阅读说明：本技术 来源于淋巴细胞具有降血糖的33种生物活性肽及其应用 (33 bioactive peptides derived from lymphocytes and having blood sugar reducing effect, and application thereof ) 是由 张少辉 占文静 张伯宇 于 2021-01-19 设计创作，主要内容包括：本发明涉及蛋白领域,具体涉及来源于淋巴细胞具有降血糖功效的生物活性肽及其应用,共包含33种生物活性肽。本发明依据生物信息学数据库BIOPEP对上述33种生物活性肽的生物活性进行检索,发现这33种生物活性肽均具有降血糖(DPP-IV抑制活性)功效,能够延长胰高血糖素肽-1的作用时间,促进胰岛素的分泌,从而具有降低血糖功效,对开发具有降血糖的食品、保健品和药物具有十分重要的意义。(The invention relates to the field of protein, in particular to bioactive peptide which is derived from lymphocyte and has the efficacy of reducing blood sugar and application thereof, and the bioactive peptide comprises 33 bioactive peptides. According to the invention, the biological activities of the 33 biological active peptides are searched according to a BIOPEP (bioinformatics database), and the 33 biological active peptides are found to have the effect of reducing blood sugar (DPP-IV inhibitory activity), can prolong the action time of glucagon-like peptide-1 and promote the secretion of insulin, so that the biological active peptides have the effect of reducing blood sugar and have very important significance for developing foods, health-care products and medicines with the effect of reducing blood sugar.)

1. A bioactive peptide derived from lymphocytes and having hypoglycemic activity, characterized by being selected from one of the following amino acid sequences:

Ala-Lys-Arg-Thr-Lys-Lys-Val-Gly-Ile-Val-Gly-Lys-Tyr，

Ala-Leu-Lys-Ala-Lys-Lys-Ala-Val-Leu-Lys-Gly-Val-His-Ser-His，

Glu-Val-Glu-Gln-Lys-Lys-Lys-Arg-Thr-Phe-Arg，

His-Leu-Lys-Ile-Val-Tyr-Arg-Arg，

Lys-Gly-Asn-Leu-Arg-Arg-Thr-Gly-Gln-Glu-Val-Ala-Gln-Ala-Gln-Glu-Ser，

Lys-Gln-Leu-Asp-Asp-Leu-Lys-Val-Glu-Leu-Ser-Gln-Leu-Arg，

Lys-Val-Thr-Gln-Tyr-Lys-Lys-Gly-Lys-Asp-Ser-Leu-Tyr，

Asn-Glu-Leu-Arg-Val-Ala-Pro-Glu-Glu-His-Pro-Val-Leu-Leu-Thr-Glu-Ala-Pro-Leu-Asn-Pro-Lys，

Asn-Ile-Arg-Ala-Ser-Met-Gln-Gln-Gln-Gln-Gln-Leu-Ala-Ser-Ala-Arg-Asn-Arg，

Asn-Leu-Val-Ile-Val-Lys-Lys-Gly-Glu-Lys-Asp-Ile-Pro-Gly-Leu-Thr-Asp-Thr-Thr-Val-Pro-Arg，

Asn-Pro-Phe-Val-Thr-Ser-Asp-Arg，

Asn-Val-Leu-Lys-Val-Thr-Lys-Ala-Ala-Gly-Thr-Lys，

Pro-Ala-Val-Thr-Gln-His-Ala-Pro-Tyr-Phe-Lys-Gly-Thr-Ala-Val，

Arg-Asp-Thr-Lys-Ser-Ile-Thr-Phe-Ser-Lys-Leu，

Arg-Ile-Gly-Val-Pro-Ser-Ala-Thr-Glu-Ile-Ile-Lys-Ala-Ser-Ser-Lys-Asp-Ala-Ile-Arg-Leu，

Arg-Lys-Ala-Val-Ile-Val-Lys-Asn-Ile-Asp-Asp-Gly-Thr-Ser-Asp-Arg-Pro-Tyr，

Arg-Val-Thr-Ile-Ala-Gln-Gly-Gly-Val-Leu-Pro-Asn-Ile-Gln-Ala-Val，

Ser-Leu-Val-Ser-Lys-Gly-Ile-Leu-Val-Gln-Thr-Lys-Gly-Thr-Gly-Ala-Ser-Gly-Ser-Phe，

Ser-Thr-Ile-Thr-Ser-Arg-Glu-Ile-Gln-Thr-Ala-Val，

Thr-Lys-Val-Lys-Ala-Ala-Pro-Glu-Glu-Ser-Glu-Ala-Gln-Ala-Glu-Gly-Cys-Ser-Glu-Glu-Arg-Thr-Tyr，

Thr-Leu-Val-Ile-Thr-Asp-Lys-Glu-Lys-Ala-Glu-Lys-Leu-Lys，

Thr-Tyr-Val-Pro-Val-Thr-Thr-Phe-Lys，

Val-Glu-Lys-Gly-Ile-Ile-Asp-Pro-Thr-Lys-Val-Val-Arg-Thr，

Val-Lys-Leu-Thr-Ala-Glu-Leu-Ile-Glu-Gln-Ala-Ala-Gln-Tyr，

Val-Arg-Tyr-Ser-Leu-Asp-Pro-Glu-Asn-Pro-Thr-Lys-Ser-Cys-Lys，

Asn-Gln-Ile-Leu-Arg-Ile-Glu-Glu-Glu-Leu-Gly-Ser-Lys-Ala-Lys-Phe-Ala-Gly-Arg-Ser-Phe，

Ser-Leu-Ala-Asp-Leu-Gln-Asn-Asp-Glu-Val-Ala-Phe-Arg-Lys-Phe，

Ile-Gly-Pro-Glu-Tyr-Gln-Gln-Asp-Leu-Asp-Arg-Glu-Leu-Tyr，

Lys-Lys-Glu-Lys-Val-Leu-Ala-Thr-Val-Thr-Lys-Thr-Val-Gly-Gly-Asp-Lys，

Leu-Leu-Pro-Lys-Lys-Ser-Ser-Ala-Thr-Val-Gly-Pro-Lys-Ala-Pro-Ala-Val-Gly-Lys，

Pro-Lys-Arg-Lys-Val-Ser-Ala-Asp-Gly-Ala-Ala-Lys-Ala-Glu-Pro-Lys-Arg-Arg，

Arg-Ser-Ala-Ile-Asn-Glu-Val-Val-Thr-Arg-Glu-Tyr，

Ser-Glu-Gly-Val-Phe-Leu-Leu-Thr-Thr-Thr-Pro-Arg-Pro-Val。

2. the bioactive peptide derived from lymphocytes and having hypoglycemic effect according to claim 1, wherein said bioactive peptide is mouse spleen derived lymphocyte peptide.

3. A polynucleotide encoding the biologically active peptide of any one of claims 1.

4. The use of bioactive peptides derived from lymphocytes and having hypoglycemic effect as claimed in claim 1, wherein any one or combination of 33 bioactive peptides is used in the preparation of food, health product, medicine or cosmetic with hypoglycemic effect.

5. The use of bioactive peptides derived from lymphocytes and having hypoglycemic effect according to claim 4, wherein any one or combination of several of said 33 bioactive peptides is used for preparing the medicine with hypoglycemic effect.

6. The use of bioactive peptides derived from lymphocytes and having hypoglycemic effect as claimed in claim 4, wherein any one or combination of several of said 33 bioactive peptides is used for preparing food, health product, pharmaceutical or cosmetic with DPP-IV inhibitory activity.

7. A hypoglycemic product comprising any one or a combination of 33 bioactive peptides as claimed in claim 1, said hypoglycemic product comprising a hypoglycemic food product or hypoglycemic drug.

8. A hypoglycemic product comprising any one or a combination of several of the derivatives of the 33 bioactive peptides of claim 1, wherein said hypoglycemic product comprises a hypoglycemic food, a hypoglycemic health product or a hypoglycemic drug; the derivatives of the 33 bioactive peptides are meant to have the same or better activity as the specific bioactive peptide.

9. The product of claim 8, wherein the derivatives of 33 bioactive peptides are derivatives of bioactive peptides obtained by modifying side chain groups, amino-terminal or carboxyl-terminal of amino acids of 33 specific bioactive peptides by hydroxylation, carboxylation, carbonylation, methylation, acetylation, phosphorylation, esterification or glycosylation.

10. The process for producing a bioactive peptide as claimed in claim 1, wherein the bioactive peptide is artificially synthesized by genetic engineering, or is directly obtained from cells by isolation and purification, or is directly produced by chemical synthesis.

Technical Field

The invention relates to the field of protein, in particular to 33 bioactive peptides with hypoglycemic activity derived from lymphocytes and application thereof.

Background

In recent years, bioactive peptides have become a word of great energy in the ear. Because of its many potential biological functions, it attracts more and more attention and becomes one of the hot spots of scientific research. The beneficial effects of many bioactive peptides, such as anti-cancer, blood pressure lowering, antibacterial, cholesterol lowering, anti-diabetic, etc., are well documented. Currently, over 3000 different bioactive peptides have been reported in the most authoritative bioactive peptide database, bipep.

In daily metabolism, ingestion of food causes the release of secretin (GLP-1 and GIP), which increases the concentration of insulin in blood, thereby lowering blood sugar and maintaining blood sugar balance in the body. DPP-IV degrades the incretin, so that the content of insulin in blood is reduced and the blood sugar is increased.

With the increasing number of people with diabetes, the development of safer, more effective and natural hypoglycemic products is a concern of people, especially for the prevention of type II diabetes, which is metabolic disorder caused by blood sugar elevation, DPP-IV inhibitor is a common medicine for treating type II diabetes, and can maintain the effect of promoting insulin secretion of incretin to achieve the purpose of reducing blood sugar. As early as 1988, Liddle et al found that protein digestion could promote secretion and expression of gut hormones in mice. Carcon et al bovine hemoglobin digest has a significant effect on the release of intestinal hormones and the inhibition of DPP-IV activity, and the genes of these hormones also show a phenomenon of high expression. In recent years, Fernando et al found that polypeptides isolated from sardines by enzymatic hydrolysis have higher DPP-IV inhibitory activity in the range of 400-1400Da, and that endoprotease and exopeptidase are synergistic to produce higher activity polypeptide products.

However, there are not many bioactive peptides with definite hypoglycemic activity, and therefore, bioactive peptides with hypoglycemic activity are one of the directions to be studied in the field of proteins.

Disclosure of Invention

The invention aims to provide biological activity with hypoglycemic activity in lymphocytes and application thereof. Further, biological activities derived from lymphocytes and having DPP-IV inhibitory activity and applications thereof are provided.

The purpose of the invention can be realized by the following technical scheme:

in a first aspect of the present invention, there is provided a bioactive peptide having hypoglycemic activity derived from lymphocytes, wherein the bioactive peptide is selected from one or more of the following bioactive peptides:

AKRTKKVGIVGKY、ALKAKKAVLKGVHSH、EVEQKKKRTFR、HLKIVYRR、KGNLRRTGQEVAQAQES、KQLDDLKVELSQLR、KVTQYKKGKDSLY、NELRVAPEEHPVLLTEAPLNPK、NIRASMQQQQQLASARNR、NLVIVKKGEKDIPGLTDTTVPR、NPFVTSDR、NVLKVTKAAGTK、PAVTQHAPYFKGTAV、RDTKSITFSKL、RIGVPSATEIIKASSKDAIRL、RKAVIVKNIDDGTSDRPY、RVTIAQGGVLPNIQAV、SLVSKGILVQTKGTGASGSF、STITSREIQTAV、TKVKAAPEESEAQAEGCSEERTY、TLVITDKEKAEKLK、TYVPVTTFK、VEKGIIDPTKVVRT、VKLTAELIEQAAQY、VRYSLDPENPTKSCK、NQILRIEEELGSKAKFAGRSF、SLADLQNDEVAFRKF、IGPEYQQDLDRELY、KKEKVLATVTKTVGGDK、LLPKKSSATVGPKAPAVGK、PKRKVSADGAAKAEPKRR、RSAINEVVTREY、SEGVFLLTTTPRPV，

the amino acid sequence thereof corresponds to the following:

Ala-Lys-Arg-Thr-Lys-Lys-Val-Gly-Ile-Val-Gly-Lys-Tyr，

Ala-Leu-Lys-Ala-Lys-Lys-Ala-Val-Leu-Lys-Gly-Val-His-Ser-His，

Glu-Val-Glu-Gln-Lys-Lys-Lys-Arg-Thr-Phe-Arg，

His-Leu-Lys-Ile-Val-Tyr-Arg-Arg，

Lys-Gly-Asn-Leu-Arg-Arg-Thr-Gly-Gln-Glu-Val-Ala-Gln-Ala-Gln-Glu-Ser，

Lys-Gln-Leu-Asp-Asp-Leu-Lys-Val-Glu-Leu-Ser-Gln-Leu-Arg，

Lys-Val-Thr-Gln-Tyr-Lys-Lys-Gly-Lys-Asp-Ser-Leu-Tyr，

Asn-Glu-Leu-Arg-Val-Ala-Pro-Glu-Glu-His-Pro-Val-Leu-Leu-Thr-Glu-Ala-Pro-Leu-Asn-Pro-Lys，

Asn-Ile-Arg-Ala-Ser-Met-Gln-Gln-Gln-Gln-Gln-Leu-Ala-Ser-Ala-Arg-Asn-Arg，

Asn-Leu-Val-Ile-Val-Lys-Lys-Gly-Glu-Lys-Asp-Ile-Pro-Gly-Leu-Thr-Asp-Thr-Thr-Val-Pro-Arg，

Asn-Pro-Phe-Val-Thr-Ser-Asp-Arg，

Asn-Val-Leu-Lys-Val-Thr-Lys-Ala-Ala-Gly-Thr-Lys，

Pro-Ala-Val-Thr-Gln-His-Ala-Pro-Tyr-Phe-Lys-Gly-Thr-Ala-Val，

Arg-Asp-Thr-Lys-Ser-Ile-Thr-Phe-Ser-Lys-Leu，

Arg-Ile-Gly-Val-Pro-Ser-Ala-Thr-Glu-Ile-Ile-Lys-Ala-Ser-Ser-Lys-Asp-Ala-Ile-Arg-Leu，

Arg-Lys-Ala-Val-Ile-Val-Lys-Asn-Ile-Asp-Asp-Gly-Thr-Ser-Asp-Arg-Pro-Tyr，

Arg-Val-Thr-Ile-Ala-Gln-Gly-Gly-Val-Leu-Pro-Asn-Ile-Gln-Ala-Val，

Ser-Leu-Val-Ser-Lys-Gly-Ile-Leu-Val-Gln-Thr-Lys-Gly-Thr-Gly-Ala-Ser-Gly-Ser-Phe，

Ser-Thr-Ile-Thr-Ser-Arg-Glu-Ile-Gln-Thr-Ala-Val，

Thr-Lys-Val-Lys-Ala-Ala-Pro-Glu-Glu-Ser-Glu-Ala-Gln-Ala-Glu-Gly-Cys-Ser-Glu-Glu-Arg-Thr-Tyr，

Thr-Leu-Val-Ile-Thr-Asp-Lys-Glu-Lys-Ala-Glu-Lys-Leu-Lys，

Thr-Tyr-Val-Pro-Val-Thr-Thr-Phe-Lys，

Val-Glu-Lys-Gly-Ile-Ile-Asp-Pro-Thr-Lys-Val-Val-Arg-Thr，

Val-Lys-Leu-Thr-Ala-Glu-Leu-Ile-Glu-Gln-Ala-Ala-Gln-Tyr，

Val-Arg-Tyr-Ser-Leu-Asp-Pro-Glu-Asn-Pro-Thr-Lys-Ser-Cys-Lys，

Asn-Gln-Ile-Leu-Arg-Ile-Glu-Glu-Glu-Leu-Gly-Ser-Lys-Ala-Lys-Phe-Ala-Gly-Arg-Ser-Phe，

Ser-Leu-Ala-Asp-Leu-Gln-Asn-Asp-Glu-Val-Ala-Phe-Arg-Lys-Phe，

Ile-Gly-Pro-Glu-Tyr-Gln-Gln-Asp-Leu-Asp-Arg-Glu-Leu-Tyr，

Lys-Lys-Glu-Lys-Val-Leu-Ala-Thr-Val-Thr-Lys-Thr-Val-Gly-Gly-Asp-Lys，

Leu-Leu-Pro-Lys-Lys-Ser-Ser-Ala-Thr-Val-Gly-Pro-Lys-Ala-Pro-Ala-Val-Gly-Lys，

Pro-Lys-Arg-Lys-Val-Ser-Ala-Asp-Gly-Ala-Ala-Lys-Ala-Glu-Pro-Lys-Arg-Arg，

Arg-Ser-Ala-Ile-Asn-Glu-Val-Val-Thr-Arg-Glu-Tyr，

Ser-Glu-Gly-Val-Phe-Leu-Leu-Thr-Thr-Thr-Pro-Arg-Pro-Val。

respectively shown in SEQ ID NO: 1 to SEQ ID NO: shown at 33.

Preferably, the 33 bioactive peptides are mouse spleen derived lymphocyte peptides.

Preferably, the 33 bioactive peptides have DPP-IV inhibitory activity.

The present invention also provides polynucleotides encoding the 33 bioactive peptides.

In the second aspect of the present invention, the biological activities of the 33 bioactive peptides were determined by research, and database search and amino acid sequence matching determination can be performed by a computer bioinformatics method.

In a third aspect of the invention, the application of the 33 bioactive peptides in preparing foods, health products, medicines or cosmetics with the function of reducing blood sugar is provided.

In particular, the 33 bioactive peptides of the present invention can be used for preparing drugs with hypoglycemic activity.

Specifically, the 33 bioactive peptides of the present invention can be used for preparing foods, health products, medicines or cosmetics with DPP-IV inhibitory activity. Namely, the application of 33 bioactive peptides in preparing foods, health products, medicines or cosmetics with DPP-IV inhibitory activity is provided.

In a fourth aspect of the present invention, a hypoglycemic product is provided, which includes any one or a combination of several of the 33 bioactive peptides and derivatives of the 33 bioactive peptides, and the hypoglycemic product includes a hypoglycemic food, a hypoglycemic health product or a hypoglycemic drug.

The derivatives of the 33 bioactive peptides are meant to have the same or better activity as the specific bioactive peptide.

The derivatives of the 33 bioactive peptides refer to bioactive peptide derivatives obtained by modifying amino acid side chain groups, amino terminals or carboxyl terminals of 33 specific bioactive peptides by hydroxylation, carboxylation, carbonylation, methylation, acetylation, phosphorylation, esterification or glycosylation.

In the fifth aspect of the present invention, there is provided a method for preparing the 33 bioactive peptides, which can be artificially synthesized by genetic engineering methods, can be directly obtained from cells by separation and purification methods, and can be directly prepared by chemical synthesis.

The artificial synthesis of the 33 bioactive peptides by genetic engineering methods is a technical solution that can be realized by those skilled in the art, and for example, the sequence synthesis of the polypeptide can be controlled by a suitable DNA template based on DNA recombination technology.

The method for directly obtaining the cell by the separation and purification method can be as follows: based on the given amino acid sequences of the 33 bioactive peptides, the 33 bioactive peptides are obtained from mouse spleen-derived lymphocytes by adopting a conventional enzymolysis and purification method in the biological technology.

The beneficial effects of the 33 bioactive peptides of the invention are as follows: the 33 bioactive peptides of the mouse spleen-derived lymphocytes have good hypoglycemic activity (DPP-IV inhibitory activity); the 33 bioactive peptides can prolong the action time of the glucagon-like peptide-1 and promote the secretion of insulin, thereby reducing blood sugar, and having very important significance for developing foods, health care products and medicines with the function of reducing blood sugar.

Detailed Description

Before the present embodiments are further described, it is to be understood that the scope of the invention is not limited to the particular embodiments described below; it is also to be understood that the terminology used in the examples is for the purpose of describing particular embodiments only, and is not intended to limit the scope of the present invention.

When numerical ranges are given in the examples, it is understood that both endpoints of each of the numerical ranges and any value therebetween can be selected unless the invention otherwise indicated. Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. In addition to the specific methods, devices, and materials used in the examples, any methods, devices, and materials similar or equivalent to those described in the examples may be used in the practice of the invention in addition to the specific methods, devices, and materials used in the examples, in keeping with the knowledge of one skilled in the art and with the description of the invention.

Unless otherwise indicated, the experimental methods, detection methods, and preparation methods disclosed herein all employ techniques conventional in the art of molecular biology, biochemistry, chromatin structure and analysis, analytical chemistry, cell culture, recombinant DNA technology, and related arts. These techniques are well described in the literature, and may be found in particular in the study of the MOLECULAR CLONING, Sambrook et al: a LABORATORY MANUAL, Second edition, Cold Spring Harbor LABORATORY Press, 1989and Third edition, 2001; ausubel et al, Current PROTOCOLS IN MOLECULAR BIOLOGY, John Wiley & Sons, New York, 1987and periodic updates; the series METHODS IN ENZYMOLOGY, Academic Press, San Diego; wolffe, CHROMATIN STRUCTURE AND FUNCTION, Third edition, Academic Press, San Diego, 1998; (iii) METHODS IN ENZYMOLOGY, Vol.304, Chromatin (P.M.Wassarman and A.P.Wolffe, eds.), Academic Press, San Diego, 1999; and METHODS IN MOLECULAR BIOLOGY, Vol.119, chromatography Protocols (P.B.Becker, ed.) Humana Press, Totowa, 1999, etc.

The present invention will be described in detail below with reference to specific examples.

Example 1

BIOPEP predicts the biological activity of polypeptides derived from lymphocytes

1. The analysis method comprises the following steps:

the polypeptides within the lymphocytes were analyzed using NanoHPLC-MS/MS. The samples were subjected to pooling analysis using PEAKS X software. The mouse protein database was downloaded from the UniProt website (item 16992). The analysis parameters were: mass tolerance of parent ion: 10ppm, mass tolerance of secondary spectrum: 0.020u, fixed modified Carbammidomethyl (C), variable modified Deamidino (NQ), oximation (M) and non-enzyme cutting mode. And obtaining the polypeptide sequence in the sample through library searching analysis. Approximately 50 biological activities were reported in BIOPEP (http:// www.uwm.edu.pl/biochemia/index. php/en/BIOPEP), a total of 4042 peptides (9 months of 2020). The known polypeptide sequence is input by utilizing the 'PROFILES OF polypeptide BIOLOGICALs ACTIVITY' in BIOPEP, so that possible bioactive fragments OF the polypeptide can be determined, and the bioactivity OF the polypeptide can be predicted.

2. Experimental results and analysis:

the DPP-IV inhibitory active fragments searched by the bipep database are shown in table 1.

DPP-IV inhibitory active fragments of Table 133 bioactive peptides

As is clear from Table 1, since the bioactive peptides derived from lymphocytes mainly exhibit DPP-IV inhibitory activity, 33 kinds of bioactive peptides derived from lymphocytes were judged to have DPP-IV inhibitory activity.

Based on this, any one or more of the 33 bioactive peptides and the derivatives of the 33 bioactive peptides are combined to have the hypoglycemic activity, so that any one or more of the 33 bioactive peptides and the derivatives of the 33 bioactive peptides can be combined to obtain the hypoglycemic product.

The embodiments described above are described to facilitate an understanding and use of the invention by those skilled in the art. It will be readily apparent to those skilled in the art that various modifications to these embodiments may be made, and the generic principles described herein may be applied to other embodiments without the use of the inventive faculty. Therefore, the present invention is not limited to the above embodiments, and those skilled in the art should make improvements and modifications within the scope of the present invention based on the disclosure of the present invention.

Sequence listing

<110> Zhejiang ghui peptide Life health science and technology Limited

<120> 33 kinds of bioactive peptides derived from lymphocytes and having hypoglycemic effect and application thereof

<160> 33

<170> SIPOSequenceListing 1.0

<210> 1

<211> 13

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<400> 1

Ala Lys Arg Thr Lys Lys Val Gly Ile Val Gly Lys Tyr

1 5 10

<210> 2

<211> 15

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<400> 2

Ala Leu Lys Ala Lys Lys Ala Val Leu Lys Gly Val His Ser His

1 5 10 15

<210> 3

<211> 11

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<400> 3

Glu Val Glu Gln Lys Lys Lys Arg Thr Phe Arg

1 5 10

<210> 4

<211> 8

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<400> 4

His Leu Lys Ile Val Tyr Arg Arg

1 5

<210> 5

<211> 17

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<400> 5

Lys Gly Asn Leu Arg Arg Thr Gly Gln Glu Val Ala Gln Ala Gln Glu

1 5 10 15

Ser

<210> 6

<211> 14

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<400> 6

Lys Gln Leu Asp Asp Leu Lys Val Glu Leu Ser Gln Leu Arg

1 5 10

<210> 7

<211> 13

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<400> 7

Lys Val Thr Gln Tyr Lys Lys Gly Lys Asp Ser Leu Tyr

1 5 10

<210> 8

<211> 22

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<400> 8

Asn Glu Leu Arg Val Ala Pro Glu Glu His Pro Val Leu Leu Thr Glu

1 5 10 15

Ala Pro Leu Asn Pro Lys

20

<210> 9

<211> 18

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<400> 9

Asn Ile Arg Ala Ser Met Gln Gln Gln Gln Gln Leu Ala Ser Ala Arg

1 5 10 15

Asn Arg

<210> 10

<211> 22

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<400> 10

Asn Leu Val Ile Val Lys Lys Gly Glu Lys Asp Ile Pro Gly Leu Thr

1 5 10 15

Asp Thr Thr Val Pro Arg

20

<210> 11

<211> 8

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<400> 11

Asn Pro Phe Val Thr Ser Asp Arg

1 5

<210> 12

<211> 12

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<400> 12

Asn Val Leu Lys Val Thr Lys Ala Ala Gly Thr Lys

1 5 10

<210> 13

<211> 15

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<400> 13

Pro Ala Val Thr Gln His Ala Pro Tyr Phe Lys Gly Thr Ala Val

1 5 10 15

<210> 14

<211> 11

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<400> 14

Arg Asp Thr Lys Ser Ile Thr Phe Ser Lys Leu

1 5 10

<210> 15

<211> 21

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<400> 15

Arg Ile Gly Val Pro Ser Ala Thr Glu Ile Ile Lys Ala Ser Ser Lys

1 5 10 15

Asp Ala Ile Arg Leu

20

<210> 16

<211> 18

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<400> 16

Arg Lys Ala Val Ile Val Lys Asn Ile Asp Asp Gly Thr Ser Asp Arg

1 5 10 15

Pro Tyr

<210> 17

<211> 16

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<400> 17

Arg Val Thr Ile Ala Gln Gly Gly Val Leu Pro Asn Ile Gln Ala Val

1 5 10 15

<210> 18

<211> 20

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<400> 18

Ser Leu Val Ser Lys Gly Ile Leu Val Gln Thr Lys Gly Thr Gly Ala

1 5 10 15

Ser Gly Ser Phe

20

<210> 19

<211> 12

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<400> 19

Ser Thr Ile Thr Ser Arg Glu Ile Gln Thr Ala Val

1 5 10

<210> 20

<211> 23

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<400> 20

Thr Lys Val Lys Ala Ala Pro Glu Glu Ser Glu Ala Gln Ala Glu Gly

1 5 10 15

Cys Ser Glu Glu Arg Thr Tyr

20

<210> 21

<211> 14

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<400> 21

Thr Leu Val Ile Thr Asp Lys Glu Lys Ala Glu Lys Leu Lys

1 5 10

<210> 22

<211> 9

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<400> 22

Thr Tyr Val Pro Val Thr Thr Phe Lys

1 5

<210> 23

<211> 14

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<400> 23

Val Glu Lys Gly Ile Ile Asp Pro Thr Lys Val Val Arg Thr

1 5 10

<210> 24

<211> 14

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<400> 24

Val Lys Leu Thr Ala Glu Leu Ile Glu Gln Ala Ala Gln Tyr

1 5 10

<210> 25

<211> 15

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<400> 25

Val Arg Tyr Ser Leu Asp Pro Glu Asn Pro Thr Lys Ser Cys Lys

1 5 10 15

<210> 26

<211> 21

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<400> 26

Asn Gln Ile Leu Arg Ile Glu Glu Glu Leu Gly Ser Lys Ala Lys Phe

1 5 10 15

Ala Gly Arg Ser Phe

20

<210> 27

<211> 15

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<400> 27

Ser Leu Ala Asp Leu Gln Asn Asp Glu Val Ala Phe Arg Lys Phe

1 5 10 15

<210> 28

<211> 14

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<400> 28

Ile Gly Pro Glu Tyr Gln Gln Asp Leu Asp Arg Glu Leu Tyr

1 5 10

<210> 29

<211> 17

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<400> 29

Lys Lys Glu Lys Val Leu Ala Thr Val Thr Lys Thr Val Gly Gly Asp

1 5 10 15

Lys

<210> 30

<211> 19

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<400> 30

Leu Leu Pro Lys Lys Ser Ser Ala Thr Val Gly Pro Lys Ala Pro Ala

1 5 10 15

Val Gly Lys

<210> 31

<211> 18

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<400> 31

Pro Lys Arg Lys Val Ser Ala Asp Gly Ala Ala Lys Ala Glu Pro Lys

1 5 10 15

Arg Arg

<210> 32

<211> 12

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<400> 32

Arg Ser Ala Ile Asn Glu Val Val Thr Arg Glu Tyr

1 5 10

<210> 33

<211> 14

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<400> 33

Ser Glu Gly Val Phe Leu Leu Thr Thr Thr Pro Arg Pro Val

1 5 10