile Thr His Pro Asn Phe Asn Pro Asn Thr

65 70 75 80

Leu Asp Asn Asp Ile Met Leu Ile Lys Leu Ser Ser Pro Ala Thr Leu

85 90 95

Asn Ser Ala Val Ala Thr Val Ser Leu Pro Thr Ser Cys Ala Ala Ala

100 105 110

Gly Thr Glu Cys Leu Ile Ser Gly Trp Gly Asn Thr Ser Ser Ser Gly

115 120 125

Ser Ser Met Pro Ser Leu Leu Gln Cys Leu Lys Ala Pro Val Leu Ser

130 135 140

Asp Ser Ser Cys Lys Ser Ser Tyr Pro Gly Gln Ile Thr Gly Asn Met

145 150 155 160

Ile Cys Val Gly Phe Leu Glu Gly Gly Lys Asp Ala Cys Gln Gly Asp

165 170 175

Ser Gly Gly Pro Val Val Cys Asn Gly Gln Leu Gln Gly Ile Val Ser

180 185 190

Ser Gly Tyr Gly Cys Ala Gln Lys Asn Lys Pro Gly Val Tyr Thr Lys

195 200 205

Val Cys Asn Tyr Val Asn Trp Ile Gln Gln Thr Ile Ala Ala Asn

210 215 220

<210> 92

<211> 223

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Porcine trypsin variant #92

<400> 92

Ile Val Gly Gly Tyr Thr Cys Ala Ala Asn Ser Ile Pro Tyr Gln Val

1 5 10 15

Ser Leu Asn Ser Gly Ser His Arg Cys Gly Gly Ser Leu Ile Asn Ser

20 25 30

Gln Trp Val Val Ser Ala Ala His Cys Tyr Lys Pro Arg Ile Gln Val

35 40 45

Arg Leu Gly Glu His Asn Ile His Val Leu Glu Gly Asn Glu Gln Phe

50 55 60

Ile Asn Ala Ala Lys Ile Ile Thr His Pro Asn Phe Asn Pro Asn Thr

65 70 75 80

Leu Asp Asn Asp Ile Met Leu Ile Lys Leu Ser Ser Pro Ala Thr Leu

85 90 95

Asn Ser Ala Val Ala Thr Val Ser Leu Pro Thr Ser Cys Ala Ala Ala

100 105 110

Gly Thr Glu Cys Leu Ile Ser Gly Trp Gly Asn Thr Cys Ser Ser Gly

115 120 125

Ser Ser Met Pro Ser Leu Leu Gln Cys Leu Lys Ala Pro Val Leu Ser

130 135 140

Asp Ser Ser Cys Lys Ser Ser Tyr Pro Gly Gln Ile Thr Gly Asn Met

145 150 155 160

Ile Cys Val Gly Phe Leu Glu Gly Gly Lys Asp Ala Cys Gln Gly Asp

165 170 175

Ser Gly Gly Pro Val Val Cys Asn Gly Gln Leu Gln Gly Ile Val Ser

180 185 190

Ser Gly Tyr Gly Cys Ala Gln Lys Asn Lys Pro Gly Val Tyr Thr Lys

195 200 205

Val Cys Asn Tyr Val Asn Trp Ile Gln Gln Thr Ile Ala Ala Asn

210 215 220

<210> 93

<211> 223

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Porcine trypsin variant #93

<400> 93

Ile Val Gly Gly Tyr Thr Cys Ala Ala Asn Ser Ile Pro Tyr Gln Val

1 5 10 15

Ser Leu Asn Ser Gly Ser His Arg Cys Gly Gly Ser Leu Ile Asn Ser

20 25 30

Gln Trp Val Val Ser Ala Ala His Cys Tyr Lys Ser Arg Ile Gln Val

35 40 45

Arg Leu Gly Glu His Asn Ile Asp Val Leu Glu Gly Asn Glu Gln Phe

50 55 60

Ile Asn Ala Ala Lys Ile Ile Thr His Pro Asn Phe Asn Pro Asn Thr

65 70 75 80

Leu Asp Asn Asp Ile Met Leu Ile Lys Leu Ser Ser Pro Ala Thr Leu

85 90 95

Asn Ser Asn Val Ala Thr Val Ser Leu Pro Thr Ser Cys Ala Ala Ala

100 105 110

Gly Thr Glu Cys Leu Ile Ser Gly Trp Gly Asn Thr Cys Ser Ser Gly

115 120 125

Ser Ser Met Pro Ser Leu Leu Gln Cys Leu Lys Ala Pro Val Leu Ser

130 135 140

Asp Ser Ser Cys Lys Ser Ser Tyr Pro Gly Gln Ile Thr Gly Asn Met

145 150 155 160

Ile Cys Val Gly Phe Leu Glu Gly Gly Ala Asp Ala Cys Gln Gly Asp

165 170 175

Ser Gly Gly Pro Val Val Cys Asn Gly Gln Leu Gln Gly Ile Val Ser

180 185 190

Ser Gly Tyr Gly Cys Ala Gln Lys Asn Lys Pro Gly Val Tyr Thr Lys

195 200 205

Val Cys Asn Tyr Val Asn Trp Ile Gln Gln Thr Ile Ala Ala Asn

210 215 220

<210> 94

<211> 223

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Porcine trypsin variant #94

<400> 94

Ile Val Gly Gly Tyr Thr Cys Ala Ala Asn Ser Ile Pro Tyr Gln Val

1 5 10 15

Ser Leu Asn Ser Gly Ser His Arg Cys Gly Gly Ser Leu Ile Asn Ser

20 25 30

Gln Trp Val Val Ser Ala Ala His Cys Tyr Lys Pro Arg Ile Gln Val

35 40 45

Arg Leu Gly Glu His Asn Ile His Val Leu Glu Gly Asn Glu Gln Phe

50 55 60

Ile Asn Ala Ala Lys Ile Ile Thr His Pro Asn Phe Asn Gly Asn Thr

65 70 75 80

Leu Asp Asn Asp Ile Met Leu Ile Lys Leu Ser Ser Pro Ala Thr Leu

85 90 95

Asn Ser Asn Val Ala Thr Val Ser Leu Pro Arg Ser Cys Ala Ala Ala

100 105 110

Gly Thr Glu Cys Leu Ile Ser Gly Trp Gly Asn Thr Cys Ser Ser Gly

115 120 125

Ser Ser Met Pro Ser Leu Leu Gln Cys Leu Lys Ala Pro Val Leu Ser

130 135 140

Asp Ser Ser Cys Lys Ser Ser Tyr Pro Gly Gln Ile Thr Gly Asn Met

145 150 155 160

Ile Cys Val Gly Phe Leu Glu Gly Gly Ala Asp Ala Cys Gln Gly Asp

165 170 175

Ser Gly Gly Pro Val Val Cys Asn Gly Gln Leu Gln Gly Ile Val Ser

180 185 190

Ser Gly Tyr Gly Cys Ala Gln Lys Asn Lys Pro Gly Val Tyr Thr Lys

195 200 205

Val Cys Asn Tyr Val Asn Trp Ile Gln Gln Thr Ile Ala Ala Asn

210 215 220

<210> 95

<211> 223

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Porcine trypsin variant #95

<400> 95

Ile Val Gly Gly Tyr Thr Cys Ala Ala Asn Ser Ile Pro Tyr Gln Val

1 5 10 15

Ser Leu Asn Ser Gly Ser His Arg Cys Gly Gly Ser Leu Ile Asn Ser

20 25 30

Gln Trp Val Val Ser Ala Ala His Cys Tyr Lys Pro Arg Ile Gln Val

35 40 45

Arg Leu Gly Glu His Asn Ile His Val Leu Glu Gly Asn Glu Gln Phe

50 55 60

Ile Asn Ala Ala Lys Ile Ile Thr His Pro Asn Phe Asn Pro Asn Thr

65 70 75 80

Leu Asp Asn Asp Ile Met Leu Ile Lys Leu Ser Ser Pro Ala Thr Leu

85 90 95

Asn Ser His Val Ala Thr Val Ser Leu Pro Arg Ser Cys Ala Ala Ala

100 105 110

Gly Thr Glu Cys Leu Ile Ser Gly Trp Gly Asn Thr Ser Ser Ser Gly

115 120 125

Ser Ser Met Pro Ser Leu Leu Gln Cys Leu Lys Ala Pro Val Leu Ser

130 135 140

Asp Ser Ser Cys Lys Ser Ser Tyr Pro Gly Gln Ile Thr Gly Asn Met

145 150 155 160

Ile Cys Val Gly Phe Leu Glu Gly Gly Ala Asp Ala Cys Gln Gly Asp

165 170 175

Ser Gly Gly Pro Val Val Cys Asn Gly Gln Leu Gln Gly Ile Val Ser

180 185 190

Ser Gly Tyr Gly Cys Ala Gln Lys Asn Lys Pro Gly Val Tyr Thr Lys

195 200 205

Val Cys Asn Tyr Val Asn Trp Ile Gln Gln Thr Ile Ala Ala Asn

210 215 220

<210> 96

<211> 223

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Porcine trypsin variant #96

<400> 96

Ile Val Gly Gly Tyr Thr Cys Ala Ala Asn Ser Ile Pro Tyr Gln Val

1 5 10 15

Ser Leu Asn Ser Gly Ser His Arg Cys Gly Gly Ser Leu Ile Asn Ser

20 25 30

Gln Trp Val Val Ser Ala Ala His Cys Tyr Lys Pro Arg Ile Gln Val

35 40 45

Arg Leu Gly Glu His Asn Ile Asp Val Leu Glu Gly Asn Glu Gln Phe

50 55 60

Ile Asn Ala Ala Lys Ile Ile Thr His Pro Asn Phe Asn Pro Asn Thr

65 70 75 80

Leu Asp Asn Asp Ile Met Leu Ile Lys Leu Ser Ser Pro Ala Thr Leu

85 90 95

Asn Ser His Val Ala Thr Val Ser Leu Pro Arg Ser Cys Ala Ala Ala

100 105 110

Gly Thr Glu Cys Leu Ile Ser Gly Trp Gly Asn Thr Ser Ser Ser Gly

115 120 125

Ser Ser Met Pro Ser Leu Leu Gln Cys Leu Lys Ala Pro Val Leu Ser

130 135 140

Asp Ser Ser Cys Lys Ser Ser Tyr Pro Gly Gln Ile Thr Gly Asn Met

145 150 155 160

Ile Cys Val Gly Phe Leu Glu Gly Gly Ala Asp Ala Cys Gln Gly Asp

165 170 175

Ser Gly Gly Pro Val Val Cys Asn Gly Gln Leu Gln Gly Ile Val Ser

180 185 190

Ser Gly Tyr Gly Cys Ala Gln Lys Asn Lys Pro Gly Val Tyr Thr Lys

195 200 205

Val Cys Asn Tyr Val Asn Trp Ile Gln Gln Thr Ile Ala Ala Asn

210 215 220

<210> 97

<211> 223

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Porcine trypsin variant #97

<400> 97

Ile Val Gly Gly Tyr Thr Cys Ala Ala Asn Ser Ile Pro Tyr Gln Val

1 5 10 15

Ser Leu Asn Ser Gly Ser His Arg Cys Gly Gly Ser Leu Ile Asn Ser

20 25 30

Gln Trp Val Val Ser Ala Ala His Cys Tyr Lys Pro Arg Ile Gln Val

35 40 45

Arg Leu Gly Glu His Asn Ile His Val Leu Glu Gly Asn Glu Gln Phe

50 55 60

Ile Asn Ala Ala Lys Ile Ile Thr His Pro Asn Phe Asn Pro Asn Thr

65 70 75 80

Leu Asp Asn Asp Ile Met Leu Ile Lys Leu Ser Ser Pro Ala Thr Leu

85 90 95

Asn Ser Ala Val Ala Thr Val Ser Leu Pro Thr Ser Cys Ala Ala Ala

100 105 110

Gly Thr Glu Cys Leu Ile Ser Gly Trp Gly Asn Thr Ala Ser Ser Gly

115 120 125

Ser Ser Met Pro Ser Leu Leu Gln Cys Leu Lys Ala Pro Val Leu Ser

130 135 140

Asp Ser Ser Cys Lys Ser Ser Tyr Pro Gly Gln Ile Thr Gly Asn Met

145 150 155 160

Ile Cys Val Gly Phe Leu Glu Gly Gly Ala Asp Ala Cys Gln Gly Asp

165 170 175

Ser Gly Gly Pro Val Val Cys Asn Gly Gln Leu Gln Gly Ile Val Ser

180 185 190

Ser Gly Tyr Gly Cys Ala Gln Lys Asn Lys Pro Gly Val Tyr Thr Lys

195 200 205

Val Cys Asn Tyr Val Asn Trp Ile Gln Gln Thr Ile Ala Ala Asn

210 215 220

<210> 98

<211> 223

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Porcine trypsin variant #98

<400> 98

Ile Val Gly Gly Tyr Thr Cys Ala Ala Asn Ser Ile Pro Tyr Gln Val

1 5 10 15

Ser Leu Asn Ser Gly Ser His Arg Cys Gly Gly Ser Leu Ile Asn Ser

20 25 30

Gln Trp Val Val Ser Ala Ala His Cys Tyr Lys Pro Arg Ile Gln Val

35 40 45

Arg Leu Gly Glu His Asn Ile His Val Leu Glu Gly Asn Glu Gln Phe

50 55 60

Ile Asn Ala Ala Lys Ile Ile Thr His Pro Asn Phe Asn Pro Asn Thr

65 70 75 80

Leu Asp Asn Asp Ile Met Leu Ile Lys Leu Ser Ser Pro Ala Thr Leu

85 90 95

Asn Ser Ala Val Ala Thr Val Ser Leu Pro Thr Ser Cys Ala Ala Ala

100 105 110

Gly Thr Glu Cys Leu Ile Ser Gly Trp Gly Asn Thr Ala Ser Ser Gly

115 120 125

Ser Ser Met Pro Ser Leu Leu Gln Cys Leu Lys Ala Pro Val Leu Ser

130 135 140

Asp Ser Ser Cys Lys Ser Ser Tyr Pro Gly Gln Ile Thr Gly Asn Met

145 150 155 160

Ile Cys Val Gly Phe Leu Glu Gly Gly Ala Asp Ser Cys Gln Gly Asp

165 170 175

Ser Gly Gly Pro Val Val Cys Asn Gly Gln Leu Gln Gly Ile Val Ser

180 185 190

Ser Gly Tyr Gly Cys Ala Gln Lys Asn Lys Pro Gly Val Tyr Thr Lys

195 200 205

Val Cys Asn Tyr Val Asn Trp Ile Gln Gln Thr Ile Ala Ala Asn

210 215 220

<210> 99

<211> 223

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Porcine trypsin variant #99

<400> 99

Ile Val Gly Gly Tyr Thr Cys Ala Ala Asn Ser Ile Pro Tyr Gln Val

1 5 10 15

Ser Leu Asn Ser Gly Ser His Arg Cys Gly Gly Ser Leu Ile Asn Ser

20 25 30

Gln Trp Val Val Ser Ala Ala His Cys Tyr Lys Ser Arg Ile Gln Val

35 40 45

Arg Leu Gly Glu His Asn Ile His Val Leu Glu Gly Asn Glu Gln Phe

50 55 60

Ile Asn Ala Ala Lys Ile Ile Thr His Pro Asn Phe Asn Pro Asn Thr

65 70 75 80

Leu Asp Asn Asp Ile Met Leu Ile Lys Leu Ser Ser Pro Ala Thr Leu

85 90 95

Asn Ser Asn Val Ala Thr Val Ser Leu Pro Thr Ser Cys Ala Ala Ala

100 105 110

Gly Thr Glu Cys Leu Ile Ser Gly Trp Gly Asn Thr Ser Ser Ser Gly

115 120 125

Ser Ser Tyr Pro Ser Leu Leu Gln Cys Leu Lys Ala Pro Val Leu Ser

130 135 140

Asp Ser Ser Cys Lys Ser Ser Tyr Pro Gly Gln Ile Thr Gly Asn Met

145 150 155 160

Ile Cys Val Gly Phe Leu Glu Gly Gly Ala Asp Ala Cys Gln Gly Asp

165 170 175

Ser Gly Gly Pro Val Val Cys Asn Gly Gln Leu Gln Gly Ile Val Ser

180 185 190

Ser Gly Tyr Gly Cys Ala Gln Lys Asn Lys Pro Gly Val Tyr Thr Lys

195 200 205

Val Cys Asn Tyr Val Asn Trp Ile Gln Gln Thr Ile Ala Ala Asn

210 215 220

<210> 100

<211> 223

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Porcine trypsin variant #100

<400> 100

Ile Val Gly Gly Tyr Thr Cys Ala Ala Asn Ser Ile Pro Tyr Gln Val

1 5 10 15

Ser Leu Asn Ser Gly Ser His Arg Cys Gly Gly Ser Leu Ile Asn Ser

20 25 30

Gln Trp Val Val Ser Ala Ala His Cys Tyr Lys Pro Arg Ile Gln Val

35 40 45

Arg Leu Gly Glu His Asn Ile Asp Val Leu Glu Gly Asn Glu Gln Phe

50 55 60

Ile Asn Ala Ala Lys Ile Ile Thr His Pro Asn Phe Asn Pro Asn Thr

65 70 75 80

Leu Asp Asn Asp Ile Met Leu Ile Lys Leu Ser Ser Pro Ala Thr Leu

85 90 95

Asn Ser His Val Ala Thr Val Ser Leu Pro Thr Ser Cys Ala Ala Ala

100 105 110

Gly Thr Glu Cys Leu Ile Ser Gly Trp Gly Asn Thr Cys Ser Ser Gly

115 120 125

Ser Ser Met Pro Ser Leu Leu Gln Cys Leu Lys Ala Pro Val Leu Ser

130 135 140

Asp Ser Ser Cys Lys Ser Ser Tyr Pro Gly Gln Ile Thr Gly Asn Met

145 150 155 160

Ile Cys Val Gly Phe Leu Glu Gly Gly Ala Asp Ala Cys Gln Gly Asp

165 170 175

Ser Gly Gly Pro Val Val Cys Asn Gly Gln Leu Gln Gly Ile Val Ser

180 185 190

Ser Gly Tyr Gly Cys Ala Gln Lys Asn Lys Pro Gly Val Tyr Thr Lys

195 200 205

Val Cys Asn Tyr Val Asn Trp Ile Gln Gln Thr Ile Ala Ala Asn

210 215 220

<210> 101

<211> 223

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Porcine trypsin variant #101

<400> 101

Ile Val Gly Gly Tyr Thr Cys Ala Ala Asn Ser Ile Pro Tyr Gln Val

1 5 10 15

Ser Leu Asn Ser Gly Ser His Arg Cys Gly Gly Ser Leu Ile Asn Ser

20 25 30

Gln Trp Val Val Ser Ala Ala His Cys Tyr Lys Ser Arg Ile Gln Val

35 40 45

Arg Leu Gly Glu His Asn Ile His Val Leu Glu Gly Asn Glu Gln Phe

50 55 60

Ile Asn Ala Ala Lys Ile Ile Thr His Pro Asn Phe Asn Pro Asn Thr

65 70 75 80

Leu Asp Asn Asp Ile Met Leu Ile Lys Leu Ser Ser Pro Ala Thr Leu

85 90 95

Asn Ser Asn Val Ala Thr Val Ser Leu Pro Thr Ser Cys Ala Ala Ala

100 105 110

Gly Thr Glu Cys Leu Ile Ser Gly Trp Gly Asn Thr Ser Ser Ser Gly

115 120 125

Ser Ser Met Pro Ser Leu Leu Gln Cys Leu Lys Ala Pro Val Leu Ser

130 135 140

Asp Ser Ser Cys Lys Ser Ser Tyr Pro Gly Gln Ile Thr Gly Asn Met

145 150 155 160

Ile Cys Val Gly Phe Leu Glu Gly Gly Ala Asp Ala Cys Gln Gly Asp

165 170 175

Ser Gly Gly Pro Val Val Cys Asn Gly Gln Leu Gln Gly Ile Val Ser

180 185 190

Ser Gly Tyr Gly Cys Ala Gln Lys Asn Lys Pro Gly Val Tyr Thr Lys

195 200 205

Val Cys Asn Tyr Val Asn Trp Ile Gln Gln Thr Ile Ala Ala Asn

210 215 220

<210> 102

<211> 223

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Porcine trypsin variant #102

<400> 102

Ile Val Gly Gly Tyr Thr Cys Ala Ala Asn Ser Ile Pro Tyr Gln Val

1 5 10 15

Ser Leu Asn Ser Gly Ser His Arg Cys Gly Gly Ser Leu Ile Asn Ser

20 25 30

Gln Trp Val Val Ser Ala Ala His Cys Tyr Lys Pro Arg Ile Gln Val

35 40 45

Arg Leu Gly Glu His Asn Ile His Val Leu Glu Gly Asn Glu Gln Phe

50 55 60

Ile Asn Ala Ala Lys Ile Ile Thr His Pro Asn Phe Asn Pro Asn Thr

65 70 75 80

Leu Asp Asn Asp Ile Met Leu Ile Lys Leu Ser Ser Pro Ala Thr Leu

85 90 95

Asn Ser His Val Ala Thr Val Ser Leu Pro Thr Ser Cys Ala Ala Ala

100 105 110

Gly Thr Glu Cys Leu Ile Ser Gly Trp Gly Asn Thr Ser Ser Ser Gly

115 120 125

Ser Ser Met Pro Ser Leu Leu Gln Cys Leu Lys Ala Pro Val Leu Ser

130 135 140

Asp Ser Ser Cys Lys Ser Ser Tyr Pro Gly Gln Ile Thr Gly Asn Met

145 150 155 160

Ile Cys Val Gly Phe Leu Glu Gly Gly Ala Asp Ala Cys Gln Gly Asp

165 170 175

Ser Gly Gly Pro Val Val Cys Asn Gly Gln Leu Gln Gly Ile Val Ser

180 185 190

Ser Gly Tyr Gly Cys Ala Gln Lys Asn Lys Pro Gly Val Tyr Thr Lys

195 200 205

Val Cys Asn Tyr Val Asn Trp Ile Gln Gln Thr Ile Ala Ala Asn

210 215 220

<210> 103

<211> 223

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Porcine trypsin variant #103

<400> 103

Ile Val Gly Gly Tyr Thr Cys Ala Ala Asn Ser Ile Pro Tyr Gln Val

1 5 10 15

Ser Leu Asn Ser Gly Ser His Arg Cys Gly Gly Ser Leu Ile Asn Ser

20 25 30

Gln Trp Val Val Ser Ala Ala His Cys Tyr Lys Pro Arg Ile Gln Val

35 40 45

Arg Leu Gly Glu His Asn Ile His Val Leu Glu Gly Asn Glu Gln Phe

50 55 60

Ile Asn Ala Ala Lys Ile Ile Thr His Pro Asn Phe Asn Pro Asn Thr

65 70 75 80

Leu Asp Asn Asp Ile Met Leu Ile Lys Leu Ser Ser Pro Ala Thr Leu

85 90 95

Asn Ser His Val Ala Thr Val Ser Leu Pro Thr Ser Cys Ala Ala Ala

100 105 110

Gly Thr Glu Cys Leu Ile Ser Gly Trp Gly Asn Thr Ser Ser Ser Gly

115 120 125

Ser Ser Met Pro Ser Leu Leu Gln Cys Leu Lys Ala Pro Val Leu Ser

130 135 140

Asp Ser Ser Cys Lys Ser Ser Tyr Pro Gly Gln Ile Thr Gly Asn Met

145 150 155 160

Ile Cys Val Gly Phe Leu Glu Gly Gly Ala Asp Ser Cys Gln Gly Asp

165 170 175

Ser Gly Gly Pro Val Val Cys Asn Gly Gln Leu Gln Gly Ile Val Ser

180 185 190

Ser Gly Tyr Gly Cys Ala Gln Lys Asn Lys Pro Gly Val Tyr Thr Lys

195 200 205

Val Cys Asn Tyr Val Asn Trp Ile Gln Gln Thr Ile Ala Ala Asn

210 215 220

<210> 104

<211> 223

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Porcine trypsin variant #104

<400> 104

Ile Val Gly Gly Tyr Thr Cys Ala Ala Asn Ser Ile Pro Tyr Gln Val

1 5 10 15

Ser Leu Asn Ser Gly Ser His Arg Cys Gly Gly Ser Leu Ile Asn Ser

20 25 30

Gln Trp Val Val Ser Ala Ala His Cys Tyr Lys Ser Arg Ile Gln Val

35 40 45

Arg Leu Gly Glu His Asn Ile His Val Leu Glu Gly Asn Glu Gln Phe

50 55 60

Ile Asn Ala Ala Lys Ile Ile Thr His Pro Asn Phe Asn Pro Asn Thr

65 70 75 80

Leu Asp Asn Asp Ile Met Leu Ile Lys Leu Ser Ser Pro Ala Thr Leu

85 90 95

Asn Ser Asn Val Ala Thr Val Ser Leu Pro Thr Ser Cys Ala Ala Ala

100 105 110

Gly Thr Glu Cys Leu Ile Ser Gly Trp Gly Asn Thr Cys Ser Ser Gly

115 120 125

Ser Ser Met Pro Ser Leu Leu Gln Cys Leu Lys Ala Pro Val Leu Ser

130 135 140

Asp Ser Ser Cys Lys Ser Ser Tyr Pro Gly Gln Ile Thr Gly Asn Met

145 150 155 160

Ile Cys Val Gly Phe Leu Glu Gly Gly Ala Asp Ala Cys Gln Gly Asp

165 170 175

Ser Gly Gly Pro Val Val Cys Asn Gly Gln Leu Gln Gly Ile Val Ser

180 185 190

Ser Gly Tyr Gly Cys Ala Gln Lys Asn Lys Pro Gly Val Tyr Thr Lys

195 200 205

Val Cys Asn Tyr Val Asn Trp Ile Gln Gln Thr Ile Ala Ala Asn

210 215 220

<210> 105

<211> 223

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Porcine trypsin variant #105

<400> 105

Ile Val Gly Gly Tyr Thr Cys Ala Ala Asn Ser Ile Pro Tyr Gln Val

1 5 10 15

Ser Leu Asn Ser Gly Ser His Arg Cys Gly Gly Ser Leu Ile Asn Ser

20 25 30

Gln Trp Val Val Ser Ala Ala His Cys Tyr Lys Ser Arg Ile Gln Val

35 40 45

Arg Leu Gly Glu His Asn Ile His Val Leu Glu Gly Asn Glu Gln Phe

50 55 60

Ile Asn Ala Ala Lys Ile Ile Thr His Pro Asn Phe Asn Pro Asn Thr

65 70 75 80

Leu Asp Asn Asp Ile Met Leu Ile Lys Leu Ser Ser Pro Ala Thr Leu

85 90 95

Asn Ser Asn Val Ala Thr Val Ser Leu Pro Thr Ser Cys Ala Ala Ala

100 105 110

Gly Thr Glu Cys Leu Ile Ser Gly Trp Gly Asn Thr Ala Ser Ser Gly

115 120 125

Ser Ser Tyr Pro Ser Leu Leu Gln Cys Leu Lys Ala Pro Val Leu Ser

130 135 140

Asp Ser Ser Cys Lys Ser Ser Tyr Pro Gly Gln Ile Thr Gly Asn Met

145 150 155 160

Ile Cys Val Gly Phe Leu Glu Gly Gly Ala Asp Ala Cys Gln Gly Asp

165 170 175

Ser Gly Gly Pro Val Val Cys Asn Gly Gln Leu Gln Gly Ile Val Ser

180 185 190

Ser Gly Tyr Gly Cys Ala Gln Lys Asn Lys Pro Gly Val Tyr Thr Lys

195 200 205

Val Cys Asn Tyr Val Asn Trp Ile Gln Gln Thr Ile Ala Ala Asn

210 215 220

<210> 106

<211> 223

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Porcine trypsin variant #106

<400> 106

Ile Val Gly Gly Tyr Thr Cys Ala Ala Asn Ser Ile Pro Tyr Gln Val

1 5 10 15

Ser Leu Asn Ser Gly Ser His Arg Cys Gly Gly Ser Leu Ile Asn Ser

20 25 30

Gln Trp Val Val Ser Ala Ala His Cys Tyr Lys Ser Arg Ile Gln Val

35 40 45

Arg Leu Gly Glu His Asn Ile His Val Leu Glu Gly Asn Glu Gln Phe

50 55 60

Ile Asn Ala Ala Lys Ile Ile Thr His Pro Asn Phe Asn Pro Asn Thr

65 70 75 80

Leu Asp Asn Asp Ile Met Leu Ile Lys Leu Ser Ser Pro Ala Thr Leu

85 90 95

Asn Ser Ala Val Ala Thr Val Ser Leu Pro Thr Ser Cys Ala Ala Ala

100 105 110

Gly Thr Glu Cys Leu Ile Ser Gly Trp Gly Asn Thr Cys Ser Ser Gly

115 120 125

Ser Ser Met Pro Ser Leu Leu Gln Cys Leu Lys Ala Pro Val Leu Ser

130 135 140

Asp Ser Ser Cys Lys Ser Ser Tyr Pro Gly Gln Ile Thr Gly Asn Met

145 150 155 160

Ile Cys Val Gly Phe Leu Glu Gly Gly Ala Asp Ala Cys Gln Gly Asp

165 170 175

Ser Gly Gly Pro Val Val Cys Asn Gly Gln Leu Gln Gly Ile Val Ser

180 185 190

Ser Gly Tyr Gly Cys Ala Gln Lys Asn Lys Pro Gly Val Tyr Thr Lys

195 200 205

Val Cys Asn Tyr Val Asn Trp Ile Gln Gln Thr Ile Ala Ala Asn

210 215 220

<210> 107

<211> 223

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Porcine trypsin variant #107

<400> 107

Ile Val Gly Gly Tyr Thr Cys Ala Ala Asn Ser Ile Pro Tyr Gln Val

1 5 10 15

Ser Leu Asn Ser Gly Ser His Arg Cys Gly Gly Ser Leu Ile Asn Ser

20 25 30

Gln Trp Val Val Ser Ala Ala His Cys Tyr Lys Pro Arg Ile Gln Val

35 40 45

Arg Leu Gly Glu His Asn Ile His Val Leu Glu Gly Asn Glu Gln Phe

50 55 60

Ile Asn Ala Ala Lys Ile Ile Thr His Pro Asn Phe Asn Pro Asn Thr

65 70 75 80

Leu Asp Asn Asp Ile Met Leu Ile Lys Leu Ser Ser Pro Ala Thr Leu

85 90 95

Asn Ser His Val Ala Thr Val Ser Leu Pro Thr Ser Cys Ala Ala Ala

100 105 110

Gly Thr Glu Cys Leu Ile Ser Gly Trp Gly Asn Thr Cys Ser Ser Gly

115 120 125

Ser Ser Met Pro Ser Leu Leu Gln Cys Leu Lys Ala Pro Val Leu Ser

130 135 140

Asp Ser Ser Cys Lys Ser Ser Tyr Pro Gly Gln Ile Thr Gly Asn Met

145 150 155 160

Ile Cys Val Gly Phe Leu Glu Gly Gly Ala Asp Ala Cys Gln Gly Asp

165 170 175

Ser Gly Gly Pro Val Val Cys Asn Gly Gln Leu Gln Gly Ile Val Ser

180 185 190

Ser Gly Tyr Gly Cys Ala Gln Lys Asn Lys Pro Gly Val Tyr Thr Lys

195 200 205

Val Cys Asn Tyr Val Asn Trp Ile Gln Gln Thr Ile Ala Ala Asn

210 215 220

<210> 108

<211> 223

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Porcine trypsin variant #108

<400> 108

Ile Val Gly Gly Tyr Thr Cys Ala Ala Asn Ser Ile Pro Tyr Gln Val

1 5 10 15

Ser Leu Asn Ser Gly Ser His Arg Cys Gly Gly Ser Leu Ile Asn Ser

20 25 30

Gln Trp Val Val Ser Ala Ala His Cys Tyr Lys Pro Arg Ile Gln Val

35 40 45

Arg Leu Gly Glu His Asn Ile Asp Val Leu Glu Gly Asn Glu Gln Phe

50 55 60

Ile Asn Ala Ala Lys Ile Ile Thr His Pro Asn Phe Asn Pro Asn Thr

65 70 75 80

Leu Asp Asn Asp Ile Met Leu Ile Lys Leu Ser Ser Pro Ala Thr Leu

85 90 95

Asn Ser Ala Val Ala Thr Val Ser Leu Pro Thr Ser Cys Ala Ala Ala

100 105 110

Gly Thr Glu Cys Leu Ile Ser Gly Trp Gly Asn Thr Cys Ser Ser Gly

115 120 125

Ser Ser Met Pro Ser Leu Leu Gln Cys Leu Lys Ala Pro Val Leu Ser

130 135 140

Asp Ser Ser Cys Lys Ser Ser Tyr Pro Gly Gln Ile Thr Gly Asn Met

145 150 155 160

Ile Cys Val Gly Phe Leu Glu Gly Gly Ala Asp Ala Cys Gln Gly Asp

165 170 175

Ser Gly Gly Pro Val Val Cys Asn Gly Gln Leu Gln Gly Ile Val Ser

180 185 190

Ser Gly Tyr Gly Cys Ala Gln Lys Asn Lys Pro Gly Val Tyr Thr Lys

195 200 205

Val Cys Asn Tyr Val Asn Trp Ile Gln Gln Thr Ile Ala Ala Asn

210 215 220

<210> 109

<211> 223

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Porcine trypsin variant #109

<400> 109

Ile Val Gly Gly Tyr Thr Cys Ala Ala Asn Ser Ile Pro Tyr Gln Val

1 5 10 15

Ser Leu Asn Ser Gly Ser His Arg Cys Gly Gly Ser Leu Ile Asn Ser

20 25 30

Gln Trp Val Val Ser Ala Ala His Cys Tyr Lys Ser Arg Ile Gln Val

35 40 45

Arg Leu Gly Glu His Asn Ile His Val Leu Glu Gly Asn Glu Gln Phe

50 55 60

Ile Asn Ala Ala Lys Ile Ile Thr His Pro Asn Phe Asn Gly Asn Thr

65 70 75 80

Leu Asp Asn Asp Ile Met Leu Ile Lys Leu Ser Ser Pro Ala Thr Leu

85 90 95

Asn Ser Asn Val Ala Thr Val Ser Leu Pro Thr Ser Cys Ala Ala Ala

100 105 110

Gly Thr Glu Cys Leu Ile Ser Gly Trp Gly Asn Thr Cys Ser Ser Gly

115 120 125

Ser Ser Met Pro Ser Leu Leu Gln Cys Leu Lys Ala Pro Val Leu Ser

130 135 140

Asp Ser Ser Cys Lys Ser Ser Tyr Pro Gly Gln Ile Thr Gly Asn Met

145 150 155 160

Ile Cys Val Gly Phe Leu Glu Gly Gly Ala Asp Ala Cys Gln Gly Asp

165 170 175

Ser Gly Gly Pro Val Val Cys Asn Gly Gln Leu Gln Gly Ile Val Ser

180 185 190

Ser Gly Tyr Gly Cys Ala Gln Lys Asn Lys Pro Gly Val Tyr Thr Lys

195 200 205

Val Cys Asn Tyr Val Asn Trp Ile Gln Gln Thr Ile Ala Ala Asn

210 215 220

<210> 110

<211> 223

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Porcine trypsin variant #110

<400> 110

Ile Val Gly Gly Tyr Thr Cys Ala Ala Asn Ser Ile Pro Tyr Gln Val

1 5 10 15

Ser Leu Asn Ser Gly Ser His Arg Cys Gly Gly Ser Leu Ile Asn Ser

20 25 30

Gln Trp Val Val Ser Ala Ala His Cys Tyr Lys Ser Arg Ile Gln Val

35 40 45

Arg Leu Gly Glu His Asn Ile His Val Leu Glu Gly Asn Glu Gln Phe

50 55 60

Ile Asn Ala Ala Lys Ile Ile Thr His Pro Asn Phe Asn Pro Asn Thr

65 70 75 80

Leu Asp Asn Asp Ile Met Leu Ile Lys Leu Ser Ser Pro Ala Thr Leu

85 90 95

Asn Ser His Val Ala Thr Val Ser Leu Pro Arg Ser Cys Ala Ala Ala

100 105 110

Gly Thr Glu Cys Leu Ile Ser Gly Trp Gly Asn Thr Cys Ser Ser Gly

115 120 125

Ser Ser Met Pro Ser Leu Leu Gln Cys Leu Lys Ala Pro Val Leu Ser

130 135 140

Asp Ser Ser Cys Lys Ser Ser Tyr Pro Gly Gln Ile Thr Gly Asn Met

145 150 155 160

Ile Cys Val Gly Phe Leu Glu Gly Gly Ala Asp Ala Cys Gln Gly Asp

165 170 175

Ser Gly Gly Pro Val Val Cys Asn Gly Gln Leu Gln Gly Ile Val Ser

180 185 190

Ser Gly Tyr Gly Cys Ala Gln Lys Asn Lys Pro Gly Val Tyr Thr Lys

195 200 205

Val Cys Asn Tyr Val Asn Trp Ile Gln Gln Thr Ile Ala Ala Asn

210 215 220

<210> 111

<211> 223

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Porcine trypsin variant #111

<400> 111

Ile Val Gly Gly Tyr Thr Cys Ala Ala Asn Ser Ile Pro Tyr Gln Val

1 5 10 15

Ser Leu Asn Ser Gly Ser His Arg Cys Gly Gly Ser Leu Ile Asn Ser

20 25 30

Gln Trp Val Val Ser Ala Ala His Cys Tyr Lys Pro Arg Ile Gln Val

35 40 45

Arg Leu Gly Glu His Asn Ile His Val Leu Glu Gly Asn Glu Gln Phe

50 55 60

Ile Asn Ala Ala Lys Ile Ile Thr His Pro Asn Phe Asn Pro Asn Thr

65 70 75 80

Leu Asp Asn Asp Ile Met Leu Ile Lys Leu Ser Ser Pro Ala Thr Leu

85 90 95

Asn Ser His Val Ala Thr Val Ser Leu Pro Thr Ser Cys Ala Ala Ala

100 105 110

Gly Thr Glu Cys Leu Ile Ser Gly Trp Gly Asn Thr Ser Ser Ser Gly

115 120 125

Ser Ser Tyr Pro Ser Leu Leu Gln Cys Leu Lys Ala Pro Val Leu Ser

130 135 140

Asp Ser Ser Cys Lys Ser Ser Tyr Pro Gly Gln Ile Thr Gly Asn Met

145 150 155 160

Ile Cys Val Gly Phe Leu Glu Gly Gly Ala Asp Ala Cys Gln Gly Asp

165 170 175

Ser Gly Gly Pro Val Val Cys Asn Gly Gln Leu Gln Gly Ile Val Ser

180 185 190

Ser Gly Tyr Gly Cys Ala Gln Lys Asn Lys Pro Gly Val Tyr Thr Lys

195 200 205

Val Cys Asn Tyr Val Asn Trp Ile Gln Gln Thr Ile Ala Ala Asn

210 215 220

<210> 112

<211> 223

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Porcine trypsin variant #112

<400> 112

Ile Val Gly Gly Tyr Thr Cys Ala Ala Asn Ser Ile Pro Tyr Gln Val

1 5 10 15

Ser Leu Asn Ser Gly Ser His Arg Cys Gly Gly Ser Leu Ile Asn Ser

20 25 30

Gln Trp Val Val Ser Ala Ala His Cys Tyr Lys Pro Arg Ile Gln Val

35 40 45

Arg Leu Gly Glu His Asn Ile His Val Leu Glu Gly Asn Glu Gln Phe

50 55 60

Ile Asn Ala Ala Lys Ile Ile Thr His Pro Asn Phe Asn Pro Asn Thr

65 70 75 80

Leu Asp Asn Asp Ile Met Leu Ile Lys Leu Ser Ser Pro Ala Thr Leu

85 90 95

Asn Ser His Val Ala Thr Val Ser Leu Pro Thr Ser Cys Ala Ala Ala

100 105 110

Gly Thr Glu Cys Leu Ile Ser Gly Trp Gly Asn Thr Ser Ser Ser Gly

115 120 125

Ser Ser Tyr Pro Ser Leu Leu Gln Cys Leu Lys Ala Pro Val Leu Ser

130 135 140

Asp Ser Ser Cys Lys Ser Ser Tyr Pro Gly Gln Ile Thr Gly Asn Met

145 150 155 160

Ile Cys Val Gly Phe Leu Glu Gly Gly Ala Asp Ser Cys Gln Gly Asp

165 170 175

Ser Gly Gly Pro Val Val Cys Asn Gly Gln Leu Gln Gly Ile Val Ser

180 185 190

Ser Gly Tyr Gly Cys Ala Gln Lys Asn Lys Pro Gly Val Tyr Thr Lys

195 200 205

Val Cys Asn Tyr Val Asn Trp Ile Gln Gln Thr Ile Ala Ala Asn

210 215 220

<210> 113

<211> 223

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Porcine trypsin variant #113

<400> 113

Ile Val Gly Gly Tyr Thr Cys Ala Ala Asn Ser Ile Pro Tyr Gln Val

1 5 10 15

Ser Leu Asn Ser Gly Ser His Arg Cys Gly Gly Ser Leu Ile Asn Ser

20 25 30

Gln Trp Val Val Ser Ala Ala His Cys Tyr Lys Pro Arg Ile Gln Val

35 40 45

Arg Leu Gly Glu His Asn Ile His Val Leu Glu Gly Asn Glu Gln Phe

50 55 60

Ile Asn Ala Ala Lys Ile Ile Thr His Pro Asn Phe Asn Gly Asn Thr

65 70 75 80

Leu Asp Asn Asp Ile Met Leu Ile Lys Leu Ser Ser Pro Ala Thr Leu

85 90 95

Asn Ser Ala Val Ala Thr Val Ser Leu Pro Thr Ser Cys Ala Ala Ala

100 105 110

Gly Thr Glu Cys Leu Ile Ser Gly Trp Gly Asn Thr Ala Ser Ser Gly

115 120 125

Ser Ser Tyr Pro Ser Leu Leu Gln Cys Leu Lys Ala Pro Val Leu Ser

130 135 140

Asp Ser Ser Cys Lys Ser Ser Tyr Pro Gly Gln Ile Thr Gly Asn Met

145 150 155 160

Ile Cys Val Gly Phe Leu Glu Gly Gly Ala Asp Ala Cys Gln Gly Asp

165 170 175

Ser Gly Gly Pro Val Val Cys Asn Gly Gln Leu Gln Gly Ile Val Ser

180 185 190

Ser Gly Tyr Gly Cys Ala Gln Lys Asn Lys Pro Gly Val Tyr Thr Lys

195 200 205

Val Cys Asn Tyr Val Asn Trp Ile Gln Gln Thr Ile Ala Ala Asn

210 215 220

<210> 114

<211> 223

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Porcine trypsin variant #114

<400> 114

Ile Val Gly Gly Tyr Thr Cys Ala Ala Asn Ser Ile Pro Tyr Gln Val

1 5 10 15

Ser Leu Asn Ser Gly Ser His Arg Cys Gly Gly Ser Leu Ile Asn Ser

20 25 30

Gln Trp Val Val Ser Ala Ala His Cys Tyr Lys Pro Arg Ile Gln Val

35 40 45

Arg Leu Gly Glu His Asn Ile His Val Leu Glu Gly Asn Glu Gln Phe

50 55 60

Ile Asn Ala Ala Lys Ile Ile Thr His Pro Asn Phe Asn Gly Asn Thr

65 70 75 80

Leu Asp Asn Asp Ile Met Leu Ile Lys Leu Ser Ser Pro Ala Thr Leu

85 90 95

Asn Ser Ala Val Ala Thr Val Ser Leu Pro Thr Ser Cys Ala Ala Ala

100 105 110

Gly Thr Glu Cys Leu Ile Ser Gly Trp Gly Asn Thr Ala Ser Ser Gly

115 120 125

Ser Ser Tyr Pro Ser Leu Leu Gln Cys Leu Lys Ala Pro Val Leu Ser

130 135 140

Asp Ser Ser Cys Lys Ser Ser Tyr Pro Gly Gln Ile Thr Gly Asn Met

145 150 155 160

Ile Cys Val Gly Phe Leu Glu Gly Gly Ala Asp Ser Cys Gln Gly Asp

165 170 175

Ser Gly Gly Pro Val Val Cys Asn Gly Gln Leu Gln Gly Ile Val Ser

180 185 190

Ser Gly Tyr Gly Cys Ala Gln Lys Asn Lys Pro Gly Val Tyr Thr Lys

195 200 205

Val Cys Asn Tyr Val Asn Trp Ile Gln Gln Thr Ile Ala Ala Asn

210 215 220

<210> 115

<211> 223

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Porcine trypsin variant #115

<400> 115

Ile Val Gly Gly Tyr Thr Cys Ala Ala Asn Ser Ile Pro Tyr Gln Val

1 5 10 15

Ser Leu Asn Ser Gly Ser His Arg Cys Gly Gly Ser Leu Ile Asn Ser

20 25 30

Gln Trp Val Val Ser Ala Ala His Cys Tyr Lys Pro Arg Ile Gln Val

35 40 45

Arg Leu Gly Glu His Asn Ile His Val Leu Glu Gly Asn Glu Gln Phe

50 55 60

Ile Asn Ala Ala Lys Ile Ile Thr His Pro Asn Phe Asn Gly Asn Thr

65 70 75 80

Leu Asp Asn Asp Ile Met Leu Ile Lys Leu Ser Ser Pro Ala Thr Leu

85 90 95

Asn Ser Asn Val Ala Thr Val Ser Leu Pro Thr Ser Cys Ala Ala Ala

100 105 110

Gly Thr Glu Cys Leu Ile Ser Gly Trp Gly Asn Thr Ser Ser Ser Gly

115 120 125

Ser Ser Met Pro Ser Leu Leu Gln Cys Leu Lys Ala Pro Val Leu Ser

130 135 140

Asp Ser Ser Cys Lys Ser Ser Tyr Pro Gly Gln Ile Thr Gly Asn Met

145 150 155 160

Ile Cys Val Gly Phe Leu Glu Gly Gly Lys Asp Ala Cys Gln Gly Asp

165 170 175

Ser Gly Gly Pro Val Val Cys Asn Gly Gln Leu Gln Gly Ile Val Ser

180 185 190

Ser Gly Tyr Gly Cys Ala Gln Lys Asn Lys Pro Gly Val Tyr Thr Lys

195 200 205

Val Cys Asn Tyr Val Asn Trp Ile Gln Gln Thr Ile Ala Ala Asn

210 215 220

<210> 116

<211> 223

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Porcine trypsin variant #116

<400> 116

Ile Val Gly Gly Tyr Thr Cys Ala Ala Asn Ser Ile Pro Tyr Gln Val

1 5 10 15

Ser Leu Asn Ser Gly Ser His Arg Cys Gly Gly Ser Leu Ile Asn Ser

20 25 30

Gln Trp Val Val Ser Ala Ala His Cys Tyr Lys Pro Arg Ile Gln Val

35 40 45

Arg Leu Gly Glu His Asn Ile His Val Leu Glu Gly Asn Glu Gln Phe

50 55 60

Ile Asn Ala Ala Lys Ile Ile Thr His Pro Asn Phe Asn Gly Asn Thr

65 70 75 80

Leu Asp Asn Asp Ile Met Leu Ile Lys Leu Ser Ser Pro Ala Thr Leu

85 90 95

Asn Ser Asn Val Ala Thr Val Ser Leu Pro Thr Ser Cys Ala Ala Ala

100 105 110

Gly Thr Glu Cys Leu Ile Ser Gly Trp Gly Asn Thr Ser Ser Ser Gly

115 120 125

Ser Ser Met Pro Ser Leu Leu Gln Cys Leu Lys Ala Pro Val Leu Ser

130 135 140

Asp Ser Ser Cys Lys Ser Ser Tyr Pro Gly Gln Ile Thr Gly Asn Met

145 150 155 160

Ile Cys Val Gly Phe Leu Glu Gly Gly Ala Asp Ala Cys Gln Gly Asp

165 170 175

Ser Gly Gly Pro Val Val Cys Asn Gly Gln Leu Gln Gly Ile Val Ser

180 185 190

Ser Gly Tyr Gly Cys Ala Gln Lys Asn Lys Pro Gly Val Tyr Thr Lys

195 200 205

Val Cys Asn Tyr Val Asn Trp Ile Gln Gln Thr Ile Ala Ala Asn

210 215 220

<210> 117

<211> 223

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Porcine trypsin variant #117

<400> 117

Ile Val Gly Gly Tyr Thr Cys Ala Ala Asn Ser Ile Pro Tyr Gln Val

1 5 10 15

Ser Leu Asn Ser Gly Ser His Arg Cys Gly Gly Ser Leu Ile Asn Ser

20 25 30

Gln Trp Val Val Ser Ala Ala His Cys Tyr Lys Pro Arg Ile Gln Val

35 40 45

Arg Leu Gly Glu His Asn Ile His Val Leu Glu Gly Asn Glu Gln Phe

50 55 60

Ile Asn Ala Ala Lys Ile Ile Thr His Pro Asn Phe Asn Gly Asn Thr

65 70 75 80

Leu Asp Asn Asp Ile Met Leu Ile Lys Leu Ser Ser Pro Ala Thr Leu

85 90 95

Asn Ser Asn Val Ala Thr Val Ser Leu Pro Thr Ser Cys Ala Ala Ala

100 105 110

Gly Thr Glu Cys Leu Ile Ser Gly Trp Gly Asn Thr Ala Ser Ser Gly

115 120 125

Ser Ser Met Pro Ser Leu Leu Gln Cys Leu Lys Ala Pro Val Leu Ser

130 135 140

Asp Ser Ser Cys Lys Ser Ser Tyr Pro Gly Gln Ile Thr Gly Asn Met

145 150 155 160

Ile Cys Val Gly Phe Leu Glu Gly Gly Lys Asp Ala Cys Gln Gly Asp

165 170 175

Ser Gly Gly Pro Val Val Cys Asn Gly Gln Leu Gln Gly Ile Val Ser

180 185 190

Ser Gly Tyr Gly Cys Ala Gln Lys Asn Lys Pro Gly Val Tyr Thr Lys

195 200 205

Val Cys Asn Tyr Val Asn Trp Ile Gln Gln Thr Ile Ala Ala Asn

210 215 220

<210> 118

<211> 223

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Porcine trypsin variant #118

<400> 118

Ile Val Gly Gly Tyr Thr Cys Ala Ala Asn Ser Ile Pro Tyr Gln Val

1 5 10 15

Ser Leu Asn Ser Gly Ser His Arg Cys Gly Gly Ser Leu Ile Asn Ser

20 25 30

Gln Trp Val Val Ser Ala Ala His Cys Tyr Lys Pro Arg Ile Gln Val

35 40 45

Arg Leu Gly Glu His Asn Ile His Val Leu Glu Gly Asn Glu Gln Phe

50 55 60

Ile Asn Ala Ala Lys Ile Ile Thr His Pro Asn Phe Asn Gly Asn Thr

65 70 75 80

Leu Asp Asn Asp Ile Met Leu Ile Lys Leu Ser Ser Pro Ala Thr Leu

85 90 95

Asn Ser His Val Ala Thr Val Ser Leu Pro Thr Ser Cys Ala Ala Ala

100 105 110

Gly Thr Glu Cys Leu Ile Ser Gly Trp Gly Asn Thr Ala Ser Ser Gly

115 120 125

Ser Ser Met Pro Ser Leu Leu Gln Cys Leu Lys Ala Pro Val Leu Ser

130 135 140

Asp Ser Ser Cys Lys Ser Ser Tyr Pro Gly Gln Ile Thr Gly Asn Met

145 150 155 160

Ile Cys Val Gly Phe Leu Glu Gly Gly Ala Asp Ala Cys Gln Gly Asp

165 170 175

Ser Gly Gly Pro Val Val Cys Asn Gly Gln Leu Gln Gly Ile Val Ser

180 185 190

Ser Gly Tyr Gly Cys Ala Gln Lys Asn Lys Pro Gly Val Tyr Thr Lys

195 200 205

Val Cys Asn Tyr Val Asn Trp Ile Gln Gln Thr Ile Ala Ala Asn

210 215 220

<210> 119

<211> 223

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Porcine trypsin variant #119

<400> 119

Ile Val Gly Gly Tyr Thr Cys Ala Ala Asn Ser Ile Pro Tyr Gln Val

1 5 10 15

Ser Leu Asn Ser Gly Ser His Arg Cys Gly Gly Ser Leu Ile Asn Ser

20 25 30

Gln Trp Val Val Ser Ala Ala His Cys Tyr Lys Pro Arg Ile Gln Val

35 40 45

Arg Leu Gly Glu His Asn Ile His Val Leu Glu Gly Asn Glu Gln Phe

50 55 60

Ile Asn Ala Ala Lys Ile Ile Thr His Pro Asn Phe Asn Gly Asn Thr

65 70 75 80

Leu Asp Asn Asp Ile Met Leu Ile Lys Leu Ser Ser Pro Ala Thr Leu

85 90 95

Asn Ser Ala Val Ala Thr Val Ser Leu Pro Thr Ser Cys Ala Ala Ala

100 105 110

Gly Thr Glu Cys Leu Ile Ser Gly Trp Gly Asn Thr Ala Ser Ser Gly

115 120 125

Ser Ser Met Pro Ser Leu Leu Gln Cys Leu Lys Ala Pro Val Leu Ser

130 135 140

Asp Ser Ser Cys Lys Ser Ser Tyr Pro Gly Gln Ile Thr Gly Asn Met

145 150 155 160

Ile Cys Val Gly Phe Leu Glu Gly Gly Lys Asp Ala Cys Gln Gly Asp

165 170 175

Ser Gly Gly Pro Val Val Cys Asn Gly Gln Leu Gln Gly Ile Val Ser

180 185 190

Ser Gly Tyr Gly Cys Ala Gln Lys Asn Lys Pro Gly Val Tyr Thr Lys

195 200 205

Val Cys Asn Tyr Val Asn Trp Ile Gln Gln Thr Ile Ala Ala Asn

210 215 220

<210> 120

<211> 223

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Porcine trypsin variant #120

<400> 120

Ile Val Gly Gly Tyr Thr Cys Ala Ala Asn Ser Ile Pro Tyr Gln Val

1 5 10 15

Ser Leu Asn Ser Gly Ser His Arg Cys Gly Gly Ser Leu Ile Asn Ser

20 25 30

Gln Trp Val Val Ser Ala Ala His Cys Tyr Lys Ser Arg Ile Gln Val

35 40 45

Arg Leu Gly Glu His Asn Ile Asp Val Leu Glu Gly Asn Glu Gln Phe

50 55 60

Ile Asn Ala Ala Lys Ile Ile Thr His Pro Asn Phe Asn Gly Asn Thr

65 70 75 80

Leu Asp Asn Asp Ile Met Leu Ile Lys Leu Ser Ser Pro Ala Thr Leu

85 90 95

Asn Ser Arg Val Ala Thr Val Ser Leu Pro Arg Ser Cys Ala Ala Ala

100 105 110

Gly Thr Glu Cys Leu Ile Ser Gly Trp Gly Asn Thr Pro Ser Ser Gly

115 120 125

Ser Ser Tyr Pro Ser Leu Leu Gln Cys Leu Lys Ala Pro Val Leu Ser

130 135 140

Asp Ser Ser Cys Lys Ser Ser Tyr Pro Gly Gln Ile Thr Gly Asn Met

145 150 155 160

Ile Cys Val Gly Phe Leu Glu Gly Gly Lys Asp Ala Cys Gln Gly Asp

165 170 175

Ser Gly Gly Pro Val Val Cys Asn Gly Gln Leu Gln Gly Ile Val Ser

180 185 190

Ser Gly Tyr Gly Cys Ala Gln Lys Asn Lys Pro Gly Val Tyr Thr Lys

195 200 205

Val Cys Asn Tyr Val Asn Trp Ile Gln Gln Thr Ile Ala Ala Asn

210 215 220

<210> 121

<211> 223

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Porcine trypsin variant #121

<400> 121

Ile Val Gly Gly Tyr Thr Cys Ala Ala Asn Ser Ile Pro Tyr Gln Val

1 5 10 15

Ser Leu Asn Ser Gly Ser His Arg Cys Gly Gly Ser Leu Ile Asn Ser

20 25 30

Gln Trp Val Val Ser Ala Ala His Cys Tyr Lys Ser Arg Ile Gln Val

35 40 45

Arg Leu Gly Glu His Asn Ile Asp Val Leu Glu Gly Asn Glu Gln Phe

50 55 60

Ile Asn Ala Ala Lys Ile Ile Thr His Pro Asn Phe Asn Gly Asn Thr

65 70 75 80

Leu Asp Asn Asp Ile Met Leu Ile Lys Leu Ser Ser Pro Ala Thr Leu

85 90 95

Asn Ser Arg Val Ala Thr Val Ser Leu Pro Arg Ser Cys Ala Ala Ala

100 105 110

Gly Thr Glu Cys Leu Ile Ser Gly Trp Gly Asn Thr Lys Ser Ser Gly

115 120 125

Ser Ser Tyr Pro Ser Leu Leu Gln Cys Leu Lys Ala Pro Val Leu Ser

130 135 140

Asp Ser Ser Cys Lys Ser Ser Tyr Pro Gly Gln Ile Thr Gly Asn Met

145 150 155 160

Ile Cys Val Gly Phe Leu Glu Gly Gly Ile Asp Ala Cys Gln Gly Asp

165 170 175

Ser Gly Gly Pro Val Val Cys Asn Gly Gln Leu Gln Gly Ile Val Ser

180 185 190

Ser Gly Tyr Gly Cys Ala Gln Lys Asn Lys Pro Gly Val Tyr Thr Lys

195 200 205

Val Cys Asn Tyr Val Asn Trp Ile Gln Gln Thr Ile Ala Ala Asn

210 215 220

<210> 122

<211> 223

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Porcine trypsin variant #122

<400> 122

Ile Val Gly Gly Tyr Thr Cys Ala Ala Asn Ser Ile Pro Tyr Gln Val

1 5 10 15

Ser Leu Asn Ser Gly Ser His Arg Cys Gly Gly Ser Leu Ile Asn Ser

20 25 30

Gln Trp Val Val Ser Ala Ala His Cys Tyr Lys Ser Arg Ile Gln Val

35 40 45

Arg Leu Gly Glu His Asn Ile Asp Val Leu Glu Gly Asn Glu Gln Phe

50 55 60

Ile Asn Ala Ala Lys Ile Ile Thr His Pro Asn Phe Asn Gly Asn Thr

65 70 75 80

Leu Asp Asn Asp Ile Met Leu Ile Lys Leu Ser Ser Pro Ala Thr Leu

85 90 95

Asn Ser Arg Val Ala Thr Val Ser Leu Pro Arg Ser Cys Ala Ala Ala

100 105 110

Gly Thr Glu Cys Leu Ile Ser Gly Trp Gly Asn Thr Lys Ser Ser Gly

115 120 125

Ser Ser Tyr Pro Ser Leu Leu Gln Cys Leu Lys Ala Pro Val Leu Ser

130 135 140

Asp Ser Ser Cys Lys Ser Ser Tyr Pro Gly Gln Ile Thr Gly Asn Met

145 150 155 160

Ile Cys Val Gly Phe Leu Glu Gly Gly Phe Asp Ala Cys Gln Gly Asp

165 170 175

Ser Gly Gly Pro Val Val Cys Asn Gly Gln Leu Gln Gly Ile Val Ser

180 185 190

Ser Gly Tyr Gly Cys Ala Gln Lys Asn Lys Pro Gly Val Tyr Thr Lys

195 200 205

Val Cys Asn Tyr Val Asn Trp Ile Gln Gln Thr Ile Ala Ala Asn

210 215 220

<210> 123

<211> 223

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Porcine trypsin variant #123

<400> 123

Ile Val Gly Gly Tyr Thr Cys Ala Ala Asn Ser Ile Pro Tyr Gln Val

1 5 10 15

Ser Leu Asn Ser Gly Ser His Gly Cys Gly Gly Ser Leu Ile Asn Ser

20 25 30

Gln Trp Val Val Ser Ala Ala His Cys Tyr Lys Ser Arg Ile Gln Val

35 40 45

Arg Leu Gly Glu His Asn Ile Asp Val Leu Glu Gly Asn Glu Gln Phe

50 55 60

Ile Asn Ala Ala Lys Ile Ile Thr His Pro Asn Phe Asn Gly Asn Thr

65 70 75 80

Leu Asp Asn Asp Ile Met Leu Ile Lys Leu Ser Ser Pro Ala Thr Leu

85 90 95

Asn Ser Arg Val Ala Thr Val Ser Leu Pro Arg Ser Cys Ala Ala Ala

100 105 110

Gly Thr Glu Cys Leu Ile Ser Gly Trp Gly Asn Thr Lys Ser Ser Gly

115 120 125

Ser Ser Tyr Pro Ser Leu Leu Gln Cys Leu Lys Ala Pro Val Leu Ser

130 135 140

Asp Ser Ser Cys Lys Ser Ser Tyr Pro Gly Gln Ile Thr Gly Asn Met

145 150 155 160

Ile Cys Val Gly Phe Leu Glu Gly Gly Lys Asp Ala Cys Gln Gly Asp

165 170 175

Ser Gly Gly Pro Val Val Cys Asn Gly Gln Leu Gln Gly Ile Val Ser

180 185 190

Ser Gly Tyr Gly Cys Ala Gln Lys Asn Lys Pro Gly Val Tyr Thr Lys

195 200 205

Val Cys Asn Tyr Val Asn Trp Ile Gln Gln Thr Ile Ala Ala Asn

210 215 220

<210> 124

<211> 223

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Porcine trypsin variant #124

<400> 124

Ile Val Gly Gly Tyr Thr Cys Ala Ala Asn Ser Ile Pro Tyr Gln Val

1 5 10 15

Ser Leu Asn Ser Gly Ser His Arg Cys Gly Gly Ser Leu Ile Asn Ser

20 25 30

Gln Trp Val Val Ser Ala Ala His Cys Tyr Lys Ser Arg Ile Gln Val

35 40 45

Arg Leu Gly Glu His Asn Ile Asp Val Leu Glu Gly Asn Glu Gln Phe

50 55 60

Ile Asn Ala Ala Lys Ile Ile Thr His Pro Asn Phe Asn Gly Asn Thr

65 70 75 80

Leu Asp Asn Asp Ile Met Leu Ile Lys Leu Ser Ser Pro Ala Thr Leu

85 90 95

Asn Ser Arg Val Ala Thr Val Ser Leu Pro Arg Ser Cys Ala Ala Ala

100 105 110

Gly Thr Glu Cys Leu Ile Ser Gly Trp Gly Asn Thr Lys Ser Ser Gly

115 120 125

Ser Ser Lys Pro Ser Leu Leu Gln Cys Leu Lys Ala Pro Val Leu Ser

130 135 140

Asp Ser Ser Cys Lys Ser Ser Tyr Pro Gly Gln Ile Thr Gly Asn Met

145 150 155 160

Ile Cys Val Gly Phe Leu Glu Gly Gly Lys Asp Ala Cys Gln Gly Asp

165 170 175

Ser Gly Gly Pro Val Val Cys Asn Gly Gln Leu Gln Gly Ile Val Ser

180 185 190

Ser Gly Tyr Gly Cys Ala Gln Lys Asn Lys Pro Gly Val Tyr Thr Lys

195 200 205

Val Cys Asn Tyr Val Asn Trp Ile Gln Gln Thr Ile Ala Ala Asn

210 215 220

<210> 125

<211> 223

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Porcine trypsin variant #125

<400> 125

Ile Val Gly Gly Tyr Thr Cys Ala Ala Asn Ser Ile Pro Tyr Gln Val

1 5 10 15

Ser Leu Asn Ser Gly Ser His Asp Cys Gly Gly Ser Leu Ile Asn Ser

20 25 30

Gln Trp Val Val Ser Ala Ala His Cys Tyr Lys Ser Arg Ile Gln Val

35 40 45

Arg Leu Gly Glu His Asn Ile Asp Val Leu Glu Gly Asn Glu Gln Phe

50 55 60

Ile Asn Ala Ala Lys Ile Ile Thr His Pro Asn Phe Asn Gly Asn Thr

65 70 75 80

Leu Asp Asn Asp Ile Met Leu Ile Lys Leu Ser Ser Pro Ala Thr Leu

85 90 95

Asn Ser Arg Val Ala Thr Val Ser Leu Pro Arg Ser Cys Ala Ala Ala

100 105 110

Gly Thr Glu Cys Leu Ile Ser Gly Trp Gly Asn Thr Lys Ser Ser Gly

115 120 125

Ser Ser Tyr Pro Ser Leu Leu Gln Cys Leu Lys Ala Pro Val Leu Ser

130 135 140

Asp Ser Ser Cys Lys Ser Ser Tyr Pro Gly Gln Ile Thr Gly Asn Met

145 150 155 160

Ile Cys Val Gly Phe Leu Glu Gly Gly Lys Asp Ala Cys Gln Gly Asp

165 170 175

Ser Gly Gly Pro Val Val Cys Asn Gly Gln Leu Gln Gly Ile Val Ser

180 185 190

Ser Gly Tyr Gly Cys Ala Gln Lys Asn Lys Pro Gly Val Tyr Thr Lys

195 200 205

Val Cys Asn Tyr Val Asn Trp Ile Gln Gln Thr Ile Ala Ala Asn

210 215 220

<210> 126

<211> 223

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Porcine trypsin variant #126

<400> 126

Ile Val Gly Gly Tyr Thr Cys Ala Ala Asn Ser Ile Pro Tyr Gln Val

1 5 10 15

Ser Leu Asn Ser Gly Ser His Glu Cys Gly Gly Ser Leu Ile Asn Ser

20 25 30

Gln Trp Val Val Ser Ala Ala His Cys Tyr Lys Ser Arg Ile Gln Val

35 40 45

Arg Leu Gly Glu His Asn Ile Asp Val Leu Glu Gly Asn Glu Gln Phe

50 55 60

Ile Asn Ala Ala Lys Ile Ile Thr His Pro Asn Phe Asn Gly Asn Thr

65 70 75 80

Leu Asp Asn Asp Ile Met Leu Ile Lys Leu Ser Ser Pro Ala Thr Leu

85 90 95

Asn Ser Arg Val Ala Thr Val Ser Leu Pro Arg Ser Cys Ala Ala Ala

100 105 110

Gly Thr Glu Cys Leu Ile Ser Gly Trp Gly Asn Thr Lys Ser Ser Gly

115 120 125

Ser Ser Tyr Pro Ser Leu Leu Gln Cys Leu Lys Ala Pro Val Leu Ser

130 135 140

Asp Ser Ser Cys Lys Ser Ser Tyr Pro Gly Gln Ile Thr Gly Asn Met

145 150 155 160

Ile Cys Val Gly Phe Leu Glu Gly Gly Lys Asp Ala Cys Gln Gly Asp

165 170 175

Ser Gly Gly Pro Val Val Cys Asn Gly Gln Leu Gln Gly Ile Val Ser

180 185 190

Ser Gly Tyr Gly Cys Ala Gln Lys Asn Lys Pro Gly Val Tyr Thr Lys

195 200 205

Val Cys Asn Tyr Val Asn Trp Ile Gln Gln Thr Ile Ala Ala Asn

210 215 220

<210> 127

<211> 223

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Porcine trypsin variant #127

<400> 127

Ile Val Gly Gly Tyr Thr Cys Ala Ala Asn Ser Ile Pro Tyr Gln Val

1 5 10 15

Ser Leu Asn Ser Gly Ser His Arg Cys Gly Gly Ser Leu Ile Asn Ser

20 25 30

Gln Trp Val Val Ser Ala Ala His Cys Tyr Lys Tyr Arg Ile Gln Val

35 40 45

Arg Leu Gly Glu His Asn Ile Asp Val Leu Glu Gly Asn Glu Gln Phe

50 55 60

Ile Asn Ala Ala Lys Ile Ile Thr His Pro Asn Phe Asn Gly Asn Thr

65 70 75 80

Leu Asp Asn Asp Ile Met Leu Ile Lys Leu Ser Ser Pro Ala Thr Leu

85 90 95

Asn Ser Arg Val Ala Thr Val Ser Leu Pro Arg Ser Cys Ala Ala Ala

100 105 110

Gly Thr Glu Cys Leu Ile Ser Gly Trp Gly Asn Thr Lys Ser Ser Gly

115 120 125

Ser Ser Tyr Pro Ser Leu Leu Gln Cys Leu Lys Ala Pro Val Leu Ser

130 135 140

Asp Ser Ser Cys Lys Ser Ser Tyr Pro Gly Gln Ile Thr Gly Asn Met

145 150 155 160

Ile Cys Val Gly Phe Leu Glu Gly Gly Lys Asp Ala Cys Gln Gly Asp

165 170 175

Ser Gly Gly Pro Val Val Cys Asn Gly Gln Leu Gln Gly Ile Val Ser

180 185 190

Ser Gly Tyr Gly Cys Ala Gln Lys Asn Lys Pro Gly Val Tyr Thr Lys

195 200 205

Val Cys Asn Tyr Val Asn Trp Ile Gln Gln Thr Ile Ala Ala Asn

210 215 220

<210> 128

<211> 223

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Porcine trypsin variant #128

<400> 128

Ile Val Gly Gly Tyr Thr Cys Ala Ala Asn Ser Ile Pro Tyr Gln Val

1 5 10 15

Ser Leu Asn Ser Gly Ser His Arg Cys Gly Gly Ser Leu Ile Asn Ser

20 25 30

Gln Trp Val Val Ser Ala Ala His Cys Tyr Lys Ser Arg Ile Gln Val

35 40 45

Arg Leu Gly Glu His Asn Ile Asp Val Leu Glu Gly Asn Glu Gln Phe

50 55 60

Ile Asn Ala Ala Lys Ile Ile Thr His Pro Asn Phe Asn Gly Asn Thr

65 70 75 80

Leu Asp Asn Asp Ile Met Leu Ile Lys Leu Ser Ser Pro Ala Thr Leu

85 90 95

Asn Ser Arg Val Ala Thr Val Ser Leu Pro Arg Ser Cys Ala Ala Ala

100 105 110

Gly Thr Glu Cys Leu Ile Ser Gly Trp Gly Asn Thr Lys Ser Ser Gly

115 120 125

Ser Ser Pro Pro Ser Leu Leu Gln Cys Leu Lys Ala Pro Val Leu Ser

130 135 140

Asp Ser Ser Cys Lys Ser Ser Tyr Pro Gly Gln Ile Thr Gly Asn Met

145 150 155 160

Ile Cys Val Gly Phe Leu Glu Gly Gly Lys Asp Ala Cys Gln Gly Asp

165 170 175

Ser Gly Gly Pro Val Val Cys Asn Gly Gln Leu Gln Gly Ile Val Ser

180 185 190

Ser Gly Tyr Gly Cys Ala Gln Lys Asn Lys Pro Gly Val Tyr Thr Lys

195 200 205

Val Cys Asn Tyr Val Asn Trp Ile Gln Gln Thr Ile Ala Ala Asn

210 215 220

<210> 129

<211> 223

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Porcine trypsin variant #129

<400> 129

Ile Val Gly Gly Tyr Thr Cys Ala Ala Asn Ser Ile Pro Tyr Gln Val

1 5 10 15

Ser Leu Asn Ser Gly Ser His Arg Cys Gly Gly Ser Leu Ile Asn Ser

20 25 30

Gln Trp Val Val Ser Ala Ala His Cys Tyr Lys Ser Arg Ile Gln Val

35 40 45

Arg Leu Gly Glu His Asn Ile Asp Val Leu Glu Gly Asn Glu Gln Phe

50 55 60

Ile Asn Ala Ala Lys Ile Ile Thr His Pro Asn Phe Asn Gly Asn Thr

65 70 75 80

Leu Asp Asn Asp Ile Met Leu Ile Lys Leu Ser Ser Pro Ala Thr Leu

85 90 95

Asn Ser Arg Val Ala Thr Val Ser Leu Pro Arg Ser Cys Ala Ala Ala

100 105 110

Gly Thr Glu Cys Leu Ile Ser Gly Trp Gly Asn Thr Lys Ser Ser Gly

115 120 125

Ser Ser Tyr Pro Ser Leu Leu Gln Cys Leu Lys Ala Pro Val Leu Ser

130 135 140

Asp Ser Ser Cys Lys Ser Ser Tyr Pro Gly Gln Ile Thr Gly Asn Met

145 150 155 160

Ile Cys Val Gly Phe Leu Glu Gly Gly Lys Asp Ala Cys Gln Gly Asp

165 170 175

Ser Gly Gly Pro Val Val Cys Asn Gly Gln Leu Gln Gly Ile Val Ser

180 185 190

Asn Gly Tyr Gly Cys Ala Gln Lys Asn Lys Pro Gly Val Tyr Thr Lys

195 200 205

Val Cys Asn Tyr Val Asn Trp Ile Gln Gln Thr Ile Ala Ala Asn

210 215 220

<210> 130

<211> 223

<212> PROTEIN

<213> Artificial sequence

<220>

<223> Porcine trypsin variant #130

<400> 130

Ile Val Gly Gly Tyr Thr Cys Ala Ala Asn Ser Ile Pro Tyr Gln Val

1 5 10 15

Ser Leu Asn Ser Gly Ser His Arg Cys Gly Gly Ser Leu Ile Asn Ser

20 25 30

Gln Trp Val Val Ser Ala Ala His Cys Tyr Lys Ser Arg Ile Gln Val

35 40 45

Arg Leu Gly Glu His Asn Ile Asp Val Leu Glu Gly Asn Glu Gln Phe

50 55 60

Ile Asn Ala Ala Lys Ile Ile Thr His Pro Asn Phe Asn Gly Asn Thr

65 70 75 80

Leu Asp Asn Asp Ile Met Leu Ile Lys Leu Ser Ser Pro Ala Thr Leu

85 90 95

Asn Ser Arg Val Ala Thr Val Ser Leu Pro Arg Ser Cys Ala Ala Ala

100 105 110

Gly Thr Glu Cys Leu Ile Ser Gly Trp Gly Asn Thr Lys Ser Ser Gly

115 120 125

Ser Ser Tyr Pro Ser Leu Leu Gln Cys Leu Lys Ala Pro Val Leu Ser

130 135 140

Asp Ser Ser Cys Lys Ser Ser Tyr Pro Gly Gln Ile Thr Gly Asn Met

145 150 155 160

Ile Cys Val Gly Phe Leu Glu Gly Gly Lys Asp Ala Cys Gln Gly Asp

165 170 175

Ser Gly Gly Pro Val Val Cys Asn Gly Gln Leu Gln Gly Ile Val Ser

180 185 190

Cys Gly Tyr Gly Cys Ala Gln Lys Asn Lys Pro Gly Val Tyr Thr Lys

195 200 205

Val Cys Asn Tyr Val Asn Trp Ile Gln Gln Thr Ile Ala Ala Asn

210 215 220

<---

Claims (54)

1. A variant of porcine trypsin that retains porcine trypsin activity, comprising or consisting of an amino acid sequence that shares at least 90% sequence identity with SEQ ID NO: 1, wherein said amino acid sequence differs from SEQ ID NO: 1 by at least one or more amino acid substitutions at one or more positions corresponding to F24, S44, D56, G78, Y131, S172 and W193 of native porcine trypsin according to SEQ ID NO: 1 , provided that the specified amino acid sequence is not a native porcine trypsin in accordance with SEQ ID NO: 1; and provided that said amino acid sequence is not the porcine trypsin S172A variant according to SEQ ID NO: 2. 2. Pork trypsin variant according to claim 1, where the amino acid at the position corresponding to F24 has been replaced by an amino acid selected from the group consisting of Ala, Asn, Arg, Gln, Ile, Leu, Lys, Met, Ser, Thr and Val; where the amino acid at the position corresponding to S44 is replaced by an amino acid selected from the group consisting of Leu and Pro; where the amino acid at the position corresponding to D56 is replaced by an amino acid selected from the group consisting of Ala, Asn, His and Trp; where the amino acid at the position corresponding to G78 is replaced by an amino acid selected from the group consisting of Ala, Glu; Pro, Ser and Tyr; where the amino acid at the position corresponding to Y131 has been replaced by an amino acid selected from the group consisting of Ala, Asn, Asp, Cys, Gln; Glu, Gly, His, Ile, Leu, Met, Ser, Thr, Trp, Val; where the amino acid at the position corresponding to S172 has been replaced by an amino acid selected from the group consisting of Ala, Cys and Thr; and/or where the amino acid at the position corresponding to W193 has been replaced by an amino acid selected from the group consisting of Phe, Ser, Thr and Tyr. 3. Variant pork trypsin according to any one of paragraphs. 1, 2, wherein said amino acid sequence further differs from SEQ ID NO: 1 by at least one or more amino acid substitutions at one or more positions corresponding to R99, R107, K125 and K170 of native porcine trypsin according to SEQ ID NO: 1. 4. Pork trypsin variant according to claim 3, where the amino acid at the position corresponding to R99 has been replaced by an amino acid selected from the group consisting of Ala, Asn, Asp, Glu, Gly, His, Leu, Phe, Thr, Trp and Tyr; where the amino acid at the position corresponding to R107 has been replaced by an amino acid selected from the group consisting of Asp, Gly, Pro, Ser and Thr; where the amino acid at the position corresponding to K125 is replaced by an amino acid selected from the group consisting of Ala, Cys, Gln, Glu, Gly, His, Leu, Ser and Tyr; and/or where the amino acid at the position corresponding to K170 has been replaced by an amino acid selected from the group consisting of Ala, Asn, Gly and Tyr. 5. Variant pork trypsin according to any one of paragraphs. 1-4, where the specified amino acid sequence is SEQ ID NO: 3, where - Xaa24 is an amino acid selected from the group consisting of Ala, Asn, Arg, Gln, Ile, Leu, Lys, Met, Phe, Ser, Thr and Val; - Xaa44 is an amino acid selected from the group consisting of Leu, Pro and Ser; - Xaa56 is an amino acid selected from the group consisting of Ala, Asn, Asp, His and Trp; - Xaa78 is an amino acid selected from the group consisting of Ala, Glu, Gly, Pro, Ser and Tyr; - Xaa99 is an amino acid selected from the group consisting of Ala, Arg, Asn, Asp, Glu, Gly, His, Leu, Phe, Thr, Trp and Tyr; - Xaa107 is an amino acid selected from the group consisting of Arg, Asp, Gly, Pro, Ser and Thr; - Xaa125 is an amino acid selected from the group consisting of Ala, Cys, Gln, Glu, Gly, His, Leu, Lys, Ser and Tyr; - Xaa131 is an amino acid selected from the group consisting of Ala, Asn, Asp, Cys, Gln; Glu, Gly, His, Ile, Leu, Met, Ser, Thr, Trp, Tyr and Val; - Xaa170 is an amino acid selected from the group consisting of Ala, Asn, Gly, Lys and Tyr; - Xaa172 is an amino acid selected from the group consisting of Ala, Cys, Ser and Thr; and/or - Xaa193 is an amino acid selected from the group consisting of Phe, Ser, Thr, Trp and Tyr; provided that SEQ ID NO: 3 is not wild-type porcine trypsin according to SEQ ID NO: 1; and provided that SEQ ID NO: 3 is not the S172A porcine trypsin variant of SEQ ID NO: 2. 6. Variant pork trypsin according to any one of paragraphs. 1-5, wherein said amino acid sequence is selected from the group consisting of SEQ ID NO: 4, SEQ ID NO: 5, SEQ ID NO: 6, SEQ ID NO: 7, SEQ ID NO: 8, SEQ ID NO: 9, SEQ ID NO: 10, SEQ ID NO: 11, SEQ ID NO: 12, SEQ ID NO: 13, SEQ ID NO: 14, SEQ ID NO: 15, SEQ ID NO: 16, SEQ ID NO: 17, SEQ ID NO: 18, SEQ ID NO: 19, SEQ ID NO: 20, SEQ ID NO: 21, SEQ ID NO: 22, SEQ ID NO: 23, SEQ ID NO: 24, SEQ ID NO: 25, SEQ ID NO: 26, SEQ ID NO: 27, SEQ ID NO: 28, SEQ ID NO: 29, SEQ ID NO: 30, SEQ ID NO: 31, SEQ ID NO: 32, SEQ ID NO: 33, SEQ ID NO: 34, SEQ ID NO: 35, SEQ ID NO: 36, SEQ ID NO: 37, SEQ ID NO: 38, SEQ ID NO: 39, SEQ ID NO: 40, SEQ ID NO: 41, SEQ ID NO: 42, SEQ ID NO: 43, SEQ ID NO: 44, SEQ ID NO: 45, SEQ ID NO: 46, SEQ ID NO: 47, SEQ ID NO: 48, SEQ ID NO: 49, SEQ ID NO: 50, SEQ ID NO: 51, SEQ ID NO: 52, SEQ ID NO: 53, SEQ ID NO: 54, SEQ ID NO: 55, SEQ ID NO: 56, SEQ ID NO: 57, SEQ ID NO: 58, SEQ ID NO: 59, SEQ ID NO: 60, SEQ ID NO: 61, SEQ ID NO: 62, SEQ ID NO: 63, SEQ ID NO: 64, SEQ I D NO: 65, SEQ ID NO: 66, SEQ ID NO: 67, SEQ ID NO: 68, SEQ ID NO: 69, SEQ ID NO: 70, SEQ ID NO: 71, SEQ ID NO: 72, SEQ ID NO : 73, SEQ ID NO: 74, SEQ ID NO: 75, SEQ ID NO: 76, SEQ ID NO: 77, SEQ ID NO: 78, SEQ ID NO: 79, SEQ ID NO: 80, SEQ ID NO: 81 , SEQ ID NO: 82, SEQ ID NO: 83, SEQ ID NO: 84, SEQ ID NO: 85, SEQ ID NO: 86, SEQ ID NO: 87, SEQ ID NO: 88, SEQ ID NO: 89, SEQ ID NO: 90, SEQ ID NO: 91, SEQ ID NO: 92, SEQ ID NO: 93, SEQ ID NO: 94, SEQ ID NO: 95, SEQ ID NO: 96, SEQ ID NO: 97, SEQ ID NO : 98, SEQ ID NO: 99, SEQ ID NO: 100, SEQ ID NO: 101, SEQ ID NO: 102, SEQ ID NO: 103, SEQ ID NO: 104, SEQ ID NO: 105, SEQ ID NO: 106 , SEQ ID NO: 107, SEQ ID NO: 108, SEQ ID NO: 109, SEQ ID NO: 110, SEQ ID NO: 111, SEQ ID NO: 112, SEQ ID NO: 113, SEQ ID NO: 114, SEQ ID NO: 115, SEQ ID NO: 116, SEQ ID NO: 117, SEQ ID NO: 118 and SEQ ID NO: 119. 7. Variant pork trypsin according to any one of paragraphs. 1-6, where (i) said variant is capable of cleaving a peptide with the general formula A-Lys-Thr-Arg-Arg-B to produce a cleavage product with the general formula A-Lys-Thr-Arg-Arg in a yield of at least 80%, where A is an amino acid sequence consisting of one or more amino acids; and where B is an amino acid sequence consisting of one or more amino acids; and/or (ii) the variant exhibits increased selectivity for cleavage of human preproinsulin, preproinsulin glargine, preproinsulin lispro, preproinsulin aspart, or preproinsulin glulisine at the C-terminal position relative to the B32-Arg position compared to the porcine trypsin variant S172A according to SEQ ID NO: 2. 8. A nucleic acid molecule encoding a variant of porcine trypsin according to any one of paragraphs. 1-7, wherein said nucleic acid molecule is optionally contained in a vector or optionally integrated into a genome. 9. A host cell comprising the nucleic acid molecule of claim 8 to produce a porcine trypsin variant that retains porcine trypsin activity. 10. The method of obtaining a variant of pork trypsin according to any one of paragraphs. 1-7, including stages: culturing the host cell according to claim 9; and isolating the trypsinogen variant porcine trypsin precursor molecule from the culture medium or from the host cell; and activation of the precursor molecule, which is trypsinogen, thus obtaining the specified variant of porcine trypsin. 11. Variant pork trypsin according to any one of paragraphs. 1-7, or a nucleic acid molecule according to claim 8, or a host cell according to claim 9 for use in a method for manufacturing a food ingredient or feed ingredient. 12. The use of variant pork trypsin according to any one of paragraphs. 1-7 in a process for the preparation of human insulin, insulin analogue or insulin derivative. 13. Use according to claim 12, wherein the insulin analog is selected from the group consisting of insulin aspart, insulin lispro, insulin glulisine, and insulin glargine. 14. The use of variant pork trypsin according to any one of paragraphs. 1-7 for cleavage of a protein or peptide with the general formula A-Lys-Thr-Arg-Arg-B, where A is an amino acid sequence consisting of one or more amino acids; and where B is an amino acid sequence consisting of one or more amino acids. 15. Use according to claim 14, wherein cleavage results in a cleavage product of A-Lys-Thr-Arg-Arg. 16. Use according to claim 15, wherein the cleavage results in a cleavage product of A-Lys-Thr-Arg-Arg in a yield of at least 80.

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