Improved peptides for converting the substrate 3-keto-deoxynivalenol into 3-epi-deoxynivalenol

Abstract

The present invention relates to a composition comprising one or more polypeptides and a carrier, as well as uses and methods of these polypeptides, wherein the polypeptides comprise or consist of the following sequence: a sequence of SEQ ID NO.1-14 or a sequence having at least 75% sequence identity with any one of SEQ ID NO.1-14.

Description

Technical Field

[0001] This application contains a Sequence Listing in computer readable form, which is incorporated herein by reference.

[0002] The present invention relates to compositions comprising one or more polypeptides and a carrier, as well as uses and methods of these polypeptides, wherein the polypeptides comprise or consist of the following sequences: sequences of SEQ ID NOs. 1-14 or sequences having at least 75% sequence identity with any of SEQ ID NOs. 1-14. Background Art

[0003] Mycotoxins are secondary metabolites produced by filamentous fungi. A representative mycotoxin is deoxynivalenol (DON), a B-type trichothecene or vomitoxin produced by various Fusarium species and found worldwide. These fungi infect cultivated plants, particularly cereals, where infection typically occurs before harvest, when fungal growth and / or mycotoxin production can occur before storage, or even after harvest, before storage or under inappropriate storage conditions. The Food and Agriculture Organization of the United Nations (FAO) has estimated that 25% of agricultural products worldwide are contaminated by fungi, causing significant economic losses. In an eight-year international study, a total of 19,757 samples were analyzed from January 2004 to December 2011; 72% tested positive for at least one fungus, 39% were found to be contaminated, and 56% tested positive for DON (Schatzmayr and Streit (2013)). Trichothecenes and DON have been found in all regions of the world and in all types of cereals and feed crops tested, such as corn, soybean meal, wheat, wheat bran, DDGS (distillers dried grains with solubles), and in finished animal feed mixtures, with an incidence of 100%.

[0004] The primary strategy for reducing trichothecene contamination of food and animal feed products is to limit fungal growth, for example, by maintaining "good agricultural practices." This includes measures such as keeping seeds free from insects and fungi or promptly removing agricultural waste from fields. Furthermore, fungal growth in fields can be reduced through the use of fungicides. After harvest, harvested material should be stored at a residual humidity below 15% and at low temperatures to prevent fungal growth. Similarly, fungal-infected material should be removed before further processing. Despite these preventative measures, between 2004 and 2011, even in regions with the highest agricultural standards, such as North America and Central Europe, up to 68% of samples tested were found to be contaminated with DON (a representative trichothecene) (Schatzmayr and Streit (2013)).

[0005] It is known that the toxicity of trichothecenes is (at least) partly due to the hydroxyl (-OH) group present at the C-3 atom. One of the most abundant trichothecenes having such a 3-hydroxyl group is deoxynivalenol (DON).

[0006] However, the hydroxyl group of DON and other trichothecenes can exist in two isomeric states, namely the S configuration (DON) or the R configuration (3-epi-DON). Several publications include He et al. (2015), Payros et al. (2016) and Pierron et al. (2016).

[0007] Hassan et al. (2017) claimed to use a two-step process to epimerize DON to 3-epi-DON by forming 3-keto-DON. 3-Keto-DON includes a 3-oxy group in place of the 3-hydroxyl group present in the isomer. Finally, Carere et al. (2018) identified an enzyme called DmDepB from Streptococcus mutans 17-2-E-8 that can reduce 3-keto-DON to 3-epi-DON and DON. WO2019 / 046954 describes an enzyme identical to that of Hassan et al. (2017) and also obtained another DepB enzyme from Rhizobium leguminosarum (RlDepB).

[0008] However, there is still a need to provide enzymes that can more efficiently convert 3-keto-DON into the non-toxic isomer 3-epi-DON or convert trichothecenes comprising a 3-oxo group into trichothecenes comprising a 3-hydroxyl group in the S configuration. Summary of the Invention

[0009] The technical solutions of the present invention are described below, illustrated by embodiments, explained by drawings and embodied in the claims.

[0010] The present invention relates to a composition comprising:

[0011] a) one or more polypeptides comprising or consisting of a sequence of SEQ ID NOs. 1-14 or a sequence having at least 75% sequence identity to any one of SEQ ID NOs. 1-14, and / or a sequence of SEQ ID NOs. 63-101, wherein the polypeptide is capable of converting the substrate 3-keto-DON (3-keto-DON) into products 3-epi-DON and DON, with a 3-epi-DON:DON ratio of at least 25:1; and

[0012] b) Carrier.

[0013] The present invention also relates to a composition comprising:

[0014] a) one or more polypeptides comprising or consisting of a sequence of SEQ ID NOs. 1-14 or a sequence having at least 75% sequence identity to any one of SEQ ID NOs. 1-14, and / or a sequence of SEQ ID NOs. 63-101, wherein the polypeptide is capable of converting the substrate 3-keto-DON to the product 3-epi-DON, and optionally also to the product DON, totaling 100% of the total product, wherein at least 96% of the total product is 3-epi-DON; and

[0015] b) Carrier.

[0016] The present invention further relates to a polypeptide comprising the sequence of SEQ ID NO. 1 or a sequence having at least 75% sequence identity to SEQ ID NO. 1, wherein the polypeptide is capable of converting 3-keto-DON into 3-epi-DON. The present invention also relates to a polypeptide comprising any one of SEQ ID NOs. 63-101.

[0017] Furthermore, the present invention relates to the use of one or more polypeptides for converting 3-keto-DON into 3-epi-DON, wherein the one or more polypeptides comprise or consist of the following sequences: a sequence in SEQ ID NO. 1-14 or a sequence having at least 75% sequence identity with any one of SEQ ID NO. 1-14, and / or a sequence in SEQ ID NO. 63-101.

[0018] In addition, the present invention relates to the use of one or more polypeptides in the preparation of a feed additive or a feed composition, wherein the one or more polypeptides comprise or consist of the following sequences: a sequence in SEQ ID NO. 1-14 or a sequence having at least 75% sequence identity with any one of SEQ ID NO. 1-14, and / or a sequence in SEQ ID NO. 63-101, wherein the polypeptide is capable of converting the substrate 3-keto-DON into products 3-epi-DON and DON, and the ratio of 3-epi-DON:DON is at least 25:1.

[0019] Furthermore, the present invention relates to the use of one or more polypeptides in the preparation of a feed additive or feed composition, wherein the one or more polypeptides comprise or consist of the following sequences: a sequence in SEQ ID NO. 1-14 or a sequence having at least 75% sequence identity with any one of SEQ ID NO. 1-14, and / or a sequence in SEQ ID NO. 63-101, wherein the polypeptide is capable of converting the substrate 3-keto-DON into the product 3-epi-DON, and optionally also into the product DON, totaling 100% of the total product, wherein at least 96% of the total product is 3-epi-DON.

[0020] The present invention relates to the use of one or more polypeptides in the production of biogas, bioethanol or sugar, preferably produced from sugarcane or sugar beet, wherein the one or more polypeptides comprise or consist of the following sequences: a sequence in SEQ ID NO. 1-14 or a sequence having at least 75% sequence identity with any one of SEQ ID NO. 1-14, and / or a sequence in SEQ ID NO. 63-101, wherein the polypeptide is capable of converting the substrate 3-keto-DON into products 3-epi-DON and DON in a ratio of at least 25:1 (3-epi-DON:DON).

[0021] Furthermore, the present invention relates to the use of one or more polypeptides in the production of biogas, bioethanol or sugar, preferably produced from sugar cane or sugar beet, wherein the one or more polypeptides comprise or consist of the following sequences: a sequence in SEQ ID NO. 1-14 or a sequence having at least 75% sequence identity with any one of SEQ ID NO. 1-14, and / or a sequence in SEQ ID NO. 63-101, wherein the polypeptide is capable of converting the substrate 3-keto-DON into the product 3-epi-DON, and optionally also into the product DON, totaling 100% of the total product, wherein at least 96% of the total product is 3-epi-DON.

[0022] The present invention also encompasses a feed additive or food additive and a feed or food comprising one or more polypeptides comprising or consisting of a sequence of SEQ ID NOs. 1-14 or a sequence having at least 75% sequence identity to any one of SEQ ID NOs. 1-14, and / or a sequence of SEQ ID NOs. 63-101, wherein the polypeptide is capable of converting the substrate 3-keto-DON into products 3-epi-DON and DON in a 3-epi-DON:DON ratio of at least 25:1.

[0023] Furthermore, the present invention relates to a feed additive or food additive or feed or food comprising one or more polypeptides comprising or consisting of a sequence of SEQ ID NO. 1-14 or a sequence having at least 75% sequence identity to any one of SEQ ID NO. 1-14, and / or a sequence of SEQ ID NO. 63-101, wherein the polypeptide is capable of converting the substrate 3-keto-DON into the product 3-epi-DON, and optionally also into the product DON, totaling 100% of the total product, wherein at least 96% of the total product is 3-epi-DON.

[0024] The present invention also encompasses an additive for use in a composition, preferably a tillage composition, comprising 3-keto-DON or 3-DON, the additive comprising one or more polypeptides comprising or consisting of a sequence of SEQ ID NOs. 1-14 or a sequence having at least 75% sequence identity to any of SEQ ID NOs. 1-14, and / or a sequence of SEQ ID NOs. 63-101, wherein the polypeptide is capable of converting the substrate 3-keto-DON into products 3-epi-DON and DON in a 3-epi-DON:DON ratio of at least 25:1.

[0025] The present invention also relates to an additive for a composition, preferably a tillage composition, comprising 3-keto-DON or 3-DON, the additive comprising one or more polypeptides comprising or consisting of a sequence in SEQ ID NOs. 1-14 or a sequence having at least 75% sequence identity to any of SEQ ID NOs. 1-14, and / or a sequence in SEQ ID NOs. 63-101, wherein the polypeptide is capable of converting the substrate 3-keto-DON into the product 3-epi-DON, and optionally also into the product DON, totaling 100% of the total product, wherein at least 96% of the total product is 3-epi-DON.

[0026] Furthermore, the present invention relates to a method for converting a trichothecene comprising a 3-oxo group into a trichothecene comprising a 3-hydroxy group, the method comprising contacting one or more polypeptides with the trichothecene comprising a 3-oxo group, the one or more polypeptides comprising or consisting of the following sequences: a sequence in SEQ ID NOs. 1-14 or a sequence having at least 75% sequence identity to any one of SEQ ID NOs. 1-14, and / or a sequence in SEQ ID NOs. 63-101.

[0027] Furthermore, the present invention relates to a method for converting 3-keto-DON into 3-epi-DON, the method comprising contacting one or more polypeptides with 3-keto-DON, the one or more polypeptides comprising or consisting of a sequence in SEQ ID NOs. 1-14 or a sequence having at least 75% sequence identity to any one of SEQ ID NOs. 1-14, and / or a sequence in SEQ ID NOs. 63-101.

[0028] Furthermore, the present invention relates to a method for reducing the DON content in a composition comprising DON or reducing the toxicity of a composition comprising DON by converting DON into 3-epi-DON, the method comprising:

[0029] a) contacting the composition with an enzyme capable of converting DON to 3-keto-DON; and

[0030] b) subsequently or simultaneously contacting the composition with one or more polypeptides comprising or consisting of a sequence in SEQ ID NOs. 1-14 or a sequence having at least 75% sequence identity to any of SEQ ID NOs. 1-14, and / or a sequence in SEQ ID NOs. 63-101.

[0031] Furthermore, the present invention relates to a host cell comprising one or more polypeptides comprising or consisting of the following sequences: a sequence in SEQ ID NO. 1-14 or a sequence having at least 75% sequence identity with any one of SEQ ID NO. 1-14, and / or a sequence in SEQ ID NO. 63-101.

[0032] The present invention also encompasses plants genetically modified to express one or more polypeptides comprising or consisting of a sequence of SEQ ID NOs. 1-14 or a sequence having at least 75% sequence identity to any of SEQ ID NOs. 1-14, and / or a sequence of SEQ ID NOs. 63-101.

[0033] The present invention relates to seeds of the plant of the present invention.

[0034] The present invention further relates to a polypeptide comprising or consisting of the following sequences: a sequence in SEQ ID NOs. 1-14 or a sequence having at least 75% sequence identity with any one of SEQ ID NOs. 1-14, and / or a sequence in SEQ ID NOs. 63-101, wherein the polypeptide is capable of converting the substrate 3-keto-DON into products 3-epi-DON and DON in a ratio of at least 25:1 (3-epi-DON / DON), for preventing and / or treating fungal poisoning.

[0035] In addition, the present invention relates to a polypeptide comprising or consisting of the following sequence: a sequence in SEQ ID NO. 1-14 or a sequence having at least 75% sequence identity with any one of SEQ ID NO. 1-14, and / or a sequence in SEQ ID NO. 63-101, wherein the polypeptide is capable of converting the substrate 3-keto-DON into the product 3-epi-DON, and optionally also into the product DON, totaling 100% of the total product, wherein at least 96% of the total product is 3-epi-DON, for preventing and / or treating mycotoxicosis. BRIEF DESCRIPTION OF THE DRAWINGS

[0036] The attached figure shows:

[0037] Figure 1A Sequence identity over the entire protein length is shown as percent identity to SEQ ID NOs. 1-16 (100% coverage of protein sequence). SEQ ID NOs. 1-14 are sequences of 3-keto DON reductases described herein, whereas SEQ ID NOs. 15 and 16 relate to prior art DepB enzyme sequences. Figure 1B Sequence identity over the entire length of the protein is shown as percent identity to SEQ ID NOs. 1, 3, 6, 63-101 (100% coverage of protein sequence).

[0038] Figure 2 Data for 3-epi-DON production by various enzymes are shown. Data for SEQ ID NO. 16 are not shown, as it was not possible to produce sufficient quantities for the required analytical assays. However, without wishing to be bound by theory, it is expected that SEQ ID NO. 16 will behave similarly to SEQ ID NO. 15. This is due to the high sequence similarity between SEQ ID NOs. 15 and 16, particularly in the Loop A and Loop B regions.

[0039] Figure 3 Data for DON production by various enzymes are shown. Data for SEQ ID NO. 16 are not shown, as it was not possible to produce sufficient quantities for the required analytical assays. However, without wishing to be bound by theory, it is expected that SEQ ID NO. 16 will behave similarly to SEQ ID NO. 15. This is due to the high sequence similarity between SEQ ID NOs. 15 and 16, particularly in the Loop A and Loop B regions.

[0040] Figure 4 The substrate-specific loops A and B are depicted throughout the sequences of the various enzymes described herein. Black ribbons represent α-helices, and white ribbons represent β-sheets. Dashed boxes indicate the positions of residues involved in the predicted active site, while the residues that comprise the catalytic tetramer are outlined by dotted lines.

[0041] These are also summarized in the Sequence Listing shown herein.

[0042] Figure 5 is a sequence alignment performed using the Clustal W program. A) Alignment of the loop A sequences of SEQ ID NOs. 1-14 with those of SEQ ID NOs. 15 and 16. B) Alignment of the loop B sequences of SEQ ID NOs. 1-14 with those of SEQ ID NOs. 15 and 16. C) Alignment of a portion of the loop B sequences of SEQ ID NOs. 1-14 with those of SEQ ID NOs. 15 and 16, which, without wishing to be bound by theory, is believed to mediate substrate contact. DETAILED DESCRIPTION

[0043] Unexpectedly, the reductase enzymes of any of SEQ ID NOs. 1-14 and 63-101 were found to be more efficient at converting 3-keto-DON to 3-epi-DON than prior art enzymes. In this conversion, the 3-oxy group of a trichothecene (e.g., DON) was more efficiently converted to a 3-hydroxy group than the prior art enzymes of SEQ ID NOs. 15 and 16. This higher efficiency is demonstrated in the Examples. Since most trichothecenes have a characteristic of containing a 3-hydroxy group at the 3-position (discussed below), this group can be converted to a 3-oxy group similar to the reaction observed for DON. Therefore, the enzymes of any of SEQ ID NOs. 1-14 and 63-101 are capable of converting any trichothecene containing a 3-oxy group to a trichothecene containing a hydroxy group.

[0044] In particular, the enzymes of the present invention are capable of highly stereoselective conversions. This means that they can convert a 3-oxyl group to a specific isomer of the 3-hydroxyl group at a higher efficiency than other 3-hydroxyl isomers. Specifically, as shown in the Examples, the polypeptides used in the present invention are capable of converting a 3-oxyl group to a hydroxyl group in the S configuration (3-epi-DON) at a higher efficiency than the conversion to a hydroxyl group in the R configuration (DON).

[0045] It was also unexpectedly discovered that the enzymes of any of SEQ ID NOs. 1-14, 63-101 have sequence identity in two loops, referred to below as Loop A and Loop B, which differ from the enzymes known in the prior art shown in the sequence listing herein. Furthermore, the polypeptides used in the present invention have a common motif for interacting with the substrate, also shown in the sequence listing herein.

[0046] The present invention relates to a composition comprising:

[0047] a) one or more polypeptides comprising or consisting of a sequence of SEQ ID NOs. 1-14 or a sequence having at least 75% sequence identity to any one of SEQ ID NOs. 1-14, and / or a sequence of SEQ ID NOs. 63-101, wherein the polypeptide is capable of converting the substrate 3-keto-DON into products 3-epi-DON and DON in a ratio of 3-epi-DON:DON of at least 25:1; and

[0048] b) Carrier.

[0049] Alternatively or additionally, the present invention relates to a composition comprising:

[0050] a) one or more polypeptides comprising or consisting of a sequence of SEQ ID NOs. 1-14 or a sequence having at least 75% sequence identity to any one of SEQ ID NOs. 1-14, and / or a sequence of SEQ ID NOs. 63-101, wherein the polypeptide is capable of converting the substrate 3-keto-DON to the product 3-epi-DON, and optionally also to the product DON, totaling 100% of the total product, wherein at least 96% of the total product is 3-epi-DON; and

[0051] b) Carrier.

[0052] The composition can be any composition. For example, the composition can be a composition that reduces the amount of 3-keto-DON. The composition can also be a composition that reduces the amount of trichothecenes including 3-oxo-DON. The composition can be, for example, an additive for feed or food. The composition can also be feed or food. The composition can also be an additive for a farming composition or a farming composition. The farming composition can also be a composition that includes a trichothecene such as DON.

[0053] The term "polypeptide" as used herein means peptide, protein, or polypeptide, which are used interchangeably and encompass an amino acid chain of a given length in which the amino acid residues are linked by covalent peptide bonds. The present invention also encompasses amino acids other than the 20 proteinogenic amino acids of the standard genetic code known to those skilled in the art, such as selenocysteine. Such polypeptides include any of the sequences in SEQ ID NOs. 1-14 and / or SEQ ID NOs. 63-101.

[0054] The term polypeptide also refers to, but does not exclude, modifications of the polypeptide. Modifications include glycosylation, acetylation, acylation, phosphorylation, ADP-ribosylation, amidation, covalent attachment of flavin, covalent attachment of a heme moiety, covalent attachment of a nucleotide or nucleotide derivative, covalent attachment of a lipid or lipid derivative, covalent attachment of phosphatidylinositol, cross-linking, cyclization, disulfide bond formation, demethylation, formation of covalent cross-links, formation of cysteine, formation of pyroglutamate, formulation, gamma-carboxylation, glycosylation, GPI anchor formation, hydroxylation, iodination, methylation, myristoylation, oxidation, pegylation, proteolytic processing, phosphorylation, prenylation, racemization, selenoylation, sulfation, transfer RNA-mediated addition of amino acids to proteins such as arginylation, and ubiquitination; see, e.g., PROTEINS-STRUCTURE AND MOLECULAR PROPERTIES, 2nd Ed., T. Creighton, W. H. Freeman and Company, New York (1993); POST-TRANSLATIONAL COVALENT MODIFICATION OF PROTEINS, BC Johnson, Ed., Academic Press, New York (1983), pgs.1-12; Seifter, Meth.Enzymol.182 (1990); 626-646, Rattan, Ann.NY Acad. Sci. 663 (1992); 48-62.

[0055] It is contemplated that one or more polypeptides described herein comprise or consist of a sequence having at least 80%, at least 83%, at least 85%, at least 87%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or more sequence identity to any one of SEQ ID NOs. 1-14. It is also contemplated that one or more polypeptides described herein comprise or consist of a sequence having at least 80%, at least 83%, at least 85%, at least 87%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or more sequence identity to any one of SEQ ID NOs. 1. For example, one or more polypeptides described herein comprise or consist of the sequences of SEQ ID NOs. 63-75.

[0056] It is also contemplated that one or more polypeptides described herein comprise or consist of a sequence having at least 80%, at least 83%, at least 85%, at least 87%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or more sequence identity to any one of SEQ ID NO. 3. For example, one or more polypeptides described herein comprise or consist of the sequences of SEQ ID NOs. 76-88.

[0057] It is also contemplated that one or more polypeptides described herein comprise or consist of a sequence having at least 80%, at least 83%, at least 85%, at least 87%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or more sequence identity to any one of SEQ ID NO. 6. For example, one or more polypeptides described herein comprise or consist of the sequences of SEQ ID NOs. 89-101.

[0058] The present invention also encompasses one or more polypeptides described herein comprising or consisting of a sequence having at least 75% sequence identity to any of SEQ ID NOs. 1-14, including one or more amino acid substitutions compared to any of SEQ ID NOs. 1-14. To determine the sequence identity of two sequences, for example, the sequence of interest is compared to any of SEQ ID NOs. 1-14. For example, one or more polypeptides described herein may comprise or consist of a sequence as set forth in SEQ ID NOs. 63-101, or a sequence having at least 75%, at least 80%, at least 83%, at least 85%, at least 87%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or more sequence identity to any of SEQ ID NOs. 63-101.

[0059] According to the present invention, the term "identical" or "percent identity" of two or more polypeptide sequences such as SEQ ID NO.1-14 and / or 63-101 in the context refers to two or more sequences or subsequences that are identical, or, when comparing and aligning to determine maximum correspondence using sequence comparison algorithms known in the art or by manual alignment and visual inspection over a comparison window or in a specified area of ​​measurement, two or more sequences or subsequences with a specific nucleotide similarity percentage (e.g., at least 85%, 90%, 95%, 96%, 97%, 98% or 99% identity). Sequences with, for example, 80% to 95% or greater sequence identity are considered to be substantially identical. This definition also applies to the complement of a test sequence. Those skilled in the art know how to determine the percent identity between sequences using algorithms, such as those based on the CLUSTALW computer program (Thompson Nucl. Acids Res. 2 (1994), 4673-4680) or FASTDB (Brutlag Comp. App. Biosci. 6 (1990), 237-245), known in the art.

[0060] Those skilled in the art can also utilize the BLAST and BLAST 2.6 algorithms (Altschul Nucl. Acids Res. 25 (1977), 3389-3402). The BLASTP program for amino acid sequences uses the default values ​​(default), a word length (W) of 6, an expectation of 10, and a comparison of both strands. In addition, the BLOSUM62 scoring matrix (Henikoff Proc. Natl. Acad. Sci., USA, 89, (1989), 10915; Henikoff and Henikoff (1992) 'Aminoacid substitution matrices from protein blocks.' Proc Natl Acad Sci USA. 1992 Nov 15; 89(22): 10915-9) can be used.

[0061] For example, BLAST 2.6, which stands for Basic Local Alignment Search Tool (Altschul, Nucl. Acids Res. 25 (1997), 3389-3402; Altschul, J. Mol. Evol. 36 (1993), 290-300; Altschul, J. Mol. Biol. 215 (1990), 403-410), can be used for local sequence alignment searches.

[0062] As described herein, one or more polypeptides may have a specific sequence identity with any one of SEQ ID NOs. 1-14. This indicates that these polypeptides may also be fragments and thus include amino acid deletions with respect to any one of SEQ ID NOs. 1-14.

[0063] As described herein, one or more polypeptides may include amino acid substitutions relative to any of SEQ ID NOs. 1-14. For example, a polypeptide may include or consist of one or more conservative amino acid substitutions compared to any of SEQ ID NOs. 1-14. It is also contemplated that a polypeptide may include or consist of one or more highly conservative amino acid substitutions compared to any of SEQ ID NOs. 1-14.

[0064] As used herein, "conservative" substitutions are those listed below under "Exemplary Substitutions" in Table 1. As used herein, "highly conservative" substitutions are those shown below under the heading "Preferred Substitutions" in Table 1.

[0065]

[0066]

[0067]

[0068]

[0069] Table 1. Conservative amino acid substitutions

[0070] The polypeptide comprising conservative amino acid substitutions may be any of SEQ ID NOs. 63-101.

[0071] The polypeptides of the present invention are capable of converting the substrate 3-keto-DON into the products 3-epi-DON (referred to herein as 3-epi-DON) and DON (referred to herein as DON) in a ratio of at least 25:1 (3-epi-DON / DON). The reaction described herein is described in the following reaction scheme, which is derived from Hassan et al. (2017) "The enzymatic epimerization of DON by Devosia mutans proceeds through the formation of 3-keto-DON intermediate." Scientific Reports 7, article number 6929.

[0072]

[0073] Reaction scheme: Conversion of 3-keto DON to its two isomers 3-epi-DON and DON, adapted from Hassan et al. (2017).

[0074] As can be seen from the reaction scheme above, 3-keto-DON can be converted into two isomers, namely DON (R configuration) and 3-epi-DON (S configuration). A ratio of at least 25:1 indicates that the polypeptide of the present invention produces a higher amount of isomer 3-epi-DON from 3-keto-DON than isomer DON. Therefore, this ratio indicates that 50 times more isomer 3-epi-DON is produced than isomer DON. This ratio is calculated by dividing the amount of isomer 3-epi-DON produced by the amount of isomer DON produced.

[0075] The invention encompasses the ability of the polypeptides described herein to convert the substrate 3-keto-DON into the products 3-epi-DON and DON in a 3-epi-DON:DON ratio of at least 25:1, at least 30:1, at least 40:1, at least 50:1, at least 60:1, at least 70:1, at least 80:1, at least 90:1, at least 100:1, at least 150:1, at least 200:1, 250:1, 300:1, 350:1, 400:1, 500:1, 600:1, 700:1, 800:1, 900:1, 1000:1 or greater.

[0076] Obviously, if DON is absent and only 3-epi-DON is obtained, this ratio cannot be calculated. This is because division by the number 0 is mathematically impossible. However, the present invention also relates to polypeptides in which 3-epi-DON is 100% stereoselective. Therefore, even if the division is by the number 0, such polypeptides are still included in the present invention. However, if the division is 0 because only DON is obtained, such polypeptides are not included in the present invention.

[0077] A method for measuring whether a polypeptide is able to convert the substrate 3-keto-DON into the products 3-epi-DON and DON in a 3-epi-DON:DON ratio of at least 25:1 is described in detail in the Examples.

[0078] The present invention also contemplates that the polypeptides used in the present invention are capable of converting the substrate 3-keto-DON into the product 3-epi-DON, and optionally also into the product DON, for a total of 100% total product, of which at least 96% is 3-epi-DON. As can be seen from the reaction scheme above, 3-keto-DON can be converted into two isomers, namely 3-epi-DON and DON. Therefore, these two isomers represent the total product. By definition, the total product equals 100%. Because the polypeptides of the present invention are highly stereoselective, one isomer (herein, 3-epi-DON) is obtained with greater efficiency than the other (herein, DON). Specifically, at least 96% of the total product (equal to 100%) obtained is 3-epi-DON.

[0079] The present invention also contemplates that at least 96.5%, 97%, 97.5%, 98%, 98.5%, 99%, 99.5%, 99.9% or 100% of the total product (equal to 100%) is 3-epi-DON.

[0080] Thus, the polypeptides of any one of SEQ ID NOs. 1-14 and / or any one of SEQ ID NOs. 63-101 are highly efficient in converting 3-keto-DON to 3-epi-DON. This is also evident from the fact that only a small amount of polypeptide is required to convert 1 μmol of 3-epi-DON per minute. This measurement demonstrates that the polypeptides described herein are highly active.

[0081] Thus, the present invention encompasses an amount of polypeptide that is at most 7 mg, at most 6 mg, at most 5 mg, at most 3 mg, at most 2 mg, at most 1 mg, or less when 3-epi-DON is produced from 3-keto-DON at a rate of 1 μmol 3-epi-DON per minute. Similarly, the polypeptides of the present invention are capable of converting 3-keto-DON to 3-epi-DON with a specific activity of at least 0.1 μmol / min / mg, at least 0.2 μmol / min / mg, at least 0.3 μmol / min / mg, at least 0.4 μmol / min / mg, at least 0.5 μmol / min / mg, at least 0.6 μmol / min / mg, at least 0.8 μmol / min / mg, at least 1.0 μmol / min / mg, at least 1.2 μmol / min / mg, at least 1.4 μmol / min / mg, or at least 1.6 μmol / min / mg.

[0082] On the other hand, the polypeptides described herein have low activity in converting 3-epi-DON to DON. Therefore, a large amount of polypeptide is required to obtain DON. Specifically, when DON is obtained from 3-keto-DON at a rate of 1 μmol DON per minute, the amount of polypeptide is at least 600 mg, at least 700 mg, at least 800 mg, at least 900 mg, at least 1000 mg, at least 1100 mg, at least 1300 mg, at least 1500 mg, at least 1600 mg, at least 1700 mg, at least 1800 mg, at least 1900 mg, at least 2000 mg, or more.

[0083] The present invention also encompasses the ability of the polypeptides described herein to convert 3-keto-DON to DON at a specific activity of at most 0.0015 μmol / min / mg, at most 0.0010 μmol / min / mg, at most 0.0009 μmol / min / mg, at most 0.0008 μmol / min / mg, at most 0.0007 μmol / min / mg, at most 0.0006 μmol / min / mg, or less.

[0084] The polypeptide of any one of SEQ ID NOs. 1-14 and / or any one of SEQ ID NOs. 63-101 capable of converting 3-keto-DON to 3-epi-DON and optionally also to DON may be a reductase. For example, the polypeptide may be an aldehyde-ketone reductase.

[0085] The polypeptides described herein not only have the same functional characteristics but also share structural features. Furthermore, these structural features are different from the structural features of the enzymes known in the prior art depicted in the sequence listing herein. Without being bound by theory, the inventors anticipate that these structural features (ring A and / or ring B) are important for the substrate specificity and activity of the enzyme because rings A and B comprise the active site portion. Since the amino acid histidine (H) of the catalytic tetrad is located outside ring A, this ring is also very likely to contribute to the activity of the enzyme of the present invention. Furthermore, without wishing to be bound by theory, it is believed that ring B has a greater effect on substrate specificity than ring A. In particular, the motif X8-G-X9-X10-X11-X12-X13-X14-X15-X16 in ring B described herein interacts with the substrate. The substrate can be, in particular, a 3-oxy-trichothecene, such as 3-keto DON or a redox factor.

[0086] It is therefore plausible that these common structural features found in all polypeptides used in the present invention are responsible for the high stereoselectivity and, for example, the high specific activity of the conversion of 3-ketoDON to 3-epi-DON.

[0087] For example, the present invention encompasses polypeptides for use herein that include or consist of more than 353, 350, 348, or 351 amino acids. This is in contrast to the prior art enzymes of SEQ ID NOs. 15 and 16, which have 343 amino acids. It is also contemplated that polypeptides include or consist of more than 345, 346, 347, 348, 349, 350, 351, 352, 353, or more amino acids. Polypeptides include or consist of more than 348, 350, 351, 353 amino acids. Polypeptides include or consist of 344-353 amino acids (preferably 348-353 amino acids).

[0088] The polypeptide used in the present invention may include one or more loops in the amino acid sequence. As used herein, a "loop" is an irregular structure in a protein / polypeptide that is responsible for the direction change and connection of the main secondary structure α-helix, β-pleated strand. The loop can be an unstructured and randomly constructed coil, turn, and strand. The difference from the main secondary structure is the lack of regular geometric repetition of ψ (psi) angles and φ (phi) angles in the residues along the structure. In enzymes, they are part of or construct substrate binding sites and active sites. Typically, the sequences of these structures are not conserved. The structural features of the loop are also described in Kundert K, Kortemme T. (2019).

[0089] The present invention also encompasses polypeptides described herein that include loops in the amino acid positions corresponding to the following amino acid positions:

[0090] Positions 125 to 133 of SEQ ID NO. 1-10 or 63-101;

[0091] Positions 127 to 135 of SEQ ID NO. 13 or 14;

[0092] Positions 124 to 132 of SEQ ID NO. 11 or 12. This loop is also described herein as loop A. Loop A disclosed herein includes a sequence located downstream of the motif ID / E-LYQ contained in β-sheet 4 and upstream of the motif I / M / LE / DE / DTL / M contained in α-helix 4, for example, SEQ ID NO. 1-14, 63-101.

[0093] It is also contemplated that polypeptides in the amino acid positions corresponding to the following amino acid positions include or consist of sequences that do not contain alanine (Ala / A):

[0094] Positions 125 to 133 of SEQ ID NO. 1-10 or 63-101;

[0095] Positions 127 to 135 of SEQ ID NO. 13 or 14;

[0096] Positions 124 to 132 of SEQ ID NO. 11 or 12.

[0097] Thus, the polypeptides described herein do not include alanine in the loop A region.

[0098] The present invention also contemplates that the polypeptides used in the present invention may include the sequence X1-H-X2-WD-X3-X4-TP in the amino acid positions corresponding to the following amino acid positions:

[0099] Positions 125 to 133 of SEQ ID NO. 1-10 or 63-101;

[0100] Positions 127 to 135 of SEQ ID NO. 13 or 14;

[0101] Positions 124 to 132 of SEQ ID NO. 11 or 12.

[0102] in,

[0103] X1 is not M;

[0104] X2 is not A;

[0105] X3 is not A;

[0106] X4 is not V.

[0107] Thus, a polypeptide described herein may include a sequence comprising the sequence X1-H-X2-WD-X3-X4-TP, wherein

[0108] X1 is not M;

[0109] X2 is not A;

[0110] X3 is not A;

[0111] X4 is not V.

[0112] For example, the polypeptides described herein comprise or consist of a sequence comprising the sequence X1-H-X2-WD-X3-X4-TP at amino acid positions corresponding to amino acid positions 125 to 133 of SEQ ID NOs. 1-10 or 63-101, 127 to 135 of SEQ ID NOs. 13 or 14, or 124 to 132 of SEQ ID NOs. 11 or 12, wherein

[0113] X1 is V, I, L, M, F or A, preferably V;

[0114] X2 is E, D, Q or N, preferably E or Q;

[0115] X3 is G or A, preferably G;

[0116] X4 is M, L, F, I, R, K, Q, N, E, V, A or norleucine, preferably M, R, Q or L.

[0117] Thus, a polypeptide described herein may include a sequence comprising the sequence X1-H-X2-WD-X3-X4-TP, wherein

[0118] X1 is V, I, L, M, F or A, preferably V;

[0119] X2 is E, D, Q or N, preferably E or Q;

[0120] X3 is G or A, preferably G;

[0121] X4 is M, L, F, I, R, K, Q, N, E, V, A or norleucine, preferably M, R, Q or L.

[0122] It is also contemplated that the polypeptides described herein may comprise or consist of a sequence of any one of SEQ ID NOs. 20-24 or a sequence having at least 80%, 85%, 90%, 95%, 98%, 99% or greater sequence identity to any one of SEQ ID NOs. 20-24.

[0123] Additionally or alternatively, the polypeptides described herein include loops at amino acid positions corresponding to the following amino acid positions:

[0124] Positions 211 to 244 of SEQ ID NO. 1-8, 10, or 63-101;

[0125] Positions 211 to 245 of SEQ ID NO. 9;

[0126] Positions 210 to 243 of SEQ ID NO. 11 or 12;

[0127] Positions 213 to 247 of SEQ ID NO. 13 or 14. This loop is also described herein as loop B. Loop B disclosed herein starts from a serine located in the common motif WSPL-XG, ends before an α-helix having the sequence XXR / K / HT / LW / YXI / V / LI / V / L, and consists of 35 residues or less, for example, SEQ ID NO. 1-14, 63-101.

[0128] It is also contemplated that the polypeptides described herein may include sequences that do not include methionine (Met / M) in the amino acid positions corresponding to the following amino acid positions:

[0129] Positions 211 to 244 of SEQ ID NO. 1-8, 10, or 63-101;

[0130] Positions 211 to 245 of SEQ ID NO. 9;

[0131] Positions 210 to 243 of SEQ ID NO. 11 or 12;

[0132] Positions 213 to 247 of SEQ ID NO. 13 or 14.

[0133] For example, the polypeptides described herein may include a sequence comprising the sequence SPL-X5-GG-X6-L-X7-GK at amino acid positions corresponding to amino acid positions 211 to 244 of SEQ ID NO. 1-8, 10, or 63-101, positions 211 to 245 of SEQ ID NO. 9, positions 210 to 243 of SEQ ID NO. 11 or 12, or positions 213 to 247 of SEQ ID NO. 13 or 14, wherein

[0134] X5 is not G;

[0135] X6 is L, I, V, M, A, Y, F, norleucine, or W;

[0136] The X7 is not an A or a T.

[0137] Thus, a polypeptide described herein may include a sequence comprising the sequence SPL-X5-GG-X6-L-X7-GK, wherein:

[0138] X5 is not G;

[0139] X6 is L, I, V, M, A, Y, F, norleucine, or W;

[0140] The X7 is not an A or a T.

[0141] The polypeptides described herein may include a sequence comprising the sequence SPL-X5-GG-X6-L-X7-GK at amino acid positions corresponding to amino acid positions 211 to 244 of SEQ ID NO. 1-8, 10, or 63-101, positions 211 to 245 of SEQ ID NO. 9, positions 210 to 243 of SEQ ID NO. 11 or 12, or positions 213 to 247 of SEQ ID NO. 13 or 14, wherein

[0142] X5 is A, V, L, G or I, preferably A;

[0143] X6 is L, I, V, M, A, Y, F, norleucine or W, preferably L or W;

[0144] X7 is S or T, preferably S.

[0145] Thus, a polypeptide described herein may include a sequence comprising the sequence SPL-X5-GG-X6-L-X7-GK, wherein:

[0146] X5 is A, V, L or I, preferably A;

[0147] X6 is L, I, V, M, A, Y, F, norleucine or W, preferably L or W;

[0148] X7 is S or T, preferably S.

[0149] It is also contemplated that the polypeptides described herein may comprise or consist of a sequence of any one of SEQ ID NOs. 33-46 or a sequence having at least 80%, 85%, 90%, 95%, 98%, 99% or greater sequence identity to any one of SEQ ID NOs. 33-46.

[0150] For example, a polypeptide described herein can include a sequence comprising the sequence X8-G-X9-X10-X11-X12-X13-X14-X15-X16 in the amino acid positions corresponding to the following amino acid positions:

[0151] Positions 211 to 244 of SEQ ID NO. 1-8, 10, or 63-101;

[0152] Positions 211 to 245 of SEQ ID NO. 9;

[0153] Positions 210 to 243 of SEQ ID NO. 11 or 12;

[0154] SEQ ID NO. 13 or 14, 213 to 247, wherein

[0155] X8 is G, R, K, Q, N, A, V, L, I, S, T, E, D;

[0156] X9 is not M or G;

[0157] X10 is not E;

[0158] X11 is not S;

[0159] X12 is not Y;

[0160] X13 is not G or A;

[0161] X14 is not P;

[0162] X15 is R, K, Q, N, P, A, Y, W, F, T, S, H;

[0163] X16 is not N.

[0164] Thus, the polypeptides described herein may include a sequence comprising the sequence X8-G-X9-X10-X11-X12-X13-X14-X15-X16, wherein

[0165] X8 is G, R, K, Q, N, A, V, L, I, S, T, E, D;

[0166] X9 is not E;

[0167] X10 is not E;

[0168] X11 is not S;

[0169] X12 is not Y;

[0170] X13 is not G or A;

[0171] X14 is not P;

[0172] X15 is R, K, Q, N, P, A, Y, W, F, T, S, H; and

[0173] X16 is not N.

[0174] It is also contemplated that the polypeptides described herein may include a sequence comprising the sequence X8-G-X9-X10-X11-X12-X13-X14-X15-X16 in the amino acid positions corresponding to the following amino acid positions:

[0175] Positions 211 to 244 of SEQ ID NO. 1-8, 10, or 63-101;

[0176] Positions 211 to 245 of SEQ ID NO. 9;

[0177] Positions 210 to 243 of SEQ ID NO. 11 or 12;

[0178] SEQ ID NO. 13 or 14, 213 to 247, wherein

[0179] X8 is G, R, K, Q, N, A, V, L, I, S, T, E, D, preferably G, R, A, E, more preferably G;

[0180] X9 is W, S, T, F, L, V, I, A or Y, preferably W, S, F, more preferably F;

[0181] X10 is T, S, R, Q, N or K, preferably T, R or K, more preferably R;

[0182] X11 is E, D or Q, preferably E;

[0183] X12 is P or A, preferably P;

[0184] X13 is P or A, preferably P;

[0185] X14 is I, L, M, A, F or V, preferably I or V, more preferably V;

[0186] X15 is R, K, Q, N, P, A, H, W, F, T, S or Y, preferably R, P, H or Y, more preferably P;

[0187] X16 is D, E or N, preferably D.

[0188] Thus, the polypeptides described herein may include a sequence comprising the sequence X8-G-X9-X10-X11-X12-X13-X14-X15-X16, wherein

[0189] X8 is G, R, K, Q, N, A, V, L, I, S, T, E, D, preferably G, R, A, E, more preferably G;

[0190] X9 is W, S, T, F, L, V, I, A or Y, preferably W, S, F, more preferably F;

[0191] X10 is T, S, R, Q, N or K, preferably T, R or K, more preferably R;

[0192] X11 is E, D or Q, preferably E;

[0193] X12 is P or A, preferably P;

[0194] X13 is P or A, preferably P;

[0195] X14 is I, L, M, A, F or V, preferably I or V, more preferably V;

[0196] X15 is R, K, Q, N, P, A, H, W, F, T, S or Y, preferably R, P, H or Y, more preferably P;

[0197] X16 is D, E or N, preferably D.

[0198] It is also contemplated that the polypeptides described herein may comprise or consist of a sequence of any one of SEQ ID NOs. 25-32 or a sequence having at least 80%, 85%, 90%, 95%, 98%, 99% or greater sequence identity to any one of SEQ ID NOs. 25-32. These sequences represent sequences capable of contacting a substrate.

[0199] It is also contemplated that the polypeptides described herein may include a sequence comprising the sequence of any one of SEQ ID NOs. 25-32 and / or a sequence as shown in any one of SEQ ID NOs. 33-46.

[0200] The present invention also encompasses that the polypeptides described herein may include a sequence comprising any one of SEQ ID NOs. 25-32 and / or 33-46, and alternatively, a sequence as shown in any one of SEQ ID NOs. 20-24.

[0201] It is also contemplated that the polypeptides described herein comprise or consist of the sequence shown in any one of SEQ ID NOs. 1-14, 63-101.

[0202] The polypeptides described herein can be present in a composition having a carrier. The carrier can be any suitable carrier. Further, the composition can include 1, 2, 3, 4, 5, 6 or more carriers. The carrier can be an edible component, preferably a non-toxic component and / or a component that provides a texture.

[0203] For example, the carrier can be a solid or a liquid. Typical solids include food / feed supplements, dietary supplements, health products and / or pharmaceuticals. Typical food / feed supplements include, among others, food / feed additives, vitamins, minerals, amino acids, essential fatty acids, dietary fiber, trace elements, minerals, antioxidants, plant extracts and herbal extracts.

[0204] The carrier can also be a carrier for the enzyme. The carrier for the enzyme can be of inorganic and organic origin. Potential inorganic materials for enzyme immobilization are silica (sol-gel silica, fumed silica, colloidal silica nanoparticles and silica gel) and different oxides such as titanium oxide, aluminum oxide and zirconium oxide. In addition, clay materials such as bentonite, halloysite, kaolinite, montmorillonite, sepiolite and calcium apatite can be used. In addition, carbon-based materials such as activated carbon and charcoal are known to be efficient enzyme immobilizers. Organic enzyme carriers can be not only biopolymers but also synthetic polymers. Biopolymers include carbohydrates and proteins. Typical examples are maltodextrin, trehalose, inulin, collagen, cellulose, keratin, carrageenan, chitin, chitosan and sodium alginate. As examples of synthetic polymers, polyaniline, polyamide, polystyrene, polyurethane, polypropylene, polyvinyl alcohol and ion exchange resins should be mentioned. The enzyme carrier can also be the carrier described in Zdarta et al. (2018).

[0205] Carrier can also be liquid.Typical liquid comprises H o, aqueous solution, saline solution (for example buffer solution), gel, viscous preparation, fat or oil.Preferred aqueous solution contains H o and other substances such as buffer substances and / or polyols such as polyethylene oxide (PAO), polyvinyl alcohol (PVA), polyethylene-maleic anhydride, polystyrene-malic anhydride, dextran, cellulose, chitosan hydrolyzate, starch, glycogen, sorbitol, agarose and derivatives thereof, guar gum, pullulan, inulin, xanthan gum, carrageenan, pectin, alginic acid hydrolyzate, biopolymer, sorbitol, glycerol, cellobiose and monopropylene glycol (MPG).

[0206] The composition can be a composition for reducing the amount of DON. In this case, the composition further includes an enzyme capable of converting DON into 3-keto-DON. The composition can also be a composition for reducing the amount of trichothecenes. In this case, the composition further includes a trichothecenes capable of converting trichothecenes into trichothecenes comprising 3-oxygen groups. The composition described herein can further include one or more enzymes capable of converting DON into 3-keto-DON and / or converting trichothecenes comprising 3-oxygen groups into trichothecenes comprising 3-hydroxyl groups. Typical enzymes that can be used for the composition are described in particular in WO2016 / 154640 or WO2019 / 046954. Therefore, the enzyme for converting DON into 3-keto-DON can include the sequence disclosed in SEQ ID No.1 of WO2016 / 154640 (SEQ ID NO:61 herein or SEQ ID NO.7 of WO2019 / 046954 (SEQ ID NO.62 herein)) or consist of the sequence. Therefore, the compositions described herein further include an enzyme comprising or consisting of SEQ ID NO. 61 and / or 62.

[0207] The present invention also relates to a polypeptide comprising the sequence of SEQ ID NO.1 or a sequence having at least 75% sequence identity to SEQ ID NO.1. Further, the present invention relates to a polypeptide comprising the sequence of SEQ ID NO.3 or a sequence having at least 75% sequence identity to SEQ ID NO.3. In addition, the present invention relates to a polypeptide comprising the sequence of SEQ ID NO.6 or a sequence having at least 75% sequence identity to SEQ ID NO.6. Further, the present invention relates to a polypeptide comprising the sequence of any one of SEQ ID NOs.63-101 or a sequence having at least 75% sequence identity to any one of SEQ ID NOs.63-101.

[0208] The present invention also relates to the use of one or more polypeptides described herein in converting 3-keto-DON to 3-epi-DON, wherein the one or more polypeptides comprise or consist of the following sequences: a sequence of SEQ ID NOs. 1-14 or a sequence having at least 75% sequence identity to any of SEQ ID NOs. 1-14, and / or a sequence of SEQ ID NOs. 63-101.

[0209] Furthermore, the present invention relates to the use of one or more polypeptides in the preparation of a feed / food additive or a feed / food composition, wherein the one or more polypeptides comprise or consist of the following sequences: a sequence in SEQ ID NO. 1-14 or a sequence having at least 75% sequence identity with any one of SEQ ID NO. 1-14, and / or a sequence in SEQ ID NO. 63-101, wherein the polypeptide is capable of converting the substrate 3-keto-DON into products 3-epi-DON and DON in a ratio of at least 25:1 (3-epi-DON:DON).

[0210] Additionally or alternatively, the present invention relates to the use of one or more polypeptides in the preparation of a feed / food additive or feed / food composition, wherein the one or more polypeptides comprise or consist of a sequence of SEQ ID NO. 1-14 or a sequence having at least 75% sequence identity to any one of SEQ ID NO. 1-14, and / or a sequence of SEQ ID NO. 63-101, wherein the polypeptide is capable of converting the substrate 3-keto-DON into the product 3-epi-DON, and optionally also into the product DON, totaling 100% of the total product, wherein at least 96% of the total product is 3-epi-DON.

[0211] The feed / food additive or feed / food composition of the present invention may further comprise an enzyme as described herein, which is capable of converting DON to 3-keto-DON and / or converting a trichothecene comprising a 3-oxol group to a trichothecene comprising a 3-hydroxyl group.

[0212] Methods for preparing such feed / food additives or feed / food compositions are known to the person skilled in the art and are described, inter alia, in WO 99 / 35240.

[0213] The present invention also relates to the use of one or more polypeptides comprising or consisting of the following sequences: SEQ ID NOs. 1-14, or sequences having at least 75% sequence identity to any of SEQ ID NOs. 1-14, and / or SEQ ID NOs. 63-101, in the production of biogas, bioethanol, or sugar, preferably from sugarcane or sugar beet, wherein the polypeptides are capable of converting the substrate 3-keto-DON into products 3-epi-DON and DON in a 3-epi-DON:DON ratio of at least 25:1. In such uses, the compositions or polypeptides of the present invention can be added to a composition comprising a trichothecene, such as DON.

[0214] Additionally or alternatively, the present invention relates to the use of one or more polypeptides comprising or consisting of a sequence of SEQ ID NOs. 1-14 or a sequence having at least 75% sequence identity to any of SEQ ID NOs. 1-14, and / or a sequence of SEQ ID NOs. 63-101, in the production of biogas, bioethanol or sugar, preferably from sugar cane or sugar beet, wherein the polypeptide is capable of converting the substrate 3-keto-DON to the product 3-epi-DON, and optionally also to the product DON, totaling 100% of the total product, wherein at least 96% of the total product is 3-epi-DON. In such use, the composition or polypeptide of the present invention may be added to a composition comprising a trichothecene, such as DON.

[0215] The present invention also relates to a feed additive or food additive or feed or food comprising one or more polypeptides comprising or consisting of a sequence of SEQ ID NOs. 1-14 or a sequence having at least 75% sequence identity to any one of SEQ ID NOs. 1-14, and / or a sequence of SEQ ID NOs. 63-101, wherein the polypeptide is capable of converting the substrate 3-keto-DON into products 3-epi-DON and DON in a 3-epi-DON:DON ratio of at least 25:1.

[0216] Additionally or alternatively, the feed additive or food additive or feed or food comprises one or more polypeptides comprising or consisting of a sequence of SEQ ID NO. 1-14 or a sequence having at least 75% sequence identity to any one of SEQ ID NO. 1-14, and / or a sequence in SEQ ID NO. 63-101, wherein the polypeptide is capable of converting the substrate 3-keto-DON into the product 3-epi-DON, and optionally also into the product DON, totaling 100% of the total product, wherein at least 96% of the total product is 3-epi-DON.

[0217] The feed / food additive of the present invention may also comprise one or more enzymes as described herein, which are capable of converting DON to 3-keto-DON and / or converting a trichothecene comprising a 3-oxo group to a trichothecene comprising a 3-hydroxy group.

[0218] The present invention also relates to an additive for a composition, preferably a tillage composition, comprising 3-keto-DON or DON, the additive comprising one or more polypeptides comprising or consisting of the following sequences: a sequence of SEQ ID NOs. 1-14 or a sequence having at least 75% sequence identity to any of SEQ ID NOs. 1-14, and / or a sequence of SEQ ID NOs. 63-101, wherein the polypeptide is capable of converting the substrate 3-keto-DON into products 3-epi-DON and DON in a 3-epi-DON:DON ratio of at least 25:1.

[0219] Additionally or alternatively, the present invention also relates to an additive for a composition, preferably a tillage composition, comprising 3-keto-DON and / or DON, the additive comprising one or more polypeptides comprising or consisting of the following sequences: a sequence of SEQ ID NO. 1-14 or a sequence having at least 75% sequence identity to any of SEQ ID NO. 1-14, and / or a sequence of SEQ ID NO. 63-101, wherein the polypeptide is capable of converting the substrate 3-keto-DON into the product 3-epi-DON, and optionally also into the product DON, totaling 100% of the total product, wherein at least 96% of the total product is 3-epi-DON.

[0220] Agrarian compositions can be any composition comprising a plant or a part of a plant, such as a seed or wood. Such agrarian composition can include 3-keto-DON and / or DON. The additive according to the present invention can further include one or more enzymes as described herein that are capable of converting DON into 3-keto-DON and / or converting a trichothecene containing a 3-oxy group into a trichothecene containing a 3-hydroxy group.

[0221] The present invention relates to a method for converting a trichothecene comprising a 3-oxyl group into a trichothecene comprising a 3-hydroxyl group, the method comprising contacting one or more polypeptides comprising or consisting of a sequence of SEQ ID NOs. 1-14 or a sequence having at least 75% sequence identity to any one of SEQ ID NOs. 1-14, and / or a sequence of SEQ ID NOs. 63-101 with the trichothecene comprising a 3-oxyl group.

[0222] Trichothecenes have the following common structure:

[0223]

[0224] Those skilled in the art will know which substitutions may be present at R1, R2, R3, R4, and R5. It is known that the toxicity of trichothecenes is attributable (at least) in part to the hydroxyl (-OH) group present at the C-3 atom. Thus, in some trichothecenes, C-3 is attached to a hydroxyl group (R1 = -OH / -hydroxy).

[0225] Trichothecenes comprising a 3-hydroxyl group (3-OH) (located at the R1 position in the above structural formula) are known to those skilled in the art. Non-limiting examples of such trichothecenes include: DON (DON, CAS No. 51481-10-8), T-2 toxin (CAS No. 21259-20-1), HT-2 toxin (CAS No. 26934-87-2), Nivalenol (CAS No. 23282-20-4), Fuseranon X (CAS No. 23255-69-8), Scirpenetriol (CAS No. 2270-41-9), 15-Acetoxyscirpenol (CAS No. 2623-22-5), 4,15-Diacetoxyscirpenol (CAS No. 2270-40-8), Deacetylneosolaniol (CAS No. 74833-39-9), Neosolaniol (CAS No. 36519-25-2), Sporotrichiol (CAS No. 101401-89-2) and Sambucinol (CAS No. 90044-33-0).

[0226] These trichothecenes containing a 3-hydroxyl group (3-OH) can be oxidized to trichothecenes containing a 3-oxyl group (=O; at the R1 position). Enzymes capable of catalyzing this conversion are described herein. Thus, the polypeptides are capable of preferentially converting these trichothecenes containing a 3-oxyl group into one of the two isomers (i.e., the R configuration and the S configuration) of the trichothecenes containing a 3-hydroxyl group (3-OH).

[0227]

[0228] The present invention contemplates that the trichothecenes comprising a 3-hydroxyl group are present in the S configuration.

[0229] Additionally or alternatively, the present invention also relates to a method for converting 3-keto-DON to 3-epi-DON, the method comprising contacting one or more polypeptides comprising or consisting of the following sequence: a sequence of SEQ ID NO. 1-14 or a sequence having at least 75% sequence identity to any one of SEQ ID NO. 1-14 with 3-keto-DON.

[0230] The present invention also relates to a method for reducing the DON content in a composition comprising DON or reducing the toxicity of a composition comprising DON by converting DON into 3-epi-DON, the method comprising:

[0231] a) contacting the composition with an enzyme capable of converting DON into 3-keto-DON, in particular an enzyme as described herein; and

[0232] b) subsequently or simultaneously contacting the composition with one or more polypeptides comprising or consisting of the following sequences: SEQ ID NOs. 1-14 or sequences having at least 75% sequence identity to any of SEQ ID NOs. 1-14. The method may further comprise contacting the composition with at least one quinone cofactor and / or at least one metal ion and / or at least one redox cofactor. The at least one quinone cofactor may be selected from the group consisting of PQQ (pyrroloquinoline quinone (CAS No. 72909-34-3)), tryptophan tryptophan quinone (TTQ, CAS No. 134645-25-3), hydroxydopaquinone (TPQ, CAS No. 64192-68-3), lysine tyrosine quinone (LTQ, CAS No. 178989-72-5), and cysteine ​​tryptophan quinone (CTQ, CAS No. 400616-72-0). A metal ion capable of rapidly and completely binding the quinone cofactor to an enzyme capable of converting DON to 3-keto-DON, preferably an alcohol dehydrogenase SEQ ID NO. 1-3 described in WO2016 / 154640, which can be selected from the following group: Li + 、Na + , K + Mg 2+ , Ca 2+ 、Zn 2+ 、Zn 3+ 、Mn 2+ 、Mn 3+ 、Fe 2+ 、Fe 3+ 、Cu 2+ 、Cu 3+ 、Co 2+ and Co 3 , preferably Ca 2+ and Mg 2+The at least one redox cofactor may be selected from the group consisting of NAD+ / NADP+, phenazine methosulfate (PMS, CAS No. 299-11-6), PMS derivatives, potassium (III) ferrocyanide, sodium (III) ferrocyanide, cytochrome C, coenzyme Q1, coenzyme Q10, methylene blue, and N,N,N',N'-tetramethyl-p-phenylenediamine (TMPD). Examples of PMS derivatives are: 1-hydroxyphenazine, 2-(pentadienyloxy)dihydrophenazine, 5,10-dihydro-9-dimethylallylphenazine-1-carboxylic acid, 5,10-dihydrophenazine-1-carboxylic acid, 5-methylphenazine methylsulfate, 6-acetylphenazine-1-carboxylic acid, benthophoenin, clofazimine, dihydromethanophenazine, esmeraldic acid, esmeraldin B, izumiphenazine AC, Janus Green B cation, methanophenazine pelagiomicin A, phenazine, phenazine-1,6-dicarboxylic acid, acid), phenazine-1-carboxamide, phenazine-1-carboxylic acid, phenosafranine, pyocyanin, saphenamycin, or saphenic acid methylester.

[0233] The present invention also relates to nucleic acid molecules encoding the polypeptides described herein. The nucleic acid can be introduced into or inserted into an expression vector. The term "expression vector" refers to a nucleic acid molecule construct capable of expressing a gene in vivo or in vitro. In particular, it can include a DNA construct suitable for transforming a nucleotide sequence encoding a polypeptide into a host cell for integration into the genome or free extrachromosomal space, and expressing the nucleotide sequence encoding the polypeptide within the cell, and optionally transporting the polypeptide outside the cell.

[0234] Furthermore, the present invention also relates to host cells (e.g., recombinantly isolated host cells) comprising one or more polypeptides comprising or consisting of the following sequences: a sequence of SEQ ID NOs. 1 to 14 or a sequence having at least 75% sequence identity to any of SEQ ID NOs. 1 to 14. The present invention also relates to host cells comprising a nucleic acid sequence encoding a polypeptide described herein.

[0235] The term "host cell" refers to all cells (e.g., recombinant host cells, isolated host cells, or isolated recombinant host cells) containing a nucleotide sequence to be expressed or an expression vector that is capable of producing an enzyme or polypeptide according to the present invention. The term also includes any progeny of a parent cell (which differs from the parent cell due to mutations occurring during replication) as well as recombinant host cells, isolated host cells (e.g., isolated recombinant host cells). In particular, this refers to prokaryotic and / or eukaryotic cells, preferably Pichia pastoris, Escherichia coli, Bacillus subtilis, Streptomyces, Hansenula, Trichoderma, lactic acid bacteria, Aspergillus, plant cells, and / or spores of Bacillus, Trichoderma, or Aspergillus. The name P. pastoris used herein has the same meaning as the name Komagataella pastoris, with P. pastoris being the older name and K. pastoris being the newer systematic name (Yamada et al. (1995)). In particular, the species of Pichia pastoris has recently been redefined as Komagataella phaffii (Kurtzman (2009)). Komagataella phaffii as used herein, for example, relates to the strain Komagataella phaffii CBS 7435, Komagataella phaffii GS115 or Komagataella phaffii JC308.

[0236] The present invention also encompasses host cells that are further capable of expressing cofactors for the polypeptides described herein. The cofactor can be any suitable cofactor. For example, the cofactor is NAD / H or NADP / H.

[0237] The present invention further relates to a plant genetically modified to express one or more polypeptides comprising or consisting of a sequence of SEQ ID NOs. 1 to 14 or a sequence having at least 75% sequence identity to any of SEQ ID NOs. 1 to 14. Typical plants include, inter alia, maize, wheat, barley, rye and oats.

[0238] The present invention also relates to seeds of the plants described herein.

[0239] The present invention also relates to a preparation comprising:

[0240] a) one or more polypeptides comprising or consisting of the following sequence: a sequence of SEQ ID NO. 1-14 or a sequence having at least 75% sequence identity to any one of SEQ ID NO. 1-14, wherein the polypeptide is capable of converting the substrate 3-keto-DON into products 3-epi-DON and DON in a ratio of 3-epi-DON:DON of at least 25:1; and

[0241] b) Carrier.

[0242] Additionally or alternatively, the present invention also relates to a formulation comprising:

[0243] a) one or more polypeptides comprising or consisting of the sequence of SEQ ID NOs. 1-14 or a sequence having at least 75% sequence identity to any one of SEQ ID NOs. 1-14, wherein the polypeptide is capable of converting the substrate 3-keto-DON to the product 3-epi-DON, and optionally also to the product DON, totaling 100% of the total product, wherein at least 96% of the total product is 3-epi-DON; and

[0244] b) Carrier.

[0245] The use of the polypeptides described herein in the compositions described herein enables the production of "preparations" according to the present invention. In one embodiment, a preparation may include a polypeptide or a portion of a polypeptide described herein and other components such as a carrier, an agricultural extract, etc. The preparation may also include other molecules and / or proteins and / or substances, such as residues of a buffer used in the methods of the present invention, due to inefficient purification of the polypeptides described herein.

[0246] The present invention further relates to a polypeptide comprising or consisting of the following sequence: a sequence of SEQ ID NO. 1-14 or a sequence having at least 75% sequence identity with any one of SEQ ID NO. 1-14, wherein the polypeptide is capable of converting the substrate 3-keto-DON into products 3-epi-DON and DON in a ratio of at least 25:1 (3-epi-DON / DON), for use in preventing and / or treating fungal poisoning.

[0247] The present invention also relates to a polypeptide comprising or consisting of the following sequence: a sequence of SEQ ID NO. 1-14 or a sequence having at least 75% sequence identity with any one of SEQ ID NO. 1-14, wherein the polypeptide is capable of converting the substrate 3-keto-DON into the product 3-epi-DON, and optionally also into the product DON, totaling 100% of the total product, wherein at least 96% of the total product is 3-epi-DON, for use in preventing and / or treating mycotoxicosis.

[0248] The present invention also relates to a pharmaceutical composition comprising a polypeptide comprising or consisting of the following sequence: a sequence of SEQ ID NO. 1-14 or a sequence having at least 75% sequence identity with any one of SEQ ID NO. 1-14, wherein the polypeptide is capable of converting the substrate 3-keto-DON into the product 3-epi-DON, and optionally also into the product DON, totaling 100% of the total product, wherein at least 96% of the total product is 3-epi-DON, for use in preventing and / or treating fungal poisoning.

[0249] The pharmaceutical composition may further include a pharmaceutically acceptable carrier.

[0250] The present invention also relates to a method for treating or preventing mold poisoning in a subject, comprising administering one or more polypeptides described herein. Preferably, a therapeutically effective amount of one or more polypeptides described herein is administered. The subject suffers from mold poisoning.

[0251] In some preferred embodiments, the methods or uses of the present invention (e.g., comprising SEQ ID NO: 1) are performed at a pH range of about 4 to about 10 (e.g., pH about 4.5 to about 10, e.g., pH about 6.5) and / or at a temperature (T) range of about 14°C to about 60°C (e.g., T is about 14.9°C to about 36°C, e.g., T is about 25°C to about 31°C), preferably, as described in the Examples section herein (e.g., Examples 4-5).

[0252] It should be noted that, as used herein, the singular forms "a," "an," and "the" include plural forms unless the context clearly dictates otherwise. Thus, for example, reference to "a polypeptide" includes one or more different polypeptides of that type, and reference to "the method" includes equivalent steps and methods known to those skilled in the art, who will be able to make modifications or substitutions to the methods described herein.

[0253] Unless otherwise indicated, the term "at least" preceding a series of elements should be understood to refer to every element in the series. Those skilled in the art will recognize or be able to ascertain using no more than routine experimentation many equivalents to the specific embodiments of the invention described herein.

[0254] Wherever used herein, the term "and / or" includes "and," "or," and "all or any other combination of the elements connected by this term."

[0255] The term "less than" or conversely "greater than" does not include a specific number.

[0256] For example, less than 20 means less than the stated number. Similarly, more than or greater than means more or greater than the stated number, for example, greater than 80% means more or greater than 80% of the stated number.

[0257] Throughout the specification and in the claims that follow, unless the context clearly indicates otherwise, the word "comprise" and variations thereof such as "comprises" and "comprising" should be understood to mean the inclusion of the specified integer or step or group of integers or steps, but not the exclusion of any other integer or step or group of integers or steps. When used herein, the term "comprise" can be replaced by the term "containing" or "including", or sometimes, when used herein, it can be replaced by the term "having". When used herein, "consisting of" excludes any element, step or element not specified.

[0258] The term "including" means "including but not limited to." "Including" and "including but not limited to" are used interchangeably.

[0259] It should be understood that the present invention is not limited to the specific methods, procedures, materials, reagents, and substances described herein, which may vary. The technical terminology used herein is intended only to describe specific embodiments and is not intended to limit the scope of the present invention, which is defined solely by the claims.

[0260] All publications cited in this specification (including all patents, patent applications, scientific publications, instructions for use, etc.) are incorporated herein by reference in their entirety, whether above or below. Nothing herein is to be construed as an admission that the present invention is not entitled to advance disclosure due to prior invention. To the extent that material included by reference conflicts or is inconsistent with this specification, this specification supersedes any such material.

[0261] The contents of all documents and patent documents cited herein are hereby incorporated by reference in their entirety.

[0262] The present invention is also characterized by the following items.

[0263] 1. A composition comprising:

[0264] a) one or more polypeptides comprising or consisting of the sequence of SEQ ID NOs. 1-14, or a sequence having at least 75% (e.g., at least 90.7%, at least 90.8%, at least 90.9%, at least 91%, at least 92%, at least 93%, at least 93.5%, at least 93.6%, at least 93.7%, at least 93.8%, at least 93.9%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100%) sequence identity to any one of SEQ ID NOs. 1-14, or the sequence of SEQ ID NOs. 63-101, wherein the polypeptide is capable of converting the substrate 3-keto-DON into the products 3-epi-DON and DON in a ratio of at least 25:1 (3-epi-DON:DON); and

[0265] b) Carrier.

[0266] 2. A composition comprising:

[0267] a) one or more polypeptides comprising or consisting of the sequence of SEQ ID NOs. 1-14, or a sequence having at least 75% (e.g., at least 90.7%, at least 90.8%, at least 90.9%, at least 91%, at least 92%, at least 93%, at least 93.5%, at least 93.6%, at least 93.7%, at least 93.8%, at least 93.9%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100%) sequence identity to any one of SEQ ID NOs. 1-14, or the sequence of SEQ ID NOs. 63-101, wherein the polypeptide is capable of converting the substrate 3-keto-DON to the product 3-epi-DON, and optionally also to the product DON, totaling 100% total product, wherein at least 96% of the total product is 3-epi-DON; and

[0268] b) Carrier.

[0269] 3. The composition of any one of the preceding items, wherein the polypeptide is a reductase.

[0270] 4. The composition of any preceding item, wherein the polypeptide comprises greater than 344 amino acids.

[0271] 5. The composition of any one of the preceding items, wherein the polypeptide comprises a loop in the amino acid position corresponding to the following amino acid position:

[0272] Positions 125 to 133 of SEQ ID NO. 1-10 or 63-101;

[0273] Positions 127 to 135 of SEQ ID NO. 13 or 14;

[0274] Positions 125 to 132 of SEQ ID NO. 11 or 12.

[0275] 6. The composition of any one of the preceding items, wherein the polypeptide comprises a sequence that does not comprise methionine (Met / M) in the amino acid positions corresponding to the following amino acid positions:

[0276] Positions 125 to 133 of SEQ ID NO. 1-10 or 63-101;

[0277] Positions 127 to 135 of SEQ ID NO. 13 or 14;

[0278] Positions 124 to 132 of SEQ ID NO. 11 or 12.

[0279] 7. The composition of any of the preceding items, wherein the polypeptide comprises a sequence comprising the sequence X1-H-X2-WD-X3-X4-TP,

[0280] in

[0281] X1 is not M;

[0282] X2 is not A;

[0283] X3 is not A;

[0284] X4 is not V.

[0285] 8. The composition of any one of the preceding items, wherein the polypeptide comprises a sequence comprising the sequence X1-H-X2-WD-X3-X4-TP,

[0286] in

[0287] X1 is V, I, L, M, F or A, preferably V;

[0288] X2 is E, D, Q or N, preferably E or Q;

[0289] X3 is G or A, preferably G;

[0290] X4 is M, L, F, I, R, K, Q, N, E, V, A, or norleucine, preferably M, R, Q or L.

[0291] 9. The composition of any one of the preceding items, wherein the polypeptide comprises a loop in the amino acid position corresponding to the following amino acid position:

[0292] Positions 211 to 244 of SEQ ID NO. 1-8, 10 or 63-101;

[0293] Positions 211 to 245 of SEQ ID NO. 9;

[0294] Positions 210 to 243 of SEQ ID NO. 11 or 12;

[0295] Positions 213 to 247 of SEQ ID NO. 13 or 14.

[0296] 10. The composition of any one of the preceding items, wherein the polypeptide comprises a sequence that does not comprise methionine (Met / M) in the amino acid positions corresponding to the following amino acid positions:

[0297] Positions 211 to 244 of SEQ ID NO. 1-8, 10 or 63-101;

[0298] Positions 211 to 245 of SEQ ID NO. 9;

[0299] Positions 210 to 243 of SEQ ID NO. 11 or 12;

[0300] Positions 213 to 247 of SEQ ID NO. 13 or 14.

[0301] 11. The composition of any one of the preceding items, wherein the polypeptide comprises a sequence comprising the sequence SPL-X5-GG-X6-L-X7-GK, wherein

[0302] X5 is not G;

[0303] X6 is L, I, V, M, A, Y, F, norleucine, or W;

[0304] The X7 is not an A or a T.

[0305] 12. The composition of any one of the preceding items, wherein the polypeptide comprises a sequence comprising the sequence SPL-X5-GG-X6-L-X7-GK, wherein

[0306] X5 is A, V, G, L or I, preferably A;

[0307] X6 is L, I, V, M, A, Y, F, norleucine W, preferably L or W;

[0308] X7 is S or T, preferably S.

[0309] 13. The composition of any of the preceding items, wherein the polypeptide comprises a sequence comprising the sequence X8-G-X9-X10-X11-X12-X13-X14-X15-X16, wherein

[0310] X8 is G, R, K, Q, N, A, V, L, I, S, T, E, D;

[0311] X9 is not M or G;

[0312] X10 is not E;

[0313] X11 is not S;

[0314] X12 is not Y;

[0315] X13 is not G or A;

[0316] X14 is not P;

[0317] X15 is R, K, Q, N, P, A, Y, W, F, T, S, H;

[0318] X16 is not N.

[0319] 14. The composition of any of the preceding items, wherein the polypeptide comprises a sequence comprising the sequence X8-G-X9-X10-X11-X12-X13-X14-X15-X16, wherein

[0320] X8 is G, R, K, Q, N, A, V, L, I, S, T, E, D, preferably G, R, A, E, more preferably G;

[0321] X9 is W, S, T, F, L, V, I, A or Y, preferably W, S, F, more preferably F;

[0322] X10 is T, S, R, Q, N or K, preferably T, R or K, more preferably R;

[0323] X11 is E, D or Q, preferably E;

[0324] X12 is P or A, preferably P;

[0325] X13 is P or A, preferably P;

[0326] X14 is I, L, M, A, F or V, preferably I or V, more preferably V;

[0327] X15 is R, K, Q, N, P, A, H, W, F, T, S or Y, preferably R, P, H or Y, more preferably P;

[0328] X16 is D, E or N, preferably D.

[0329] 15. The composition of any of the preceding items, wherein the sequence has at least 80%, at least 83%, at least 85%, at least 87%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% sequence identity or higher with any of SEQ ID NO. 1-14, preferably, the composition consists of the polypeptide of any of SEQ ID NO. 1-14 or a combination thereof.

[0330] 16. The composition of any of the preceding items, wherein the one or more polypeptides comprise one or more amino acid substitutions compared to the sequence of any of SEQ ID NOs. 1-14, wherein the one or more polypeptides comprise a sequence having at least 75% (e.g., at least 90.7%, at least 90.8%, at least 90.9%, at least 91%, at least 92%, at least 93%, at least 93.5%, at least 93.6%, at least 93.7%, at least 93.8%, at least 93.9%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100%) sequence identity to any of SEQ ID NOs. 1-14.

[0331] 17. The composition of any one of the preceding items, wherein the one or more amino acid substitutions comprise or consist of conservative amino acid substitutions.

[0332] 18. The composition of any preceding item, wherein the polypeptide is capable of converting the substrate 3-keto-DON to the products 3-epi-DON and DON, and the ratio of 3-epi-DON to DON is at least 50:1, at least 100:1, at least 150:1, at least 200:1, 250:1, 300:1, 350:1, 400:1, 500:1, 600:1, 700:1, 800:1, 900:1, 1000:1 or greater.

[0333] 19. The composition of any of the preceding items, wherein the amount of polypeptide is at most 7 mg, at most 6 mg, at most 5 mg, at most 3 mg, at most 2 mg, at most 1 mg or less when 3-epi-DON is obtained at a rate of 1 μmol 3-epi-DON per minute.

[0334] 20. The composition of any of the preceding items, wherein the polypeptide is capable of converting 3-keto-DON to 3-epi-DON with a specific activity of at least 0.1 μmol / min / mg, at least 0.2 μmol / min / mg, at least 0.3 μmol / min / mg, at least 0.4 μmol / min / mg, at least 0.5 μmol / min / mg, at least 0.6 μmol / min / mg, at least 0.8 μmol / min / mg, at least 1.0 μmol / min / mg, at least 1.2 μmol / min / mg, at least 1.4 μmol / min / mg or at least 1.6 μmol / min / mg.

[0335] 21. The composition of any of the preceding items, wherein the amount of polypeptide is at least 600 mg, at least 700 mg, at least 800 mg, at least 900 mg, at least 1000 mg, at least 1100 mg, at least 1300 mg, at least 1500 mg, at least 1600 mg, at least 1700 mg, at least 1800 mg, at least 1900 mg, at least 2000 mg or more when DON is obtained at a rate of 1 μmol DON per minute.

[0336] 22. The composition of any of the preceding items, wherein the polypeptide present in the composition is capable of converting 3-keto-DON into DON with a specific activity of at most 0.0015 μmol / min / mg, at most 0.0010 μmol / min / mg, at most 0.0009 μmol / min / mg, at most 0.0008 μmol / min / mg, at most 0.0007 μmol / min / mg, at most 0.0006 μmol / min / mg or less.

[0337] 23. The composition of any of the preceding items, wherein the carrier is a liquid, preferably H2O, or a solid, preferably a health product and / or a medicine.

[0338] 24. The composition of any of the preceding items, wherein the carrier is an edible component, preferably a non-toxic component and / or a component that provides mouthfeel.

[0339] 25. A polypeptide comprising the sequence of SEQ ID NO. 1, or a sequence having at least 75% (e.g., at least 90.7%, at least 90.8%, at least 90.9%, at least 91%, at least 92%, at least 93%, at least 93.5%, at least 93.6%, at least 93.7%, at least 93.8%, at least 93.9%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100%) sequence identity to SEQ ID NO. 1, wherein the polypeptide is capable of converting 3-keto-DON to 3-epi-DON.

[0340] 26. Use of one or more polypeptides for converting 3-keto-DON to 3-epi-DON, wherein the one or more polypeptides comprise or consist of a sequence of SEQ ID NOs. 1-14, or a sequence having at least 75% (e.g., at least 90.7%, at least 90.8%, at least 90.9%, at least 91%, at least 92%, at least 93%, at least 93.5%, at least 93.6%, at least 93.7%, at least 93.8%, at least 93.9%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100%) sequence identity to any one of SEQ ID NOs. 1-14, and / or a sequence of SEQ ID NOs. 63-101; preferably, wherein: (i) the use (e.g., comprising SEQ ID NOs. 1-14) NO.1) is carried out at a pH range of about 4 to about 10 (e.g., pH about 4.5 to about 10, e.g., pH about 6.5), and / or, (ii) the use (e.g., comprising SEQ ID NO.1) is carried out at a temperature (T) range of about 14°C to about 60°C (e.g., T is about 14.9°C to about 36°C, e.g., T is about 25°C to about 31°C).

[0341] 27. Use of one or more polypeptides for preparing a feed additive, a feed composition or a pharmaceutical composition, wherein the one or more polypeptides comprise or consist of the following sequences: a sequence of SEQ ID NOs. 1-14, or a sequence having at least 75% (e.g., at least 90.7%, at least 90.8%, at least 90.9%, at least 91%, at least 92%, at least 93%, at least 93.5%, at least 93.6%, at least 93.7%, at least 93.8%, at least 93.9%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100%) sequence identity to any one of SEQ ID NOs. 1-14, and / or a sequence of SEQ ID NOs. 63-101;

[0342] wherein the polypeptide is capable of converting the substrate 3-keto-DON into products 3-epi-DON and DON in a 3-epi-DON:DON ratio of at least 25:1;

[0343] Preferably, wherein: (i) the use (e.g., comprising SEQ ID NO. 1) is carried out at a pH range of about 4 to about 10 (e.g., pH about 4.5 to about 10, e.g., pH about 6.5), and / or, (ii) the use (e.g., comprising SEQ ID NO. 1) is carried out at a temperature (T) range of about 14°C to about 60°C (e.g., T is about 14.9°C to about 36°C, e.g., T is about 25°C to about 31°C).

[0344] 27. Use of one or more polypeptides for preparing a feed additive, a feed composition or a pharmaceutical composition, wherein the one or more polypeptides comprise or consist of the following sequences: a sequence of SEQ ID NOs. 1-14, or a sequence having at least 75% (e.g., at least 90.7%, at least 90.8%, at least 90.9%, at least 91%, at least 92%, at least 93%, at least 93.5%, at least 93.6%, at least 93.7%, at least 93.8%, at least 93.9%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100%) sequence identity to any one of SEQ ID NOs. 1-14, and / or a sequence of SEQ ID NOs. 63-101,

[0345] wherein the polypeptide is capable of converting the substrate 3-keto-DON to the product 3-epi-DON, and optionally also to the product DON, totaling 100% of the total product, wherein at least 96% of the total product is 3-epi-DON;

[0346] Preferably, wherein: (i) the use (e.g., comprising SEQ ID NO. 1) is carried out at a pH range of about 4 to about 10 (e.g., pH about 4.5 to about 10, e.g., pH about 6.5), and / or, (ii) the use (e.g., comprising SEQ ID NO. 1) is carried out at a temperature (T) range of about 14°C to about 60°C (e.g., T is about 14.9°C to about 36°C, e.g., T is about 25°C to about 31°C).

[0347] 28. Use of one or more polypeptides in the production of biogas, bioethanol or sugar, preferably from sugar cane or sugar beet, wherein the one or more polypeptides comprise or consist of the following sequence: a sequence of SEQ ID NOs. 1-14 or a sequence having at least 75% (e.g., at least 90.7%, at least 90.8%, at least 90.9%, at least 91%, at least 92%, at least 93%, at least 93.5%, at least 93.6%, at least 93.7%, at least 93.8%, at least 93.9%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100%) sequence identity to any of SEQ ID NOs. 1-14, and / or SEQ ID NOs. NO.63-101, wherein the polypeptide is capable of converting the substrate 3-keto-DON into products 3-epi-DON and DON in a 3-epi-DON:DON ratio of at least 25:1; further preferably, wherein: (i) the use (e.g., comprising SEQ ID NO. 1) is carried out at a pH range of about 4 to about 10 (e.g., pH about 4.5 to about 10, for example, pH about 6.5), and / or, (ii) the use (e.g., comprising SEQ ID NO. 1) is carried out at a temperature (T) range of about 14°C to about 60°C (e.g., T is about 14.9°C to about 36°C, for example, T is about 25°C to about 31°C).

[0348] 29. Use of one or more polypeptides in the production of biogas, bioethanol or sugar, preferably from sugar cane or sugar beet, wherein the one or more polypeptides comprise or consist of the following sequence: a sequence of SEQ ID NOs. 1-14 or a sequence having at least 75% (e.g., at least 90.7%, at least 90.8%, at least 90.9%, at least 91%, at least 92%, at least 93%, at least 93.5%, at least 93.6%, at least 93.7%, at least 93.8%, at least 93.9%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100%) sequence identity to any of SEQ ID NOs. 1-14, and / or SEQ ID NOs. Sequence Nos. 63-101, wherein the polypeptide is capable of converting the substrate 3-keto-DON to the product 3-epi-DON, and optionally also to the product DON, totaling 100% of the total product, wherein at least 96% of the total product is 3-epi-DON;

[0349] Preferably, wherein: (i) the use (e.g., comprising SEQ ID NO. 1) is carried out at a pH range of about 4 to about 10 (e.g., pH about 4.5 to about 10, e.g., pH about 6.5), and / or, (ii) the use (e.g., comprising SEQ ID NO. 1) is carried out at a temperature (T) range of about 14°C to about 60°C (e.g., T is about 14.9°C to about 36°C, e.g., T is about 25°C to about 31°C).

[0350] 30. A feed or food additive or feed or food comprising one or more polypeptides comprising or consisting of the following sequences: a sequence of SEQ ID NOs. 1-14 or a sequence having at least 75% (e.g., at least 90.7%, at least 90.8%, at least 90.9%, at least 91%, at least 92%, at least 93%, at least 93.5%, at least 93.6%, at least 93.7%, at least 93.8%, at least 93.9%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100%) sequence identity to any one of SEQ ID NOs. 1-14, and / or a sequence of SEQ ID NOs. 63-101,

[0351] The polypeptide is capable of converting the substrate 3-keto-DON into products 3-epi-DON and DON at a ratio of 3-epi-DON:DON of at least 25:1.

[0352] 31. A feed or food additive or feed or food comprising one or more polypeptides comprising or consisting of the sequences of SEQ ID NOs. 1-14 or a sequence having at least 75% (e.g., at least 90.7%, at least 90.8%, at least 90.9%, at least 91%, at least 92%, at least 93%, at least 93.5%, at least 93.6%, at least 93.7%, at least 93.8%, at least 93.9%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100%) sequence identity to any one of SEQ ID NOs. 1-14, and / or the sequences of SEQ ID NOs. 63-101, wherein the polypeptide is capable of converting the substrate 3-keto-DON to the product 3-epi-DON, and optionally further to the product DON, totaling 100% total product, wherein at least 96% of the total product is 3-epi-DON.

[0353] 32. Use of an additive in a composition comprising 3-keto-DON or DON, preferably a tillage composition, the additive comprising one or more polypeptides comprising or consisting of the sequences of SEQ ID NOs. 1-14, or a sequence having at least 75% (e.g., at least 90.7%, at least 90.8%, at least 90.9%, at least 91%, at least 92%, at least 93%, at least 93.5%, at least 93.6%, at least 93.7%, at least 93.8%, at least 93.9%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100%) sequence identity to any of SEQ ID NOs. 1-14, and / or the sequences of SEQ ID NOs. 63-101, wherein the polypeptides are capable of converting the substrate 3-keto-DON into products 3-epi-DON and DON in a 3-epi-DON:DON ratio of at least 25:1.

[0354] 33. Use of an additive in a composition (preferably a tillage composition) comprising 3-keto-DON or DON, said additive comprising one or more polypeptides comprising or consisting of the following sequences: a sequence of SEQ ID NOs. 1-14 or a sequence having at least 75% (e.g., at least 90.7%, at least 90.8%, at least 90.9%, at least 91%, at least 92%, at least 93%, at least 93.5%, at least 93.6%, at least 93.7%, at least 93.8%, at least 93.9%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100%) sequence identity to any of SEQ ID NOs. 1-14, and / or, Sequence No. 63-101, wherein the polypeptide is capable of converting substrate 3-keto-DON to product 3-epi-DON, and optionally also to product DON, totaling 100% total product, wherein at least 96% of the total product is 3-epi-DON.

[0355] 34. A method for converting a trichothecene comprising a 3-oxyl group to a trichothecene comprising a 3-hydroxyl group, the method comprising contacting one or more polypeptides comprising or consisting of a sequence of SEQ ID NOs. 1-14, or a sequence having at least 75% (e.g., at least 90.7%, at least 90.8%, at least 90.9%, at least 91%, at least 92%, at least 93%, at least 93.5%, at least 93.6%, at least 93.7%, at least 93.8%, at least 93.9%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100%) sequence identity to any one of SEQ ID NOs. 1-14, and / or a sequence of SEQ ID NOs. 63-101; preferably, wherein: (i) the method (e.g., comprising SEQ ID NOs. 1-14) comprises contacting one or more polypeptides comprising or consisting of a sequence of SEQ ID NOs. 1-14, or a sequence having at least 75% (e.g., at least 90.7%, at least 90.8%, at least 90.9%, at least 91%, at least 92%, at least 93%, at least 93.5%, at least 93.6%, at least 93.7%, at least 93.8%, at least 93.9%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100%) sequence identity to any one of SEQ ID NOs. 1-14; NO.1) is carried out at a pH range of about 4 to about 10 (e.g., pH about 4.5 to about 10, e.g., pH about 6.5), and / or, (ii) the method (e.g., comprising SEQ ID NO.1) is carried out at a temperature (T) range of about 14°C to about 60°C (e.g., T is about 14.9°C to about 36°C, e.g., T is about 25°C to about 31°C).

[0356] 35. The method according to item 29, wherein the trichothecene comprising a 3-hydroxyl group is present in the S configuration.

[0357] 36. A method for converting 3-keto-DON to 3-epi-DON, the method comprising contacting one or more polypeptides comprising or consisting of a sequence of SEQ ID NOs. 1-14, or a sequence having at least 75% (e.g., at least 90.7%, at least 90.8%, at least 90.9%, at least 91%, at least 92%, at least 93%, at least 93.5%, at least 93.6%, at least 93.7%, at least 93.8%, at least 93.9%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100%) sequence identity to any one of SEQ ID NOs. 1-14, and / or a sequence of SEQ ID NOs. 63-101; preferably, wherein: (i) the method (e.g., comprising SEQ ID NOs. 1-14) comprises contacting one or more polypeptides comprising or consisting of a sequence of SEQ ID NOs. 1-14, or a sequence having at least 75% (e.g., at least 90.7%, at least 90.8%, at least 90.9%, at least 91%, at least 92%, at least 93%, at least 93.5%, at least 93.6%, at least 93.7%, at least 93.8%, at least 93.9%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100%) sequence identity to any one of SEQ ID NOs. 1-14, and / or a sequence of SEQ ID NOs. 63-101; NO.1) is carried out at a pH range of about 4 to about 10 (e.g., pH about 4.5 to about 10, e.g., pH about 6.5), and / or, (ii) the method (e.g., comprising SEQ ID NO.1) is carried out at a temperature (T) range of about 14°C to about 60°C (e.g., T is about 14.9°C to about 36°C, e.g., T is about 25°C to about 31°C).

[0358] 37. A method for reducing the amount of DON in a composition comprising DON or reducing the toxicity of a composition comprising DON by converting DON into 3-epi-DON, the method comprising:

[0359] a) contacting the composition with an enzyme capable of converting DON to 3-keto-DON; and

[0360] b) subsequently or simultaneously contacting the composition with one or more polypeptides comprising or consisting of the following sequences: a sequence of SEQ ID NO. 1-14 or a sequence having at least 75% sequence identity to any one of SEQ ID NO. 1-14, and / or a sequence in SEQ ID NO. 63-101; preferably, wherein: (i) the method (e.g., comprising SEQ ID NO. 1) is performed at a pH range of about 4 to about 10 (e.g., pH about 4.5 to about 10, e.g., pH about 6.5), and / or, (ii) the method (e.g., comprising SEQ ID NO. 1) is performed at a temperature (T) range of about 14° C. to about 60° C. (e.g., T is about 14.9° C. to about 36° C., e.g., T is about 25° C. to about 31° C.).

[0361] 38. A host cell comprising one or more polypeptides comprising or consisting of the following sequences: a sequence of SEQ ID NOs. 1-14, or a sequence having at least 75% (e.g., at least 90.7%, at least 90.8%, at least 90.9%, at least 91%, at least 92%, at least 93%, at least 93.5%, at least 93.6%, at least 93.7%, at least 93.8%, at least 93.9%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100%) sequence identity to any one of SEQ ID NOs. 1-14, and / or a sequence of SEQ ID NOs. 63-101.

[0362] 39. The host cell according to item 14, further expressing a cofactor for said polypeptide, preferably NADH or NADPH.

[0363] 40. A plant genetically modified to express one or more polypeptides comprising or consisting of the sequences of SEQ ID NOs. 1-14, or a sequence having at least 75% (e.g., at least 90.7%, at least 90.8%, at least 90.9%, at least 91%, at least 92%, at least 93%, at least 93.5%, at least 93.6%, at least 93.7%, at least 93.8%, at least 93.9%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100%) sequence identity to any one of SEQ ID NOs. 1-14, and / or the sequences of SEQ ID NOs. 63-101.

[0364] 41. Seeds of the plants defined in item 40.

[0365] 42. A preparation comprising:

[0366] a) one or more polypeptides comprising or consisting of a sequence of SEQ ID NOs. 1-14, or a sequence having at least 75% (e.g., at least 90.7%, at least 90.8%, at least 90.9%, at least 91%, at least 92%, at least 93%, at least 93.5%, at least 93.6%, at least 93.7%, at least 93.8%, at least 93.9%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100%) sequence identity to any one of SEQ ID NOs. 1-14, and / or a sequence of SEQ ID NOs. 63-101, wherein the polypeptide is capable of converting the substrate 3-keto-DON into products 3-epi-DON and DON in a 3-epi-DON:DON ratio of at least 25:1;

[0367] b) Carrier.

[0368] 43. A preparation comprising:

[0369] a) one or more polypeptides comprising or consisting of a sequence of SEQ ID NOs. 1-14, or a sequence having at least 75% (e.g., at least 90.7%, at least 90.8%, at least 90.9%, at least 91%, at least 92%, at least 93%, at least 93.5%, at least 93.6%, at least 93.7%, at least 93.8%, at least 93.9%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100%) sequence identity to any one of SEQ ID NOs. 1-14, and / or a sequence of SEQ ID NOs. 63-101, wherein the polypeptide is capable of converting the substrate 3-keto-DON to the product 3-epi-DON, and optionally also to the product DON, totaling 100% total product, wherein at least 96% of the total product is 3-epi-DON;

[0370] b) Carrier.

[0371] 43. A polypeptide for preventing and / or treating mold poisoning, comprising or consisting of a sequence of SEQ ID NOs. 1-14, or a sequence having at least 75% (e.g., at least 90.7%, at least 90.8%, at least 90.9%, at least 91%, at least 92%, at least 93%, at least 93.5%, at least 93.6%, at least 93.7%, at least 93.8%, at least 93.9%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100%) sequence identity to any one of SEQ ID NOs. 1-14, and / or a sequence of SEQ ID NOs. 63-101, wherein the polypeptide is capable of converting the substrate 3-keto-DON into products 3-epi-DON and DON at a 3-epi-DON:DON ratio of at least 25:1.

[0372] 44. A polypeptide for preventing and / or treating mold poisoning, the polypeptide comprising or consisting of the following sequence: a sequence of SEQ ID NOs. 1-14, or a sequence having at least 75% (e.g., at least 90.7%, at least 90.8%, at least 90.9%, at least 91%, at least 92%, at least 93%, at least 93.5%, at least 93.6%, at least 93.7%, at least 93.8%, at least 93.9%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100%) sequence identity to any one of SEQ ID NOs. 1-14, and / or SEQ ID NOs. Sequence No. 63-101, wherein the polypeptide is capable of converting substrate 3-keto-DON to product 3-epi-DON, and optionally also to product DON, totaling 100% total product, wherein at least 96% of the total product is 3-epi-DON.

[0373] 45. The composition of any of the preceding items, wherein the polypeptide comprises loop B, and loop B is at most 39 amino acids, at most 38 amino acids, at most 37 amino acids, at most 35 amino acids, at most 34 amino acids or fewer in length.

[0374] 46. ​​A polypeptide comprising a sequence of SEQ ID NOs. 63-101 or a sequence having at least 75% sequence identity to any one of SEQ ID NOs. 63-101, wherein the polypeptide is capable of converting 3-keto-DON to 3-epi-DON.

[0375] The following sequences were used in this application.

[0376]

[0377]

[0378]

[0379]

[0380]

[0381]

[0382]

[0383]

[0384]

[0385]

[0386]

[0387]

[0388]

[0389]

[0390]

[0391]

[0392]

[0393]

[0394]

[0395]

[0396] Table 2: depicts the sequences described herein. For some sequence examples, the positions of loop A and loop B are directly described in the table. Sequences 63-101 include amino acid substitutions according to the description in Table 1 herein.

[0397] Example

[0398] Example 1: Cloning, expression and enzyme purification

[0399] The inventors have discovered various candidate genes encoding enzymes capable of reducing 3-oxotrichothecenes such as keto-DON to 3-hydroxylated trichothecenes such as epi-DON. For expression and protein purification of all candidate genes, the following sequences were incorporated into plasmids for expression in a suitable host organism (such as E. coli): nucleotide sequences encoding polypeptides of enzymes SEQ ID NOs. 1–14 (i.e., SEQ ID NOs. 47-60), as well as sequences encoding prior art enzymes SEQ ID NOs. 15 and 16, and sequences encoding variants of any of the enzymes SEQ ID NOs. 1 (SEQ ID NOs. 63-75), SEQ ID NOs. 3 (SEQ ID NOs. 76-88), and SEQ ID NOs. 6 (SEQ ID NOs. 89-101), comprising the amino acid substitutions disclosed in Table 1 herein. The recombinant vectors containing each coding sequence were assembled into the E. coli expression vector pET28a by TWIST bioscience.com (US). The nucleotide sequence includes the restriction enzyme site NcoI at the 5' end and the restriction enzyme site XhoI at the 3' end, as well as a 6xHis tag at the C-carboxyl end or N-amino end. The sequence identity of SEQ ID NO. 1-16, 63-101 is shown in Figure 1A and Figure 1B and the sequence listing herein.

[0400] For expression, a pET28a-based recombinant vector was transformed into E. coli or any other suitable host organism. IPTG was added to induce protein expression. The cells were harvested by centrifugation and then disrupted by ultrasound, a French press, or any other suitable method to open the bacterial cells and produce a crude cell lysate. The cell lysate was cleaned, and the soluble recombinant protein was used to determine the catalytic properties of the reductase produced. To determine the specific properties of the reductase, the enzyme was selectively purified using nickel-based affinity chromatography using a nickel-agarose column. Unfortunately, the amount of SEQ ID NO. 16 produced was too small to allow for purity analysis and enzyme analysis. To determine the purity of the recombinant enzyme, they were separated on an SDS-PAGE protein gel so that the protein could be visualized using standard Coomassie blue staining. The concentrations of the different proteins were determined photometrically using Bradford reagent (Sigma #B6916) at a wavelength of 595 nm and / or using the Qubit protein assay (Thermofischer Life technologies #Q33211) with fluorescence measurement in a plate reader (Biotek, Synergy HT). For the Bradford assay, the concentrations were quantified from 0 μg ml -1 Up to 1500 μg ml -1 A standard curve of bovine serum albumin (BSA, Sigma #A4919) was used to determine protein concentrations. For the qubit analysis, 0 μg ml -1 Up to 400 μg ml -1 The protein concentration of the samples was determined by constructing a standard curve.

[0401] Example 2: In vitro conversion of 3-keto DON

[0402] The ability of the reductase prepared in Example 1 to use 3-keto DON as a substrate was determined, and the reductases having SEQ ID NOs. 15 and 16 were used as examples of the prior art. All enzymes were prepared according to Example 1 and used in the following activity assays.

[0403] The following enzyme assay should be defined as a reference method for determining activity (particularly the specific activity of the enzymes claimed in the present invention). The activity of 3-ketoDON reductase was measured in a standard 3-ketoDON degradation assay consisting of 100 mM Tris buffer pH 8.0 containing 0.5 mM NADPH, 0.5 mM NADH, 25 ppm 3-ketoDON, and 100 nM of each enzyme. All reaction components except NADH and NADPH were mixed on ice, and the 0-minute sample (100% methanol was mixed with the sample in a 1:1 (v / v) ratio) was removed to prevent any reaction. The reaction mixture was then immediately transferred to a hot plate and the cofactor mixture (i.e., NADH and NADPH) was added to initiate the reaction, which was carried out at 30°C for 2 hours. Samples were removed periodically to monitor the progress of the reaction and immediately mixed with 100% methanol in a 1:1 (v / v) ratio to stop the reaction. Subsequently, they were analyzed using the following HPLC-MS / MS method.

[0404] For the HPLC analysis of DON, 3-keto DON, and 3-epi DON, a 150 mm × 4.6 mm Phenomenex Gemini-NX C18 column with a particle size of 5 μm was used. A mixture of acetonitrile and ultrapure water was used as the mobile phase, and the UV signal at 220 nm was recorded and evaluated to determine the concentration of the analytes.

[0405] For LC-MS / MS analysis, DON, 3-keto-DON, and 3-epi-DON were separated using a 150 mm × 2.1 mm Phenomenex Kinetex biphenyl column with a 2.6 μm particle size. The mobile phase consisted of a mixture of methanol and ultrapure water with 0.1% (v / v) acetic acid. Electrospray ionization (ESI) was used in negative mode to generate ions. Quantification was performed using a QTrap and / or triple-quadrupole mass spectrometer. Samples were diluted to within the linear range of the method, meaning that the analyte concentration in the injected sample ranged from 1 ppb to 500 ppb. The limit of detection (LOQ) is the limit of reliable detection; concentrations below 1 ppb of a compound in a sample are considered unreliable; therefore, the LOQ is 1 ppb. If the analyte concentration is below the LOQ, all further calculations (e.g., determination of the specific activity or ratio of the analyte) are performed based on 1 ppb.

[0406] Prior to HPLC-MS / MS analysis, all samples were diluted with 40% aqueous methanol to a theoretical final concentration of 200 ppb 3-keto DON in the sample.

[0407] Based on the linear range of the reaction in μmol 产物 / min / mg 酶The 3-epi-DON (or DON) production was calculated and the results were used to calculate the specific activity of the enzyme (μmol 产物 / min / mg 酶 This is achieved by converting the absolute amount of 3-epi-DON (or DON) produced in the reaction up to the selected time point (i.e., 10 minutes for 3-epi-DON and 120 minutes for DON) from ppm to μmol (i.e., the concentration of 3-epi-DON in ppm (= μg / ml) is multiplied by the reaction volume in ml and divided by the molecular weight of 3-epi-DON in μg / μmol to obtain μmol of 3-epi-DON in the reaction at the selected time point). The result is then divided by the selected time range in minutes. This value is then divided by the total amount of enzyme in the reaction in mg to calculate the specific activity (μmol). 产物 / min / mg 酶 ), which corresponds to enzyme units per milligram (U / mg). The results of the activity assay are shown in Figure 2 (Conversion of 3-keto-DON to 3-epi-DON) and Figure 3 (Conversion of 3-ketoDON to DON).

[0408] Example 3: Sequence analysis of conserved regions and unique structural features

[0409] Sequences SEQ ID NOs. 1-14 and 63-101, as well as prior art enzymes (SEQ ID NOs. 15 and 16), belong to the class of oxidoreductases and are often predicted to be members of the aldo-keto reductase family, which catalyzes the reduction of aldehydes and ketones to produce primary and secondary alcohols, respectively, using NAD(P)(H). Aldo-keto reductases share a common (α / β)8 structure and a conserved catalytic mechanism with a catalytic tetramer composed of the residues tyrosine (Tyr, Y), aspartic acid (Asp, D), histidine (His, H), and lysine (Lys, K). In SEQ ID NOs. 15 and 16 (both known in the prior art), the catalytic tetramer is composed of Asp48, Tyr53, Lys81, and His122. In the reductase SEQ ID NO. 1, the catalytic tetramer is located at Asp53, Tyr58, Lys85, and His126. However, they differ significantly at least in loops A and B, which are located at the rear of the barrel structure. Variation in these loops, along with the substrate binding pocket, determines substrate specificity (Jez and Penning (1998)).

[0410] This superfamily is characterized by feature cd06660 in the reference database of the National Center for Biotechnology Information (NCBI; http: / / www.ncbi.nlm.nih.gov / ). To further analyze the domain structure of the sequences, a batch search was performed in the CDD Conserved Domain Database (NCBI; https: / / www.ncbi.nlm.nih.gov / cdd / ), specifically applied to multiple sequences https: / / www.ncbi.nlm.nih.gov / Structure / bwrpsb / bwrpsb.cgi. The search was performed using standard parameters: automatic search mode against the database CDD-52910 PSSMs, an expectation level of 0.01, and the application of composition-corrected scoring. PSSM = Position-Specific Scoring Matrix (PSSM), which calculates the range of consensus sequences. The PSSM curve describes the frequency with which a specific residue is detected at a given position in a multiple sequence alignment of a domain model. All analyzed sequences belong to superfamily cl30127 (Marchler-Bauer et al. (2011), Marchler-Bauer and Bryant (2004), Marchler-Bauer et al. (2017)).

[0411] Aldo-keto reductases differ in their substrate-specific ring structures. Therefore, rings A and B are defined as substrate-specific rings A and B. The substrate-specific rings A and B in enzymes SEQ ID NOs. 1-14 and 63-101 differ significantly from those in prior art enzymes (particularly SEQ ID NOs. 15 and 16). Figure 4 In all cases, loop A consists of nine residues and contains the histidine of the catalytic tetramer. However, instead of a nonpolar alanine located downstream of histidine 122 in the prior art enzymes (position 123 in the prior art enzymes), the residue immediately downstream of the catalytic histidine in SEQ ID NOs. 1-14 and 63-101 is not an alanine. The substrate-specific loop A includes sequences downstream of the consensus motif ID / E-LYQ and upstream of the motif I / M / LE / DE / DTL / M. The substrate-specific loop B begins at a serine within the consensus motif WSPL-XG and ends upstream of the consensus motif XXR / K / HT / LW / YXI / V / LI / V / L. Loop B varies significantly in length, being 35 residues or fewer.

[0412] Variability of loops A and B in terms of residues and length resulted in nearly 100% stereoselectivity towards 3-epi-DON and improved specific activity of SEQ ID NOs. 1-14 and 63-101 compared to SEQ ID NOs. 15-16.

[0413] Figure 5. Substrate specificity loops A and B. Start residue, end residue and amino acid length. These are also summarized in the sequence listing herein.

[0414] Example 4: Determination of optimal pH

[0415] In order to determine the optimal pH value for the catalytic activity of the polypeptides mentioned herein, reactions for the conversion of 3-keto-DON at different pH values ​​were prepared. To this end, 16 reactions were prepared with different pH values, each with a final volume of 200 μL. A polypeptide having an amino acid sequence of SEQ ID NO: 1 was used at a final concentration of 200 nM. The substrate 3-keto-DON was used at a final concentration of 30 ppm, and NADPH was used at a final concentration of 1 mM. As for the buffer, Teorell Stenhagen (TS) buffer (Stenhagen & Teorell. 1938. Nature 141, 415) was used, and its pH was set to any value of 2.5, 3.0, 3.5, 4.0, 4.5, 5.0, 5.5, 6.0, 6.5, 7.0, 7.5, 8.0, 8.5, 9.0, 9.5, or 10.0. The conversion reaction was incubated at 30° C. for 120 minutes, and the conversion reaction was started by adding NADPH. During the incubation, 20 μL samples were collected at 0 min, 10 min, 20 min, 30 min, 60 min, and 120 min. The 0 min sample was collected before the addition of NADPH. The 20 μL sample was immediately mixed with 20 μL of pure methanol to stop the conversion reaction and then kept on ice until the end of the incubation.

[0416] Prior to LC-MS / MS analysis, all samples were diluted with 40% methanol in water to a theoretical final concentration of 0.3 ppm 3-keto-DON. For LC-MS / MS analysis, DON, 3-keto-DON, and 3-epi-DON were separated on a 150 mm × 2.1 mm Phenomenex Kinetex biphenyl column with a particle size of 2.6 μm. The mobile phase consisted of a mixture of methanol and ultrapure water with 0.1% (v / v) acetic acid. Ions were generated using electrospray ionization (ESI) in negative mode. Quantification was performed using a QTrap and / or triple- and quadrupole mass spectrometer detector. If necessary, samples were diluted to within the linear range of the method, i.e., the concentration range of the analyte in the injected sample was 1 ppb to 500 ppb. The limit of detection (LOQ) is the limit of reliable detection; detection of compounds in the sample at concentrations below 1 ppb is considered unreliable; therefore, the LOQ was 1 ppb. If the analyte concentration was below the LOQ, all further calculations (e.g., determination of the specific activity or ratio of the analyte) were performed at 1 ppb. Kinetic parameters were calculated based on the measured amounts of 3-epi-DON and / or DON at individual sample time points. Specific activity was calculated from the maximum linear product formation rate for each reaction. Table 3 below shows the specific activity of 3-epi-DON formation at various pH values.

[0417] Table 3: Specific activity of the polypeptide having the amino acid sequence of SEQ ID NO. 1 at different pH values, expressed as μmol of 3-epi-DON formed per minute and per mg of polypeptide.

[0418]

[0419]

[0420] The polypeptide was found to be active in the pH range of pH 4.5 to pH 10, with optimal 3-epi-DON formation at pH 6.5.

[0421] Example 5: Determination of the optimum temperature

[0422] To determine the optimal catalytic temperature for the peptides described herein, reactions were prepared for the conversion of 3-keto-DON at various temperatures. Twenty-four reactions were prepared, each with a final volume of 180 μL. The peptide with the amino acid sequence of SEQ ID NO: 1 was used at a final concentration of 200 nM. The substrate 3-keto-DON was used at a final concentration of 30 ppm, and NADPH was used at a final concentration of 1 mM. TS buffer at pH 7.0 was used as the buffer. The conversion reactions were incubated for 120 minutes at various temperatures in a laboratory PCR thermocycler, and the conversion reactions were initiated by the addition of NADPH. During the incubation period, 20 μL samples were taken at 0 min, 10 min, 20 min, 30 min, 60 min, and 120 min. A 0 min sample was taken from the reaction before the addition of NADPH. The 20 μL sample was immediately mixed with 180 μL of pure methanol to stop the conversion reaction and then stored on ice until the end of the incubation period. LC-MS / MS analysis was performed as described in Example 4. Table 4 below shows the specific activity of 3-epi-DON formation at different temperatures.

[0423] Table 4: Specific activity of the polypeptide having the amino acid sequence of SEQ ID NO. 1 at different temperatures, expressed as μmol of 3-epi-DON formed per minute and per mg of polypeptide.

[0424]

[0425]

[0426] Most of the 3-epi-DON was formed by polypeptides at temperatures between 14.9°C and 36°C, with the optimum temperature being between 25°C and 31°C.

[0427] References

[0428] Altschul,J.Mol.Evol.36(1993),290-300

[0429] Altschul,J.Mol.Biol.215(1990),403-410

[0430] Altschul et al., 1997 (Nucleic Acids Res. (1997) 25:3389-3402

[0431] Carere et al.2018(Carere(2018)"The identification of DepB:An enzymeresponsible for the final detroxification Step in the deoxynivalenolepimerization pathway in Devosia mutans 17-2-E-8."Frontiers inMicrobiology.9:1573Cregg,Pichia Protocols,second Edition,ISBN-10:1588294293,2007

[0432] Ellis(2002)“Microbial aldo-keto reductases”FEMS Microbiol Lett.5;216(2):123-31

[0433] Hassan et al.(2017)“The enzymatic epimerization of deoxynivalenol byDevosia mutans proceeds through the formation of 3-keto-DON intermediate.”Scientific Reports 7,article number 6929

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Toxins.9(4):130 Sequence Listing <110> Elber AG <120> Improved peptides for converting the substrate 3-keto-deoxynivalenol into 3-epi-deoxynivalenol <130> ERB16790PCT <150> EP 19194632 <151> 2019-08-30 <160> 101 <170> PatentIn version 3.5 <210> 1 <211> 353 <212> PRT <213> Artificial sequence <220> <223> enzymes <400> 1 Met Asp Tyr Arg Tyr Leu Gly Arg Ser Gly Leu Arg Val Ser Val Leu 1 5 10 15 Thr Met Gly Thr Met Thr Phe Gly Gly Ser Glu Lys Val Gly His Thr 20 25 30 Ala Gln Ala Asp Ala Thr Arg Gln Ile Asp Leu Cys Leu Asp His Gly 35 40 45 Ile Asn Leu Leu Asp Thr Ala Asn Val Tyr Asn Ala Gly Val Ser Glu 50 55 60 Glu Met Ile Gly Val Ala Leu Ser Glu His Gly Arg Arg Gln Lys Ala 65 70 75 80 Leu Ile Ala Thr Lys Val Arg Phe Arg Met Gly Asn Gly Pro Asn Glu 85 90 95 Val Gly Leu Ser Arg His His Ile Ile Ala Gln Ala Glu Ala Ser Leu 100 105 110 Lys Arg Leu Lys Thr Asp Val Ile Asp Leu Tyr Gln Val His Glu Trp 115 120 125 Asp Gly Met Thr Pro Ile Glu Glu Thr Met Glu Ala Leu Asp Thr Leu 130 135 140 Val Lys Gln Gly Lys Val Arg Tyr Val Gly Cys Ser Asn Tyr Ser Gly 145 150 155 160 Trp His Ile Met Lys Ala Leu Ala Ala Ala Asp Lys His His Gly Gln 165 170 175 Arg Phe Ile Ser Gln Gln Ile His Tyr Thr Leu His Ser Arg Glu Ala 180 185 190 Glu Tyr Glu Leu Val Pro Ile Ser Gln Asp Gln Gly Leu Gly Ile Leu 195 200 205 Ile Trp Ser Pro Leu Ala Gly Gly Leu Leu Ser Gly Lys Tyr Arg Arg 210 215 220 Asp Gly Gly Pro Asp Ala Gly Arg His Val Gly Gly Phe Arg Glu Pro 225 230 235 240 Pro Val Tyr Asp Trp Asp Lys Leu Tyr Asp Ile Ile Asp Val Ile Val 245 25​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​Met Asp Tyr Arg Tyr Leu Gly Arg Ser Gly Leu Lys Val Ser Ser Leu 1 5 10 15 Thr Met Gly Thr Met Thr Phe Gly Gly Ser Glu Lys Val Gly His Thr 20 25 30 Pro Gln Ala Glu Ala Thr Arg Gln Ile Asp Leu Cys Leu Asp His Gly 35 40 45 Ile Asn Leu Leu Asp Thr Ala Asn Val Tyr Asn Ala Gly Val Ser Glu 50 55 60 Glu Met Ile Gly Val Ala Leu Ala Glu Asn Gly Arg Arg Gln Lys Ala 65 70 75 80 Leu Val Ala Thr Lys Val Arg Phe Arg Met Gly Asp Gly Pro Asn Glu 85 90 95 Val Gly Leu Ser Arg His His Ile Val Ala Gln Ala Glu Ala Ser Leu 100 105 110 Lys Arg Leu Lys Thr Asp Val Ile Asp Leu Tyr Gln Val His Glu Trp 115 120 125 Asp Gly Met Thr Pro Ile Glu Glu Thr Met Glu Ala Leu Asp Thr Leu 130 135 140 Val Arg Gln Gly Lys Val Arg Tyr Ile Gly Cys Ser Asn Tyr Ser Gly 145 150 155 160 Trp His Ile Met Lys Ala Leu Ala Ala Ala Asp Lys Asn His Gly Gln 165 170 175 Arg Phe Ile Ser Gln Gln Ile His Tyr Thr Leu His Ser Arg Glu Ala 180 185 190 Glu Tyr Glu Leu Val Pro Ile Ser Gln Asp Gln Gly Leu Gly Ile Leu 195 200 205 Ile Trp Ser Pro Leu Ala Gly Gly Leu Leu Ser Gly Lys Tyr Arg Arg 210 215 220 Asp Gly Gly Pro Glu Ser Gly Arg His Val Gly Gly Ser Arg Glu Pro 225 230 235 240 Pro Val Pro Asp Trp Gly Lys Leu Tyr Asp Ile Val Glu Val Ile Val 245 250 255 Ala Ile Ala Glu Ala Arg Gly Val Ser Gly Ala Gln Val Ala Leu Ala 260 265 270 Trp Ala Leu Gly Arg Pro Gly Val Thr Ser Val Ile Ile Gly Gly Arg 275 280 285 Ser Glu Ala Gln Phe Lys Asp Asn Leu Ala Ala Ala Asp Leu Lys Leu 290 295 300 Thr Pro Glu Glu Arg Gln Lys Leu Asp Ala Val Ser Gln Pro Pro Leu 305 310 315 320 Leu Tyr Pro Tyr Trp His Gln Ser Phe Thr Ala Gln Asp Arg Leu Ser 325 330 335 Arg Ile Asp Leu Asp Leu Ile Gly Pro Tyr Ala Glu Glu Phe Lys Arg 340 345 350 Gly <210> 3 <211> 353 <212> PRT <213> Artificial Sequence <220> <223> Enzyme <400> 3 Met Asp Tyr Arg Tyr Leu Gly Arg Ser Gly Leu Arg Val Ser Thr Leu 1 5 10 15 Thr Met Gly Thr Met Thr Phe Gly Gly Ser Glu Lys Val Gly His Thr​​​​​​​​​​​​​​​​​​​Val Gly Leu Ser Arg His His Ile Ile Ala Gln Ala Glu Ala Ser Leu 100 105 110 Lys Arg Leu Lys Thr Asp Val Ile Asp Leu Tyr Gln Val His Glu Trp 115 120 125 Asp Gly Met Thr Pro Ile Glu Glu Thr Met Glu Ala Leu Asp Thr Leu 130 135 140 Val Lys Gln Gly Lys Val Arg Tyr Ile Gly Cys Ser Asn Tyr Ser Gly 145 150 155 160 Trp His Ile Met Lys Ala Leu Ala Ala Ala Asp Lys Ser His Gly Gln 165 170 175 Arg Phe Ile Ser Gln Gln Ile His Tyr Thr Leu His Ser Arg Glu Ala 180 185 190 Glu Tyr Glu Leu Val Pro Ile Ser Gln Asp Gln Gly Leu Gly Ile Leu 195 200 205 Ile Trp Ser Pro Leu Ala Gly Gly Leu Leu Ser Gly Lys Tyr Arg Arg 210 215 220 Asp Gly Gly Pro Glu Ser Gly Arg His Val Gly Gly Ser Arg Glu Pro 225 230 235 240 Pro Val Pro Asp Trp Asp Lys Leu Tyr Asp Ile Val Asp Val Ile Val 245 250 255 Ala Ile Ala Glu Glu Arg Gly Val Ser Gly Ala Gln Val Ala Leu Ala 260 265 270 Trp Ala Leu Gly Arg Pro Gly Val Thr Ser Val Ile Ile Gly Gly Arg 275 280 285 Ser Glu Ala Gln Phe Lys Asp Asn Leu Ala Ala Ala Asp Leu Lys Leu 290 295 300 Ser Asp Glu Glu Arg Gln Lys Leu Asp Ala Val Ser Arg Pro Pro Leu 305 310 315 320 Leu Tyr Pro Tyr Trp His Gln Thr Phe Thr Ala Asn Asp Arg Leu Gly 325 330 335 Pro Ala Asp Gln Asp Leu Leu Ser Thr Tyr Val Glu Asp Phe Lys Arg 340 345 350 Gly <210> 4 <211> 353 <212> PRT <213> Artificial Sequence <220> <223> Enzyme <400> 4 Met Glu Tyr Arg Tyr Leu Gly Arg Ser Gly Leu Lys Val Ser Thr Leu 1 5 10 15 Thr Met Gly Thr Met Thr Phe Gly Gly Ser Glu Lys Val Gly Ala Thr 20 25 30 Ala Gln Asp Asp Ala Thr Arg Gln Ile Asp Leu Cys Leu Asp Ala Gly 35 40 45 Ile Asn Leu Leu Asp Thr Ala Asn Val Tyr Asn Ala Gly Val Ser Glu 50 55 60 Glu Met Ile Gly Val Ala Leu Ala Glu Asn Gly Arg Arg Gln Lys Ala 65 70 75 80 Leu Val Ala Thr Lys Val Arg Phe Lys Met Gly Asp Gly Pro Asn Glu 85 90 95 Ile Gly Leu Ser Arg His His Ile Ile Ala Gln Ala Glu Ala Ser Leu 100 105 110 Lys Arg Leu Lys Thr Asp Val Ile Asp Leu Tyr Gln Val His Glu Trp 115 120 125 Asp Gly Met Thr Pro Ile Glu Glu Thr Met Glu Ala Leu Asp Thr Leu 130 135 140 Ile Lys Gln Gly Lys Val Arg Tyr Ile Gly Cys Ser Asn Tyr Ser Gly 145 150 155 160 Trp His Ile Met Lys Ala Leu Gln Ala Ala Asp Lys Asn His Gly Gln 165 170 175 Arg Phe Ile Ser Gln Gln Ile His Tyr Thr Leu His Ser Arg Glu Ala 180 185 190 Glu Tyr Glu Leu Ile Pro Ile Thr Gly Asp Gln Gly Leu Gly Val Leu 195 200 205 Ile Trp Ser Pro Leu Ala Gly Gly Leu Leu Ser Gly Lys Tyr Arg Arg 210 215 220 Asn Gly Gly Pro Asp Ser Gly Arg His Val Gly Gly Phe Arg Glu Pro 225 230 235 240 Pro Val Tyr Asp Trp Glu Lys Leu Tyr Asp Ile Val Asp Val Ile Val 245 250 255 Asp Ile Ala Asp Asp Arg Gly Val Ser Gly Ala Gln Val Ala Leu Ala 260 265 270 Trp Ser Leu Gly Arg Pro Gly Val Thr Ser Val Ile Ile Gly Gly Arg 275 280 285 Asn Asp Ala Gln Phe Lys Asp Asn Leu Ala Ala Ala Glu Leu Lys Leu 290 295 300 Thr Ala Glu Glu Arg Ala Arg Leu Asp Ala Val Ser Arg Pro Pro Leu 305 310 315 320 Leu Tyr Pro Tyr Trp His Gln Thr Phe Thr Ala Gln Asp Arg Leu Gly 325 330 335 Pro Val Asp Arg Asn Leu Ile Glu Pro Tyr Ala Glu Glu Leu Asn Arg 340 345 350 Gly <210> 5 <211> 353 <212> PRT <213> Artificial Sequence <220> <223> Enzyme <400> 5 Met Gln Tyr Arg Tyr Leu Gly Arg Ser Gly Leu Lys Val Ser Thr Leu 1 5 10 15 Thr Met Gly Thr Met Thr Phe Gly Gly Gly Glu Lys Ile Gly Gly Thr [[ID=2�]]20 25 30 Gly Gln Asp Glu Ala Asn Arg Gln Ile Asp Leu Cys Leu Asp His Gly 35 40 45 Ile Asn Leu Leu Asp Thr Ala Asn Val Tyr Asn Ala Gly Val Ser Glu 50 55 60 Glu Met Ile Gly Val Ala Leu Ser Glu Asn Gly Arg Arg Gln Lys Ala 65 70 75 80 Leu Ile Ala Thr Lys Val Arg Phe Lys Met Gly Glu Gly Pro Asn Glu 85 90 95 Val Gly Leu Ser Arg His His Ile Ile Ala Gln Ala Glu Ala Ser Leu 100 105 110 Lys Arg Leu Lys Thr Asp Val Ile Asp Leu Tyr Gln Val His Glu Trp 115 120 125 Asp Gly Met Thr Pro Ile Glu Glu Thr Met Glu Ala Leu Asp Thr Leu 130 135 140 Val Lys Gln Gly Lys Val Arg Tyr Ile Gly Cys Ser Asn Tyr Ser Gly 145 150 155 160 Trp His Ile Met Lys Ala Leu Ala Ala Ala Asp Ser Arg His Gln Gln 165 170 175 Arg Phe Val Ser Gln Gln Ile His Tyr Thr Leu His Ser Arg Glu Ala 180 185 190 Glu Tyr Glu Leu Val Pro Ile Thr Gln Asp Gln Gly Leu Gly Ile Leu 195 200 205 Val Trp Ser Pro Leu Ala Gly Gly Leu Leu Ser Gly Lys Tyr Arg Arg 210 215 220 Asp Gly Gly Pro Asp Ala Gly Arg His Val Gly Gly Phe Arg Glu Pro 225 230 235 240 Pro Val Pro Asp Trp Asn Arg Leu Tyr Asp Ile Val Asp Ala Leu Val 245 250 255 Ala Ile Ala Glu Ala Arg Gly Val Ser Gly Ala Gln Val Ala Leu Ala 260 265 270 Trp Leu Leu Gly Arg Pro Gly Val Thr Ser Val Ile Ile Gly Gly Arg 275 280 285 Ser Glu Ala Gln Phe Arg Asp Asn Leu Ala Ala Ala Asp Leu Gln Leu 290 295 300 Thr Ala Asp Glu Arg Gln Arg Leu Asp Ala Val Ser Gln Pro Pro Leu 305 310 315 320 Leu Tyr Pro Tyr Trp His Gln Thr Phe Thr Ala Asn Asp Arg Leu Gly ​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​Asp Ile Leu Gly Gln Val Leu Ala Glu Asn Gly Arg Arg Gln Arg Ala 65 70 75 80 Leu Val Ala Thr Lys Val Arg Phe Lys Met Gly Asn Gly Pro Asn Glu 85 90 95 Ala Gly Leu Ser Arg His His Ile Ile Ala Gln Ala Glu Ala Ser Leu 100 105 110 Lys Arg Leu Arg Thr Asp Ala Ile Asp Leu Tyr Gln Val His Glu Trp 115 120 125 Asp Gly Met Thr Pro Ile Glu Glu Thr Met Glu Ala Leu Asp Thr Leu 130 135 140 Val Arg Gln Gly Lys Val Arg Tyr Val Gly Cys Ser Asn Tyr Ser Gly 145 150 155 160 Trp His Ile Met Lys Ala Leu Met Ala Ala Asp Lys Arg Gln Gly Gln 165 170 175 Arg Phe Val Ser Gln Gln Ile His Tyr Ser Leu His Ser Arg Asp Ala 180 185 190 Glu Tyr Glu Leu Val Pro Ile Ser Gln Asp Gln Gly Leu Gly Ile Leu 195 200 205 Val Trp Ser Pro Leu Ala Gly Gly Leu Leu Ser Gly Lys Tyr Arg Arg 210 215 220 Thr Gly Gly Pro Glu Ser Gly Arg His Val Gly Gly Phe Arg Glu Pro 225 230 235 240 Pro Val Tyr Asp Trp Asp Lys Leu Tyr Asp Leu Ile Asp Val Ile Val 245 250 255 Asp Ile Ala Ala Gln Arg Gly Val Ser Gly Ala Gln Val Ala Leu Ser 260 265 270 Trp Leu Leu Glu Arg Pro Gly Val Thr Ser Leu Ile Ile Gly Gly Arg 275 280 285 Asn Glu Ala Gln Phe Ala Asp Asn Ile Ala Ala Ile His Leu Lys Leu 290 295 300 Ser Ser Glu Glu Arg Ala Arg Leu Asp Ala Ala Ser Arg Pro Pro Leu 305 310 315 320 Leu Tyr Pro Tyr Trp His Gln Ser Phe Thr Ala Gln Asp Arg Leu Ser 325 330 335 Arg Ile Asp Leu Asp Leu Ile Glu Pro Tyr Ala Glu Glu Phe Lys Arg 340 345 350 Gly <210> 7 <211> 353 <212> PRT <​​​​​​​Met Glu Tyr Arg Tyr Leu Gly Arg Ser Gly Leu Arg Val Ser Val Leu 1 5 10 15 Thr Met Gly Thr Met Thr Phe Gly Gly Ser Glu Lys Ile Gly Asn Thr 20 25 30 Ser Pro Ala Asp Ala Ala Arg Gln Val Asp Met Cys Leu Asp Ala Gly 35 40 45 Ile Asn Leu Tyr Asp Thr Ala Asn Val Tyr Asn Ala Gly Val Ser Glu 50 55 60 Glu Ile Leu Gly Ala Ile Leu Ala Glu Asn Gly Arg Arg Gln Lys Ala 65 70 75 80 Leu Ile Ala Thr Lys Val Arg Phe Lys Met Gly Asp Gly Pro Asn Glu 85 90 95 Ala Gly Leu Ser Arg His His Ile Ile Glu Gln Cys Lys Ala Ser Leu 100 105 110 Lys Arg Leu Lys Thr Asp Val Ile Asp Leu Tyr Gln Val His Glu Trp 115 120 125 Asp Gly Arg Thr Pro Ile Glu Glu Thr Met Glu Ala Leu Asp Thr Leu 130 135 140 Val Arg His Gly His Val Arg Tyr Ile Gly Cys Ser Asn Tyr Ser Gly 145 150 155 160 Trp His Ile Met Lys Ala Leu Ala Ser Ala Asp Gln Arg His Asn Ala 165 170 175 Arg Phe Val Ser Gln Gln Ile His Tyr Ser Leu His Ser Arg Glu Ala 180 185 190 Glu Tyr Glu Leu Val Pro Ile Ser Gln Asp Gln Gly Leu Gly Ile Leu 195 200 205 Val Trp Ser Pro Leu Ala Gly Gly Leu Leu Ser Gly Lys Tyr Arg Arg 210 215 220 Ser Gly Gly Pro Glu Thr Gly Arg His Val Gly Gly Phe Arg Glu Pro 225 230 235 240 Pro Val Tyr Asp Gln Glu Lys Leu Tyr Asp Val Ile Glu Glu Ile Val 245 250 255 Ala Ile Ala Glu Asp Arg Gly Val Ser Gly Ala Gln Val Ala Leu Ala 260 265 270 Trp Leu Leu Gly Arg Pro Gly Val Thr Ser Leu Ile Ile Gly Gly Arg 275 280 285 Asn Glu Lys Gln Phe Ala Asp Asn Ile Ala Ala Ala Asp Phe Lys Leu 290 295 300 Ser Ala Glu Glu Arg Ala Arg Leu Asp Lys Val Ser Arg Pro Pro Leu 305 310 315 320 Leu Tyr Pro Tyr Trp His Gln Ser Phe Ser Ser Gln Asp Arg Leu Ser 325 330 335 Pro Val Asp Met Asp Leu Ile Gly Pro Tyr Ala Glu Glu Phe Lys Arg 340 345 350 Gly <210> 8 <211> 353 <212> PRT <213> Artificial Sequence <220> <223> Enzyme <400> 8 Met Asp Tyr Arg Tyr Leu Gly Arg Ser Gly Leu Lys Val Ser Ser Met 1 5 10 15 Thr Met Gly Thr Met Thr Phe Gly Gly Ser Glu Lys Ile Gly Asn Thr 20 25 30 Ser Arg Gln Asp Ala Ala Arg Gln Val Asp Met Cys Leu Asp Ala Gly 35 40 45 Ile Asn Leu Tyr Asp Ser Ala Asn Val Tyr Asn Ala Gly Val Ser Glu 50 55 60 Glu Ile Leu Gly Asp Ile Leu Ala Glu Asn Gly Arg Arg Gln Lys Ala 65 70 75 80 Leu Ile Ala Thr Lys Val Arg Phe Lys Met Gly Asp Gly Pro Asn Glu 85 90 95 Ala Gly Leu Ser Arg His His Ile Ile Glu Gln Cys Lys Ala Ser Leu 100 105 110 Lys Arg Leu Lys Thr Asp Val Ile Asp Leu Tyr Gln Val His Glu Trp 115 120 125 Asp Gly Gln Thr Pro Ile Glu Glu Thr Met Glu Ala Leu Asp Ile Leu 130 135 140 Val Arg His Gly His Val Arg Tyr Ile Gly Cys Ser Asn Tyr Ser Gly 145 150 155 160 Trp His Ile Ser Lys Ala Leu Ala Ala Ala Asp Gln Arg His Asn Ala 165 170 175 Arg Phe Val Ser Gln Gln Ile His Tyr Ser Leu His Ser Arg Asp Ala 180 185 190 Glu Tyr Glu Leu Val Pro Ile Ser Gln Asp Gln Gly Leu Gly Ile Leu 195 200 205 Val Trp Ser Pro Leu Ala Gly Gly Leu Leu Ser Gly Lys Tyr Arg Arg 210 215 220 Asp Ser Ala Pro Asp Ser Gly Arg His Val Gly Gly Phe Arg Glu Pro 225 230 235 240 Pro Val Tyr Asp Trp Asp Lys Leu Tyr Ser Val Ile Asp Val Leu Val 245 250 255 Ala Val Ala Glu Glu Arg Gly Val Ser Gly Ala Gln Val Ala Leu Ser 260 265 270 Trp Leu Met Gly Arg Pro Gly Val Thr Ser Val Ile Ile Gly Gly Arg 275 280 285 Asn Glu Lys Gln Phe Ala Asp Asn Ile Ala Ala Ala Gln Leu Val Leu 290 295 300 Ser Asp Glu Glu Arg Ala Arg Leu Asp Ala Ala Ser Arg Pro Pro Leu 305 310 315 320 Leu Tyr Pro Tyr Trp His Gln Ser Phe Thr Ala Gln Asp Arg Leu Gly 325 330 335 Lys Ile Asp Met Asp Leu Ile Gly Pro Tyr Ala Glu Glu Phe Lys Arg 340 345 350 Gly <210> 9 <211> 350 <212> PRT <213> Artificial Sequence <220> <223> Enzyme <400> 9 Met Glu Tyr Arg Leu Leu Gly Arg Ser Gly Leu Lys Val Ser Thr Leu 1 5 10 15 Thr Met Gly Thr Met Thr Phe Gly Gly Asn Pro Lys Val Gly Asn Thr 20 25 30 Ser Leu Lys Asp Ala Gln Arg Gln Ile Asp Met Cys Leu Glu Ala Gly 35 40 45 Ile Asn Met Leu Asp Thr Ala Asn Val Tyr Thr Arg Gly Val Ser Glu 50 55 60 Ser Ile Ile Gly Glu Ala Leu Thr Asp Gly Arg Arg Gln Arg Thr Leu 65 70 75 80 Leu Ala Thr Lys Val Arg Phe Pro Met Asp Asp Thr Asp Pro Asn Glu 85 90 95 Arg Gly Leu Ser Arg Tyr His Ile Val Asn Gln Cys Glu Ala Ser Leu 100 105 110 Lys Arg Leu Lys Thr Asp Val Ile Asp Leu Tyr Gln Val His Glu Trp 115 120 125 Asp Gly Gln Thr Pro Leu Glu Glu Thr Met Glu Ala Leu Asp Met Leu 130 135 140 Val Arg His Gly Lys Val Arg Tyr Ile Gly Cys Ser Asn Tyr Ser Gly 145 150 155 160 Trp His Ile Met Lys Ala Leu Gly Val Ser Glu Lys Asp Ala Arg Gln 165 170 175 Arg Phe Val Ser Gln Gln Ile His Tyr Thr Leu Phe Ser Arg Glu Ala 180 185 190 Glu Asn Glu Leu Val Pro Ile Ser Leu Asp Gln Gly Leu Gly Ile Leu 195 200 205 Ile Trp Ser Pro Leu Ala Gly Gly Leu Leu Ser Gly Lys Tyr Arg Arg 210 215 220 Gly Gln Thr Ala Pro Glu Gly Ala Arg His Ala Gly Gly Phe Lys Glu 225 230 235 240 Pro Pro Val His Asp Trp Glu Arg Leu Tyr Asp Ile Val Glu Val Leu 245 250 255 Val Ser Ile Ala Glu Val Arg Gly Val Ser Gly Ala Gln Ile Ala Leu 260 265 270 Ala Trp Leu Leu Gly Arg Pro Gly Val Thr Ser Val Ile Ile Gly Gly 275 280 285 Arg Thr Glu Asp Gln Phe Arg Asp Asn Leu Ala Ala Ala Glu Leu Lys 290 295 300 Leu Thr Glu His Glu Arg Lys Arg Leu Asp Asp Val Ser Leu Leu Pro 305 310 315 320 Leu Gln Tyr Pro Tyr Trp His Gln Ser Phe Thr Ala Ser Asp Arg Leu 325 330 335 Gly Pro Ala Asp Leu Ala Leu His Ala Pro His Leu Glu Lys 340 345 350 <210> 10 <211> 353 <212> PRT <213> Artificial sequence <220> <223> Enzyme <400> 10 Met Asp Tyr Arg Tyr Leu Gly Arg Ser Gly Leu Lys Val Ser Val Leu 1 5 10 15 Ser Val Gly Thr Val Thr Phe Gly Gly Asp Gly Leu Trp Gly Asp Thr 20 25 30 Asp Val Thr Gly Ala Ala Arg Gln Ile Asp Met Cys Leu Asp Ala Gly 35 40 45 Ile Asn Leu Ile Asp Thr Ala Asn Val Tyr Asn Lys Gly Val Ser Glu 50 55 60 Glu Ile Thr Gly Glu Val Leu Arg Asp Gly Gly Arg Arg Gln Arg Ala 65 70 75 80 Leu Ile Ala Thr Lys Val Arg Phe Pro Phe Gly Asp Gly Pro Asn Asp 85 90 95 Arg Gly Leu Ser Arg His His Ile Ile Ala Gln Cys Glu Gly Ser Leu 100 105 110 Lys Arg Leu Gln Thr Asp Val Ile Asp Leu Tyr Gln Val His Gln Trp 115 120 125 Asp Gly Gln Thr Pro Leu Glu Glu Thr Met Glu Ala Leu Asp Thr Leu 130 135 140 Val Arg Gln Gly Lys Val Arg Tyr Ile Gly Cys Ser Asn Tyr Ser Gly 145 150 155 160 Trp His Leu Met Lys Ala Leu Ala Val Ala Asp Ser Arg Gly Tyr Gln 165 170 175 Arg Phe Val Ala Gln Gln Ile His Tyr Thr Leu Tyr Ser Arg Glu Ala 180 185 190 Glu Tyr Glu Leu Gln Pro Ile Thr Val Asp Gln Gly Leu Gly Ile Leu 195 200 205 Val Trp Ser Pro Leu Ala Gly Gly Trp Leu Ser Gly Lys Tyr Thr Arg 210 215 220 Ala Ser Gln Pro Glu Asp Gly Arg His Val Arg Gly Phe Arg Glu Pro 225 230 235 240 Pro Val Pro Asp Pro Glu Arg Leu Tyr Asp Ile Val Asp Val Ile Asn 245 250 255 Ala Val Ala Ala Glu Arg Gly Val Ser Gly Ala Val Val Ser Leu Ala 260 265 270 Trp Ala Leu Gln Arg Lys Gly Val Ser Ser Val Ile Val Gly Ala Arg 275 280 285 Thr Asp Ala Gln Phe Ala Asp Asn Leu Lys Ala Ala Asp Leu Lys Leu 290 295 300 Ser Asp Asp Glu Val Ala Arg Leu Asp Ala Val Ser Gln Arg Pro Leu 305 310 315 320 Pro Tyr Pro Tyr Trp His Gln Ser Phe Thr Ala Gln Asp Arg Leu Ser 325 330 335 Glu Ala Asp Leu Val Leu Leu Ala Pro Tyr Leu Glu Glu Tyr Lys Arg 340 345 350 Gly <210> 11 <211> 348 <212> PRT <213> Artificial sequence <220> <223> Enzyme[[ID=2⑨]] <400> 11 Met Asp Tyr Arg Leu Leu Gly Arg Ser Gly Leu Lys Val Ser Ser Leu 1 5 10 15 Ser Ile Gly Thr Ala Thr Phe Gly Gly Asp Gly Leu Trp Gly Asp Thr 20 25 30[[ID=3⑨]] Asn Val Lys Glu Ala Gln Arg Gln Ile Asp Leu Cys Leu Asp His Gly 35 40 45 Val Asn Leu Ile Asp Thr Ala Asn Val Tyr Ser Gly Ser Val Ser Glu 50 55 60 Thr Ile Ile Gly Glu Ala Leu Thr Asp Gly Arg Arg Asp Arg Leu Leu 65 70 75 80 Ile Ala Thr Lys Val Arg Phe Pro Arg Gly Lys Gly Pro Asn Asp Arg 85 90 95 Gly Leu Ser Arg Trp His Ile Ile Arg Glu Ala Glu Ala Ser Leu Lys 100 105 110 Arg Met Lys Thr Asp Val Ile Asp Leu Tyr Gln Val His Glu Trp Asp 115 120 125 Gly Gln Thr Pro Leu Glu Glu Thr Leu Glu Ala Leu Asp Thr Leu Val 130 135 140 Arg Ser Gly Lys Val Arg Tyr Ile Gly Cys Ser Asn Tyr Ser Ala Trp 145 150 155 160 His Leu Met Lys Ala Leu Ala Val Ser Asn Glu Arg Asn Tyr Gln Arg 165 170 175 Phe Val Ser Gln Gln Ile His Tyr Thr Leu His Ser Arg Glu Ala Glu 180 185 190 Phe Glu Leu Ile Pro Ala Gly Leu Asp Gln Gly Val Ser Val Leu Val 195 200 205 Trp Ser Pro Leu Ala Gly Gly Trp Leu Ser Gly Lys Tyr Thr Arg Asp 210 215 220 Ser Lys Pro Ser Asp Gly Arg Gln Val Arg Gly Phe Lys Glu Pro Pro 225 230 235 240 Ile Tyr Asp Pro Glu His Leu Trp Lys Ile Val Asp Val Val Asn Asp 245 250 255 Val Ala Gly Ala Arg Gly Val Ser Gly Ala Gln Val Ser Leu Ala Trp 260 265 270 Leu Leu Ser Lys Pro Ile Val Ala Ser Val Ile Val Gly Gly Arg Asn 275 280 285 Ile Gly Gln Phe Glu Asp Asn Leu Lys Ala Ala Glu Leu Thr Leu Thr 290 295 300 Thr Glu Glu Leu Gln Lys Leu Asp Ala Val Ser Gln Pro Asn Leu Pro 305 310 315 320 Tyr Pro Tyr Trp His Gln Ser Phe Thr Ala Gln Asp Arg Leu Gly Glu 325 330 335 Pro Asp Leu Ala Leu His Arg Pro Tyr Leu Asp Lys 340 345 <210> 12 <211> 348 <212> PRT <213> Synthetic Sequence <220> <223> Enzyme [[ID=4,4]]<400> 12 Met Asp Tyr Arg Leu Leu Gly Arg Ser Gly Leu Lys Val Ser Ser Leu 1 5 10 15 Ser Ile Gly Thr Ala Thr Phe Gly Gly Asp Gly Leu Trp Gly Asp Thr 20 25 30 Asn Val Lys Glu Ala Gln Arg Gln Ile Asp Leu Cys Leu Asp His Gly 35 40 45 Val Asn Leu Ile Asp Thr Ala Asn Val Tyr Ser Gly Ser Val Ser Glu 50 55 60 Thr Ile Ile Gly Glu Ala Leu Thr Asp Gly Arg Arg Asp Arg Leu Leu 65 70 75 80 Ile Ala Thr Lys Val Arg Phe Pro Arg Gly Lys Gly Pro Asn Asp Arg 85 90 95 Gly Leu Ser Arg Trp His Ile Ile Arg Glu Ala Glu Ala Ser Leu Lys 100 105 110 Arg Met Lys Thr Asp Val Ile Asp Leu Tyr Gln Val His Glu Trp Asp 115 120 125 Gly Gln Thr Pro Leu Glu Glu Thr Leu Glu Ala Leu Asp Thr Leu Val 130 135 140 Arg Ser Gly Lys Val Arg Tyr Ile Gly Cys Ser Asn Tyr Ser Ala Trp 145 150 155 160 His Leu Met Lys Ala Leu Ala Thr Ser Asn Glu Arg Asn Tyr Gln Arg 165 170 175 Phe Val Ser Gln Gln Ile His Tyr Thr Leu His Ser Arg Glu Ala Glu 180 185 190 Phe Glu Leu Ile Pro Ala Gly Leu Asp Gln Gly Val Ser Val Leu Val 195 200 205 Trp Ser Pro Leu Ala Gly Gly Trp Leu Ser Gly Lys Tyr Thr Arg Asn 210 215 220 Ser Lys Pro Ser Asp Gly Arg Gln Val Arg Gly Phe Lys Glu Pro Pro 225 230 235 240 Ile Tyr Asp Pro Glu His Leu Trp Lys Ile Val Asp Val Val Asn Glu 245 250 255 Val Ala Ser Ala His Gly Val Ser Gly Ala Gln Val Ser Leu Ala Trp 260 265 270 Leu Leu Ser Lys Pro Ile Val Ala Ser Val Ile Val Gly Gly Arg Asn 275 280 285 Ile Glu Gln Phe Glu Asp Asn Leu Lys Ala Ala Glu Leu Gln Leu Thr 290 295 300 Ala Glu Glu Leu Gly Arg Leu Asp Ala Val Ser Gln Pro Asn Leu Pro 305 310 315 320 Tyr Pro Tyr Trp His Gln Asn Phe Thr Ala Gln Asp Arg Leu Ser Glu 325 330 335 Pro Asp Leu Ala Leu His Arg Pro Tyr Leu Asp Lys 340 345 <210> 13 <211> 351 <212> PRT <213> Artificial Sequence <220> <223> Enzyme <400> 13 Met Glu Tyr Arg Leu Leu Gly Arg Ser Gly Leu Lys Val Ser Thr Leu 1 5 10 15 Thr Met Gly Thr Met Thr Phe Gly Gly Gln Gly Lys Phe Ala Ser Val 20 25 30 Gly Asn Ser Gly Val Asp Glu Ala Arg Arg Leu Ile Asp Met Cys Ile 35 40 45 Asp Ala Gly Val Asn Leu Ile Asp Thr Ala Asn Val Tyr Ser Asp Gly 50 55 60 Arg Ser Glu Glu Ile Ile Gly Glu Val Leu Gly Gly Lys Arg Pro Lys 65 70 75 80 Asp Val Leu Ile Ala Thr Lys Ala Arg Phe Pro Met Gly Asp Gly Pro 85 90 95 Asn Asn Gly Gly Leu Ser Arg Tyr His Leu Ile Arg Glu Cys Glu Ala 100 105 110 Ser Leu Lys Arg Leu Arg Thr Asp Val Ile Asp Leu Tyr Gln Val His 115 120 125 Gln Trp Asp Gly Leu Thr Pro Leu Glu Glu Thr Leu Glu Ala Leu Asp 130 135 140 Thr Leu Val Arg Asp Gly Lys Val Arg Tyr Leu Gly Cys Ser Asn Tyr 145 150 155 160 Ser Ala Trp His Leu Met Lys Ala Leu Gly Ile Ser Glu Arg Glu His 165 170 175 Arg Gln Arg Phe Val Ser Gln Gln Ile His Tyr Thr Leu Glu Ala Arg 180 185 190 Glu Ala Glu Tyr Glu Leu Leu Pro Ile Ser Ile Asp Gln Gly Leu Gly 195 200 205 Val Leu Val Trp Ser Pro Leu Ala Gly Gly Leu Leu Ser Gly Lys His 210 215 220 Arg Arg Asp Gly Lys Ala Pro Glu Gly Ser Arg Gln Leu Glu Gly Trp 225 230 235 240 Thr Glu Pro Pro Ile Arg Asp Glu Glu Arg Leu Trp Arg Ile Val Asp 245 250 255 Val Leu Val Ala Ile Ala Glu Ala Arg Gly Val Ser Gly Ala Gln Val 260 265 270 Ala Leu Ala Trp Leu Leu Glu Arg Pro Ala Val Thr Ser Leu Val Ile 275 280 285 Gly Gly Arg Thr Glu Ala Gln Phe Lys Asp Asn Leu Ala Ala Ala Glu 290 295 300 Leu Lys Leu Thr Ala Glu Glu Arg Gln Arg Leu Asp Glu Val Ser Ala 305 310 315 320 Pro Pro Val Leu Tyr Pro Tyr Trp His Gln Leu Met Thr Ala Ser Asp 325 330 335 Arg Leu Gly Pro Ala Asp Met Pro Leu Leu Ala Pro His Leu Lys 340 345 350 <210> 14 <211> 351 <212> PRT <213> Artificial Sequence <220> <223> Enzyme <400> 14 Met Glu Tyr Arg Leu Leu Gly Arg Ser Gly Leu Lys Val Ser Thr Leu 1 5 10 15 Thr Met Gly Thr Met Thr Phe Gly Gly Gln Gly Lys Phe Ala Ser Val 20 25 30 Gly Ser Ser Gly Val Asp Glu Ala Arg Arg Leu Ile Asp Met Cys Ile 35 40 45 Asp Ala Gly Val Asn Leu Ile Asp Thr Ala Asn Val Tyr Ser Asp Gly 50 55 60 Arg Ser Glu Glu Ile Ile Gly Glu Val Leu Gly Gly Lys Arg Pro Lys 65 70 75 80 Asp Val Leu Ile Ala Thr Lys Ala Arg Phe Pro Met Gly Asp Gly Pro 85 90 95 Asn His Gly Gly Leu Ser Arg Tyr His Leu Ile Arg Glu Cys Glu Ala 100 105 110 Ser Leu Lys Arg Leu Arg Thr Asp Val Ile Asp Leu Tyr Gln Val His 115 120 125 Gln Trp Asp Gly Leu Thr Pro Leu Glu Glu Thr Leu Glu Ala Leu Asp 130 135 140 Thr Leu Val Arg Asp Gly Lys Val Arg Tyr Leu Gly Cys Ser Asn Tyr 145 150 155 160 Ser Ala Trp His Leu Met Lys Ala Leu Gly Val Ser Glu Arg Glu His 165 170 175 Arg Gln Arg Phe Ile Ser Gln Gln Ile His Tyr Thr Leu Glu Ala Arg 180 185 190 Glu Ala Glu Tyr Glu Leu Leu Pro Ile Ser Ile Asp Gln Gly Leu Gly 195 200 205 Val Leu Val Trp Ser Pro Leu Ala Gly Gly Leu Leu Ser Gly Lys His 210 215 220 Arg Arg Asp Thr Lys Ala Pro Glu Gly Ser Arg Gln Leu Ala Gly Trp 225 230 235 240 Thr Glu Pro Pro Ile Arg Asp Glu Glu Arg Leu Trp Arg Val Val Asp 245 250 255 Val Leu Val Ser Ile Ala Gln Ser Arg Gly Val Ser Gly Ala Gln Val 260 265 270 Ala Leu Ala Trp Leu Leu Glu Arg Pro Ala Val Thr Ser Leu Val Ile 275 280 285 Gly Gly Arg Thr Glu Ala Gln Phe Lys Asp Asn Leu Ala Ala Ala Glu 290 295 300 Leu Lys Leu Thr Ala Glu Glu Arg Gln Arg Leu Asp Glu Val Ser Ala 305 310 315 320 Pro Pro Val Leu Tyr Pro Tyr Trp His Gln Leu Met Thr Ala Ser Asp 325 330 335 Arg Leu Gly Pro Ala Asp Met Pro Leu Leu Ala Pro His Leu Lys 340 345 350 <210> 15 <211> 343 <212> PRT <213> artificial sequence <220> <223> Enzyme <400> 15 Met Glu Tyr Arg Lys Leu Gly Asn Ser Gly Thr Val Val Thr Ser Tyr 1 5 10 15 Cys Leu Gly Thr Met Thr Phe Gly Gln Glu Thr Asp Glu Ala Thr Ser 20 25 30 His Leu Ile Met Asp Asp Tyr Ile Lys Ala Gly Gly Asn Phe Ile Asp 35 40 45 Thr Ala Asn Val Tyr Ser Ala Gly Val Ser Glu Glu Ile Val Gly Arg 50 55 60 Trp Leu Lys Ala Arg Pro Ser Glu Ala Arg Gln Val Val Val Ala Thr 65 70 75 80 Lys Gly Arg Phe Pro Met Gly Ala Gly Pro Asn Asp Leu Gly Leu Ser 85 90 95 Arg Thr Asn Leu Asn Arg Ala Leu Asn Asp Ser Leu Arg Arg Leu Gly 100 105 110 Val Glu Gln Ile Asp Leu Tyr Gln Met His Ala Trp Asp Ala Val Thr 115 120 125 Pro Ile Glu Glu Thr Leu Arg Phe Leu Asp Asp Ala Val Ser Ala Gly 130 135 140 [[ID= 145 150 155 160 Lys Ala Val His Val Ala Arg Ala Asn His Trp Thr Ala Pro Val Thr 165 170 175 Leu Gln Pro Gln Tyr Asn Leu Leu Val Arg Asp Ile Glu His Glu Ile 180 185 190 Val Pro Ala Cys Gln Asp Ala Ala Met Gly Leu Leu Pro Trp Ser Pro 195 200 205 Leu Gly Gly Gly Trp Leu Ala Gly Lys Tyr Gln Arg Asp Val Met Pro 210 215 220 Ser Gly Ala Thr Arg Leu Gly Glu Asn Pro Asn Arg Gly Met Glu Ser 225 230 235 240 Tyr Gly Pro Arg Asn Ala Gln Glu Arg Thr Trp Gln Ile Ile Asp Met 245 250 255 Val Ala Glu Ile Ala Lys Glu Arg Gly Val Ser Ala Ala Gln Val Ala 260 265 270 Leu Ala Trp Val Val Ala Arg Pro Ala Val Thr Ala Val Ile Leu Gly 275 280 285 Ala Arg Thr Arg Glu Gln Leu Ala Asp Asn Leu Gly Ala Val Ala Val 290 295 300 Thr Leu Ser Thr Glu Glu Met Glu Arg Leu Asn Arg Val Ser Ala Pro 305 310 315 320 Ala Met Ala Asp Tyr Pro Tyr Gly Glu Arg Gly Val Ser Gln Arg His 325 330 335 Arg Lys Met Asp Gly Gly Arg 340 <210> 16 <211> 342 <212> PRT <213> Artificial Sequence <220> <223> Enzyme <400> 16 Met Asp Tyr Arg Lys Leu Gly Pro Ser Gly Thr Val Val Thr Ala Tyr 1 5 10 15 Cys Leu Gly Thr Met Thr Gly Ala Glu Ala Asp Glu Ala Ala Ser His 20 25 30 Lys Leu Leu Asp Asp Tyr Phe Ala Trp Gly Gly Asn Phe Ile Asp Thr 35 40 45 Ala Asp Val Tyr Ser Ala Gly Lys Ser Glu Glu Ile Ile Gly Arg Trp 50 55 60 Leu Lys Ala Arg Pro Thr Glu Ala Arg Gln Ala Ile Val Ala Thr Lys 65 70 75 80 Gly Arg Phe Pro Met Gly Asn Gly Pro Asn Asp Ile Gly Leu Ser Arg 85 90 95 Arg His Leu Ser Gln Ala Leu Asp Asp Ser Leu Arg Arg Leu Gly Leu 100 105 110 Glu Gln Ile Asp Leu Tyr Gln Met His Ala Trp Asp Ala Leu Thr Pro 115 120 125 Ile Glu Glu Thr Leu Arg Phe Leu Asp Asp Ala Val Ser Ser Gly Lys 130 135 140 Ile Gly Tyr Tyr Gly Phe Ser Asn Tyr Val Gly Trp His Ile Ala Lys 145 150 155 160 Ala Ser Glu Ile Ala Lys Ala Arg Gly Tyr Thr Arg Pro Val Thr Leu 165 170 175 Gln Pro Gln Tyr Asn Leu Leu Met Arg Asp Ile Glu Leu Glu Ile Val 180 185 190 Ala Ala Cys Gln Asp Ala Gly Met Gly Leu Leu Pro Trp Ser Pro Leu 195 200 205 Gly Gly Gly Trp Leu Thr Gly Lys Tyr Lys Arg Asp Glu Met Pro Thr 210 215 220 Gly Ala Thr Arg Leu Gly Glu Asn Pro Asn Arg Gly Gly Glu Ser Tyr 225 230 235 240 Ala Pro Arg Asn Ala Gln Glu Arg Thr Trp Ala Ile Ile Gly Thr Val 245 250 255 Glu Glu Ile Ala Lys Ala Arg Gly Val Ser Met Ala Gln Val Ala Leu 260 265 270 Ala Trp Thr Ala Ala Arg Pro Ala Ile Thr Ser Val Ile Leu Gly Ala 275 280 285 Arg Thr Pro Glu Gln Leu Ala Asp Asn Leu Gly Ala Met Lys Val Glu 290 295 300 Leu Ser Gly Glu Glu Met Ala Arg Leu Asn Glu Val Ser Ala Pro Gln 305 310 315 320 Pro Leu Asp Tyr Pro Tyr Gly Lys Gly Gly Ile Asn Gln Arg His Arg 325 330 335 Lys Ile Glu Gly Gly Arg 340 <210> 17 <211> 9 <212> PRT <213> Artificial sequence <220> <223> motif <220> <221> MISC_FEATURE <222> (1)..(1) <223> X is not M or X is V, I, L, M, F or A. <220> <221> MISC_FEATURE <222> (3) <223> X is not A or X is E, D, Q or N. <220> <221> MISC_FEATURE <222> (6) <223> X is not A or X is G or A. <220> <221> MISC_FEATURE <222> (7)..(7) <223> X is not V or X is M, L, F, I, R, K, Q, N, E, V, A, or norleucine. <220> <221> MISC_FEATURE <222> (7)..(7) <223> X is not V and / or X is M, L, F, I, R, K, Q, N, E, V, A, or norleucine. <400> 17 Xaa His Xaa Trp Asp Xaa Xaa Thr Pro 1 5 <210> 18 <400> 18 000 <210> 19 <211> 10 <212> PRT <213> Artificial sequence <220> <223> motif <220> <221> MISC_FEATURE <222> (1)..(1) <223> X is G, R, K, Q, N, A, V, L, I, S, T, E, D <220> <221> MISC_FEATURE <222> (3) <223> X is not M or G or X is W, S, T, F, L, V, I, A or Y <220> <221> MISC_FEATURE <222> (4)..(4) <223> X is not E or X is T, S, R, Q, N or K. <220> <221> MISC_FEATURE <222> (4)..(4) <223> X is not E and / or X is T, S, R, Q, N or K. <220> <221> MISC_FEATURE <222> (5)..(5) <223> X is not S and / or X is E, D or Q. <220> <221> MISC_FEATURE <222> (5)..(5) <223> X is not S and / or X is E, D or Q. <220> <221> MISC_FEATURE <222> (5)..(5) <223> X is not S and / or X is P or A. <220> <221> misc_feature <222> (6)..(7) <223> Xaa can be any naturally occurring amino acid <220> <221> MISC_FEATURE <222> (8) <223> X is not P and / or X is I, L, M, A, F or V. <220> <221> MISC_FEATURE <222> (9)..(9) <223> X is R, K, Q, N, P, A, Y, W, F, T, S, H. <220> <221> MISC_FEATURE <222> (10)..(10) <223> X is not N and / or X is D, E or N. <400> 19 Xaa Gly Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa 1 5 10 <210> 20 <211> 9 <212> PRT <213> Artificial sequence <220> <223> motif <400> 20 Val His Glu Trp Asp Gly Met Thr Pro 1 5 <210> twenty one <211> 9 <212> PRT <213> Artificial sequence <220> <223> motif <400> twenty one Val His Glu Trp Asp Gly Arg Thr Pro 1 5 <210> twenty two <211> 9 <212> PRT <213> Artificial sequence <220> <223> motif <400> twenty two Val His Glu Trp Asp Gly Gln Thr Pro 1 5 <210> twenty three <211> 9 <212> PRT <213> Artificial sequence <220> <223> motif <400> twenty three Val His Gln Trp Asp Gly Leu Thr Pro 1 5 <210> twenty four <211> 9 <212> PRT <213> Artificial sequence <220> <223> motif <400> twenty four Val His Gln Trp Asp Gly Gln Thr Pro 1 5 <210> 25 <211> 10 <212> PRT <213> Artificial sequence <220> <223> motif <400> 25 Gly Gly Phe Arg Glu Pro Pro Val Tyr Asp 1 5 10 <210> 26 <211> 10 <212> PRT <213> Artificial sequence <220> <223> motif <400> 26 Gly Gly Ser Arg Glu Pro Pro Val Pro Asp 1 5 10 <210> 27 <211> 10 <212> PRT <213> Artificial sequence <220> <223> motif <400> 27 Gly Gly Phe Arg Glu Pro Pro Val Pro Asp 1 5 10 <210> 28 <211> 10 <212> PRT <213> Artificial sequence <220> <223> motif <400> 28 Gly Gly Phe Lys Glu Pro Pro Val His Asp 1 5 10 <210> 29 <211> 10 <212> PRT <213> Artificial sequence <220> <223> motif <400> 29 Arg Gly Phe Arg Glu Pro Pro Val Pro Asp 1 5 10 <210> 30 <211> 10 <212> PRT <213> Artificial sequence <220> <223> motif <400> 30 Glu Gly Trp Thr Glu Pro Pro Ile Arg Asp 1 5 10 <210> 31 <211> 10 <212> PRT <213> Artificial sequence <220> <223> motif <400> 31 Ala Gly Trp Thr Glu Pro Pro Ile Arg Asp 1 5 10 <210> 32 <211> 10 <212> PRT <213> Artificial sequence <220> <223> motif <400> 32 Arg Gly Phe Lys Glu Pro Pro Ile Tyr Asp 1 5 10 <210> 33 <211> 34 <212> PRT <213> Artificial sequence <220> <223> Motif <400> 33 Ser Pro Leu Ala Gly Gly Leu Leu Ser Gly Lys Tyr Arg Arg Asp Gly 1 5 10 15 Gly Pro Asp Ala Gly Arg His Val Gly Gly Phe Arg Glu Pro Pro Val 20 25 30 Tyr Asp <210> 34 <211> 34 <212> PRT <213> Artificial sequence <220> <223> Motif <400> 34 Ser Pro Leu Ala Gly Gly Leu Leu Ser Gly Lys Tyr Arg Arg Asp Gly 1 5 10 15 Gly Pro Glu Ser Gly Arg His Val Gly Gly Ser Arg Glu Pro Pro Val s20 25 30 Pro Asp [[ID=4B]]<210> 35 <211> 34 <212> PRT <213> Artificial sequence <220> <223> Motif <400> 35 Ser Pro Leu Ala Gly Gly Leu Leu Ser Gly Lys Tyr Arg Arg Asp Gly 1 5 10 15 Gly Pro Glu Ser Gly Arg His Val Gly Gly Ser Arg Glu Pro Pro Val[[ID=B2]] 20 25 30 Pro Asp <210> 36 <211> 34 <212> PRT <213> Artificial sequence <220> <223> motif <400> 36 Ser Pro Leu Ala Gly Gly Leu Leu Ser Gly Lys Tyr Arg Arg Asn Gly 1 5 10 15 Gly Pro Asp Ser Gly Arg His Val Gly Gly Phe Arg Glu Pro Pro Val 20 25 30 Tyr Asp <210> 37 <211> 34 <212> PRT <213> Artificial sequence <220> <223> motif <400> 37 Ser Pro Leu Ala Gly Gly Leu Leu Ser Gly Lys Tyr Arg Arg Asp Gly 1 5 10 15 Gly Pro Asp Ala Gly Arg His Val Gly Gly Phe Arg Glu Pro Pro Val 20 25 30 Pro Asp <210> 38 <211> 34 <212> PRT <213> Artificial sequence <220> <223> motif <400> 38 Ser Pro Leu Ala Gly Gly Leu Leu Ser Gly Lys Tyr Arg Arg Thr Gly 1 5 10 15 Gly Pro Glu Ser Gly Arg His Val Gly Gly Phe Arg Glu Pro Pro Val 20 25 30 Tyr Asp <210> 39 <211> 34 <212> PRT <213> Artificial sequence <220> <223> motif <400> 39 Ser Pro Leu Ala Gly Gly Leu Leu Ser Gly Lys Tyr Arg Arg Ser Gly 1 5 10 15 Gly Pro Glu Thr Gly Arg His Val Gly Gly Phe Arg Glu Pro Pro Val 20 25 30 Tyr Asp <210> 40 <211> 34 <212> PRT <213> Artificial sequence <220> <223> motif <400> 40 Ser Pro Leu Ala Gly Gly Leu Leu Ser Gly Lys Tyr Arg Arg Asp Ser 1 5 10 15 Ala Pro Asp Ser Gly Arg His Val Gly Gly Phe Arg Glu Pro Pro Val 20 25 30 Tyr Asp <210> 41 <211> 35 <212> PRT <213> Artificial sequence <220> <223> motif <400> 41 Ser Pro Leu Ala Gly Gly Leu Leu Ser Gly Lys Tyr Arg Arg Gly Gln 1 5 10 15 Thr Ala Pro Glu Gly Ala Arg His Ala Gly Gly Phe Lys Glu Pro Pro 20 25 30 Val His Asp 35 <210> 42 <211> 34 <212> PRT <213> Synthetic sequence <220> <223> Motif <400> 42 Ser Pro Leu Ala Gly Gly Trp Leu Ser Gly Lys Tyr Thr Arg Ala Ser 1 5 10 15 Gln Pro Glu Asp Gly Arg His Val Arg Gly Phe Arg Glu Pro Pro Val 20 25 30 Pro Asp <210> 43 <211> 34 <212> PRT <213> Synthetic sequence <220> <223> Motif <400> 43 Ser Pro Leu Ala Gly Gly Trp Leu Ser Gly Lys Tyr Thr Arg Asp Ser 1 5 10 15 Lys Pro Ser Asp Gly Arg Gln Val Arg Gly Phe Lys Glu Pro Pro Ile 20 25 30 Tyr Asp <210> 44 <211> 34 <212> PRT <213> Artificial sequence <220> <223> motif <400> 44 Ser Pro Leu Ala Gly Gly Trp Leu Ser Gly Lys Tyr Thr Arg Asn Ser 1 5 10 15 Lys Pro Ser Asp Gly Arg Gln Val Arg Gly Phe Lys Glu Pro Pro Ile 20 25 30 Tyr Asp <210> 45 <211> 35 <212> PRT <213> Artificial sequence <220> <223> motif <400> 45 Ser Pro Leu Ala Gly Gly Leu Leu Ser Gly Lys His Arg Arg Asp Gly 1 5 10 15 Lys Ala Pro Glu Gly Ser Arg Gln Leu Glu Gly Trp Thr Glu Pro Pro 20 25 30 Ile Arg Asp 35 <210> 46 <211> 35 <212> PRT <213> Artificial sequence <220> <223> motif <400> 46 Ser Pro Leu Ala Gly Gly Leu Leu Ser Gly Lys His Arg Arg Asp Thr 1 5 10 15 Lys Ala Pro Glu Gly Ser Arg Gln Leu Ala Gly Trp Thr Glu Pro Pro 20 25 30 Ile Arg Asp 35 <210> 47 <211> 771 <212> DNA <213> Artificial Sequence <220> <223> Enzyme <400> 47 atgacccgat ttgcccatta tcccagcctg accgatatgc ccgtcgtcat ctcaggcggt 60 gcctcgggca tcggcgaaag cctggtgcga aactttgccg cccagggcgc cagggtcggt 120 ttcgtcgata tcgccgtcga cgccggtgaa cgcctcgccg ccgaattgac ggggcagggg 180 caaagcgtag ccttcaccgc ctgcgacatc accgataccc aggcctatca atccgccatt 240 gccggcttcg ccgccaccca tggcgatgcc ctggccctgg tcaacaatgc cgcccatgac 300 cagcgccacg attgggctga ggtcacccct gcctattggg acgatcgcat ggcggtcaac 360 ctcaagcatg cctttttcgc catccaggcc gtggcgccgg gcatggtccg cgccgggcgc 420 ggctccatca tcaataccgg ctcgatcagc tggatgatca tgtcctcccg cattcccgtc 480 tatgaaaccg ccaaggccgc catccatggc ctcacccgcg gcatggcgcg cgaactgggg 540 cagtcgggca ttcgcgtcaa ttccctggtg cccggctggg tcatgaccga gcggcaattg 600 acccattggc tcgatgccga gggcgagcgg gccatcgcac aaaaccaggt gctggccggc 660 cgggtctatc ccgacgatgt cgcgcgcatg gccctgttcc ttgccgccga cgacagcgcc 720 atgatttcgg cccagcaatt tgtcgtcgat ggcggctggg ccaatgcctg a 771 <210> 48 <211> 1086 <212> DNA <213> Artificial Sequence <220> <223> Enzyme <400> 48 atgagccgca ttttcaacgt cgccatcgtc ggggtcggca tcggccgcag ccacctggtc 60 gagggctatg tgcccaatgc cgacaaatac aaggtcgtcg cgctgtgcga tctcaatgtc 120 gagcgcatga cgcccattgc cgacgaattc ggcgtcgagc gccgcatcac caatttcgac 180 gatatcctcg ccatggacga tatcgacatc atcgatatct gcaccccgcc gggcctgcat 240 tatcccatgg tcatggcggc gctgaaggcc ggcaagcatg taatctgcga aaagccgctg 300 gtcggctcgc tcgcccaggt cgatgaagtc attgcccagg aaaagatctc caagggcaag 360 ctgatgccgg tgttccagta tcgctttggc gacggcatcg aacaggccaa ggccatcatc 420 gatgccggca ttgccggcaa gccattcgtc ggcacggtgg aaaccatgtg gcgccgtggc 480 cccgattatt atgccgtgcc atggcgcggc aaatgggcca ccgaactggg cggcgtgctg 540 atgacccatg ccatccacca gcacgatctc ttcacctatc tcatgggcga tgtgacgcgg 600 ctcttcggcc gggtcgccac ccgcgtcaac gccattgaag tggaagactg cgttaccgcc 660 agccttgagc tcgaaaatgg cgccctgggc agctttaccg ccacgctggg cgcagccgag 720 gaaatcacaa ggctgttcct gaccttcgag aacgtcacct tcgaaagcga ttacgaggcc 780 tataatcccg gcgccaagcc gtggcgcatc cagccgcgca atcccgagac ggcggcgaaa 840 atcgacgccc tgctcaagga ctggaagcat gtgccctcgc gtttcgaaac gcagatggcc 900 cgcttccatg cggccctctt gggtgaagcg ccgatgccgg tgacctcggc cgattcgcgc 960 cgggcgctgg aaatcgtcac cgccttctac cattcctcga ctacccacga agatgtggcc 1020 ctgcccatcg cgtccgacca tccgcgttat cagagctggg ttccgcccca gttccgcgct 1080 gcctga 1086 <210> 49 <211> 1062 <212> DNA <213> Artificial Sequence <220> <223> Enzyme <400> 49 atggactatc gctatctcgg ccgctcgggc cttagggttt cggtgctgac catgggcaca 60 atgacctttg gcggctcgga aaaggtgggg catacagccc aggccgatgc gacgcgccag 120 attgatctct gccttgatca cggcatcaac ctgctcgaca ccgccaatgt ctataatgcc 180 ggggtctccg aggagatgat cggggtggcg ctgagcgaac atggccggcg gcaaaaggct 240 ttgattgcca ccaaggtgcg cttcaggatg ggtaacgggc ccaatgaagt ggggctgagc 300 cggcatcata tcattgccca ggccgaggcg agcctcaagc ggctcaagac cgatgtcatc 360 gatctctacc aggtgcatga atgggatggg atgacgccca tcgaggagac gatggaagcg 420 ctcgatacgc tcgtgaaaca gggcaaggtg cgctatgtcg gctgctcgaa ttattcgggc 480 tggcacatca tgaaggcgct ggctgcggcc gacaaacatc atggccagcg cttcatctcc 540 cagcagatcc actacacgct gcattcgcgc gaagccgaat atgaactggt gccgatcagc 600 caggaccagg ggctgggtat tctgatttgg tcgccgctgg cgggcggact gctctcgggc 660 aagtatcgtc gcgatggcgg gcccgatgcc ggccgccatg ttggaggctt ccgcgagccg 720 ccggtttatg actgggacaa gctctacgac attatcgacg tcatcgtcgc cattgcccgg 780 gagcgcgatg tgtcgggcgc gcaggtggcg ttggcctggg cattggggcg gccgggcgtg 840 acctcggcca ttatcggtgg ccgtagcgag gcccagttcc gcgacaatct ggcggcggcc 900 gaactcaagc tgaccgatga cgagcgcgcc cggctcgatg cggccagccg gccgccattg 960 ctttatccct attggcacca gtcttttacc gcccaggacc ggctggacca gatcgacctg 1020 gacctgatcg ggccctatgc cgaggagttt aaacgtggct ga 1062 <Type> 50 <Length> 786 <Molecule> DNA <Sequence> Synthetic sequence <Description> <Function> Enzyme <Database cross-reference> 50 atggacttga atttgcgcga caaggttgcc gtcattaccg ggggaacggt ggggatcggc 60<ctggccattg ccgaaggcct ggcggcggag ggcgtcaatc tggtgctggt ggggcgcgac 120 aagctgcggg ctgacgatgc ggcagcgctt gtggccgaca agttcgggat tatcgccacg 180 gcgatcagcg cggatgtcgc gactgccgaa gggtgcgatg ccgtgatcaa gggcacggcc 240 aaggcctttg gtggcgccga tatcctgatc aacaatgccg ggaccgggtc gaacgagacc 300 attgccgagg ccgatgatgc caaatggcaa tattattggg acctgcatgt gatggcggcc 360 gtgcggctgg cgcgggggct ggtgccggga atgaaaaagc ggggcggcgg cgtcgtgctg 420 cacaatgcct cgatctgcgc ggtgcagccg ctctggtatg agccgatcta caacaccacc 480 aaggccgcct tgatgatgtt ttccaagacg ctggccaatg aggtggtggg cgacaatatt 540 cgcgtcaata ccatcaatcc cgggctggtg ctgacgcccg attgggtcaa gaccgccaag 600 cagattgccg gcgaagatgg ctggcaggcg catctgcaag gggtggccga cgaggcgggc 660 ggcatgaagc gcttcgcgac gcccgaggaa ttggccaatt tcttcgtctt catgtgttcg 720 gacagagcca gctattccac cggctcgacc tattttgtcg atggcggctg gctcaagacg 780 gtttga 786 <210> 51 <211> 1038 <212> DNA <213> Artificial sequence <220> <223> Enzyme <400> 51 atggaactcg gactctattc cttcgccgaa aacacgcccg atcccctgaa cggccgccgc 60 ctgcaaagcc cggccgaacg tctcaaggac ctgctggaag aaatcgaact ggccgaccag 120 ctcggcctat ccttttacgg cctgggcgaa caccaccgcc ccgactacac cgcttccgcg 180 cccgtcacca tcctggccgc cgccgccgcg cgcaccaagt cgatccgcct ctctaccgcg 240 gtcaccgtgc tcagctcgga agaccccatc cgcgtctatc agcaattcgc cacgctcgat 300 aatctcagca atggccgcgc cgagatcatg gccggccgcg gctccttcat cgagagcttc 360 cccctcttcg ggctcgacct caacgattac gacgcgctgt tcgaggaaaa gctcgaaatg 420 ctgctcaaaa tccgcgaagg cggcaaggtc acctggcccg gcagcaccca taccaagccc 480 atccccggca ttgccatcta tcccatcccg gtccagcagc ccctgccgct ctggatcgcc 540 gtcggcggca cgccaaattc cgttgcccgc gcggcctatt acggcctgcc cctgatgatc 600 gccatcatcg gcggtcagcc gcgccagttt gcgccgctgg tcgatttcta ccgcgaaacc 660 gctgaaaagg tcggccgcga ccccaaatcg ctgcccgtcg gcatcagctc gcacggcttc 720 atcgccgccg acagccagga cgcccgcgac atcgccttcc ccgcccatag cgaggccatg 780 agccgcatcg gcaaggaacg cggttggccg cccaccaccc gcgcccaatt cgacgccggc 840 gccacgccca acggcgccta tttcatgggg tctccgcagg aagtcatcga caagattctg 900 atgcagcacg aatggttcaa acacgaccgc ttcggcctgc aattcagcgt cggcaccctg 960 ccccacgaca aggtcatgaa agccatcgag ctttacggca ccgtggtcaa gccggcggtg 1020 gacaaggcgc taggctaa 1038 <210> 52 <211> XXXX <212> DNA <213> Artificial Sequence <220> <223> Enzyme <400> 52 atgagcactg aaatcgtcac cgatgtcgtt gtgattggcg cgggcccctg cgggctgttt 60 gcggccttcg aactcgggct gctcgacctc aaatgccatt tcatcgacat tctggaccgc 120 cccggcgggc agtgcgccga gctttatccc gaaaagccga tctacgatat tcccggcttc 180 It should be noted that the "XXXX" in the translation of <211> is because the original "1026" seems to be an incorrect or incomplete representation in the context of a proper translation. It might need further clarification depending on the actual meaning it is supposed to convey. ccggtggtga ccggccagga tctgaccgac aatctgatgg cccagatcgc gcccttcgcc 240 cccgaattcc acttcaaccg catggtcaat tcgatcgaga agcaggaaga cggctcgttt 300 aagctcgaaa ccgacgccga cgagattttc cacgcgaagg tcgtggtgat cgcggccggc 360 ggcggctcgt tccagcccaa gcgcccgccg gtcgacaata tcgaagccta tgaaaacaag 420 tcggttttct attcggtccg caagatggac gatttccgtg accaggacgt ggtgatcgtc 480 ggcggcggcg attccgccct cgactggacg ctcaacctgc agcccatcgt gcgctcgctg 540 accctggtac accgccgcga tgccttcaag gccgcgcccg cttcggtcaa caagatgaag 600 gaaatggtcg ccgagggtaa gatcaatttc ctgctcggcc aggtggccaa gctcgatggc 660 gaaaacggcc agatcaacca cgtgcatctg accaccgatg ccggcgatct ctcggtgccc 720 gccacccgcc tgctgccctt cttcggcctc accatgaagc tgggtccggt ggccgattgg 780 ggccttgagc tcaatgacaa tacgatcaag gtcgataccg aaaagttcga gacctcggtg 840 cccggcattt tcgccattgg cgacatcaac tggtatccgg gcaagctcaa gctgatcctc 900 tccggcttcc acgaggctgc cctcatggcc caggccgcca agaagattgt gtcgcccgac 960 gagcgcgtgg tgttccaata caccaccagc tccaccaagc tgcagaagaa gctgggcgtc 1020 gcctag 1026 <210> 53 <211> 1062 <212> DNA <213> Artificial sequence <220> <223> Enzyme <400> 53 atggactatc gctatctcgg gcgctcgggc ctgaaagtct cgagcatgac catgggcacc 60 atgacctttg gcggcagcga aaagatcggc aataccagcc gacaggacgc ggcgcggcag 120 4]gtggatatgt gtctcgatgc gggcatcaac ctctatgaca gcgccaatgt ctataatgcg 180 ggggtgtccg aggaaatcct gggcgacatt ctcgccgaaa acgggcggcg gcagaaagcg 240 ctgatcgcca ccaaggtacg cttcaagatg ggcgatggcc ccaacgaggc cgggctcagc 300 cgacaccata tcatcgagca gtgcaaggcg agcctcaaac gcctcaagac cgatgtgatc 360 gacctctatc aggtgcatga atgggatggg cagaccccca tcgaggaaac gatggaagcg 420 ctcgacattc tcgtgcgcca cggccatgtg cgctatatcg ggtgctccaa ttattcgggc 480 tggcatattt ccaaggccct tgccgccgcc gaccagcggc acaatgcgcg ctttgtgtcc 540 cagcagatcc attattcgct gcatagccgc gacgccgaat atgagctggt gccgatttcc 600 caggaccagg ggctgggcat tctcgtctgg tcgccgctgg cgggcgggtt gctttcgggc 660 aagtatcgcc gggacagcgc gccggacagc gggcggcatg tcggcgggtt ccgcgagccg 720 ccggtctatg actgggacaa gctctattcg gtcatcgacg tgctggtggc ggtggccgag 780 gagcgtggcg tttcgggggc gcaggtggcg ctgagctggc tgatgggacg cccgggcgtc 840 acttccgtca tcatcggcgg acgtaacgaa aagcagtttg ccgacaatat cgcagcggcc 900 cagctcgtgc tcagtgacga ggaacgcgcc cggctcgatg cggccagccg accgccgttg 960 ctctatccct actggcacca gtccttcacc gcgcaggatc ggctgggcaa gatcgacatg 1020 gatttgatcg gtccttatgc cgaggagttc aagcgtggct ga 1062 <210> 54 <211> 1053 <212> DNA <213> Artificial Sequence <220> <223> Enzyme <400> 54 atggaatacc gcttgctcgg gcgctcggga ctgaaggtct cgacccttac catgggcacg 60 atgacgttcg gggggaaccc gaaggtcggc aataccagcc tcaaggacgc gcagcgccag 120 atcgacatgt gtctggaggc gggcatcaac atgctcgaca cggccaatgt ctatacgcgg 180 ggcgtgtcgg aaagcattat cggggaggca ctgaccgatg ggcggcgcca gcggacgctg 240 ctggcgacca aggtgcgctt cccgatggac gacaccgacc ccaacgagcg gggactgtcg 300 cgctaccaca tcgtcaacca gtgcgaggca agcctcaagc ggctcaagac cgatgtgatc 360 gacctctacc aggtgcatga atgggacggg cagaccccgc tcgaggagac catggaagcg 420 ctcgacatgc tcgtgcgcca cggcaaggtc cgctatatcg gctgctcgaa ctattcgggc 480 tggcacatca tgaaggcgct cggcgtttcc gagaaggacg cccggcagcg cttcgtctcc 540 cagcagatcc actacacgct gttttcgcgc gaggcggaga acgagctggt gccgatcagc 600 ctcgaccagg ggctgggcat tctcatctgg agcccgctgg cgggcggcct gctttcgggc 660 aagtatcgcc gcggccagac ggcgccggaa ggggcgcgcc atgccggggg cttcaaggag 720 ccgccggtgc atgactggga gcggctctac gatatcgtgg aagtgctggt ttcgatcgcc 780 gaggtgcgcg gggtttcggg cgcgcagatc gcgctggcct ggctgctggg gcgcccgggg 840 gtgacctcgg tgatcatcgg cgggcgcacg gaagaccagt tcagggataa cctggcggcg 900 gccgaactca agctgaccga gcatgagcgc aagcggctag acgacgtgtc gctgctgccg 960 ctgcaatatc cctactggca ccagagcttt acggcctcgg accggctggg gccggcggac 1020 ctggcgctgc atgcacccca tctcgaaaaa tag 1053 <210> 55 <211> 1062 <212> DNA <213> Artificial Sequence <220> <223> Enzyme <400> 55 atggactacc gctatctcgg ccggtcgggc ctcaaggttt cggtgctgag cgtcgggacg 60 gtgacgttcg gcggcgacgg gctgtggggc gatacggacg tcaccggcgc cgcccggcag 120 atcgacatgt gcctcgatgc aggcatcaac ctcatcgata ccgccaatgt ctacaacaag 180 ggcgtttcgg aagagatcac cggcgaggtg ctgcgcgacg gcggtcggcg gcagcgcgcg 240 ctgatcgcca ccaaggtgcg ctttccgttc ggcgacggtc ccaatgaccg cgggctgtcg 300 cgccaccaca tcatcgccca gtgcgagggc agcctcaaac gcctccagac cgacgtcatc 360 gatctctacc aggtccatca gtgggacggg cagacgcccc tcgaggagac gatggaagcc 420 ctcgataccc tggtgcggca ggggaaggtc cgctacatcg gctgctcgaa ctattcgggc 480 tggcacctga tgaaggcgct cgccgtcgcc gacagccgcg gctaccagcg gttcgtcgcc 540 cagcagatcc actacaccct ctattcgcgc gaggccgaat acgagctgca gccgatcacc 600 gtcgaccagg ggctcggcat actcgtctgg agcccgctcg ccggcgggtg gctctcgggc 660 aagtacacgc gcgccagcca gcccgaggac ggccgacacg tgcgcggttt ccgcgagccg 720 ccggtgcccg atcccgagcg gctctacgac atcgtcgatg tcatcaatgc ggttgccgcc 780 gagcggggcg tttcgggcgc ggtcgtttcg ctggcttggg ctttgcagcg caagggcgtc 840 tcgtcggtga tcgtgggcgc gcgcaccgac gcgcagttcg ccgataacct caaggccgcc 900 gatctaaagc tcagcgacga cgaggtcgcc cggctcgacg ccgtcagcca gcgtccgctt 960 ccctatccct attggcacca gtccttcacc gcccaggacc gcttgagcga agccgatctc 1020 gtgctgctcg ccccttatct cgaggagtac aagcgtggct ga 1062 <210> 56 <211> 1047 <212> DNA <213> Artificial Sequence <220> <223> Enzyme <400> 56 atggactatc gattgctcgg ccgttccggc ctcaaggtgt cgtcgctgtc gatcggcacg 60 gcgacgttcg gcggcgacgg gttgtggggc gacaccaacg tcaaggaggc gcagcgccag 120 atcgacctct gcctcgacca cggcgtcaac ctgatcgaca ccgccaatgt ctattcgggc 180 agcgtgtcgg agacgatcat cggcgaggcg ctgaccgatg gccggcgcga ccgcttgctg 240 atcgccacca aggtgcgctt ccccaggggc aagggaccga acgatcgcgg gctcagccga 300 tggcacatca tccgtgaggc ggaagcctcg ctgaagcgga tgaagaccga cgtcatcgac 360 ctgtaccagg tgcatgagtg ggatgggcag acgccgctcg aggagacgct ggaagcactc 420 gacaccctcg tccgctccgg caaggtccgc tatatcggct gctcgaacta ttcggcctgg 480 cacctgatga aggcgctcgc ggtttccaac gagcgcaact accagcgctt cgtttcccag 540 cagatccact acacgctgca ctcgcgcgaa gccgagttcg agctgatccc ggccggcctc 600 gaccagggtg tctcggtgct ggtgtggtcg ccgctcgccg gcgggtggct ctcgggcaag 660 tacacccgcg attccaagcc gagcgacggc cggcaggtgc gcggcttcaa ggagccgccg 720 atctacgatc ccgagcatct gtggaagatc gtcgacgtgg tcaacgacgt ggcgggcgcg 780 cgtggcgttt ccggcgcgca ggtgtcgctt gcctggttgc tgtcgaagcc gatcgtcgct 840 tcggtgatcg tcggcggccg caatatcgga cagttcgaag acaacctcaa agccgcggaa 900 ctgactctca ccaccgagga gctgcagaag ctcgacgcgg tcagccagcc gaacctgccc 960 tacccgtact ggcaccagag ctttaccgcg caggaccggc tcggtgagcc cgacctggcg 1020 ctgcaccgcc cctatctcga caagtga 1047 <210> 57 <211> 1047 <212> DNA <213> Artificial Sequence <220> <223> Enzyme <400> 57 atggactatc gattgctcgg ccgttccggc ctcaaggtct cgtcgctgtc gatcggcacg 60 gcgacgttcg gcggcgatgg gttgtggggc gataccaacg tcaaggaggc gcagcgccag 120 atcgacctct gcctcgacca tggcgtcaac ctgatcgaca ccgccaacgt ctattcgggg 180 agcgtctcgg aaacgatcat cggcgaggcg ctgaccgacg ggcggcgtga tcggctgctg 240 atcgccacca aggtgcgctt cccacgcggc aaggggccca atgatcgcgg cctcagccgc 300 tggcacatca tccgcgaggc ggaagcgtcg ctcaaacgga tgaagaccga cgtcatcgac 360 ctctaccagg tgcatgagtg ggacgggcag acgccgctcg aggaaacgct ggaggcgctc 420 gatacgctcg tccgctcggg caaggtccgc tatatcggct gctcgaacta ttcggcctgg 480 cacctgatga aggcgctcgc cacctcgaac gagcgcaact accagcgctt cgtctcccag 540 cagatccact acacgctgca ttcgcgtgag gccgagttcg agctgatccc ggccgggctc 600 gaccagggcg tttcggtgct ggtgtggtcg ccgctggccg gcgggtggct gtcgggcaag 660 tacacccgca actcgaagcc gagcgatggc cggcaggtgc gcggcttcaa ggagccgccg 720 atctatgatc cggagcacct gtggaagatc gtcgatgtgg tcaacgaagt ggccagcgcg 780 catggcgtct ccggcgcgca ggtgtcgctg gcctggctgt tgagtaagcc gatcgttgcc 840 tcggtgatcg tcggcggccg caatatcgag cagttcgagg acaacctgaa agccgccgag 900 ctgcagctca ccgccgagga actgggcaga ctcgacgcgg tcagccagcc gaacctcccc 960 tacccctact ggcaccagaa ctttaccgcg caggatcgcc tcagcgagcc tgacctggcg 1020 ctgcaccggc cctatctcga caaatga 1047 <210> 58 <211> 1056 <212> DNA <213> Artificial Sequence <220> <223> Enzyme <400> 58 atggaatatc gactgcttgg ccgctccggc ctcaaagtct ccaccctcac gatgggcacg 60 atgaccttcg gcggccaggg caagttcgcc tcggtcggca actccggcgt cgacgaagca 120 cgccggctga tcgacatgtg catcgatgcc ggcgtcaatc tcatcgatac ggcgaacgtc 180 tactccgatg gccgttcgga agaaatcatc ggcgaagtgc tcggcggcaa gcgtccgaag 240 gacgtactga tcgcgaccaa ggcacgcttt ccgatgggcg acggccccaa caacggcggc 300 ctctctcgct atcacctgat tcgcgagtgc gaagcgagcc tgaagcgcct gcgcacggac 360 gtgatcgatc tctaccaggt tcaccagtgg gatggcctga ccccgctgga agaaaccctg 420 gaagcgctcg acaccctggt tcgcgacggc aaggtgcgtt acctgggctg ctccaactat 480 tccgcgtggc acctgatgaa ggcgctcggc atcagtgagc gcgagcatcg ccagcggttc 540 gtcagccaac agatccatta cacgctggag gcgcgcgagg ccgaatacga gctgctgccg 600 atctcgatcg accagggcct cggcgtcctc gtctggagtc cgcttgccgg tggcctgctt 660 tcgggcaagc accgtcgcga tggcaaggca ccggagggct cacgtcagct ggagggctgg 720 accgagccgc cgatccgcga cgaagagcgc ctttggcgca tcgtcgacgt gctggtggcc 780 atcgccgaag cgcgcggcgt gtcgggtgca caggtcgcgc tggcctggct gctcgagcgt 840 cccgcggtga cctccctggt catcggcggt cgtaccgagg cgcagttcaa ggacaacctc 900 gccgccgccg agctgaagct cacggccgaa gagcgtcagc ggctggacga ggtcagtgcg 960 ccgccggtgc tctacccgta ctggcaccag ctgatgacgg cgtcggaccg cctgggtccg 1020 gcggacatgc cattgctggc gccgcatctg aagtaa 1056 <210> 59 <211> 1056 <212> DNA <213> Artificial sequence <220> <223> Enzyme <400> 59 atggaatacc gactgcttgg ccgctccggc ctcaaagtct ccaccctcac gatgggcacg 60 atgaccttcg gcggccaggg caaattcgcc tcggtcggta gctccggcgt cgacgaagcg 120 cgccggctga tcgacatgtg catcgacgcc ggcgtgaatc tcatcgacac ggcgaacgtc 180 tattccgacg gtcgttcgga agagattatc ggtgaagtgc tcggcggcaa gcgtccgaag 240 gacgtcctga tcgcgaccaa ggcgcgcttt cccatgggtg atggcccgaa ccatggtggc 300 ctgtcgcgct atcacctgat tcgcgaatgc gaagccagcc tgaagcggct gcgcacggac 3,60 gtgatcgatc tctaccaggt tcaccagtgg gatggcctga cgcctctgga ggaaaccctg 420 gaagcgctcg acaccctcgt tcgcgacggc aaggtgcgtt acctgggttg ctccaactat 480 tccgcctggc acctgatgaa ggcgctcggc gtgagcgagc gtgagcaccg ccagcgtttc 540 atcagccagc agatccatta cacgctggaa gcgcgcgaag ccgaatacga actgctgccg 600 atttcgatcg accagggcct cggcgtgctg gtgtggagtc cgctcgccgg cgggctgctt 660 tcgggcaagc accgtcgcga taccaaggcc ccggaaggct cgcgccagct ggccggctgg 720 accgaaccgc cgatccgcga cgaagagcgc ctgtggcgcg tcgtcgacgt actggtttca 780 atcgcccaat cgcgtggcgt gtcgggagcg caggtcgcgc tggcctggtt gctcgagcgt 840 cctgcggtca cctcgctggt gatcggtggt cgtaccgagg cacagttcaa ggacaacctg 900 gcggccgccg aactgaagct cacggccgaa gagcgtcagc gtctggatga agtcagcgcg 960 ccgccggtgc tctatccgta ctggcaccag ttgatgaccg catcggatcg cctgggtccg 1'020 gccgacatgc ccctgctggc gccgcacctc aaataa 1056 <210> 60 <211> 1062 <212> DNA <213> Artificial Sequence <220> <223> Enzyme <400> 60 Note: There seems to be a formatting issue in the original text where '1'020' should probably be '1020'. I've translated it as '1020' in the English version.atggaatacc gctatctcgg ccgctcaggc ttgaaggtgt cgacattgac catgggcacc 60 atgacctttg gcggctcgga aaaggttggg gcgacggcgc aagacgatgc gacgcgccag 120 atcgacctgt gcctcgatgc agggatcaat ctgctcgaca cggccaatgt ctacaatgcc 180 ggggtctccg aggaaatgat tggcgtggcg ctggccgaga atggtcgccg ccagaaagca 240 ctcgtagcca ccaaggtccg cttcaagatg ggcgatgggc ccaacgagat cggcctcagc 300 cgtcaccata ttattgccca ggccgaagcc agccttaagc gcttgaaaac agacgttatt 360 gatctctatc aggtccacga atgggacggc atgacgccca tcgaggagac gatggaagcg 420 ctcgataccc tgatcaagca gggcaaagtc cgctatatcg gctgctcaaa ttattcgggc 480 tggcatatca tgaaggcgct gcaggccgcc gacaaaaacc acggccagcg cttcatcagc 540 cagcagatcc actacacgct gcattcgcgc gaagccgaat atgaactgat cccgatcact 600 ggggatcaag gccttggcgt cctcatctgg tcgccgctgg ccggtggttt gctctcgggc 660 aaataccgtc gcaacggtgg ccctgacagc gggcgccatg tcggcggctt ccgcgagccg 720 ccggtttatg actgggagaa gctctacgac atcgtcgatg tgatcgtgga tattgctgat 780 gaccgcggcg tgtcgggtgc acaggttgcg ctggcatggt cgctggggcg gccgggtgta 840 acctcggtga tcatcggtgg ccgcaatgat gcccagttca aagacaatct ggcggcagcc 900 gagttgaagc tgacggcgga agagcgcgcg cgacttgacg cggtcagccg tccaccgctg 960 ctctatcctt attggcatca gacgtttacg gcgcaggacc ggttaggccc tgtcgatcga 1020<​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​​ Asn Trp Pro Ile Leu Arg Gly Asn Tyr Gln Gly Trp Gly Tyr Ser Pro 65 70 75 80 Leu Asp Gln Ile Asn Lys Asp Asn Val Gly Asp Leu Gln Leu Val Trp 85 90 95 Ser Arg Thr Met Glu Pro Gly Ser Asn Glu Gly Ala Ala Ile Ala Tyr 100 105 110 Asn Gly Val Ile Phe Leu Gly Asn Thr Asn Asp Val Ile Gln Ala Ile 115 120 125 Asp Gly Lys Thr Gly Ser Leu Ile Trp Glu Tyr Arg Arg Lys Leu Pro 130 135 140 Ser Ala Ser Lys Phe Ile Asn Ser Leu Gly Ala Ala Lys Arg Ser Ile 145 150 155 160 Ala Leu Phe Gly Asp Lys Val Tyr Phe Val Ser Trp Asp Asn Phe Val 165 170 175 Val Ala Leu Asp Ala Lys Thr Gly Lys Leu Ala Trp Glu Thr Asn Arg 180 185 190 Gly Gln Gly Val Glu Glu Gly Val Ala Asn Ser Ser Gly Pro Ile Val 195 200 205 Val Asp Gly Val Val Ile Ala Gly Ser Thr Cys Gln Phe Ser Gly Phe 210 215 220 Gly Cys Tyr Val Thr Gly Thr Asp Ala Glu Ser Gly Glu Glu Leu Trp 225 230 235 240 Arg Asn Thr Phe Ile Pro Arg Pro Gly Glu Glu Gly Asp Asp Thr Trp 245 250 255 Gly Gly Ala Pro Tyr Glu Asn Arg Trp Met Thr Gly Ala Trp Gly Gln 260 265 270 Ile Thr Tyr Asp Pro Glu Leu Asp Leu Val Tyr Tyr Gly Ser Thr Gly 275 280 285 Ala Gly Pro Ala Ser Glu Val Gln Arg Gly Thr Glu Gly Gly Thr Leu 290 295 300 Ala Gly Thr Asn Thr Arg Phe Ala Val Lys Pro Lys Thr Gly Glu Val 305 310 315 320 Val Trp Lys His Gln Thr Leu Pro Arg Asp Asn Trp Asp Ser Glu Cys 325 330 335 Thr Phe Glu Met Met Val Val Ser Thr Ser Val Asn Pro Asp Ala Lys 340 345 350 Ala Asp Gly Met Met Ser Val Gly Ala Asn Val Pro Arg Gly Glu Thr 355 360 365 Arg Lys Val Leu Thr Gly Val Pro Cys Lys Thr Gly Val Ala Trp Gln 370 375 380 Phe Asp Ala Lys Thr Gly Asp Tyr Phe Trp Ser Lys Ala Thr Val Glu 385 390 395 400 Gln Asn Ser Ile Ala Ser Ile Asp Asp Thr Gly Leu Val Thr Val Asn 405 410 415 Glu Asp Met Ile Leu Lys Glu Pro Gly Lys Thr Tyr Asn Tyr Cys Pro 420 425 430 Thr Phe Leu Gly Gly Arg Asp Trp Pro Ser Ala Gly Tyr Leu Pro Lys 435 440 445 Ser Asn Leu Tyr Val Ile Pro Leu Ser Asn Ala Cys Tyr Asp Val Met 450 455 460 Ala Arg Thr Thr Glu Ala Thr Pro Ala Asp Val Tyr Asn Thr Asp Ala 465 470 475 480 Thr Leu Val Leu Ala Pro Gly Lys Thr Asn Met Gly Arg Val Asp Ala 485 490 495 Ile Asp Leu Ala Thr Gly Glu Thr Lys Trp Ser Tyr Glu Thr Arg Ala 500 505 510 Ala Leu Tyr Asp Pro Val Leu Thr Thr Gly Gly Asp Leu Val Phe Val 515 520 525 Gly Gly Ile Asp Arg Asp Phe Arg Ala Leu Asp Ala Glu Ser Gly Lys 530 535 540 Glu Val Trp Ser Thr Arg Leu Pro Gly Ala Val Ser Gly Tyr Thr Thr 545 550 555 560 Ser Tyr Ser Ile Asp Gly Arg Gln Tyr Val Ala Val Val Ser Gly Gly 565 570 575 Ser Leu Gly Gly Pro Thr Phe Gly Pro Thr Thr Pro Asp Val Asp Ser 580 585 590 Ala Ser Gly Ala Asn Gly Ile Tyr Val Phe Ala Leu Pro Glu Lys Lys 595 600 605 <210> 62 <211> 586 <212> PRT <213> Synthetic Sequence <220> <223> Enzyme <400> 62 Met Lys Lys Arg Thr Ser Ile Leu Leu Ala Ser Val Ala Met Leu Gly 1 5 10 15 Met Gly Ser Thr Ala Phe Ala Gln Val Asp Ile Asn Ala Leu Pro Ala 20 25 30 Val Thr Asp Ala Ile Leu Ala Asn Pro Asp Ala Gly Asp Trp Pro Ser 35 40 45 Tyr Gly Arg Asp Ile Thr Asn Tyr Arg Phe Ser Pro Leu Asp Gln Val 50 55 60 Asn Lys Asp Asn Val Gly Gln Leu Thr Leu Ala Trp Ala Arg Ala Leu 65 70 75 80 Glu Pro Gly Asn Leu Gln Ser Ala Pro Leu Glu Phe Gly Gly Val Leu 85 90 95 Phe Thr Ala Ala Pro Gly Asp Val Val Gln Ala Met Asp Ala Ala Thr 100 105 110 Gly Gln Leu Ile Trp Glu Tyr Arg Arg Gln Leu Pro Asp Arg Ala Thr 115 120 125 Leu Asn Ser Leu Gly Glu Asn Lys Arg Gly Ile Ala Leu Tyr Glu Asp 130 135 140 Lys Ile Tyr Val Ala Thr Trp Asp Asn Phe Ile Val Ala Leu Asp Ala 145 150 155 160 Lys Thr Gly Gln Val Ala Trp Glu Ser Asp Arg Gly Gly Gly Ala Asp 165 170 175 Leu Ile Ser Asn Thr Thr Gly Pro Ile Val Ala Asn Gly Val Val Val 180 185 190 Ala Gly Ser Thr Cys Gln Phe Ser Glu Phe Gly Cys Tyr Val Thr Gly 195 200 205 His Asp Ala Ala Thr Gly Glu Glu Leu Trp Arg Asn Asn Phe Ile Pro 210 215 220 Lys Lys Gly Glu Glu Gly Asp Asp Thr Trp Gly Asp Ser Thr Glu Asp 225 230 235 240 Gln Arg Trp Met Thr Gly Ala Trp Gly Gln Met Thr Tyr Asp Pro Glu 245 250 255 Leu Asp Leu Val Tyr Tyr Gly Ser Thr Gly Ala Gly Pro Ala Ala Glu 260 265 270 Phe Gln Arg Asn Thr Val Gly Gly Thr Leu Phe Gly Ser Asn Thr Arg 275 280 285 Phe Ala Val Lys Pro Lys Thr Gly Glu Ile Val Trp Arg His Gln Val 290 295 300 Leu Pro Arg Asp Asn Trp Asp Gln Glu Cys Thr Tyr Glu Met Val Pro 305 310 315 320 Val Asp Ile Asp Ser Ala Pro Ala Ala Asp Met Glu Gly Leu Leu Ala 325 330 335 Leu Gly Thr Ala Ala Pro Gly Lys Lys Arg Val Leu Thr Gly Val Pro 340 345 350 Cys Lys Thr Gly Val Met Trp Gln Phe Asp Ala Gln Thr Gly Glu Phe 355 360 365 Ile Tyr Ala Arg Asp Thr Val Gln Gln Thr Leu Ile Glu Ser Val Asp 370 375 380 Asn Thr Gly Leu Val Thr Val Asn Glu Ala Ala Ile Pro Thr Glu Val 385 390 395 400 Asp Val Ala Thr Pro Met Cys Pro Thr Tyr Leu Gly Gly Arg Asp Trp 405 410 415 Ser Pro Thr Ala Phe Asn Pro Thr Ser Lys Val Met Phe Val Pro Leu 420 425 430 Thr Asn Met Cys Ala Asp Val Thr Val Leu Asp Gln Glu Pro Thr Gly 435 440 445 Leu Asp Val Tyr Asn Thr Glu Leu Thr Tyr Lys Met Pro Glu Gly Val 450 455 460 Thr Asp Ala Gly Arg Ile Asp Ala Ile Asn Val Glu Thr Gly Lys Thr 465 470 475 480 Leu Trp Ser Trp Thr Gln Gln Thr Pro Gln Tyr Ala Ser Ile Thr Ala 485 490 495 Thr Ala Gly Gly Leu Ile Phe Thr Gly Gly Ala Asp Arg Arg Phe Lys 500 505 510 Ala Ile Asp Gln Glu Thr Gly Glu Leu Val Trp Ser Val Thr Leu Gly 515 520 525 Ser Arg Ala Thr Gly His Pro Ile Ser Tyr Glu Val Asp Gly Arg Gln 530 535 540 Tyr Ile Ala Ile Pro Ala Gly Gly Pro Gly Tyr Ala Thr Asp Leu Ile 545 550 555 560 Thr Ala Ser Gly Ser Thr Val Asp Val Val Ser Gly Ser Asn Met Leu 565 570 575 Tyr Val Phe Ala Leu Pro Glu Gln Lys Lys 580 585 <210> 63 <211> 353 <212> PRT <213> Artificial Sequence <220> <223> Enzyme <400> 63 Met Asp Tyr Arg Tyr Ile Gly Arg Ser Gly Leu Arg Val Ser Val Leu 1 5 10 15 Thr Met Gly Thr Met Thr Phe Gly Gly Ser Asp Lys Val Gly His Thr 20 25​​​​​​​​​​​​​​ 85 90 95 Val Gly Leu Ser Lys His His Ile Ile Ala Gln Ala Glu Ala Ser Leu 100 105 110 Lys Arg Leu Lys Thr Asp Val Ile Glu Leu Tyr Gln Val His Glu Trp 115 120 125 Asp Gly Met Thr Pro Ile Glu Glu Thr Met Glu Ala Leu Asp Thr Leu 130 135 140 Val Lys Gln Gly Lys Val Arg Tyr Val Ala Cys Ser Asn Tyr Ser Gly 145 150 155 160 Trp His Ile Met Lys Ala Leu Ala Ala Ala Asp Lys His His Gly Gln 165 170 175 Arg Phe Ile Ser Gln Gln Ile His Tyr Thr Leu His Ser Arg Glu Ala 180 185 190 Asp Tyr Glu Leu Val Pro Ile Ser Gln Asp Gln Gly Leu Gly Ile Leu 195 200 205 Ile Trp Ser Pro Leu Ala Gly Gly Leu Leu Ser Gly Lys Tyr Arg Arg 210 215 220 Asp Gly Gly Pro Asp Ala Gly Arg His Val Gly Gly Phe Arg Glu Pro 225 230 235 240 Pro Val Tyr Asp Trp Asp Lys Leu Tyr Glu Ile Ile Asp Val Ile Val 245 250 255 Ala Ile Ala Arg Glu Arg Asp Val Ser Gly Ala Gln Val Ala Leu Ala 260 265 270 Trp Ala Leu Gly Arg Pro Gly Val Thr Ser Ala Ile Ile Gly Gly Arg 275 280 285 Ser Glu Ala Gln Phe Arg Asp Asn Leu Ala Ala Ala Glu Leu Arg Leu 290 295 300 Thr Asp Asp Glu Arg Ala Arg Leu Asp Ala Ala Ser Arg Pro Pro Leu[[ID='17']] 305 310 315 320 Leu Tyr Pro Tyr Trp His Gln Ser Phe Thr Ala Gln Glu Arg Leu Asp 325 330 335 Gln Ile Asp Leu Asp Leu Ile Gly Pro Tyr Ala Glu Glu Phe Lys Arg 340 345 350 Gly <210> 64 <211> 353 <212> PRT[[ID='35']] <'213'> Artificial Sequence[[ID='37']] <220> <223> Enzyme <400> 64 Met Asp Tyr Lys Tyr Leu Gly Arg Ser Gly Leu Arg Val Ser Val Leu 1 5 10 15 Thr Met Gly Thr Met Thr Phe Ala Gly Ser Glu Lys Val Gly His Thr 20 25 30 Ala Gln Ala Asp Ala Thr Arg Gln Ile Asp Leu Cys Ile Asp His Gly 35 40 45 Ile Asn Leu Leu Asp Thr Ala Asn Val Tyr Asn Ala Gly Val Ser Glu 50 55 60 Glu Met Ile Gly Val Ala Leu Ser Glu His Gly Arg Arg Gln Lys Ala 65 70 75 80 Leu Ile Ala Thr Lys Val Arg Phe Arg Met Gly Asn Gly Pro Asn Glu 85 90 95 Val Gly Leu Ser Arg His His Leu Ile Ala Gln Ala Glu Ala Ser Ile 100 105 110 Lys Arg Leu Lys Thr Asp Val Ile Asp Leu Tyr Gln Val His Glu Trp 115 120 125 Asp Gly Met Thr Pro Ile Glu Glu Thr Met Glu Ala Leu Asp Thr Leu 130 135 140 Val Lys Gln Gly Lys Val Arg Tyr Val Gly Cys Ser Asn Tyr Ser Gly 145 150 155 160 Trp His Ile Met Lys Gly Leu Ala Ala Ala Asp Lys His His Gly Gln 165 170 175 Arg Phe Ile Ser Gln Gln Ile His Tyr Thr Leu His Ser Arg Glu Ala 180 185 190 Glu Tyr Glu Leu Val Pro Ile Ser Gln Asp Gln Gly Leu Gly Ile Leu 195 200 205 Ile Trp Ser Pro Leu Ala Gly Gly Leu Leu Ser Gly Lys Tyr Arg Arg 210 215 220 Asp Gly Gly Pro Asp Ala Gly Arg His Val Gly Gly Phe Arg Asp Pro 225 230 235 240 Pro Val Tyr Asp Trp Asp Lys Leu Tyr Glu Ile Leu Asp Val Ile Val 245 250 255 Ala Ile Ala Arg Glu Lys Asp Val Ser Gly Ala Gln Val Ala Leu Ala 260 265 270 Trp Ala Leu Gly Arg Pro Gly Val Thr Ser Ala Ile Ile Gly Gly Arg 275 280 285 Ser Glu Ala Gln Phe Arg Asp Asn Leu Ala Ala Ala Glu Leu Arg Leu 290 295 300 Thr Asp Asp Glu Arg Ala Arg Leu Asp Ala Ala Ser Arg Pro Pro Leu 305 310 315 320 Leu Tyr Pro Tyr Trp His Gln Ser Phe Thr Gly Gln Asp Arg Leu Asp 325 330 335 Gln Val Asp Leu Asp Leu Ile Gly Pro Tyr Ala Glu Glu Phe Lys Arg 340 345 350 Gly <210> 65 <211> 353 <212> PRT <213> Synthetic sequence <220> <223> Enzyme <400> 65 Met Asp Tyr Lys Tyr Leu Gly Arg Ser Gly Ile Arg Val Ser Val Leu 1 5 10 15 Thr Met Gly Thr Met Thr Phe Ala Gly Ser Glu Lys Val Gly His Thr 20 25 30 Ala Gln Ala Asp Ala Thr Arg Gln Ile Asp Leu Cys Leu Asp His Gly 35 40 45 Ile Asn Leu Leu Asp Thr Ala Asn Val Tyr Asn Ala Gly Val Ser Glu 50 55 60 Glu Met Ile Gly Val Ala Leu Ser Glu His Gly Arg Arg Gln Lys Ala 65 70 75 80 Leu Ile Ala Thr Lys Val Arg Phe Arg Met Gly Asn Gly Pro Asn Glu 85 90 95 Val Gly Leu Ser Arg His His Leu Ile Ala Gln Ala Glu Ala Ser Leu 100 105 110 Lys Arg Ile Lys Thr Asp Val Ile Asp Leu Tyr Gln Val His Glu Trp 115 120 125 Asp Gly Met Thr Pro Ile Glu Asp Thr Met Glu Ala Leu Asp Thr Leu 130 135 140 Val Lys Gln Ala Lys Val Arg Tyr Val Gly Cys Ser Asn Tyr Ser Gly 145 150 155 160 Trp His Ile Met Lys Ala Leu Ala Gly Ala Asp Lys His His Gly Gln 165 170 175 Arg Phe Ile Ser Gln Gln Ile His Tyr Thr Leu His Ser Arg Glu Ala 180 185 190 Glu Tyr Glu Leu Val Pro Ile Ser Gln Asp Gln Gly Leu Gly Ile Leu 195 200 205 Ile Trp Ser Pro Leu Ala Gly Gly Leu Leu Ser Gly Lys Tyr Arg Arg 210 215 220 Asp Gly Gly Pro Asp Ala Gly Arg His Val Gly Gly Phe Arg Asp Pro 225 230 235 240 Pro Val Tyr Asp Trp Asp Arg Leu Tyr Asp Ile Ile Asp Val Ile Val 245 250 255 Ala Ile Ala Arg Glu Lys Asp Val Ser Gly Ala Gln Val Ala Leu Ala 260 265 270 Tyr Ala Leu Gly Arg Pro Gly Val Thr Ser Ala Ile Ile Gly Gly Arg 275 280 285 Ser Glu Ala Gln Phe Arg Asp Asn Leu Gly Ala Ala Glu Leu Lys Leu 290 295 300 Thr Asp Asp Glu Arg Ala Arg Leu Asp Ala Ala Ser Arg Pro Pro Leu 305 310 315 320 Leu Tyr Pro Tyr Trp His Asn Ser Phe Thr Gly Gln Asp Arg Leu Asp 325 330 335 Gln Val Asp Leu Asp Leu Ile Gly Pro Tyr Ala Glu Glu Phe Lys Arg 340 345 350 Gly <210> 66 <211> 353 <212> PRT <213> Synthetic Sequence <220> <223> Enzyme <400> 66 Met Asp Tyr Arg Phe Leu Gly Arg Ser Gly Ile Arg Val Ser Val Leu 1 5 10 15 Thr Met Gly Thr Met Thr Phe Gly Gly Thr Glu Lys Val Gly His Thr 20 25 30 Ala Gln Ala Asp Ala Thr Arg Gln Leu Asp Ile Cys Leu Asp His Gly 35 40 45 Ile Asn Leu Leu Asp Thr Ala Asn Val Tyr Asn Ala Gly Val Ser Glu 50 55 60 Glu Met Ile Gly Val Ala Leu Ser Glu His Gly Lys Arg Gln Lys Ala 65 70 75 80 Leu Ile Ala Thr Lys Val Arg Phe Arg Met Ala Asn Gly Pro Asn Glu 85 90 95 Val Gly Leu Ser Arg His His Ile Ile Ala Gln Ala Glu Ala Ser Leu 100 105 110 Lys Arg Ile Lys Thr Asp Val Ile Asp Leu Tyr Gln Val His Glu Trp 115 120 125 Asp Gly Met Thr Pro Ile Asp Glu Thr Leu Glu Ala Leu Asp Thr Leu 130 135 140 Val Lys Gln Gly Lys Val Arg Tyr Val Gly Cys Ser Asn Tyr Ser Gly 145 150 155 160 Trp His Ile Met Lys Gly Leu Ala Ala Ala Asp Lys His His Gly Gln 165 170 175 Arg Phe Ile Ser Gln Gln Ile His Tyr Thr Leu His Ser Arg Glu Gly 180 185 190 Glu Tyr Glu Leu Val Pro Ile Ser Gln Asp Gln Gly Leu Ala Ile Leu 195 200 205 Ile Trp Ser Pro Leu Ala Gly Gly Leu Leu Ser Gly Lys Tyr Lys Arg 210 215 220 Asp Gly Gly Pro Asp Ala Gly Arg His Val Gly Gly Phe Arg Glu Pro 225 230 235 240 Pro Leu Tyr Asp Trp Asp Arg Leu Tyr Asp Ile Ile Asp Val Ile Val 245 250 255 Ala Ile Ala Arg Glu Arg Asp Val Ser Gly Ala Gln Val Ala Leu Ala 260 265 270 Phe Ala Leu Gly Arg Pro Gly Val Thr Ser Ala Ile Ile Gly Gly Arg 275 280 285 Ser Glu Ala Asn Phe Arg Asp Asn Leu Gly Ala Ala Glu Leu Arg Leu ​​​​​​​​​​​​​​​​​​​​​​​ <220> <223> Enzyme <400> 67 Met Asp Tyr Lys Tyr Ile Gly Arg Ser Gly Ile Arg Val Ser Val Leu 1 5 10 15 Thr Met Gly Thr Met Thr Phe Ala Gly Ser Glu Arg Val Gly His Thr 20 25 30 Gly Gln Ala Glu Ala Thr Arg Gln Leu Asp Ile Cys Leu Asp His Gly<00029​​​​​​​​​​​​​​​​​​​​​​​​​​Val Lys Gln Ala Lys Val Arg Tyr Val Ala Cys Ser Asn Tyr Ser Gly 145 150 155 160 Trp His Ile Met Lys Gly Leu Ala Gly Ala Asp Lys His His Gly Gln 165 170 175 Arg Phe Ile Ser Gln Gln Ile His Tyr Thr Leu His Ser Arg Asp Gly 180 185 190 Glu Tyr Glu Leu Val Pro Ile Ser Gln Asp Gln Gly Leu Ala Ile Leu 195 200 205 Ile Trp Ser Pro Leu Ala Gly Gly Leu Leu Ser Gly Lys Tyr Arg Arg 210 215 220 Asp Gly Gly Pro Asp Ala Gly Arg His Val Gly Gly Phe Arg Glu Pro 225 230 235 240 Pro Val Tyr Asp Trp Asp Lys Leu Tyr Glu Ile Leu Asp Val Ile Val 245 250 255 Ala Ile Ala Arg Glu Arg Asp Val Ser Gly Ala Gln Val Ala Leu Ala 260 265 270 Trp Ala Leu Gly Arg Pro Gly Val Thr Ser Ala Ile Ile Gly Gly Arg 275 280 285 Ser Glu Ala Gln Phe Arg Asp Asn Ile Gly Ala Ala Asp Leu Lys Leu 290 295 300 Ser Asp Glu Glu Arg Ala Arg Leu Asp Ala Ala Ser Arg Pro Pro Leu 305 310 315 320 Leu Tyr Pro Tyr Trp His Asn Ser Phe Thr Gly Asn Asp Arg Leu Asp 325 330 335 Gln Leu Asp Leu Asp Leu Leu Gly Pro Tyr Ala Asp Glu Phe Lys Arg 340 345 350 Gly <210> 68 <211> 353 <212> PRT <213> Artificial Sequence <220> <223> Enzyme <400> 68 Met Asp Tyr Lys Tyr Leu Gly Arg Ser Gly Ile Arg Val Ser Val Leu 1 5 10 15 Thr Met Gly Thr Met Thr Phe Ala Gly Ser Glu Arg Val Gly His Thr 20 25 30 Gly Gln Ala Glu Ala Thr Arg Gln Leu Asp Ile Cys Leu Asp His Gly 35 40 45 Ile Asn Leu Leu Asp Thr Ala Asn Val Tyr Asn Ala Gly Val Ser Glu 50 55 60 Glu Met Leu Gly Val Gly Leu Ser Glu His Gly Arg Arg Gln Lys Ala 65 70 75 80 Leu Ile Ala Thr Lys Val Arg Phe Arg Met Ala Asn Gly Pro Asn Glu 85 90 95 Val Gly Leu Ser Arg His His Ile Ile Ala Gln Ala Glu Ala Ser Leu 100 105 110 Lys Arg Leu Lys Thr Asp Val Ile Glu Leu Tyr Gln Val His Glu Trp 115 120 125 Asp Gly Met Thr Pro Ile Glu Asp Thr Met Glu Ala Leu Asp Thr Leu 130 135 140 Val Lys Gln Ala Lys Val Arg Tyr Val Gly Cys Ser Asn Tyr Ser Gly 145 150 155 160 Trp His Ile Met Lys Gly Leu Ala Gly Ala Asp Lys His His Gly Gln 165 170 175 Arg Phe Ile Ser Gln Gln Ile His Tyr Thr Leu His Ser Arg Asp Ala 180 185 190 Glu Tyr Glu Leu Val Pro Ile Ser Gln Asp Gln Gly Leu Gly Ile Leu 195 200 205 Ile Trp Ser Pro Leu Ala Gly Gly Leu Leu Ser Gly Lys Tyr Arg Arg 210 215 220 Asp Gly Gly Pro Asp Ala Gly Arg His Val Gly Gly Phe Arg Glu Pro 225 230 235 240 Pro Val Tyr Asp Trp Asp Lys Leu Tyr Glu Leu Ile Asp Val Ile Val 245 250 255 Ala Ile Ala Arg Glu Arg Asp Val Ser Gly Ala Gln Val Ala Leu Ala 260 265 270 Trp Ala Leu Gly Arg Pro Gly Val Thr Ser Ala Ile Ile Gly Gly Arg 275 280 285 Ser Glu Ala Gln Phe Arg Asp Asn Ile Ala Gly Ala Glu Leu Lys Leu 290 295 300 Ser Asp Asp Glu Arg Ala Arg Leu Asp Ala Ala Ser Arg Pro Pro Leu 305 310 315 320 Leu Tyr Pro Tyr Trp His Gln Ser Phe Thr Gly Asn Asp Arg Leu Asp 325 330 335 Gln Ile Asp Leu Asp Leu Leu Gly Pro Tyr Ala Asp Glu Phe Lys Arg 340 345 350 Gly <210> 69 <211> 353 <212> PRT <213> Artificial Sequence <220> <223> Enzyme <400> 69 Met Asp Tyr Lys Tyr Ile Gly Arg Ser Gly Ile Arg Val Ser Val Leu 1 5 10 15 Thr Met Gly Thr Met Thr Phe Gly Gly Ser Glu Lys Val Gly His Thr 20 25 30 Gly Gln Ala Asp Ala Thr Arg Gln Ile Asp Ile Cys Leu Asp His Gly 35 40 45 Ile Asn Leu Leu Asp Thr Ala Asn Val Tyr Asn Ala Gly Val Ser Glu 50 55 60 Glu Met Leu Gly Val Ala Leu Ser Glu His Gly Arg Arg Gln Lys Ala 65 70 75 80 Leu Ile Ala Thr Lys Val Arg Phe Arg Met Gly Asn Gly Pro Asn Glu 85 90 95 Val Gly Leu Ser Arg His His Ile Ile Ala Gln Ala Glu Ala Ser Leu 100 105 110 Lys Arg Leu Lys Thr Asp Val Ile Glu Leu Tyr Gln Val His Glu Trp 115 120 125 Asp Gly Met Thr Pro Ile Asp Glu Thr Met Glu Ala Leu Asp Thr Leu 130 135 140 Val Lys Gln Ala Lys Val Arg Tyr Val Ala Cys Ser Asn Tyr Ser Gly 145 150 155 160 Trp His Ile Met Lys Gly Leu Ala Ala Ala Asp Lys His His Gly Gln 165 170 175 Arg Phe Ile Ser Gln Gln Ile His Tyr Thr Leu His Ser Arg Glu Gly 180 185 190 Glu Tyr Glu Leu Val Pro Ile Ser Gln Asp Gln Gly Leu Ala Ile Leu 195 200 205 Ile Trp Ser Pro Leu Ala Gly Gly Leu Leu Ser Gly Lys Tyr Arg Arg 210 215 220 Asp Gly Gly Pro Asp Ala Gly Arg His Val Gly Gly Phe Arg Glu Pro 225 230 235 240 Pro Val Tyr Asp Trp Asp Lys Leu Tyr Asp Ile Leu Asp Val Ile Val 245 250 255 Ala Ile Ala Arg Glu Arg Asp Val Ser Gly Ala Gln Val Ala Leu Ala 260 265 270 Trp Ala Leu Gly Arg Pro Gly Val Thr Ser Ala Ile Ile Gly Gly Arg 275 280 285 Ser Glu Ala Gln Phe Arg Asp Asn Ile Gly Ala Ala Glu Leu Lys Leu 290 295 300 Thr Asp Glu Glu Arg Ala Arg Leu Asp Ala Ala Ser Arg Pro Pro Leu 305 310 315 320 Leu Tyr Pro Tyr Trp His Asn Ser Phe Thr Gly Gln Asp Arg Leu Asp 325 330 335 Gln Leu Asp Leu Asp Leu Leu Gly Pro Tyr Ala Asp Asp Phe Lys Arg 340 345 350 Gly <210> 70 <211> 353 <212> PRT <213> Artificial sequence <220> <223> Enzyme <400> 70 Met Asp Tyr Lys Tyr Leu Gly Arg Ser Gly Leu Arg Val Ser Val Leu 1 5 10 15 Thr Met Gly Thr Met Thr Phe Gly Gly Ser Glu Arg Val Gly His Thr 20 25 30 Ala Gln Ala Glu Ala Thr Arg Gln Ile Asp Leu Cys Leu Asp His Gly 35 40 45 Ile Asn Leu Leu Asp Thr Ala Asn Val Tyr Asn Ala Gly Val Ser Glu 50 55 60 Glu Met Leu Gly Val Ala Leu Ser Glu His Gly Arg Arg Gln Lys Ala 65 70 75 80 Leu Ile Ala Thr Lys Val Arg Phe Arg Met Gly Asn Gly Pro Asn Glu 85 90 95 Val Gly Leu Ser Arg His His Ile Ile Ala Gln Ala Glu Ala Ser Leu ? 100 105 110 Lys Arg Leu Lys Thr Asp Val Ile Glu Leu Tyr Gln Val His Glu Trp 115 120 125 Asp Gly Met Thr Pro Ile Glu Asp Thr Met Glu Ala Leu Asp Thr Leu 130 135 140 Val Lys Gln Gly Lys Val Arg Tyr Val Ala Cys Ser Asn Tyr Ser Gly 145 150 155 160 Trp His Ile Met Lys Gly Leu Ala Ala Ala Asp Lys His His Gly Gln 165 170 175 Arg Phe Ile Ser Gln Gln Ile His Tyr Thr Leu His Ser Arg Glu Ala 180 185 190 Glu Tyr Glu Leu Val Pro Ile Ser Gln Asp Gln Gly Leu Gly Ile Leu 195 200 205 Ile Trp Ser Pro Leu Ala Gly Gly Leu Leu Ser Gly Lys Tyr Arg Arg 210 215 220 Asp Gly Gly Pro Asp Ala Gly Arg His Val Gly Gly Phe Arg Glu Pro 225 230 235 240 Pro Val Tyr Asp Trp Asp Lys Leu Tyr Glu Ile Leu Asp Val Ile Val 245 250 255 Ala Ile Ala Arg Glu Arg Asp Val Ser Gly Ala Gln Val Ala Leu Ala 260 265 270 Trp Ala Leu Gly Arg Pro Gly Val Thr Ser Ala Ile Ile Gly Gly Arg 275 280 285 Ser Glu Ala Gln Phe Arg Asp Asn Ile Gly Ala Ala Asp Leu Lys Leu 290 295 300 Ser Asp Glu Glu Arg Ala Arg Leu Asp Ala Ala Ser Arg Pro Pro Leu 305 310 315 320 Leu Tyr Pro Tyr Trp His Gln Ser Phe Thr Ala Asn Asp Arg Leu Asp 325 330 335 Gln Leu Asp Leu Asp Leu Ile Gly Pro Tyr Ala Glu Asp Phe Lys Arg 340 345 350<00031z118>Gly <210> 71<00u3120><211> 353 <212> PRT <213> Artificial Sequence <220> <223> Enzyme<u <400> 71 Met Asp Tyr Arg Tyr Leu Gly Arg Ser Gly Leu Arg Val Ser Val Leu 1 5 10 15 Thr Met Gly Thr Met Thr Phe Ala Gly Ser Glu Lys Val Gly His Thr 20 25 30 Gly Gln Ala Asp Ala Thr Arg Gln Ile Asp Leu Cys Leu Asp His Gly 35 40 45 It should be noted that there seems to be an error in the "u0003125" in the original text. It should probably be "0003125". This translation is based on the best understanding of the text with the given errors.Ile Asn Leu Leu Asp Thr Ala Asn Val Tyr Asn Ala Gly Val Ser Glu 50 55 60 Glu Met Ile Gly Val Gly Leu Ser Glu His Gly Arg Arg Gln Lys Ala 65 70 75 80 Leu Ile Ala Thr Lys Val Arg Phe Arg Met Ala Asn Gly Pro Asn Glu 85 90 95 Val Gly Leu Ser Arg His His Ile Ile Ala Gln Ala Glu Ala Ser Ile 100 105 110 Lys Arg Leu Lys Thr Asp Val Ile Asp Leu Tyr Gln Val His Glu Trp 115 120 125 Asp Gly Met Thr Pro Ile Asp Glu Thr Met Glu Ala Leu Asp Thr Leu 130 135 140 Val Lys Gln Gly Lys Val Arg Tyr Val Ala Cys Ser Asn Tyr Ser Gly 145 150 155 160 Trp His Ile Met Lys Gly Leu Ala Gly Ala Asp Lys His His Gly Gln 165 170 175 Arg Phe Ile Ser Gln Gln Ile His Tyr Thr Leu His Ser Arg Glu Gly 180 185 190 Glu Tyr Glu Leu Val Pro Ile Ser Gln Asp Gln Gly Leu Ala Ile Leu 195 200 205 Ile Trp Ser Pro Leu Ala Gly Gly Leu Leu Ser Gly Lys Tyr Arg Arg 210 215 220 Asp Gly Gly Pro Asp Ala Gly Arg His Val Gly Gly Phe Arg Glu Pro 225 230 235 240 Pro Val Tyr Asp Trp Asp Lys Leu Tyr Asp Ile Leu Asp Val Ile Val 245 250 255 Ala Ile Ala Arg Glu Arg Asp Val Ser Gly Ala Gln Val Ala Leu Ala 260 265 270 Trp Ala Leu Gly Arg Pro Gly Val Thr Ser Ala Ile Ile Gly Gly Arg 275 280 285 Ser Glu Ala Gln Phe Arg Asp Asn Ile Ala Ala Ala Glu Leu Lys Leu 290 295 300 Ser Asp Asp Glu Arg Ala Arg Leu Asp Ala Ala Ser Arg Pro Pro Leu 305 310 315 320 Leu Tyr Pro Tyr Trp His Gln Ser Phe Thr Gly Gln Asp Arg Leu Asp 325 330 335 Gln Ile Asp Leu Asp Leu Leu Gly Pro Tyr Ala Glu Glu Phe Lys Arg 340 345 350 Gly <210> 72 <211> 353 <212> PRT <. / 213> Synthetic sequence <220> <223> Enzyme <400> 72 Met Asp Tyr Arg Tyr Ile Gly Arg Ser Gly Leu Arg Val Ser Val Leu 1 5 10 15 Thr Met Gly Thr Met Thr Phe Gly Gly Ser Glu Lys Val Gly His Thr 20 25 30 Gly Gln Ala Asp Ala Thr Arg Gln Ile Asp Ile Cys Leu Asp His Gly 35 40 45 Ile Asn Leu Leu Asp Thr Ala Asn Val Tyr Asn Ala Gly Val Ser Glu 50 55 60 Glu Met Leu Gly Val Gly Leu Ser Glu His Gly Arg Arg Gln Lys Ala 65 70 75 80 Leu Ile Ala Thr Lys Val Arg Phe Arg Met Ala Asn Gly Pro Asn Glu 85 90 95 Val Gly Leu Ser Arg His His Ile Ile Ala Gln Ala Glu Ala Ser Ile 100 105 110 Lys Arg Leu Lys Thr Asp Val Ile Asp Leu Tyr Gln Val His Glu Trp 115 120 125 Asp Gly Met Thr Pro Ile Glu Asp Thr Met Glu Ala Leu Asp Thr Leu 130 135 140 Val Lys Gln Gly Lys Val Arg Tyr Val Ala Cys Ser Asn Tyr Ser Gly 145 150 155 160 Trp His Ile Met Lys Ala Leu Ala Gly Ala Asp Lys His His Gly Gln 165 170 175 Arg Phe Ile Ser Gln Gln Ile His Tyr Thr Leu His Ser Arg Glu Ala 180 185 190 Glu Tyr Glu Leu Val Pro Ile Ser Gln Asp Gln Gly Leu Gly Ile Leu 195 200 205 Ile Trp Ser Pro Leu Ala Gly Gly Leu Leu Ser Gly Lys Tyr Arg Arg 210 215 220 Asp Gly Gly Pro Asp Ala Gly Arg His Val Gly Gly Phe Arg Glu Pro 225 230 235 240 Pro Val Tyr Asp Trp Asp Lys Leu Tyr Asp Leu Ile Asp Val Ile Val 245 250 255 Ala Ile Ala Arg Glu Arg Asp Val Ser Gly Ala Gln Val Ala Leu Ala 260 265 270 Trp Ala Leu Gly Arg Pro Gly Val Thr Ser Ala Ile Ile Gly Gly Arg 275 280 285 Ser Glu Ala Gln Phe Arg Asp Asn Ile Gly Ala Ala Asp Leu Lys Leu 290 295 300 Thr Asp Asp Glu Arg Ala Arg Leu Asp Ala Ala Ser Arg Pro Pro Leu 305 310 315 320 Leu Tyr Pro Tyr Trp His Asn Ser Phe Thr Ala Asn Asp Arg Leu Asp 325 330 335 Gln Ile Asp Leu Asp Leu Ile Gly Pro Tyr Ala Glu Asp Phe Lys Arg 340 345 350 Gly <210> 73 <211> 353 <212> PRT <213> Artificial Sequence <220> <223> Enzyme <400> 73 Met Asp Tyr Arg Tyr Leu Gly Arg Ser Gly Leu Arg Val Ser Val Leu 1 5 10 15 Thr Met Gly Thr Met Thr Phe Gly Gly Ser Glu Lys Val Gly His Thr 20 25 30 Gly Gln Ala Asp Ala Thr Arg Gln Leu Asp Leu Cys Leu Asp His Gly 35 40 45 Ile Asn Leu Leu Asp Thr Ala Asn Val Tyr Asn Ala Gly Val Ser Glu 50 55 60 Glu Met Ile Gly Val Gly Leu Ser Glu His Gly Arg Arg Gln Lys Ala 65 70 75 80 Leu Ile Ala Thr Lys Val Arg Phe Arg Met Gly Asn Gly Pro Asn Glu 85 90 95 Val Gly Leu Ser Arg His His Ile Ile Ala Gln Ala Glu Ala Ser Leu 100 105 110 Lys Arg Leu Lys Thr Asp Val Ile Glu Leu Tyr Gln Val His Glu Trp 115 120 125 Asp Gly Met Thr Pro Ile Glu Asp Thr Met Glu Ala Leu Asp Thr Leu 130 135 140 Val Lys Gln Ala Lys Val Arg Tyr Val Ala Cys Ser Asn Tyr Ser Gly 145 150 155 160 Trp His Ile Met Lys Ala Leu Ala Gly Ala Asp Lys His His Gly Gln 165 170 175 Arg Phe Ile Ser Gln Gln Ile His Tyr Thr Leu His Ser Arg Glu Ala 180 185 190 Glu Tyr Glu Leu Val Pro Ile Ser Gln Asp Gln Gly Leu Ala Ile Leu 195 200 205 Ile Trp Ser Pro Leu Ala Gly Gly Leu Leu Ser Gly Lys Tyr Arg Arg 210 215 220 Asp Gly Gly Pro Asp Ala Gly Arg His Val Gly Gly Phe Arg Glu Pro 225 230 235 240 Pro Val Tyr Asp Trp Asp Lys Leu Tyr Glu Ile Ile Asp Val Ile Val 245 250 255 Ala Ile Ala Arg Glu Arg Asp Val Ser Gly Ala Gln Val Ala Leu Ala 260 265 270 Trp Ala Leu Gly Arg Pro Gly Val Thr Ser Ala Ile Ile Gly Gly Arg 275 280 285 Ser Glu Ala Gln Phe Arg Asp Asn Leu Ala Ala Ala Glu Leu Lys Leu 290 295 300 Thr Asp Glu Glu Arg Ala Arg Leu Asp Ala Ala Ser Arg Pro Pro Leu 305 310 315 320 Leu Tyr Pro Tyr Trp His Gln Ser Phe Thr Ala Gln Asp Arg Leu Asp 325 330 335 Gln Leu Asp Leu Asp Leu Ile Gly Pro Tyr Ala Glu Asp Phe Lys Arg 340 345 350 Gly <210> 74 <211> 353 <212> PRT <213> Artificial Sequence <220> <223> Enzyme <400> 74 Met Asp Tyr Arg Tyr Ile Gly Arg Ser Gly Ile Arg Val Ser Val Leu 1 5 10 15 Thr Met Gly Thr Met Thr Phe Gly Gly Ser Glu Arg Val Gly His Thr 20 25 30 Ala Gln Ala Asp Ala Thr Arg Gln Leu Asp Leu Cys Leu Asp His Gly 35 40 45 Ile Asn Leu Leu Asp Thr Ala Asn Val Tyr Asn Ala Gly Val Ser Glu 50 55 60 Glu Met Ile Gly Val Ala Leu Ser Glu His Gly Arg Arg Gln Lys Ala 65 70 75 80 Leu Ile Ala Thr Lys Val Arg Phe Arg Met Ala Asn Gly Pro Asn Glu 85 90 95 Val Gly Leu Ser Arg His His Ile Ile Ala Gln Ala Glu Ala Ser Ile 100 105 110 Lys Arg Leu Lys Thr Asp Val Ile Glu Leu Tyr Gln Val His Glu Trp 115 120 125 Asp Gly Met Thr Pro Ile Glu Glu Thr Met Glu Ala Leu Asp Thr Leu 130 135 140 Val Lys Gln Ala Lys Val Arg Tyr Val Gly Cys Ser Asn Tyr Ser Gly 145 150 155 160 Trp His Ile Met Lys Ala Leu Ala Gly Ala Asp Lys His His Gly Gln 165 170 175 Arg Phe Ile Ser Gln Gln Ile His Tyr Thr Leu His Ser Arg Glu Gly 180 185 190 Glu Tyr Glu Leu Val Pro Ile Ser Gln Asp Gln Gly Leu Gly Ile Leu 195 200 205 Ile Trp Ser Pro Leu Ala Gly Gly Leu Leu Ser Gly Lys Tyr Arg Arg 210 215 220 Asp Gly Gly Pro Asp Ala Gly Arg His Val Gly Gly Phe Arg Glu Pro 225 230 235 240 Pro Val Tyr Asp Trp Asp Lys Leu Tyr Asp Leu Ile Asp Val Ile Val 245 250 255 Ala Ile Ala Arg Glu Arg Asp Val Ser Gly Ala Gln Val Ala Leu Ala 260 265 270 Trp Ala Leu Gly Arg Pro Gly Val Thr Ser Ala Ile Ile Gly Gly Arg 275 280 285 Ser Glu Ala Gln Phe Arg Asp Asn Leu Gly Ala Ala Glu Leu Lys Leu 290 295 300 Thr Asp Glu Glu Arg Ala Arg Leu Asp Ala Ala Ser Arg Pro Pro Leu 305 310 315 320 Leu Tyr Pro Tyr Trp His Gln Ser Phe Thr Gly Asn Asp Arg Leu Asp 325 330 335 Gln Ile Asp Leu Asp Leu Leu Gly Pro Tyr Ala Asp Glu Phe Lys Arg 340 345 350 Gly <210> 75 <211> 353 <212> PRT <213> Artificial sequence <220> <223> Enzyme <400> 75 Met Asp Tyr Arg Tyr Ile Gly Arg Ser Gly Ile Arg Val Ser Val Leu 1 5 10 15 Thr Met Gly Thr Met Thr Phe Ala Gly Ser Glu Arg Val Gly His Thr 20 25 30 Ala Gln Ala Asp Ala Thr Arg Gln Leu Asp Ile Cys Leu Asp His Gly 35 40 45 Ile Asn Leu Leu Asp Thr Ala Asn Val Tyr Asn Ala Gly Val Ser Glu 50 55 60 Glu Met Leu Gly Val Gly Leu Ser Glu His Gly Arg Arg Gln Lys Ala 65 70 75 80 Leu Ile Ala Thr Lys Val Arg Phe Arg Met Ala Asn Gly Pro Asn Glu 85 90 95 Val Gly Leu Ser Arg His His Ile Ile Ala Gln Ala Glu Ala Ser Leu 100 105 110 Lys Arg Leu Lys Thr Asp Val Ile Glu Leu Tyr Gln Val His Glu Trp 115 120 125 Asp Gly Met Thr Pro Ile Asp Glu Thr Met Glu Ala Leu Asp Thr Leu 130 135 140 Val Lys Gln Ala Lys Val Arg Tyr Val Ala Cys Ser Asn Tyr Ser Gly 145 150 155 160 Trp His Ile Met Lys Gly Leu Ala Gly Ala Asp Lys His His Gly Gln 165 170 175 Arg Phe Ile Ser Gln Gln Ile His Tyr Thr Leu His Ser Arg Asp Gly 180 185 190 Glu Tyr Glu Leu Val Pro Ile Ser Gln Asp Gln Gly Leu Ala Ile Leu 195 200 205 Ile Trp Ser Pro Leu Ala Gly Gly Leu Leu Ser Gly Lys Tyr Arg Arg 210 215 220 Asp Gly Gly Pro Asp Ala Gly Arg His Val Gly Gly Phe Arg Glu Pro 225 230 235 240 Pro Val Tyr Asp Trp Asp Lys Leu Tyr Glu Leu Ile Asp Val Ile Val 245 250 255 Ala Ile Ala Arg Glu Arg Asp Val Ser Gly Ala Gln Val Ala Leu Ala 260 265 270 Trp Ala Leu Gly Arg Pro Gly Val Thr Ser Ala Ile Ile Gly Gly Arg 275 280 285 Ser Glu Ala Gln Phe Arg Asp Asn Ile Gly Ala Ala Asp Leu Lys Leu 290 295 300 Ser Asp Asp Glu Arg Ala Arg Leu Asp Ala Ala Ser Arg Pro Pro Leu 305 310 315 320 Leu Tyr Pro Tyr Trp His Asn Ser Phe Thr Gly Asn Asp Arg Leu Asp[[ID=X]] 325 330 335 Gln Leu Asp Leu Asp Leu Leu Gly Pro Tyr Ala Asp Glu Phe Lys Arg 34********** Gly <210> 76 <211> 353 <212> PRT <213> Artificial Sequence <220> <223> Enzyme <400> 76 Met Asp Tyr Arg Tyr Leu Gly Arg Ser Gly Ile Arg Val Ser Thr Leu 1 5 10 15 Thr Met Gly Thr Met Thr Phe Ala Gly Ser Glu Lys Val Gly His Thr 20 25 30 Gly Gln Ala Glu Ala Thr Arg Gln Leu Asp Ile Cys Leu Asp His Gly 35 40 45 It seems there is an issue with the number "34**********" in the original text. I've translated it as best as possible while keeping the format intact. If this is incorrect, please provide the correct information.Ile Asn Leu Leu Asp Thr Ala Asn Val Tyr Asn Ala Gly Val Ser Glu 50 55 60 Glu Met Leu Gly Val Gly Leu Ser Glu Asn Gly Arg Arg Gln Lys Ala 65 70 75 80 Leu Ile Ala Thr Lys Val Arg Phe Arg Met Ala Asn Gly Pro Asn Glu 85 90 95 Val Gly Leu Ser Arg His His Ile Ile Ala Gln Ala Glu Ala Ser Ile 100 105 110 Lys Arg Leu Lys Thr Asp Val Ile Glu Leu Tyr Gln Val His Glu Trp 115 120 125 Asp Gly Met Thr Pro Ile Asp Glu Thr Met Glu Ala Leu Asp Thr Leu 130 135 140 Val Lys Gln Ala Lys Val Arg Tyr Ile Ala Cys Ser Asn Tyr Ser Gly 145 150 155 160 Trp His Ile Met Lys Gly Leu Ala Gly Ala Asp Lys Ser His Gly Gln 165 170 175 Arg Phe Ile Ser Gln Gln Ile His Tyr Thr Leu His Ser Arg Glu Ala 180 185 190 Glu Tyr Glu Leu Val Pro Ile Ser Gln Asp Gln Gly Leu Ala Ile Leu 195 200 205 Ile Trp Ser Pro Leu Ala Gly Gly Leu Leu Ser Gly Lys Tyr Arg Arg 210 215 220 Asp Gly Gly Pro Asp Ser Gly Arg His Val Gly Gly Ser Arg Glu Pro 225 230 235 240 Pro Val Pro Asp Trp Asp Lys Leu Tyr Glu Ile Leu Asp Val Ile Val 245 250 255 Ala Ile Ala Glu Glu Arg Gly Val Ser Gly Ala Gln Val Ala Leu Ala 260 265 270 Trp Ala Leu Gly Arg Pro Gly Val Thr Ser Ala Ile Ile Gly Gly Arg 275 280 285 Ser Glu Ala Gln Phe Arg Asp Asn Leu Gly Ala Ala Asp Leu Lys Leu 290 295 300 Ser Asp Glu Glu Arg Gln Arg Leu Asp Ala Ala Ser Arg Pro Pro Leu 305 310 315 320 Leu Tyr Pro Tyr Trp His Asn Ser Phe Thr Gly Asn Asp Arg Leu Asp 325 330 335 Pro Ala Asp Leu Asp Leu Leu Gly Thr Tyr Ala Asp Glu Phe Lys Arg 340 345 350 Gly <210> 77 <211> 353 <212> PRT <213> Artificial Sequence <220> <223> Enzyme <400> 77 Met Asp Tyr Arg Tyr Leu Gly Arg Ser Gly Ile Arg Val Ser Thr Leu 1 5 10 15 Thr Met Gly Thr Met Thr Phe Ala Gly Ser Glu Lys Val Gly His Thr 20 25 30 Pro Gln Ala Glu Ala Thr Arg Gln Leu Asp Leu Cys Leu Asp His Gly 35 40 45 Ile Asn Leu Leu Asp Thr Ala Asn Val Tyr Asn Ala Gly Val Ser Glu 50 55 60 Glu Met Ile Gly Val Ala Leu Ala Glu Asn Gly Arg Arg Gln Lys Ala 65 70 75 80 Leu Ile Ala Thr Lys Val Arg Phe Arg Met Gly Asn Gly Pro Asn Glu 85 90 95 Val Gly Leu Ser Arg His His Ile Ile Ala Gln Ala Glu Ala Ser Leu 100 105 110 Lys Arg Leu Lys Thr Asp Val Ile Asp Leu Tyr Gln Val His Glu Trp 115 120 125 Asp Gly Met Thr Pro Ile Glu Asp Thr Met Glu Ala Leu Asp Thr Leu 130 135 140 Val Lys Gln Ala Lys Val Arg Tyr Ile Gly Cys Ser Asn Tyr Ser Gly 145 150 155 160 Trp His Ile Met Lys Ala Leu Ala Ala Ala Asp Lys Ser His Gly Gln 165 170 175 Arg Phe Ile Ser Gln Gln Ile His Tyr Thr Leu His Ser Arg Glu Ala 180 185 190 Glu Tyr Glu Leu Val Pro Ile Ser Gln Asp Gln Gly Leu Gly Ile Leu 195 200 205 Ile Trp Ser Pro Leu Ala Gly Gly Leu Leu Ser Gly Lys Tyr Arg Arg 210 215 220 Asp Gly Gly Pro Glu Ser Gly Arg His Val Gly Gly Ser Arg Glu Pro 225 230 235 240 Pro Val Pro Asp Trp Asp Lys Leu Tyr Asp Ile Ile Asp Val Ile Val 245 250 255 Ala Ile Ala Glu Glu Arg Gly Val Ser Gly Ala Gln Val Ala Leu Ala 260 265 270 Trp Ala Leu Gly Arg Pro Gly Val Thr Ser Val Ile Ile Gly Gly Arg 275 280 285 Ser Glu Ala Gln Phe Arg Asp Asn Leu Ala Ala Ala Asp Leu Lys Leu 290 295 300 Ser Asp Glu Glu Arg Gln Arg Leu Asp Ala Ala Ser Arg Pro Pro Leu 305 310 315 320 Leu Tyr Pro Tyr Trp His Gln Thr Phe Thr Gly Asn Asp Arg Leu Gly 325 330 335 Pro Ala Asp Gln Asp Leu Leu Ser Thr Tyr Val Glu Asp Phe Lys Arg 340 345 350 Gly <210> 78 <211> 354 <212> PRT <213> Artificial Sequence <220> <223> Enzyme <400> 78 Met Asp Tyr Arg Tyr Leu Gly Arg Ser Gly Ile Arg Val Ser Thr Leu 1 5 10 15 Thr Met Gly Thr Met Thr Phe Gly Gly Ser Glu Lys Val Gly His Thr 20 25 30 Gly Gln Ala Asp Ala Thr Arg Gln Ile Asp Ile Cys Leu Asp His Gly 35 40 45 Ile Asn Leu Leu Asp Thr Ala Asn Val Tyr Asn Ala Gly Val Ser Glu 50 55 6​​​​​Leu Ile Ala Thr Lys Val Arg Phe Arg Met Gly Asn Gly Pro Asn Glu 85 90 95 Val Gly Leu Ser Arg His His Ile Ile Ala Gln Ala Glu Ala Ser Leu 100 105 110 Lys Arg Leu Lys Thr Asp Val Ile Glu Leu Tyr Gln Val His Glu Trp 115 120 125 Asp Gly Met Thr Pro Ile Asp Glu Thr Met Glu Ala Leu Asp Thr Leu 130 135 140 Val Lys Gln Ala Lys Val Arg Tyr Ile Ala Cys Ser Asn Tyr Ser Gly 145 150 155 160 Trp His Ile Met Lys Gly Leu Ala Ala Ala Asp Lys Ser His Gly Gln 165 170 175 Arg Phe Ile Ser Gln Gln Ile His Tyr Thr Leu His Ser Arg Glu Ala 180 185 190 Glu Tyr Glu Leu Val Pro Ile Ser Gln Asp Gln Gly Leu Ala Ile Leu 195 200 205 Ile Trp Ser Pro Leu Ala Gly Gly Leu Leu Ser Gly Lys Tyr Arg Arg 210 215 220 Asp Gly Gly Pro Asp Ser Gly Arg His Val Gly Gly Ser Arg Glu Pro 225 230 235 240 Pro Val Pro Asp Trp Asp Lys Leu Tyr Asp Ile Leu Asp Val Ile Val 245 250 255 Ala Ile Ala Glu Glu Arg Gly Val Ser Gly Ala Gln Val Ala Leu Ala 260 265 270 Trp Ala Leu Gly Arg Pro Gly Val Thr Ser Ala Ile Ile Gly Gly Arg 275 280 285 Ser Glu Ala Gln Phe Arg Asp Asn Leu Gly Ala Ala Asp Leu Lys Leu 290 295 300 Thr Asp Glu Glu Arg Gln Arg Leu Asp Ala Ala Ser Arg Pro Pro Leu 305 310 315 320 Leu Tyr Pro Tyr Trp His Asn Ser Phe Thr Gly Gln Asp Arg Leu Gly 325 330 335 Pro Ala Asp Gln Asp Leu Leu Leu Ser Thr Tyr Val Glu Asp Phe Lys 340 345 350 Arg Gly <210> 79 <211> 353 <212> PRT <213> Artificial Sequence <220> <223> Enzyme <400> 79 Met Asp Tyr Arg Tyr Leu Gly Arg Ser Gly Leu Arg Val Ser Thr Leu 1 5 10 15 Thr Met Gly Thr Met Thr Phe Gly Gly Ser Glu Lys Val Gly His Thr 20 25 30 Ala Gln Ala Glu Ala Thr Arg Gln Ile Asp Leu Cys Leu Asp His Gly 35 40 45 Ile Asn Leu Leu Asp Thr Ala Asn Val Tyr Asn Ala Gly Val Ser Glu 50 55 60 Glu Met Ile Gly Val Ala Leu Ala Glu Asn Gly Arg Arg Gln Lys Ala 65 70 75 80 Leu Ile Ala Thr Lys Val Arg Phe Arg Met Gly Asn Gly Pro Asn Glu 85 90 95 Val Gly Leu Ser Arg His His Ile Ile Ala Gln Ala Glu Ala Ser Leu 100 105 110 Lys Arg Leu Lys Thr Asp Val Ile Glu Leu Tyr Gln Val His Glu Trp 115 120 125 Asp Gly Met Thr Pro Ile Glu Asp Thr Met Glu Ala Leu Asp Thr Leu 130 135 140 Val Lys Gln Gly Lys Val Arg Tyr Ile Ala Cys Ser Asn Tyr Ser Gly 145 150 155 160 Trp His Ile Met Lys Gly Leu Ala Ala Ala Asp Lys Ser His Gly Gln 165 170 175 Arg Phe Ile Ser Gln Gln Ile His Tyr Thr Leu His Ser Arg Glu Ala 180 185 190 Glu Tyr Glu Leu Val Pro Ile Ser Gln Asp Gln Gly Leu Gly Ile Leu 195 200 205 Ile Trp Ser Pro Leu Ala Gly Gly Leu Leu Ser Gly Lys Tyr Arg Arg 210 215 220 Asp Gly Gly Pro Asp Ser Gly Arg His Val Gly Gly Ser Arg Glu Pro 225 230 235 240 Pro Val Pro Asp Trp Asp Lys Leu Tyr Glu Ile Leu Asp Val Ile Val 245 250 255 Ala Ile Ala Glu Glu Arg Gly Val Ser Gly Ala Gln Val Ala Leu Ala 260 265 270 Trp Ala Leu Gly Arg Pro Gly Val Thr Ser Ala Ile Ile Gly Gly Arg 275 280 285 Ser Glu Ala Gln Phe Arg Asp Asn Leu Gly Ala Ala Asp Leu Lys Leu 290 295 300 Ser Asp Glu Glu Arg Gln Arg Leu Asp Ala Ala Ser Arg Pro Pro Leu 305 310 315 320 Leu Tyr Pro Tyr Trp His Gln Ser Phe Thr Ala Asn Asp Arg Leu Gly 325 330 335 Pro Ala Asp Gln Asp Leu Ile Gly Thr Tyr Ala Glu Asp Phe Lys Arg 340 345 350 Gly <210> 80 <211> 353 <212> PRT <213> Artificial Sequence <220> <223> Enzyme <400> 80 Met Asp Tyr Arg Tyr Leu Gly Arg Ser Gly Leu Arg Val Ser Thr Leu 1 5 10 15 Thr Met Gly Thr Met Thr Phe Ala Gly Ser Glu Lys Val Gly His Thr 20 25 30 Gly Gln Ala Asp Ala Thr Arg Gln Ile Asp Leu Cys Leu Asp His Gly 35 40 45 Ile Asn Leu Leu Asp Thr Ala Asn Val Tyr Asn Ala Gly Val Ser Glu 50 55 60 Glu Met Ile Gly Val Gly Leu Ser Glu Asn Gly Arg Arg Gln Lys Ala 65 70 75 80 Leu Ile Ala Thr Lys Val Arg Phe Arg Met Ala Asn Gly Pro Asn Glu 85 90 95 Val Gly Leu Ser Arg His His Ile Ile Ala Gln Ala Glu Ala Ser Ile 100 105 110 Lys Arg Leu Lys Thr Asp Val Ile Asp Leu Tyr Gln Val His Glu Trp 115 120 125 Asp Gly Met Thr Pro Ile Asp Glu Thr Met Glu Ala Leu Asp Thr Leu 130 135 140 Val Lys Gln Gly Lys Val Arg Tyr Ile Ala Cys Ser Asn Tyr Ser Gly 145 150 155 160 Trp His Ile Met Lys Gly Leu Ala Gly Ala Asp Lys Ser His Gly Gln 165 170 175 Arg Phe Ile Ser Gln Gln Ile His Tyr Thr Leu His Ser Arg Glu Ala 180 185 190 Glu Tyr Glu Leu Val Pro Ile Ser Gln Asp Gln Gly Leu Ala Ile Leu 195 200 205 Ile Trp Ser Pro Leu Ala Gly Gly Leu Leu Ser Gly Lys Tyr Arg Arg 210 215 220 Asp Gly Gly Pro Asp Ser Gly Arg His Val Gly Gly Ser Arg Glu Pro 225 230 235 240 Pro Val Pro Asp Trp Asp Lys Leu Tyr Asp Ile Leu Asp Val Ile Val 245 250 255 Ala Ile Ala Glu Glu Arg Gly Val Ser Gly Ala Gln Val Ala Leu Ala 260 265 270 Trp Ala Leu Gly Arg Pro Gly Val Thr Ser Ala Ile Ile Gly Gly Arg 275 280 285 Ser Glu Ala Gln Phe Arg Asp Asn Leu Ala Ala Ala Asp Leu Lys Leu 290 295 300 Ser Asp Asp Glu Arg Gln Arg Leu Asp Ala Ala Ser Arg Pro Pro Leu 305 310 315 320 Leu Tyr Pro Tyr Trp His Gln Thr Phe Thr Gly Gln Asp Arg Leu Asp 325 330 335 Pro Ala Asp Leu Asp Leu Leu Gly Thr Tyr Ala Glu Glu Phe Lys Arg 340 345 350 Gly <210> 81 <211> 353 <212> PRT [[ID=​​​​​​​​​​​​​​​​​​​Ile Asn Leu Leu Asp Thr Ala Asn Val Tyr Asn Ala Gly Val Ser Glu 50 55 60 Glu Met Leu Gly Val Gly Leu Ser Glu Asn Gly Arg Arg Gln Lys Ala 65 70 75 80 Leu Ile Ala Thr Lys Val Arg Phe Arg Met Ala Asn Gly Pro Asn Glu 85 90 95 Val Gly Leu Ser Arg His His Ile Ile Ala Gln Ala Glu Ala Ser Ile 100 105 110 Lys Arg Leu Lys Thr Asp Val Ile Asp Leu Tyr Gln Val His Glu Trp 115 120 125 Asp Gly Met Thr Pro Ile Glu Asp Thr Met Glu Ala Leu Asp Thr Leu 130 135 140 Val Lys Gln Gly Lys Val Arg Tyr Ile Ala Cys Ser Asn Tyr Ser Gly 145 150 155 160 Trp His Ile Met Lys Ala Leu Ala Gly Ala Asp Lys Ser His Gly Gln 165 170 175 Arg Phe Ile Ser Gln Gln Ile His Tyr Thr Leu His Ser Arg Glu Ala 180 185 190 Glu Tyr Glu Leu Val Pro Ile Ser Gln Asp Gln Gly Leu Gly Ile Leu 195 200 205 Ile Trp Ser Pro Leu Ala Gly Gly Leu Leu Ser Gly Lys Tyr Arg Arg 210 215 220 Asp Gly Gly Pro Asp Ser Gly Arg His Val Gly Gly Ser Arg Glu Pro 225 230 235 240 Pro Val Pro Asp Trp Asp Lys Leu Tyr Asp Ile Ile Asp Val Ile Val 245 250 255 Ala Ile Ala Glu Glu Arg Gly Val Ser Gly Ala Gln Val Ala Leu Ala 260 265 270 Trp Ala Leu Gly Arg Pro Gly Val Thr Ser Ala Ile Ile Gly Gly Arg 275 280 285 Ser Glu Ala Gln Phe Arg Asp Asn Leu Gly Ala Ala Asp Leu Lys Leu 290 295 300 Thr Asp Glu Glu Arg Gln Arg Leu Asp Ala Ala Ser Arg Pro Pro Leu 305 310 315 320 Leu Tyr Pro Tyr Trp His Gln Thr Phe Thr Ala Asn Asp Arg Leu Asp 325 330 335 Pro Ala Asp Leu Asp Leu Leu Ser Thr Tyr Val Glu Asp Phe Lys Arg 340 345 350 Gly <210> 82 <211> 353 <212> PRT <213> Artificial Sequence <220> <223> Enzyme <400> 82 Met Asp Tyr Arg Tyr Leu Gly Arg Ser Gly Leu Arg Val Ser Thr Leu 1 5 10 15 Thr Met Gly Thr Met Thr Phe Gly Gly Ser Glu Lys Val Gly His Thr 20 25 30 Gly Gln Ala Asp Ala Thr Arg Gln Leu Asp Leu Cys Leu Asp His Gly 35 40 45 Ile Asn Leu Leu Asp Thr Ala Asn Val Tyr Asn Ala Gly Val Ser Glu 50 55 60 Glu Met Ile Gly Val Gly Leu Ser Glu Asn Gly Arg Arg Gln Lys Ala 65 70 75 80 Leu Ile Ala Thr Lys Val Arg Phe Arg Met Gly Asn Gly Pro Asn Glu 85 90 95 Val Gly Leu Ser Arg His His Ile Ile Ala Gln Ala Glu Ala Ser Leu 100 105 110 Lys Arg Leu Lys Thr Asp Val Ile Glu Leu Tyr Gln Val His Glu Trp 115 120 125 Asp Gly Met Thr Pro Ile Glu Asp Thr Met Glu Ala Leu Asp Thr Leu 130 135 140 Val Lys Gln Ala Lys Val Arg Tyr Ile Ala Cys Ser Asn Tyr Ser Gly 145 150 155 160 Trp His Ile Met Lys Ala Leu Ala Gly Ala Asp Lys Ser His Gly Gln 165 170 175 Arg Phe Ile Ser Gln Gln Ile His Tyr Thr Leu His Ser Arg Glu Ala 180 185 190 Glu Tyr Glu Leu Val Pro Ile Ser Gln Asp Gln Gly Leu Ala Ile Leu 195 200 205 Ile Trp Ser Pro Leu Ala Gly Gly Leu Leu Ser Gly Lys Tyr Arg Arg 210 215 220 Asp Gly Gly Pro Asp Ser Gly Arg His Val Gly Gly Ser Arg Glu Pro 225 230 235 240 Pro Val Pro Asp Trp Asp Lys Leu Tyr Glu Ile Ile Asp Val Ile Val 245 250 255 Ala Ile Ala Glu Glu Arg Gly Val Ser Gly Ala Gln Val Ala Leu Ala 260 265 270 Trp Ala Leu Gly Arg Pro Gly Val Thr Ser Ala Ile Ile Gly Gly Arg 275 280 285 Ser Glu Ala Gln Phe Arg Asp Asn Leu Ala Ala Ala Asp Leu Lys Leu 290 295 300 Thr Asp Glu Glu Arg Gln Arg Leu Asp Ala Ala Ser Arg Pro Pro Leu 305 310 315 320 Leu Tyr Pro Tyr Trp His Gln Thr Phe Thr Ala Gln Asp Arg Leu Gly 325 330 335 Pro Ala Asp Gln Asp Leu Ile Gly Thr Tyr Ala Glu Asp Phe Lys Arg 340 345 350 Gly <210> 83 <211> 353 <212> PRT <213> Artificial Sequence <220> <223> Enzyme <400> 83 Met Asp Tyr Arg Tyr Leu Gly Arg Ser Gly Ile Arg Val Ser Thr Leu 1 5 10 15 Thr Met Gly Thr Met Thr Phe Gly Gly Ser Glu Lys Val Gly His Thr 20 25 30 Ala Gln Ala Asp Ala Thr Arg Gln Leu Asp Leu Cys Leu Asp His Gly 35 40 45 Ile Asn Leu Leu Asp Thr Ala Asn Val Tyr Asn Ala Gly Val Ser Glu 50 55 60 Glu Met Ile Gly Val Ala Leu Ser Glu Asn Gly Arg Arg Gln Lys Ala 65 70 75 80 Leu Ile Ala Thr Lys Val Arg Phe Arg Met Ala Asn Gly Pro Asn Glu 85 90 95 Val Gly Leu Ser Arg His His Ile Ile Ala Gln Ala Glu Ala Ser Ile 100 105 110 Lys Arg Leu Lys Thr Asp Val Ile Glu Leu Tyr Gln Val His Glu Trp 115 120 125 Asp Gly Met Thr Pro Ile Glu Glu Thr Met Glu Ala Leu Asp Thr Leu 130 135 140 Val Lys Gln Ala Lys Val Arg Tyr Ile Gly Cys Ser Asn Tyr Ser Gly 145 150 155 160 Trp His Ile Met Lys Ala Leu Ala Gly Ala Asp Lys Ser His Gly Gln 165 170 175 Arg Phe Ile Ser Gln Gln Ile His Tyr Thr Leu His Ser Arg Glu Ala 180 185 190 Glu Tyr Glu Leu Val Pro Ile Ser Gln Asp Gln Gly Leu Gly Ile Leu 195 200 205 Ile Trp Ser Pro Leu Ala Gly Gly Leu Leu Ser Gly Lys Tyr Arg Arg 210 215 220 Asp Gly Gly Pro Asp Ser Gly Arg His Val Gly Gly Ser Arg Glu Pro 225 230 235 240 Pro Val Pro Asp Trp Asp Lys Leu Tyr Asp Ile Ile Asp Val Ile Val 245 250 255 Ala Ile Ala Glu Glu Arg Gly Val Ser Gly Ala Gln Val Ala Leu Ala 260 265 270 Trp Ala Leu Gly Arg Pro Gly Val Thr Ser Ala Ile Ile Gly Gly Arg 275 280 285 Ser Glu Ala Gln Phe Arg Asp Asn Leu Gly Ala Ala Asp Leu Lys Leu 290 295 300 Thr Asp Glu Glu Arg Gln Arg Leu Asp Ala Ala Ser Arg Pro Pro Leu 305 310 315 320 Leu Tyr Pro Tyr Trp His Gln Ser Phe Thr Gly Asn Asp Arg Leu Asp 325 330 335 Pro Ala Asp Leu Asp Leu Leu Gly Thr Tyr Ala Asp Glu Phe Lys Arg 340 345 350 Gly <210> 84 <211> 353 <212> PRT <213> Artificial Sequence <220> <223> Enzyme <400> 84 Met Asp Tyr Arg Tyr Leu Gly Arg Ser Gly Ile Arg Val Ser Thr Leu 1 5 10 15 Thr Met Gly Thr Met Thr Phe Ala Gly Ser Glu Lys Val Gly His Thr 20 25 30 Ala Gln Ala Asp Ala Thr Arg Gln Leu Asp Ile Cys Leu Asp His Gly 35 40 45 Ile Asn Leu Leu Asp Thr Ala Asn Val Tyr Asn Ala Gly Val Ser Glu 50 55 60 Glu Met Leu Gly Val Gly Leu Ser Glu Asn Gly Arg Arg Gln Lys Ala 65 70 75 80 Leu Ile Ala Thr Lys Val Arg Phe Arg Met Ala Asn Gly Pro Asn Glu 85 90 95 Val Gly Leu Ser Arg His His Ile Ile Ala Gln Ala Glu Ala Ser Leu 100 105 110 Lys Arg Leu Lys Thr Asp Val Ile Glu Leu Tyr Gln Val His Glu Trp 115 120 125 Asp Gly Met Thr Pro Ile Asp Glu Thr Met Glu Ala Leu Asp Thr Leu 130 135 140 Val Lys Gln Ala Lys Val Arg Tyr Ile Ala Cys Ser Asn Tyr Ser Gly 145 150 155 160 Trp His Ile Met Lys Gly Leu Ala Gly Ala Asp Lys Ser His Gly Gln 165 170 175 Arg Phe Ile Ser Gln Gln Ile His Tyr Thr Leu His Ser Arg Glu Ala 180 185 190 Glu Tyr Glu Leu Val Pro Ile Ser Gln Asp Gln Gly Leu Gly Ile Leu 195 200 205 Ile Trp Ser Pro Leu Ala Gly Gly Leu Leu Ser Gly Lys Tyr Arg Arg 210 215 220 Asp Gly Gly Pro Asp Ser Gly Arg His Val Gly Gly Ser Arg Glu Pro 225 230 235 240 Pro Val Pro Asp Trp Asp Lys Leu Tyr Glu Ile Ile Asp Val Ile Val 245 250 255 Ala Ile Ala Glu Glu Arg Gly Val Ser Gly Ala Gln Val Ala Leu Ala 260 265 270 Trp Ala Leu Gly Arg Pro Gly Val Thr Ser Ala Ile Ile Gly Gly Arg 275 280 285 Ser Glu Ala Gln Phe Arg Asp Asn Leu Gly Ala Ala Asp Leu Lys Leu 290 295 300 Ser Asp Glu Glu Arg Gln Arg Leu Asp Ala Ala Ser Arg Pro Pro Leu 305 310 315 320 Leu Tyr Pro Tyr Trp His Asn Ser Phe Thr Gly Asn Asp Arg Leu Asp 325 330 335 Pro Ala Asp Leu Asp Leu Leu Gly Thr Tyr Ala Asp Glu Phe Lys Arg 340 345 350 Gly <210> 85 <211> 353 <212> PRT <213> Artificial Sequence <220> <223> Enzyme <400> 85 Met Asp Tyr Arg Tyr Leu Gly Arg Ser Gly Leu Arg Val Ser Thr Leu 1 5 10 15 Thr Met Gly Thr Met Thr Phe Gly Gly Ser Asp Lys Val Gly His Thr 20 25 30 Pro Gln Ala Asp Ala Thr Arg Gln Ile Asp Ile Cys Leu Asp His Gly 35 40 45 Ile Asn Leu Leu Asp Thr Ala Asn Val Tyr Asn Ala Gly Val Ser Glu 50 55 60 Glu Met Ile Gly Val Ala Leu Ala Glu Gly Arg Arg Gln Gln Lys Ala 65 70 75 80 Leu Ile Ala Thr Lys Val Arg Phe Arg Gly Gly Gly Gly Pro Asn Glu 85 90 95 Val Gly Leu Ser Lys His His Ile Ile Ala Gln Ala Glu Ala Ser Leu 100 105 110 Lys Arg Leu Lys Thr Asp Val Ile Glu Leu Tyr Gln Val His Glu Trp 115 120 125 Asp Gly Met Thr Pro Ile Glu Glu Thr Met Glu Ala Leu Asp Thr Leu 130 135 140 Val Lys Gln Gly Lys Val Arg Tyr Ile Ala Cys Ser Asn Tyr Ser Gly 145 150 155 160 Trp His Ile Met Lys Ala Leu Ala Ala Ala Asp Lys Ser His Gly Gln 165 170 175 Arg Phe Ile Ser Gln Gln Ile His Tyr Thr Leu His Ser Arg Glu Ala 180 185 190 Asp Tyr Glu Leu Val Pro Ile Ser Gln Asp Gln Gly Leu Gly Ile Leu 195 200 205 Ile Trp Ser Pro Leu Ala Gly Gly Leu Leu Ser Gly Lys Tyr Arg Arg 210 215 220 Asp Gly Gly Pro Asp Ala Gly Arg His Val Gly Gly Ser Arg Glu Pro 225 230 235 240 Pro Val Pro Asp Trp Asp Lys Leu Tyr Glu Ile Ile Asp Val Ile Val 245 250 255 Ala Ile Ala Glu Glu Arg Gly Val Ser Gly Ala Gln Val Ala Leu Ala 260 265 270 Trp Ala Leu Gly Arg Pro Gly Val Thr Ser Ala Ile Ile Gly Gly Arg 275 280 285 Ser Glu Ala Gln Phe Arg Asp Asn Leu Ala Ala Ala Glu Leu Lys Leu 290 295 300 Thr Asp Asp Glu Arg Gln Arg Leu Asp Ala Ala Ser Arg Pro Pro Leu 305 310 315 320 Leu Tyr Pro Tyr Trp His Gln Ser Phe Thr Ala Gln Glu Arg Leu Gly 325 330 335 Pro Ala Asp Gln Asp Leu Leu Ser Thr Tyr Val Glu Asp Phe Lys Arg 340 345 350​​​​​​​​​​​​​​​​​​​​​​​​​​​​​Ile Asn Leu Leu Asp Thr Ala Asn Val Tyr Asn Ala Gly Val Ser Glu 50 55 60 Glu Met Ile Gly Val Ala Leu Ala Glu His Gly Arg Arg Gln Lys Ala 65 70 75 80 Leu Ile Ala Thr Lys Val Arg Phe Arg Met Gly Asp Gly Pro Asn Glu 85 90 95 Val Gly Leu Ser Arg His His Ile Ile Ala Gln Ala Glu Ala Ser Leu 100 105 110 Lys Arg Leu Lys Thr Asp Val Ile Asp Leu Tyr Gln Val His Glu Trp 115 120 125 Asp Gly Met Thr Pro Ile Glu Glu Thr Met Glu Ala Leu Asp Thr Leu 130 135 140 Val Lys Gln Gly Lys Val Arg Tyr Ile Gly Cys Ser Asn Tyr Ser Gly 145 150 155 160 Trp His Ile Met Lys Ala Leu Ala Ala Ala Asp Lys Ser His Gly Gln 165 170 175 Arg Phe Ile Ser Gln Gln Ile His Tyr Thr Leu His Ser Arg Glu Ala 180 185 190 Glu Tyr Glu Leu Val Pro Ile Ser Gln Asp Gln Gly Leu Gly Ile Leu 195 200 205 Ile Trp Ser Pro Leu Ala Gly Gly Leu Leu Ser Gly Lys Tyr Arg Arg 210 215 220 Asp Gly Gly Pro Asp Ala Gly Arg His Val Gly Gly Ser Arg Asp Pro 225 230 235 240 Pro Val Pro Asp Trp Asp Lys Leu Tyr Glu Ile Leu Asp Val Ile Val 245 250 255 Ala Ile Ala Arg Glu Lys Asp Val Ser Gly Ala Gln Val Ala Leu Ala 260 265 270 Trp Ala Leu Gly Arg Pro Gly Val Thr Ser Ala Ile Ile Gly Gly Arg 275 280 285 Ser Glu Ala Gln Phe Arg Asp Asn Leu Ala Ala Ala Glu Leu Lys Leu 290 295 300 Thr Asp Asp Glu Arg Gln Arg Leu Asp Ala Ala Ser Arg Pro Pro Leu 305 310 315 320 A Leu Tyr Pro Tyr Trp His Gln Ser Phe Thr Gly Gln Asp Arg Leu Asp 325 330 335 Gln Val Asp Leu Asp Leu Ile Gly Pro Tyr Ala Glu Glu Phe Lys Arg 340 345 350 Gly <210> 87 <211> 353 <212> PRT <213> Artificial Sequence <220> <223> Enzyme <400> 87 Met Asp Tyr Lys Tyr Leu Gly Arg Ser Gly Ile Arg Val Ser Thr Leu 1 5 10 15 Thr Met Gly Thr Met Thr Phe Gly Gly Ser Glu Lys Val Gly His Thr 20 25 30 Pro Gln Ala Asp Ala Thr Arg Gln Ile Asp Leu Cys Leu Asp His Gly 35 40 45 Ile Asn Leu Leu Asp Thr Ala Asn Val Tyr Asn Ala Gly Val Ser Glu 50 55 60 Glu Met Ile Gly Val Ala Leu Ala Glu His Gly Arg Arg Gln Lys Ala 65 70 75 80 Leu Ile Ala Thr Lys Val Arg Phe Arg Met Gly Asp Gly Pro Asn Glu 85 90 95 Val Gly Leu Ser Arg His His Ile Ile Ala Gln Ala Glu Ala Ser Leu 100 105 110 Lys Arg Ile Lys Thr Asp Val Ile Asp Leu Tyr Gln Val His Glu Trp 115 120 125 Asp Gly Met Thr Pro Ile Glu Glu Thr Met Glu Ala Leu Asp Thr Leu 130 135 140 Val Lys Gln Ala Lys Val Arg Tyr Ile Gly Cys Ser Asn Tyr Ser Gly 145 150 155 160 Trp His Ile Met Lys Ala Leu Ala Gly Ala Asp Lys Ser His Gly Gln 165 170 175 Arg Phe Ile Ser Gln Gln Ile His Tyr Thr Leu His Ser Arg Glu Ala 180 185 190 Glu Tyr Glu Leu Val Pro Ile Ser Gln Asp Gln Gly Leu Gly Ile Leu 195 200 205 Ile Trp Ser Pro Leu Ala Gly Gly Leu Leu Ser Gly Lys Tyr Arg Arg 210 215 220 Asp Gly Gly Pro Asp Ala Gly Arg His Val Gly Gly Ser Arg Asp Pro 225 230 235 240 Pro Val Pro Asp Trp Asp Arg Leu Tyr Asp Ile Ile Asp Val Ile Val 245 250 255 Ala Ile Ala Glu Glu Arg Gly Val Ser Gly Ala Gln Val Ala Leu Ala 260 265 270 Tyr Ala Leu Gly Arg Pro Gly Val Thr Ser Ala Ile Ile Gly Gly Arg 275 280 285 Ser Glu Ala Gln Phe Arg Asp Asn Leu Gly Ala Ala Glu Leu Lys Leu 290 295 300 Thr Asp Asp Glu Arg Gln Arg Leu Asp Ala Ala Ser Arg Pro Pro Leu 305 310 315 320 Leu Tyr Pro Tyr Trp His Asn Ser Phe Thr Gly Gln Asp Arg Leu Gly 325 330 335 Pro Ala Asp Gln Asp Leu Ile Gly Pro Tyr Ala Glu Glu Phe Lys Arg 340 345 350 Gly <210> 88 <211> 353 <212> PRT <213> Artificial sequence <220> <223> Enzyme <400> 88 Met Asp Tyr Arg Tyr Leu Gly Arg Ser Gly Ile Arg Val Ser Thr Leu 1 5 10 15 Thr Met Gly Thr Met Thr Phe Gly Gly Ser Glu Lys Val Gly His Thr 20 25 30 Pro Gln Ala Asp Ala Thr Arg Gln Ile Asp Ile Cys Leu Asp His Gly 35 40 45 Ile Asn Leu Leu Asp Thr Ala Asn Val Tyr Asn Ala Gly Val Ser Glu 50 55 60 Glu Met Ile Gly Val Ala Leu Ala Glu His Gly Arg Arg Gln Lys Ala 65 70 75 80 Leu Ile Ala Thr Lys Val Arg Phe Arg Met Gly Asp Gly Pro Asn Glu 85 90 95 Val Gly Leu Ser Arg His His Ile Ile Ala Gln Ala Glu Ala Ser Leu 100 105 110 Lys Arg Ile Lys Thr Asp Val Ile Asp Leu Tyr Gln Val His Glu Trp 115 120 125 Asp Gly Met Thr Pro Ile Glu Glu Thr Met Glu Ala Leu Asp Thr Leu 130 135 140 Val Lys Gln Gly Lys Val Arg Tyr Ile Gly Cys Ser Asn Tyr Ser Gly 145 150 155 160 Trp His Ile Met Lys Ala Leu Ala Ala Ala Asp Lys Ser His Gly Gln 165 170 175 Arg Phe Ile Ser Gln Gln Ile His Tyr Thr Leu His Ser Arg Glu Gly 180 185 190 Glu Tyr Glu Leu Val Pro Ile Ser Gln Asp Gln Gly Leu Ala Ile Leu 195 200 205 Ile Trp Ser Pro Leu Ala Gly Gly Leu Leu Ser Gly Lys Tyr Lys Arg 210 215 220 Asp Gly Gly Pro Asp Ala Gly Arg His Val Gly Gly Ser Arg Glu Pro 225 230 235 240 Pro Leu Pro Asp Trp Asp Arg Leu Tyr Asp Ile Ile Asp Val Ile Val 245 250 255 Ala Ile Ala Arg Glu Arg Asp Val Ser Gly Ala Gln Val Ala Leu Ala 260 265 270 Phe Ala Leu Gly Arg Pro Gly Val Thr Ser Ala Ile Ile Gly Gly Arg 275 280 285 Ser Glu Ala Asn Phe Arg Asp Asn Leu Gly Ala Ala Glu Leu Lys Leu 290 295 300 Ser Asp Asp Glu Arg Gln Arg Leu Asp Ala Ala Ser Arg Pro Pro Leu 305 310 315 320 Leu Tyr Pro Tyr Trp His Gln Ser Phe Thr Ala Asn Asp Arg Leu Asp 325 330 335 Gln Leu Asp Leu Asp Leu Ile Gly Pro Tyr Ala Asp Glu Phe Lys Arg 340 345 350 Gly <210> 89 <211> 353 <212> PRT <213> Artificial Sequence <220> <223> Enzyme <400> 89 Met Asp Tyr Arg Tyr Ile Gly Arg Ser Gly Leu Lys Val Ser Ala Met 1 5 10 15 Thr Met Gly Thr Met Thr Phe Gly Gly Ser Glu Arg Val Gly His Thr 20 25 30 Asp Leu Ala Gly Ala Ala Arg Gln Val Asp Leu Cys Leu Asp Ala Gly 35 40 45 Ile Asn Leu Tyr Asp Thr Ala Asn Val Tyr Asn Ala Gly Val Ser Glu 50 55 60 Asp Ile Leu Gly Gln Val Leu Ala Glu His Gly Arg Arg Gln Arg Ala 65 70 75 80 Leu Val Ala Thr Lys Val Arg Phe Arg Met Gly Asn Gly Pro Asn Glu 85 90 95 Ala Gly Leu Ser Lys His His Ile Ile Ala Gln Ala Glu Ala Ser Leu 100 105 110 Lys Arg Leu Arg Thr Asp Ala Ile Glu Leu Tyr Gln Val His Glu Trp 115 120 125 Asp Gly Met Thr Pro Ile Glu Glu Thr Met Glu Ala Leu Asp Thr Leu 130 135 140 Val Arg Gln Gly Lys Val Arg Tyr Val Gly Cys Ser Asn Tyr Ser Gly 145 150 155 160 Trp His Ile Met Lys Ala Leu Met Ala Ala Asp Lys His His Gly Gln 165 170 175 Arg Phe Val Ser Gln Gln Ile His Tyr Ser Leu His Ser Arg Asp Ala 180 185 190 Glu Tyr Glu Leu Val Pro Ile Ser Gln Asp Gln Gly Leu Gly Ile Leu 195 200 205 Val Trp Ser Pro Leu Ala Gly Gly Leu Leu Ser Gly Lys Tyr Arg Arg 210 215 220 Thr Gly Gly Pro Glu Ala Gly Arg His Val Gly Gly Phe Arg Glu Pro 225 230 235 240 Pro Val Tyr Asp Trp Asp Lys Leu Tyr Glu Leu Ile Asp Val Ile Val 245 250 255 Ala Ile Ala Ala Gln Arg Gly Val Ser Gly Ala Gln Val Ala Leu Ala 260 265 270 Trp Leu Leu Gly Arg Pro Gly Val Thr Ser Leu Ile Ile Gly Gly Arg 275 280 285 Ser Glu Ala Gln Phe Arg Asp Asn Leu Ala Ala Ala His Leu Arg Leu 290 295 300 Thr Ser Glu Glu Arg Ala Arg Leu Asp Ala Ala Ser Arg Pro Pro Leu 305 310 315 320 Leu Tyr Pro Tyr Trp His Gln Ser Phe Thr Ala Gln Asp Arg Leu Ser 325 330 335 Arg Ile Asp Leu Asp Leu Ile Gly Pro Tyr Ala Glu Glu Phe Lys Arg 340 345 350 Gly <210> 90 <211> 353 <212> PRT <213> Artificial Sequence <220> <223> Enzyme <400> 90 Met Asp Tyr Arg Tyr Leu Gly Arg Ser Gly Leu Lys Val Ser Ala Met 1 5 10 15 Thr Met Gly Thr Met Thr Phe Gly Gly Ser Glu Arg Val Gly His Thr 20 25 30 Asp Leu Ala Gly Ala Ala Arg Gln Ile Asp Leu Cys Ile Asp Ala Gly 35 40 45 Ile Asn Leu Leu Asp Thr Ala Asn Val Tyr Asn Ala Gly Val Ser Glu 50 55 60 Asp Ile Ile Gly Val Ala Leu Ala Glu His Gly Arg Arg Gln Arg Ala 65 70 75 80 Leu Val Ala Thr Lys Val Arg Phe Lys Met Gly Asn Gly Pro Asn Glu 85 90 95 Val Gly Leu Ser Arg His His Ile Ile Ala Gln Ala Glu Ala Ser Leu 100 105 110 Lys Arg Leu Arg Thr Asp Ala Ile Asp Leu Tyr Gln Val His Glu Trp 115 120 125 Asp Gly Met Thr Pro Ile Glu Glu Thr Met Glu Ala Leu Asp Thr Leu 130 135 140 Val Lys Gln Gly Lys Val Arg Tyr Val Gly Cys Ser Asn Tyr Ser Gly 145 150 155 160 Trp His Ile Met Lys Ala Leu Met Ala Ala Asp Lys Arg Gln Gly Gln 165 170 175 Arg Phe Ile Ser Gln Gln Ile His Tyr Ser Leu His Ser Arg Glu Ala 180 185 190 Glu Tyr Glu Leu Val Pro Ile Ser Gln Asp Gln Gly Leu Gly Ile Leu 195 200 205 Ile Trp Ser Pro Leu Ala Gly Gly Leu Leu Ser Gly Lys Tyr Arg Arg 210 215 220 Asp Gly Gly Pro Glu Ser Gly Arg His Val Gly Gly Phe Arg Glu Pro 225 230 235 240 Pro Val Tyr Asp Trp Asp Lys Leu Tyr Glu Ile Ile Asp Val Ile Val 245 250 255 Ala Ile Ala Arg Gln Lys Gly Val Ser Gly Ala Gln Val Ala Leu Ser 260 265 270 Trp Leu Leu Glu Arg Pro Gly Val Thr Ser Leu Ile Ile Gly Gly Arg 275 280 285 Ser Glu Ala Gln Phe Ala Asp Asn Leu Ala Ala Ile His Leu Arg Leu 290 295 300 Thr Ser Asp Glu Arg Ala Arg Leu Asp Ala Ala Ser Arg Pro Pro Leu 305 310 315 320 Leu Tyr Pro Tyr Trp His Gln Ser Phe Thr Ala Gln Asp Arg Leu Ser 325 330 335 Gln Ile Asp Leu Asp Leu Ile Gly Pro Tyr Ala Glu Glu Phe Lys Arg 340 345 350 Gly <210> 91 <211> 353 <212> PRT <213> Artificial Sequence <220> <223> Enzyme <400> 91 Met Asp Tyr Arg Tyr Leu Gly Arg Ser Gly Leu Lys Val Ser Ala Met 1 5 10 15 Thr Met Gly Thr Met Thr Phe Gly Gly Ser Glu Arg Val Gly His Thr 20 25 30 Asp Leu Ala Gly Ala Ala Arg Gln Ile Asp Leu Cys Leu Asp His Gly 35 40 45 Ile Asn Leu Tyr Asp Thr Ala Asn Val Tyr Asn Ala Gly Val Ser Glu 50 55 60 Asp Ile Ile Gly Gln Ala Leu Ala Glu His Gly Arg Arg Gln Arg Ala 65 70 75 80 Leu Ile Ala Thr Lys Val Arg Phe Lys Met Gly Asn Gly Pro Asn Glu 85 90 95 Ala Gly Leu Ser Arg His His Ile Ile Ala Gln Ala Glu Ala Ser Leu 100 105 110 Lys Arg Leu Arg Thr Asp Ala Ile Glu Leu Tyr Gln Val His Glu Trp 115 120 125 Asp Gly Met Thr Pro Ile Glu Glu Thr Met Glu Ala Leu Asp Thr Leu 130 135 140 Val Lys Gln Ala Lys Val Arg Tyr Val Gly Cys Ser Asn Tyr Ser Gly 145 150 155 160 Trp His Ile Met Lys Ala Leu Ala Ala Ala Asp Lys Arg Gln Gly Gln 165 170 175 Arg Phe Ile Ser Gln Gln Ile His Tyr Ser Leu His Ser Arg Asp Ala 180 185 190 Glu Tyr Glu Leu Val Pro Ile Ser Gln Asp Gln Gly Leu Gly Ile Leu 195 200 205 Ile Trp Ser Pro Leu Ala Gly Gly Leu Leu Ser Gly Lys Tyr Arg Arg 210 215 220 Asp Gly Gly Pro Glu Ser Gly Arg His Val Gly Gly Phe Arg Glu Pro 225 230 235 240 Pro Val Tyr Asp Trp Asp Arg Leu Tyr Asp Leu Ile Asp Val Ile Val 245 250 255 Asp Ile Ala Ala Gln Lys Gly Val Ser Gly Ala Gln Val Ala Leu Ser 260 265 270 Tyr Ala Leu Gly Arg Pro Gly Val Thr Ser Leu Ile Ile Gly Gly Arg 275 280 285 Asn Glu Ala Gln Phe Arg Asp Asn Leu Ala Ala Ile His Leu Lys Leu 290 295 300 Thr Asp Glu Glu Arg Ala Arg Leu Asp Ala Ala Ser Arg Pro Pro Leu​​​​​​​​​​​​​​​​​​​​ <220> <223> Enzyme <400> 92 Met Asp Tyr Arg Phe Leu Gly Arg Ser Gly Leu Lys Val Ser Ala Met 1 5 10 15 Thr Met Gly Thr Met Thr Phe Gly Gly Ser Glu Arg Val Gly His Thr 20 25 30 Ala Gln Ala Gly Ala Ala Arg Gln Val Asp Leu Cys Leu Asp Ala Gly 35 40 45 Ile Asn Leu Leu Asp Thr Ala Asn Val Tyr Asn Ala Gly Val Ser Glu 50 55 60 Asp Ile Leu Gly Val Val Leu Ala Glu His Gly Arg Arg Gln Arg Ala 65 70 75 80 Leu Val Ala Thr Lys Val Arg Phe Lys Met Gly Asn Gly Pro Asn Glu 85 90 95 Ala Gly Leu Ser Arg His His Ile Ile Ala Gln Ala Glu Ala Ser Leu 100 105 110 Lys Arg Leu Arg Thr Asp Ala Ile Asp Leu Tyr Gln Val His Glu Trp 115 120 125 Asp Gly Met Thr Pro Ile Asp Glu Thr Met Glu Ala Leu Asp Thr Leu 130 135 140 Val Arg Gln Gly Lys Val Arg Tyr Val Gly Cys Ser Asn Tyr Ser Gly 145 150 155 160 Trp His Ile Met Lys Ala Leu Met Ala Ala Asp Lys Arg Gln Gly Gln 165 170 175 Arg Phe Val Ser Gln Gln Ile His Tyr Ser Leu His Ser Arg Asp Ala 180 185 190 Glu Tyr Glu Leu Val Pro Ile Ser Gln Asp Gln Gly Leu Gly Ile Leu 195 200 205 Val Trp Ser Pro Leu Ala Gly Gly Leu Leu Ser Gly Lys Tyr Lys Arg 210 215 220 Asp Gly Gly Pro Glu Ser Gly Arg His Val Gly Gly Phe Arg Glu Pro 225 230 235 240 Pro Leu Tyr Asp Trp Asp Lys Leu Tyr Asp Leu Ile Asp Val Ile Val 245 250 255 Ala Ile Ala Arg Gln Arg Gly Val Ser Gly Ala Gln Val Ala Leu Ser 260 265 270 Phe Leu Leu Glu Arg Pro Gly Val Thr Ser Leu Ile Ile Gly Gly Arg 275 280 285 Ser Glu Ala Asn Phe Ala Asp Asn Leu Ala Ala Ala His Leu Lys Leu 290 295 300 Ser Ser Glu Glu Arg Ala Arg Leu Asp Ala Ala Ser Arg Pro Pro Leu 305 310 315 320 Leu Tyr Pro Tyr Trp His Gln Ser Phe Thr Ala Gln Asp Arg Leu Ser 325 330 335 Gln Ile Asp Leu Asp Leu Ile Glu Pro Tyr Ala Glu Glu Phe Lys Arg 340 345 350 Gly <210> 93 <211> 353 <212> PRT <213> Synthetic Sequence <220> <223> Enzyme <400> 93 Met Asp Tyr Lys Tyr Ile Gly Arg Ser Gly Leu Lys Val Ser Ala Met 1 5 10 15 Thr Met Gly Thr Met Thr Phe Gly Gly Ser Glu Arg Val Gly His Thr 20 25 30 Asp Leu Ala Gly Ala Ala Arg Gln Val Asp Ile Cys Leu Asp Ala Gly 35 40 45 Ile Asn Leu Leu Asp Thr Ala Asn Val Tyr Asn Ala Gly Val Ser Glu 50 55 60 Asp Ile Leu Gly Val Val Leu Ala Glu His Gly Arg Arg Gln Arg Ala 65 70 75 80 Leu Ile Ala Thr Lys Val Arg Phe Arg Met Gly Asn Gly Pro Asn Glu 85 90 95 Gly Gly Leu Ser Arg His His Ile Ile Ala Gln Ala Glu Ala Ser Leu 100 105 110 Lys Arg Leu Arg Thr Asp Ala Ile Glu Leu Tyr Gln Val His Glu Trp 115 120 125 Asp Gly Met Thr Pro Ile Asp Glu Thr Met Glu Ala Leu Asp Thr Leu 130 135 140 Val Lys Gln Ala Lys Val Arg Tyr Val Ala Cys Ser Asn Tyr Ser Gly 145 150 155 160 Trp His Ile Met Lys Ala Leu Ala Gly Ala Asp Lys Arg Gln Gly Gln 165 170 175 Arg Phe Val Ser Gln Gln Ile His Tyr Ser Leu His Ser Arg Asp Gly 180 185 190 Glu Tyr Glu Leu Val Pro Ile Ser Gln Asp Gln Gly Leu Gly Ile Leu 195 200 205 Ile Trp Ser Pro Leu Ala Gly Gly Leu Leu Ser Gly Lys Tyr Arg Arg 210 215 220 Asp Gly Gly Pro Glu Ser Gly Arg His Val Gly Gly Phe Arg Glu Pro 225 230 235 240 Pro Val Tyr Asp Trp Asp Lys Leu Tyr Glu Ile Ile Asp Val Ile Val 245 250 255 Asp Ile Ala Arg Gln Arg Gly Val Ser Gly Ala Gln Val Ala Leu Ala 260 265 270 Trp Ala Leu Glu Arg Pro Gly Val Thr Ser Leu Ile Ile Gly Gly Arg 275 280 285 Ser Glu Ala Gln Phe Ala Asp Asn Ile Ala Ala Ile His Leu Lys Leu 290 295 300 Ser Asp Glu Glu Arg Ala Arg Leu Asp Ala Ala Ser Arg Pro Pro Leu 305 310 315 320 Leu Tyr Pro Tyr Trp His Gln Ser Phe Thr Gly Gln Asp Arg Leu Ser 325 330 335 Gln Ile Asp Leu Asp Leu Ile Glu Pro Tyr Ala Glu Glu Phe Lys Arg 340 345 350 Gly <210> 94 <211> 353 <212> PRT <213> Artificial Sequence <220> <223> Enzyme <400> 94 Met Asp Tyr Lys Tyr Leu Gly Arg Ser Gly Leu Lys Val Ser Ala Met 1 5 10 15 Thr Met Gly Thr Met Thr Phe Gly Gly Ser Glu Arg Val Gly His Thr 20 25 30 Gly Gln Ala Gly Ala Thr Arg Gln Val Asp Ile Cys Leu Asp Ala Gly 35 40 45 Ile Asn Leu Leu Asp Thr Ala Asn Val Tyr Asn Ala Gly Val Ser Glu 50 55 60 Glu Met Leu Gly Val Val Leu Ala Glu His Gly Arg Arg Gln Arg Ala 65 70 75 80 Leu Ile Ala Thr Lys Val Arg Phe Lys Met Gly Asn Gly Pro Asn Glu 85 90 95 Val Gly Leu Ser Arg His His Ile Ile Ala Gln Ala Glu Ala Ser Leu 100 105 110 Lys Arg Leu Arg Thr Asp Val Ile Glu Leu Tyr Gln Val His Glu Trp 115 120 125 Asp Gly Met Thr Pro Ile Glu Asp Thr Met Glu Ala Leu Asp Thr Leu 130 135 140 Val Arg Gln Ala Lys Val Arg Tyr Val Gly Cys Ser Asn Tyr Ser Gly 145 150 155 160 Trp His Ile Met Lys Gly Leu Ala Gly Ala Asp Lys Arg Gln Gly Gln 165 170 175 Arg Phe Ile Ser Gln Gln Ile His Tyr Thr Leu His Ser Arg Asp Ala 180 185 190 Glu Tyr Glu Leu Val Pro Ile Ser Gln Asp Gln Gly Leu Gly Ile Leu 195 200 205 Ile Trp Ser Pro Leu Ala Gly Gly Leu Leu Ser Gly Lys Tyr Arg Arg 210 215 220 Asp Gly Gly Pro Asp Ala Gly Arg His Val Gly Gly Phe Arg Glu Pro 225 230 235 240 Pro Val Tyr Asp Trp Asp Lys Leu Tyr Glu Leu Ile Asp Val Ile Val 245 250 255 Asp Ile Ala Arg Gln Arg Gly Val Ser Gly Ala Gln Val Ala Leu Ser 260 265 270 Trp Ala Leu Glu Arg Pro Gly Val Thr Ser Leu Ile Ile Gly Gly Arg 275 280 285 Ser Glu Ala Gln Phe Ala Asp Asn Ile Ala Gly Ala His Leu Lys Leu 290 295 300 Ser Ser Glu Glu Arg Ala Arg Leu Asp Ala Ala Ser Arg Pro Pro Leu 305 310 315 320 Leu Tyr Pro Tyr Trp His Gln Ser Phe Thr Gly Gln Asp Arg Leu Ser 325 330 335 Gln Ile Asp Leu Asp Leu Ile Gly Pro Tyr Ala Glu Glu Phe Lys Arg 340 345 350 Gly <210> 95 <211> 353 <212> PRT <213> Artificial Sequence <220> <223> Enzyme <400> 95[[ID=......]] Met Asp Tyr Lys Tyr Leu Gly Arg Ser Gly Leu Lys Val Ser Ala Met 1 5 10 15 Thr Met Gly Thr Met Thr Phe Gly Gly Ser Glu Arg Val Gly His Thr 20 25 30 Asp Leu Ala Gly Ala Ala Arg Gln Val Asp Leu Cys Leu Asp Ala Gly 35 40 45 Ile Asn Leu Tyr Asp Thr Ala Asn Val Tyr Asn Ala Gly Val Ser Glu 50 55 60 Asp Met Leu Gly Val Val Leu Ala Glu Asn Gly Arg Arg Gln Arg Ala 65 70 75 80 Leu Val Ala Thr Lys Val Arg Phe Lys Met Gly Asn Gly Pro Asn Glu [ 85 90 95 Ala Gly Leu Ser Arg His His Ile Ile Ala Gln Ala Glu Ala Ser Leu 100 105 110 Lys Arg Leu Arg Thr Asp Ala Ile Asp Leu Tyr Gln Val His Glu Trp 115 120 125 Asp Gly Met Thr Pro Ile Asp Glu Thr Met Glu Ala Leu Asp Thr Leu 130 135 140 Val Arg Gln Gly Lys Val Arg Tyr Val Gly Cys Ser Asn Tyr Ser Gly 145 150 155 160 Trp His Ile Met Lys Ala Leu Met Ala Ala Asp Lys Arg Gln Gly Gln 165 170 175 Arg Phe Val Ser Gln Gln Ile His Tyr Ser Leu His Ser Arg Asp Ala 180 185 190 Glu Tyr Glu Leu Val Pro Ile Ser Gln Asp Gln Gly Leu Gly Ile Leu 195 200 205 Val Trp Ser Pro Leu Ala Gly Gly Leu Leu Ser Gly Lys Tyr Arg Arg 210 215 220 Thr Gly Gly Pro Glu Ser Gly Arg His Val Gly Gly Phe Arg Glu Pro 225 230 235 240 Pro Val Tyr Asp Trp Asp Lys Leu Tyr Asp Leu Ile Asp Val Ile Val 245 250 255 Ala Ile Ala Arg Gln Arg Gly Val Ser Gly Ala Gln Val Ala Leu Ala 260 265 270 Trp Leu Leu Glu Arg Pro Gly Val Thr Ser Leu Ile Ile Gly Gly Arg 275 280 285 Ser Glu Ala Gln Phe Arg Asp Asn Ile Ala Ala Ile His Leu Lys Leu 290 295 300 Ser Ser Glu Glu Arg Ala Arg Leu Asp Ala Ala Ser Arg Pro Pro Leu 305 310 315 320 Leu Tyr Pro Tyr Trp His Asn Ser Phe Thr Gly Gln Asp Arg Leu Asp 325 330 335 Gln Ile Asp Leu Asp Leu Ile Glu Pro Tyr Ala Glu Glu Phe Lys Arg 340 345 350 Gly <210> 96 <211> 353 <212> PRT <213> Artificial Sequence <220> <223> Enzyme <400> 96 Met Asp Tyr Lys Tyr Leu Gly Arg Ser Gly Leu Lys Val Ser Ala Met 1 5 10 15 Thr Met Gly Thr Met Thr Phe Gly Gly Ser Glu Arg Val Gly His Thr 20 25 30 Ala Gln Ala Gly Ala Ala Arg Gln Val Asp Leu Cys Leu Asp His Gly 35 40 45 Ile Asn Leu Leu Asp Thr Ala Asn Val Tyr Asn Ala Gly Val Ser Glu 50 55 60 Glu Met Leu Gly Val Val Leu Ala Glu His Gly Arg Arg Gln Lys Ala 65 70 75 80 Leu Val Ala Thr Lys Val Arg Phe Lys Met Gly Asn Gly Pro Asn Glu 85 90 95 Gly Gly Leu Ser Arg His His Ile Ile Ala Gln Ala Glu Ala Ser Leu 100 105 110 Lys Arg Leu Lys Thr Asp Ala Ile Glu Leu Tyr Gln Val His Glu Trp 115 120 125 Asp Gly Met Thr Pro Ile Glu Asp Thr Met Glu Ala Leu Asp Thr Leu 130 135 140 Val Arg Gln Gly Lys Val Arg Tyr Val Ala Cys Ser Asn Tyr Ser Gly 145 150 155 160 Trp His Ile Met Lys Ala Leu Ala Ala Ala Asp Lys Arg Gln Gly Gln 165 170 175 Arg Phe Val Ser Gln Gln Ile His Tyr Ser Leu His Ser Arg Asp Ala 180 185 190 Glu Tyr Glu Leu Val Pro Ile Ser Gln Asp Gln Gly Leu Gly Ile Leu 195 200 205 Val Trp Ser Pro Leu Ala Gly Gly Leu Leu Ser Gly Lys Tyr Arg Arg 210 215 220 Asp Gly Gly Pro Glu Ser Gly Arg His Val Gly Gly Phe Arg Glu Pro 225 230 235 240 Pro Val Tyr Asp Trp Asp Lys Leu Tyr Glu Ile Leu Asp Val Ile Val 245 250 255 Ala Ile Ala Arg Gln Arg Gly Val Ser Gly Ala Gln Val Ala Leu Ala 260 265 270 Trp Leu Leu Glu Arg Pro Gly Val Thr Ser Leu Ile Ile Gly Gly Arg 275 280 285 Ser Glu Ala Gln Phe Ala Asp Asn Ile Ala Ala Ile His Leu Lys Leu 290 295 300 Ser Asp Glu Glu Arg Ala Arg Leu Asp Ala Ala Ser Arg Pro Pro Leu 305 310 315 320 Leu Tyr Pro Tyr Trp His Gln Ser Phe Thr Ala Asn Asp Arg Leu Asp 325 330 335 Gln Ile Asp Leu Asp Leu Ile Glu Pro Tyr Ala Glu Asp Phe Lys Arg 340 345 350 Gly <210> 97 <211> 353 <212> PRT <213> Artificial Sequence <220> <223> Enzyme <400> 97 Met Asp Tyr Arg Tyr Leu Gly Arg Ser Gly Leu Lys Val Ser Ala Met 1 5 10 15 Thr Met Gly Thr Met Thr Phe Gly Gly Ser Glu Arg Val Gly His Thr 20 25 30 Asp Leu Ala Gly Ala Thr Arg Gln Val Asp Leu Cys Leu Asp His Gly 35 40 45 Ile Asn Leu Leu Asp Thr Ala Asn Val Tyr Asn Ala Gly Val Ser Glu 50 55 60 Glu Met Ile Gly Val Val Leu Ser Glu His Gly Arg Arg Gln Arg Ala 65 70 75 80[[ID=K]] Leu Ile Ala Thr Lys Val Arg Phe Lys Met Gly Asn Gly Pro Asn Glu[[ID=S]] 85 90 95 Gly Gly Leu Ser Arg His His Ile Ile Ala Gln Ala Glu Ala Ser Leu 100 105 110 Lys Arg Leu Arg Thr Asp Ala Ile Asp Leu Tyr Gln Val His Glu Trp 115 120 125 Asp Gly Met Thr Pro Ile Asp Glu Thr Met Glu Ala Leu Asp Thr Leu 130 135 140 Val Arg Gln Gly Lys Val Arg Tyr Val Ala Cys Ser Asn Tyr Ser Gly 145 150 155 160 Trp His Ile Met Lys Ala Leu Met Gly Ala Asp Lys Arg Gln Gly Gln 165 170 175 Arg Phe Val Ser Gln Gln Ile His Tyr Thr Leu His Ser Arg Asp Gly 180 185 190 Glu Tyr Glu Leu Val Pro Ile Ser Gln Asp Gln Gly Leu Gly Ile Leu 195 200 205 Val Trp Ser Pro Leu Ala Gly Gly Leu Leu Ser Gly Lys Tyr Arg Arg 210 215 220 Asp Gly Gly Pro Asp Ala Gly Arg His Val Gly Gly Phe Arg Glu Pro 225 230 235 240 Pro Val Tyr Asp Trp Asp Lys Leu Tyr Asp Leu Leu Asp Val Ile Val 245 250 255 Asp Ile Ala Arg Gln Arg Gly Val Ser Gly Ala Gln Val Ala Leu Ser 260 265 270 Trp Leu Leu Gly Arg Pro Gly Val Thr Ser Leu Ile Ile Gly Gly Arg 275 280 285 Ser Glu Ala Gln Phe Arg Asp Asn Ile Ala Ala Ala His Leu Lys Leu 290 295 300 Ser Ser Asp Glu Arg Ala Arg Leu Asp Ala Ala Ser Arg Pro Pro Leu 305 310 315 320 Leu Tyr Pro Tyr Trp His Gln Ser Phe Thr Ala Gln Asp Arg Leu Asp 325 330 335 Gln Ile Asp Leu Asp Leu Leu Glu Pro Tyr Ala Glu Glu Phe Lys Arg 340 345 350 Gly [[ID=I5]]<210> 98 <211> 353 <212> PRT <213> Artificial Sequence <220> <223> Enzyme <400> 98 Met Asp Tyr Arg Tyr Ile Gly Arg Ser Gly Leu Lys Val Ser Ala Met 1 5 10 15 Thr Met Gly Thr Met Thr Phe Gly Gly Ser Glu Lys Val Gly His Thr 20 25 30 Gly Gln Ala Gly Ala Ala Arg Gln Val Asp Leu Cys Leu Asp His Gly 35 40 45 Ile Asn Leu Leu Asp Thr Ala Asn Val Tyr Asn Ala Gly Val Ser Glu 50 55 60 Asp Ile Leu Gly Val Val Leu Ala Glu His Gly Arg Arg Gln Lys Ala 65 �0 75 80 Leu Ile Ala Thr Lys Val Arg Phe Arg Met Gly Asn Gly Pro Asn Glu 85 90 95 Val Gly Leu Ser Arg His His Ile Ile Ala Gln Ala Glu Ala Ser Leu 100 105 110 Lys Arg Leu Arg Thr Asp Ala Ile Asp Leu Tyr Gln Val His Glu Trp 115 120 125 Asp Gly Met Thr Pro Ile Glu Asp Thr Met Glu Ala Leu Asp Thr Leu 130 135 140 Val Arg Gln Gly Lys Val Arg Tyr Val Ala Cys Ser Asn Tyr Ser Gly 145 150 155 160 Trp His Ile Met Lys Ala Leu Ala Gly Ala Asp Lys His His Gly Gln 165 170 175 Arg Phe Ile Ser Gln Gln Ile His Tyr Thr Leu His Ser Arg Glu Ala 180 185 190 Glu Tyr Glu Leu Val Pro Ile Ser Gln Asp Gln Gly Leu Gly Ile Leu 195 200 205 Ile Trp Ser Pro Leu Ala Gly Gly Leu Leu Ser Gly Lys Tyr Arg Arg 210 215 220 Asp Gly Gly Pro Glu Ser Gly Arg His Val Gly Gly Phe Arg Glu Pro 225 230 235 240 Pro Val Tyr Asp Trp Asp Lys Leu Tyr Asp Leu Ile Asp Val Ile Val 245 250 255 Asp Ile Ala Arg Gln Arg Gly Val Ser Gly Ala Gln Val Ala Leu Ser 260 265 270 Trp Leu Leu Glu Arg Pro Gly Val Thr Ser Leu Ile Ile Gly Gly Arg 275 280 285 Ser Glu Ala Gln Phe Arg Asp Asn Ile Ala Ala Ile His Leu Lys Leu 290 295 300 Thr Ser Glu Glu Arg Ala Arg Leu Asp Ala Ala Ser Arg Pro Pro Leu 305 310 315 320 Leu Tyr Pro Tyr Trp His Asn Ser Phe Thr Ala Gln Asp Arg Leu Ser 325 330 335 Gln Ile Asp Leu Asp Leu Ile Gly Pro Tyr Ala Glu Asp Phe Lys Arg 340 345 350 Gly <210> 99 <211> 353 <212> PRT <213> Artificial Sequence <220> <​​​​​​​Thr Met Gly Thr Met Thr Phe Gly Gly Ser Glu Arg Val Gly His Thr 20 25 30 Gly Gln Ala Gly Ala Ala Arg Gln Leu Asp Leu Cys Leu Asp Ala Gly 35 40 45 Ile Asn Leu Leu Asp Thr Ala Asn Val Tyr Asn Ala Gly Val Ser Glu 50 55 60 Asp Ile Leu Gly Val Val Leu Ser Glu His Gly Arg Arg Gln Arg Ala 65 70 75 80 Leu Val Ala Thr Lys Val Arg Phe Lys Met Gly Asn Gly Pro Asn Glu 85 90 95 Gly Gly Leu Ser Arg His His Ile Ile Ala Gln Ala Glu Ala Ser Leu 100 105 110 Lys Arg Leu Arg Thr Asp Val Ile Glu Leu Tyr Gln Val His Glu Trp 115 120 125 Asp Gly Met Thr Pro Ile Glu Asp Thr Met Glu Ala Leu Asp Thr Leu 130 135 140 Val Lys Gln Ala Lys Val Arg Tyr Val Ala Cys Ser Asn Tyr Ser Gly 145 150 155 160 Trp His Ile Met Lys Ala Leu Met Gly Ala Asp Lys His His Gly Gln 165 170 175 Arg Phe Val Ser Gln Gln Ile His Tyr Ser Leu His Ser Arg Asp Ala 180 185 190 Glu Tyr Glu Leu Val Pro Ile Ser Gln Asp Gln Gly Leu Gly Ile Leu 195 200 205 Ile Trp Ser Pro Leu Ala Gly Gly Leu Leu Ser Gly Lys Tyr Arg Arg 210 215 220 Asp Gly Gly Pro Asp Ala Gly Arg His Val Gly Gly Phe Arg Glu Pro 225 230 235 240 Pro Val Tyr Asp Trp Asp Lys Leu Tyr Glu Leu Ile Asp Val Ile Val 245 250 255 Ala Ile Ala Arg Gln Arg Gly Val Ser Gly Ala Gln Val Ala Leu Ala 260 265 270 Trp Leu Leu Glu Arg Pro Gly Val Thr Ser Leu Ile Ile Gly Gly Arg 275 280 285 Ser Glu Ala Gln Phe Arg Asp Asn Leu Ala Ala Ala His Leu Lys Leu 290 295 300 Thr Asp Glu Glu Arg Ala Arg Leu Asp Ala Ala Ser Arg Pro Pro Leu 305 310 315 320 Leu Tyr Pro Tyr Trp His Gln Ser Phe Thr Ala Gln Asp Arg Leu Ser 325 330 335 Gln Ile Asp Leu Asp Leu Ile Glu Pro Tyr Ala Glu Asp Phe Lys Arg 340 345 350 Gly <210> 100 <211> 353 <212> PRT <213> Artificial sequence <220> <223> Enzyme <400> 100 Met Asp Tyr Arg Tyr Ile Gly Arg Ser Gly Leu Lys Val Ser Ala Met 1 5 10 15 Thr Met Gly Thr Met Thr Phe Gly Gly Ser Glu Arg Val Gly His Thr 20 25 30 Asp Leu Ala Gly Ala Thr Arg Gln Val Asp Leu Cys Leu Asp Ala Gly 35 40 45 Ile Asn Leu Leu Asp Thr Ala Asn Val Tyr Asn Ala Gly Val Ser Glu 50 55 60 Glu Met Ile Gly Val Val Leu Ala Glu His Gly Arg Arg Gln Arg Ala 65 70 75 80 Leu Val Ala Thr Lys Val Arg Phe Arg Met Gly Asn Gly Pro Asn Glu 85 90 95 Val Gly Leu Ser Arg His His Ile Ile Ala Gln Ala Glu Ala Ser Leu 100 105 110 Lys Arg Leu Arg Thr Asp Ala Ile Glu Leu Tyr Gln Val His Glu Trp 115 120 125 Asp Gly Met Thr Pro Ile Glu Glu Thr Met Glu Ala Leu Asp Thr Leu 130 135 140 Val Arg Gln Ala Lys Val Arg Tyr Val Gly Cys Ser Asn Tyr Ser Gly 145 150 155 160 Trp His Ile Met Lys Ala Leu Met Gly Ala Asp Lys Arg Gln Gly Gln 165 170 175 Arg Phe Val Ser Gln Gln Ile His Tyr Ser Leu His Ser Arg Asp Gly 180 185 190 Glu Tyr Glu Leu Val Pro Ile Ser Gln Asp Gln Gly Leu Gly Ile Leu 195 200 205 Val Trp Ser Pro Leu Ala Gly Gly Leu Leu Ser Gly Lys Tyr Arg Arg 210 215 220 Asp Gly Gly Pro Asp Ala Gly Arg His Val Gly Gly Phe Arg Glu Pro 225 230 235 240 Pro Val Tyr Asp Trp Asp Lys Leu Tyr Asp Leu Ile Asp Val Ile Val 245 250 255 Asp Ile Ala Arg Gln Arg Gly Val Ser Gly Ala Gln Val Ala Leu Ala 260 265 270 Trp Ala Leu Glu Arg Pro Gly Val Thr Ser Leu Ile Ile Gly Gly Arg 275 280 285 Ser Glu Ala Gln Phe Ala Asp Asn Leu Ala Ala Ile His Leu Lys Leu 290 295 300 Thr Ser Glu Glu Arg Ala Arg Leu Asp Ala Ala Ser Arg Pro Pro Leu 305 310 315 320 Leu Tyr Pro Tyr Trp His Gln Ser Phe Thr Gly Asn Asp Arg Leu Asp 325 330 335 Gln Ile Asp Leu Asp Leu Ile Gly Pro Tyr Ala Asp Glu Phe Lys Arg 340 345 3'50 Gly <210> 101 <211> 353 <212> PRT <213> Artificial Sequence <220> <223> Enzyme <400> 101 Met Asp Tyr Arg Tyr Ile Gly Arg Ser Gly Leu Lys Val Ser Ala Met 1 5 10 15 Thr Met Gly Thr Met Thr Phe Gly Gly Ser Glu Arg Val Gly His Thr 20 25 30 Ala Gln Ala Gly Ala Ala Arg Gln Val Asp Leu Cys Leu Asp Ala Gly 35 40 45 It should be noted that there may be a small error in the original text where "3'50" in line 19 should probably be "3'50" or something else more meaningful. This might need to be double - checked in the original context.Ile Asn Leu Tyr Asp Thr Ala Asn Val Tyr Asn Ala Gly Val Ser Glu 50 55 60 Asp Ile Leu Gly Val Val Leu Ser Glu Asn Gly Arg Arg Gln Lys Ala 65 70 75 80 Leu Val Ala Thr Lys Val Arg Phe Lys Met Gly Asn Gly Pro Asn Glu 85 90 95 Ala Gly Leu Ser Arg His His Ile Ile Ala Gln Ala Glu Ala Ser Leu 100 105 110 Lys Arg Leu Lys Thr Asp Ala Ile Asp Leu Tyr Gln Val His Glu Trp 115 120 125 Asp Gly Met Thr Pro Ile Glu Glu Thr Met Glu Ala Leu Asp Thr Leu 130 135 140 Val Arg Gln Gly Lys Val Arg Tyr Val Gly Cys Ser Asn Tyr Ser Gly 145 150 155 160 Trp His Ile Met Lys Ala Leu Met Ala Ala Asp Lys Arg Gln Gly Gln 165 170 175 Arg Phe Val Ser Gln Gln Ile His Tyr Ser Leu His Ser Arg Asp Ala 180 185 190 Glu Tyr Glu Leu Val Pro Ile Ser Gln Asp Gln Gly Leu Gly Ile Leu 195 200 205 Val Trp Ser Pro Leu Ala Gly Gly Leu Leu Ser Gly Lys Tyr Arg Arg 210 215 220 Thr Gly Gly Pro Glu Ser Gly Arg His Val Gly Gly Phe Arg Glu Pro 225 230 235 240 Pro Val Tyr Asp Trp Asp Lys Leu Tyr Glu Leu Ile Asp Val Ile Val 245 250 255 Asp Ile Ala Ala Gln Arg Gly Val Ser Gly Ala Gln Val Ala Leu Ala 260 265 270 Trp Leu Leu Gly Arg Pro Gly Val Thr Ser Leu Ile Ile Gly Gly Arg 275 280 285 Asn Glu Ala Gln Phe Arg Asp Asn Ile Ala Ala Ile His Leu Lys Leu 290 295 300 Ser Ser Glu Glu Arg Ala Arg Leu Asp Ala Ala Ser Arg Pro Pro Leu 305 310 315 320 Leu Tyr Pro Tyr Trp His Gln Ser Phe Thr Gly Gln Asp Arg Leu Asp 325 330 335 Gln Ile Asp Leu Asp Leu Ile Gly Pro Tyr Ala Asp Glu Phe Lys Arg 340 345 350 Gly

Claims

1. A cultivation composition comprising a plant or a part of a plant, comprising: a) one or more polypeptides having a sequence as shown in SEQ ID NO: 1 or SEQ ID NO: 3, wherein the polypeptide is capable of converting the substrate 3-keto-DON into products 3-epi-DON and DON, and the ratio of 3-epi-DON:DON is at least 25:1; and b) Vector.

2. The composition according to claim 1, wherein The polypeptide is a reductase.

3. A composition according to claim 1 or 2, wherein The carrier is liquid or solid.

4. The composition according to claim 3, wherein The carrier is H2O, or a solid selected from solids of health products and / or medicines.

5. Use of one or more polypeptides having the sequence shown in SEQ ID NO: 1 or SEQ ID NO: 3 for converting 3-keto-DON into 3-epi-DON.

6. Use of one or more polypeptides having a sequence as shown in SEQ ID NO: 1 or SEQ ID NO: 3 in the preparation of a feed additive or a feed composition, wherein: The polypeptide is capable of converting the substrate 3-keto-DON into products 3-epi-DON and DON, and the ratio of 3-epi-DON:DON is at least 25:

1.

7. A feed additive or food additive or feed or food, comprising one or more polypeptides having a sequence as shown in SEQ ID NO: 1 or SEQ ID NO: 3, wherein: The polypeptide is capable of converting the substrate 3-keto-DON into products 3-epi-DON and DON, and the ratio of 3-epi-DON:DON is at least 25:

1.

8. An additive for a composition comprising 3-keto-DON or DON, wherein the additive comprises one or more polypeptides having a sequence as shown in SEQ ID NO: 1 or SEQ ID NO: 3, wherein: The polypeptide is capable of converting the substrate 3-keto-DON into products 3-epi-DON and DON, and the ratio of 3-epi-DON:DON is at least 25:

1.

9. The additive for a composition according to claim 8, wherein The composition is a tillage composition.

10. An in vitro method for converting a trichothecene comprising a 3-oxyl group into a trichothecene comprising a 3-hydroxyl group, the method comprising contacting one or more polypeptides having a sequence of SEQ ID NO: 1 or SEQ ID NO: 3 with one or more trichothecenes comprising a 3-oxyl group.

11. An in vitro method for reducing the content of DON in a composition comprising DON or reducing the toxicity of a composition comprising DON by converting DON into 3-epi-DON, the method comprising: a) contacting the composition with an enzyme capable of converting DON to 3-keto-DON; and b) subsequently or simultaneously contacting the composition with one or more polypeptides having the sequence shown in SEQ ID NO: 1 or SEQ ID NO: 3.

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