CN112480233A

CN112480233A - Bioactive peptide IAHPKLGKRIR, and preparation method and application thereof

Info

Publication number: CN112480233A
Application number: CN202011468779.1A
Authority: CN
Inventors: 张少辉; 王娟; 栾媛媛; 占文静; 张伯宇; 郭婷婷; 谢雨佳; 李政
Original assignee: Zhejiang Huitai Life Health Technology Co ltd; Shanghai Jiaotong University
Current assignee: Zhejiang Huitai Life Health Technology Co ltd; Shanghai Jiaotong University
Priority date: 2020-12-14
Filing date: 2020-12-14
Publication date: 2021-03-12
Anticipated expiration: 2040-12-14
Also published as: CN112480233B

Abstract

The invention relates to the field of protein, and in particular relates to a bioactive peptide IAHPKLGKRIR, a preparation method and application thereof, wherein the amino acid sequence of the bioactive peptide IAHPKLGKRIR is Ile-Ala-His-Pro-Lys-Leu-Gly-Lys-Arg-Ile-Arg. In vitro immune function regulating experiment proves that the bioactive peptide IAHPKLGKRIR has better immune regulating function. The bioactive peptide IAHPKLGKRIR can promote the in vitro proliferation capacity of lymphocytes, improve the immunity of a human body, has a remarkable promoting effect on the phagocytosis capacity of in vitro macrophages, improves the resistance of an organism against infection of external pathogens, reduces the morbidity of the organism, improves the quality of life, and has a very important significance for developing foods, health-care products and medicines with immunoregulation function.

Description

Bioactive peptide IAHPKLGKRIR, and preparation method and application thereof

Technical Field

The invention relates to the field of protein, in particular to a bioactive peptide IAHPKLGKRIR, and a preparation method 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 more than 3000 different bioactive peptides have been reported in the most authoritative bioactive peptide database BIOPEP-UMW.

Currently, studies on bioactive peptides are mostly focused on food-derived polypeptides, and studies and reports on non-food-derived polypeptides are less. And it has been confirmed from the research results that non-food-derived bioactive peptides have higher affinity and can effectively exert their bioactive functions, compared to food-derived bioactive peptides. Lymphocytes are central regulatory cells of the immune system, most of whose function is mediated by a group of small molecule polypeptides called lymphokines. Expression and secretion of these small molecule polypeptides are induced by antigen-stimulated cellular activation. Lymphocytes are therefore the primary source of immunoregulatory peptides produced in the animal body.

Immunomodulatory peptides are a class of bioactive peptides that were first obtained from milk following opioid peptide discovery and demonstrated their physiological activity. Jolles et al found in 1981 for the first time that a hexapeptide with an amino acid sequence Val-Glu-Pro-Ile-Pro-Tyr can be obtained by hydrolyzing human milk protein with trypsin, and in vitro experiments prove that the hexapeptide can enhance the phagocytosis of mouse abdominal cavity macrophages to sheep erythrocytes. Migliore-Samour et al found that the casein-derived hexapeptide Thr-Thr-Met-Pro-Leu-Trp was able to stimulate phagocytosis of murine peritoneal macrophages by sheep red blood cells and to enhance resistance to Klebsiella pneumoniae, with anti-inflammatory properties. Lemna hexandra et al, fed rats with synthetic mouse bone marrow macrophages and a source peptide (PGPIPN), found that phagocytosis of rat peritoneal macrophages and red blood cell-related anti-inflammatory function were significantly enhanced. Bowdis et al, in studying the immune function of the 13 amino acid peptide indolicidin derived from bovine neutrophils, found that the polypeptide indolicidin inhibits LPS-induced TNF- α production in a macrophage-like cell line.

Research shows that the immunoregulation peptide can enhance the immunity of the organism, stimulate the proliferation of lymphocytes of the organism, enhance the phagocytic function of macrophages, promote the release of cytokines, promote the increase of the induced amount of nitric oxide of the macrophages, improve the capability of the organism for resisting the infection of external pathogens, reduce the morbidity of the organism and cannot cause the immune rejection reaction of the organism.

Immunomodulatory peptides generally refer to small, relatively small molecular weight peptides with immunomodulatory activity. The immunomodulatory peptides presently disclosed are generally small peptides with specific immunomodulatory activity, isolated enzymatically from proteins or synthesized chemically. However, when these small peptides are not enzymatically separated from the protein, the protein itself often has no immunomodulatory activity. It is one of the directions in the field of protein research to find bioactive peptides with specific functions from a wide variety of proteins whose amino acid sequences are known, and to study the functions of these polypeptides.

The amino acid sequence of the 60S ribosomal protein L29 protein is shown in SEQ ID NO: 2, respectively. At present, no research on the related functions of the polypeptide fragment of the 60S ribosomal protein L29 protein exists in the prior art.

Disclosure of Invention

The invention aims to provide a bioactive peptide IAHPKLGKRIR, and a preparation method and application thereof.

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

in a first aspect of the present invention, a bioactive peptide IAHPKLGKRIR is provided, which has the amino acid sequence Ile-Ala-His-Pro-Lys-Leu-Gly-Lys-Arg-Ile-Arg, as shown in SEQ ID NO: 1 is shown.

Preferably, the bioactive peptide is mouse spleen derived lymphocyte peptide. The protein is specifically derived from 60S ribosol protein L29 protein and is the 97 th to 107 th amino acid residues of 60S ribosol protein L29 protein. The amino acid sequence of the 60S ribosomal protein L29 is shown in SEQ ID NO: 2, respectively.

The amino acid sequence and the corresponding nucleotide sequence of the 60S ribosomal protein L29 protein are the existing technology, and the nucleotide fragment coding the 97 th to 107 th amino acid residues of the 60S ribosomal protein L29 protein can code the mature bioactive peptide IAHPKLGKRIR.

Preferably, the bioactive peptide has anti-inflammatory and immunoregulatory functions.

The present invention also provides polynucleotides encoding the biologically active peptide IAHPKLGKRIR.

In the second aspect of the present invention, there is provided a method for preparing the bioactive peptide IAHPKLGKRIR, 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 bioactive peptide IAHPKLGKRIR by genetic engineering is a technical solution that can be realized by those skilled in the art, and for example, the synthesis of the sequence 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 amino acid sequence of the given bioactive peptide IAHPKLGKRIR, the bioactive peptide IAHPKLGKRIR is obtained from mouse spleen-derived lymphocytes by a conventional enzymolysis and purification method in biological technology.

In a third aspect of the present invention, there is provided a use of the bioactive peptide IAHPKLGKRIR in the preparation of a medicament or a cosmetic having an anti-inflammatory function.

Further, the application of the bioactive peptide IAHPKLGKRIR in preparing a medicament for promoting the ability of macrophages in vitro to phagocytose neutral red is provided.

In a fourth aspect of the invention, there is provided an anti-inflammatory product comprising said biologically active peptide IAHPKLGKRIR or a derivative of said biologically active peptide IAHPKLGKRIR; the anti-inflammatory product comprises an anti-inflammatory drug or an anti-inflammatory cosmetic; the derivative of the bioactive peptide IAHPKLGKRIR refers to a bioactive peptide derivative obtained by modifying the amino acid side chain group, amino terminal or carboxyl terminal of the bioactive peptide IAHPKLGKRIR by hydroxylation, carboxylation, carbonylation, methylation, acetylation, phosphorylation, esterification or glycosylation.

In the fifth aspect of the present invention, the application of the bioactive peptide IAHPKLGKRIR in preparing food or medicine with immunoregulation function is provided.

Further, the use of the bioactive peptide IAHPKLGKRIR in the preparation of a medicament for promoting the in vitro proliferative capacity of lymphocytes is provided.

In a sixth aspect of the present invention, there is provided a product having an immunoregulatory function, comprising said biologically active peptide IAHPKLGKRIR or a derivative of said biologically active peptide IAHPKLGKRIR; the derivative of the bioactive peptide IAHPKLGKRIR refers to a bioactive peptide derivative obtained by modifying the amino acid side chain group, amino terminal or carboxyl terminal of the bioactive peptide IAHPKLGKRIR by hydroxylation, carboxylation, carbonylation, methylation, acetylation, phosphorylation, esterification or glycosylation.

Derivatives of the bioactive peptides IAHPKLGKRIR are meant to have the same activity or better activity than the bioactive peptides IAHPKLGKRIR.

The bioactive peptide IAHPKLGKRIR has the beneficial effects that: the bioactive peptide IAHPKLGKRIR has good anti-inflammatory activity; the bioactive peptide IAHPKLGKRIR can promote the in vitro proliferation capacity of lymphocytes, improve the immunity of a human body, has a remarkable promoting effect on the ability of phagocytizing neutral red by in vitro macrophages under the condition of inflammation, improves the ability of resisting the infection of external pathogens of the body, reduces the morbidity of the body, improves the quality of life, and has very important significance for developing foods, health-care products and medicines with the immunoregulation function.

Drawings

FIG. 1: a first order mass spectrum of a fragment with a mass to charge ratio of 430.2825 (m/z 430.2825);

FIG. 2: a secondary mass spectrum of a segment with the mass-to-charge ratio of 430.2825 and the breaking conditions of the bioactive peptides az and by;

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, 1989 and Third edition, 2001; ausubel et al, Current PROTOCOLS IN MOLECULAR BIOLOGY, John Wiley & Sons, New York, 1987 and 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 invention is described in detail below with reference to the figures and specific embodiments.

Example 1 Artificial Synthesis of bioactive peptide IAHPKLGKRIR

Synthesis of bioactive peptide

1. 3g of RINK resin (degree of substitution 0.3mmol/g) was weighed into a 150ml reactor and soaked with 50ml of Dichloromethane (DCM).

After 2.2 hours, the resin was washed with 3 resin volumes of N-Dimethylformamide (DMF) and then drained, and this was repeated four times and the resin was drained until use.

3. The Fmoc protecting group on the resin was removed by adding a quantity of 20% piperidine (piperidine/DMF: 1:4, v: v) to the reactor and shaking on a decolourising shaker for 20 min. After deprotection, the resin was washed four times with 3 resin volumes of DMF and then drained.

4. And (3) detecting a small amount of resin by a ninhydrin (ninhydrin) method (detecting A and B, respectively, and reacting at 100 ℃ for 1min), wherein the resin is colored, which indicates that the deprotection is successful.

5. Weighing an appropriate amount of amino acid Ile and an appropriate amount of 1-hydroxy-benzotriazole (HOBT) into a 50ml centrifuge tube, adding 20ml of DMF to dissolve the amino acid Ile and the 1-hydroxy-benzotriazole (HOBT), then adding 3ml of N, N Diisopropylcarbodiimide (DIC) to shake and shake for 1min, adding the solution into a reactor after the solution is clarified, and then placing the reactor into a 30 ℃ shaking table to react.

After 6.2 hours, the column was capped with a suitable amount of acetic anhydride (acetic anhydride: DIEA: DCM ═ 1:1:2, v: v: v) for half an hour, then washed four times with 3 resin volumes of DMF and drained until needed.

7. The Fmoc protecting group on the resin was removed by adding a quantity of 20% piperidine (piperidine/DMF: 1:4, v: v) to the reactor and shaking on a decolourising shaker for 20 min. After deprotection was washed four times with DMF and then drained.

8. And (3) detecting a small amount of resin by a ninhydrin (ninhydrin) method (detecting A and B, respectively, and reacting at 100 ℃ for 1min), wherein the resin is colored, which indicates that the deprotection is successful.

9. Weighing a second proper amount of amino acid and a proper amount of HOBT in a 50ml centrifuge tube, adding 25ml of DMF to dissolve the amino acid and the HOBT, adding 2.5ml of DIC to shake and shake for 1min, adding the solution into a reactor after the solution is clarified, and then placing the reactor in a shaking table at 30 ℃ to react.

After 10.1 hours, a small amount of resin is taken for detection, and the detection is carried out by an indanthrone method (two drops are respectively detected A and B, and the reaction is carried out for 1min at 100 ℃), if the resin is colorless, the reaction is complete; if the resin is colored, the condensation is not complete and the reaction is continued.

11. After the reaction was completed, the resin was washed four times with DMF and then drained, and a certain amount of 20% piperidine (piperidine/DMF ═ 1:4, v: v) was added to the reactor, and the mixture was shaken on a decolorizing shaker for 20min to remove the Fmoc-protecting group from the resin. After the protection is removed, washing with DMF for four times, and then draining to detect whether the protection is removed.

12. Amino acids Ile, Ala, His, Pro, Lys, Leu, Gly, Lys, Arg, Ile, Arg are sequentially grafted according to steps 9-11.

13. After the last amino acid had been grafted, the protection was removed, washed four times with DMF and the resin was drained with methanol. The biologically active peptide was then cleaved from the resin using 95 cleavage medium (trifluoroacetic acid: 1,2 ethanedithiol: 3, isopropylsilane: water: 95:2:2:1, v: v: v) (10 ml of cleavage medium per gram of resin) and centrifuged four times with glacial ethyl ether (cleavage medium: ethyl ether: 1:9, v: v).

To this end, bioactive peptide IAHPKLGKRIR was synthesized.

Confirmation of biologically active peptides

1) UPLC analysis

UPLC conditions were as follows:

the instrument comprises the following steps: waters ACQUITY UPLC ultra-high performance liquid phase, electrospray, quadrupole and time-of-flight mass spectrometer

Specification of chromatographic column: BEH C18 chromatographic column

Flow rate: 0.4mL/min

Temperature: 50 deg.C

Ultraviolet detection wavelength: 210nm

Sample introduction amount: 2 μ L

Gradient conditions: solution A: water containing 0.1% formic acid (v/v), liquid B: acetonitrile containing 0.1% formic acid (v/v)

2) Mass spectrometric analysis

The mass spectrometry conditions were as follows:

ion mode: ES +

Mass range (m/z): 100. 1000A

Capillary voltage (Capillary) (kV): 3.0

Sampling cone (V): 35.0

Ion source temperature (. degree. C.): 115

Desolvation temperature (. degree. C.): 350

Desolventizing gas stream (L/hr): 700.0

Collision energy (eV): 4.0

Scan time (sec): 0.25

Inner scan time (sec): 0.02

According to the above analysis method, the bioactive peptide IAHPKLGKRIR was subjected to chromatographic analysis and mass spectrometric analysis using ultra high performance liquid, electrospray, quadrupole, time-of-flight mass spectrometry. The primary mass spectrum of the bioactive peptide IAHPKLGKRIR is shown in figure 1, the secondary mass spectrum of the extracted peak and the az and by breaking conditions are shown in figure 2, the mass-to-charge ratio of the bioactive peptide of the peak is 430.2825, and the retention time is 6.96 min.

3) Results

As can be seen from fig. 2, the fragments with mass-to-charge ratios 430.2825 obtained from az and by cleavage were Ile, Ala, His, Pro, Lys, Leu, Gly, Lys, Arg, Ile, Arg (IAHPKLGKRIR), and are represented by SEQ ID NO: 1. the fragment corresponds to residue sequences of 97 th to 107 th sites of 60S ribosomal protein L29 protein, the GenBank number of the amino acid sequence of the 60S ribosomal protein L29 protein is AAH82292.1, and the sequence is shown in SEQ ID NO: 2.

example 2 immunological Activity assay of bioactive peptides

First, experiment of macrophage phagocytosis neutral red promoting ability of biological active peptide IAHPKLGKRIR

1. Experimental reagents and instruments:

reagent: experimental animals balb/c mice (male 6-8 weeks old) were collected at the animal Experimental center of the college of agriculture and biology of Shanghai university of transportation; the mouse spleen lymphocyte-derived bioactive peptide IAHPKLGKRIR obtained in example 1; LPS, purchased from Sigma; neutral red staining solution, produced by Biyuntian biotechnological research institute.

The instrument equipment comprises: LRH-250F Biochemical incubator Shanghai Hengshi Co., Ltd; GL-22M high speed refrigerated centrifuge Shanghai Luxiang apparatus centrifuge Instrument Co., Ltd; hera cell 150 CO2 incubator Heraeus; dragon Wellscan MK3 microplate reader Labsystems.

2. The experimental method comprises the following steps:

the number of the added cells was 2X 10⁶100 mul/well of cell suspension per ml, adding 200 mul/well of RPMI1640 complete culture medium (10% FBS) containing bioactive peptide IAHPKLGKRIR (1mg/ml) after adherent purification as experimental group, adding R containing no bioactive peptidePMI1640 complete medium (10% FBS)200 u l/hole were cultured as blank group; and LPS is added into the experimental group and the blank group when the culture time reaches 24h to reach the final concentration of 10 mug/ml; after further culturing for 48h, the cell culture solution was aspirated. After washing the bottom of the well with PBS, 80. mu.l/well of neutral red dye solution at 37 ℃ was added, and after 10 minutes the dye solution was aspirated and washed twice with PBS, 150. mu.l of cell lysate (glacial acetic acid: absolute ethanol ═ 1:1, v/v) was added to each well. After overnight dissolution at 4 ℃ the absorbance value (OD540) was determined at a wavelength of 540 nm.

3. Experimental results and analysis:

TABLE 1 determination of the ability of the bioactive peptide IAHPKLGKRIR to promote phagocytosis of neutral Red by macrophages

Experiment grouping	Absorbance value (OD540)
		Blank group	0.1074±0.0281
Experimental group	0.1475±0.0134**

Note: significant difference compared to negative control (P <0.05)

Significant difference compared with negative control group (P < 0.01)

The experimental results are shown in table 1, compared with the blank cell group, the macrophage phagocytosis ability of the inflammatory group added with 1mg/ml bioactive peptide IAHPKLGKRIR is obviously increased, and compared with the blank cell group, the macrophage phagocytosis ability of the inflammatory group added with 1mg/ml bioactive peptide IAHPKLGKRIR has a very significant difference (P is less than 0.01). The biological active peptide IAHPKLGKRIR is proved to have obvious promotion effect on the ability of phagocytizing neutral red by macrophages in vitro under the condition of inflammation. Thus, bioactive peptide IAHPKLGKRIR has anti-inflammatory activity.

Second, in vitro lymphocyte proliferation potency assay (MTT method) for bioactive peptide IAHPKLGKRIR

1. Experimental materials and instruments:

reagents and materials: experimental animals balb/c mice (male 6-8 weeks old, animal experiment center of Shanghai university of transportation, college of agriculture and biology); the mouse spleen lymphocyte-derived bioactive peptide IAHPKLGKRIR obtained in example 1; mouse lymphocyte extract (ex solibao); RPMI1640 medium (purchased from GIBCO); 3- (4, 5-Dimethylthiazol-2) -2, 5-diphenyltetrazolium bromide salt (MTT, available from Amresco, Inc.); concanavalin (ConA, available from Sigma); bovine serum albumin (BSA, available from Genebase); pepsin (available from Sigma); pancreatin (Corolase PP, from AB).

The instrument equipment comprises: LRH-250F Biochemical incubator, Shanghai Hengshi Co., Ltd; GL-22M high speed refrigerated centrifuge, shanghai luxiang instrument centrifuge instruments ltd; hera cell 150 CO2 incubator, Heraeus; dragon Wellscan MK3 microplate reader, Labsystems Inc.; ALPHA 1-2-LD vacuum freeze drier, Christ company; ultra performance liquid chromatography-quadrupole time-of-flight mass spectrometer, waters corporation.

2. The experimental method comprises the following steps:

taking mouse spleen under aseptic condition, extracting mouse lymphocyte with lymphocyte extract, and performing primary culture. The cell density was adjusted to 2.5X 10 with complete RPMI1640 medium⁶one/mL. To a 96-well cell culture plate were added in sequence: 100 μ L mouse lymphocyte suspension, 100 μ L RPMI1640 complete medium, 20 μ L concanavalin, 100 μ L bioactive peptide sample. In addition, a blank control group (PBS with pH7.2-7.4 and 3 mol/L) and a negative control group (500 mu g/mL BSA) are arranged, and the research shows that the blank control group has no influence on the in vitro lymphocyte proliferation. Each set of 3 replicates. At 5% CO₂Culturing at 37 deg.C for 68 hr, adding 20 μ L MTT into each well under aseptic condition, culturing for 4 hr, carefully removing supernatant, adding 100 μ L dimethyl sulfoxide into each well, incubating at 37 deg.C for 10min, shaking, and usingThe microplate reader measures the absorbance at 570 nm.

The in vitro lymphocyte proliferation capacity is expressed by a stimulation index and is calculated as follows:

in the formula: a. the₁Absorbance at 570nm for the blank; a. the₂Absorbance at 570nm for the negative control, A₃The absorbance at 570nm for the experimental group.

3. Experimental results and analysis:

TABLE 2 Effect of bioactive peptide IAHPKLGKRIR on lymphocyte proliferation in vitro

Experiment grouping	Stimulation index SI
		BSA	1
Bioactive peptide samples	1.158±0.028*

Note: the number marked as significant difference (P <0.05) compared to the negative control.

The results are shown in Table 2. As can be seen from Table 2, the stimulation index of the bioactive peptide IAHPKLGKRIR was greater than that of BSA, indicating that IAHPKLGKRIR can stimulate the proliferation of mouse lymphocytes in vitro to some extent. And IAHPKLGKRIR reached a stimulation index of 1.158, which was significantly different from that of the negative control group (P < 0.05). Therefore, the bioactive peptide IAHPKLGKRIR is considered to have the capacity of remarkably promoting mouse lymphocyte proliferation, can be used as a substance with immunoregulation activity to be added into health products, and can improve the immunity of human bodies.

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> Shanghai university of transportation, Zhejiang river peptide Life health science and technology Limited

<120> bioactive peptide IAHPKLGKRIR, and preparation method and application thereof

<160> 2

<170> SIPOSequenceListing 1.0

<210> 1

<211> 11

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<400> 1

Ile Ala His Pro Lys Leu Gly Lys Arg Ile Arg

1 5 10

<210> 2

<211> 160

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<400> 2

Met Ala Lys Ser Lys Asn His Thr Thr His Asn Gln Ser Arg Lys Trp

1 5 10 15

His Arg Asn Gly Ile Lys Lys Pro Arg Ser Gln Arg Tyr Glu Ser Leu

20 25 30

Lys Gly Val Asp Pro Lys Phe Leu Arg Asn Met Arg Phe Ala Lys Lys

35 40 45

His Asn Lys Lys Gly Leu Lys Lys Met Gln Ala Asn Asn Ala Lys Ala

50 55 60

Val Ser Ala Arg Ala Glu Ala Ile Lys Ala Leu Val Lys Pro Gln Ala

65 70 75 80

Ile Lys Pro Lys Met Pro Lys Gly Pro Lys Leu Lys Arg Leu Ala Phe

85 90 95

Ile Ala His Pro Lys Leu Gly Lys Arg Ile Arg Ser Tyr Met Ala Lys

100 105 110

Gly Gln Arg Leu Cys Gln Pro Lys Pro Lys Val Gln Thr Lys Ala Gly

115 120 125

Ala Lys Ala Pro Ala Lys Ala Gln Ala Ser Ala Pro Ala Gln Ala Pro

130 135 140

Lys Gly Ala Gln Ala Pro Lys Gly Ala Gln Ala Pro Val Lys Ala Pro

145 150 155 160

Claims

1. A bioactive peptide IAHPKLGKRIR, characterized in that its amino acid sequence is Ile-Ala-His-Pro-Lys-Leu-Gly-Lys-Arg-Ile-Arg.

2. A polynucleotide encoding the biologically active peptide IAHPKLGKRIR of claim 1.

3. The method of claim 1, wherein the bioactive peptide IAHPKLGKRIR is synthesized by genetic engineering, isolated from cells, purified, or chemically synthesized.

4. The use of bioactive peptide IAHPKLGKRIR as claimed in claim 1, wherein the use of bioactive peptide IAHPKLGKRIR in the manufacture of a medicament or cosmetic product with anti-inflammatory properties.

5. The use of biologically active peptide IAHPKLGKRIR of claim 4, wherein the use of biologically active peptide IAHPKLGKRIR in the manufacture of a medicament for promoting the ability of macrophages in vitro to phagocytose neutral red.

6. The use of biologically active peptide IAHPKLGKRIR of claim 1, wherein the use of biologically active peptide IAHPKLGKRIR in the preparation of a food or a medicament having immunomodulatory properties.

7. The use of biologically active peptide IAHPKLGKRIR of claim 6, wherein said biologically active peptide IAHPKLGKRIR is used in the manufacture of a medicament for promoting the ability of lymphocytes to proliferate in vitro.

8. An anti-inflammatory product comprising the biologically active peptide IAHPKLGKRIR of claim 1 or a derivative of the biologically active peptide IAHPKLGKRIR; the anti-inflammatory product comprises an anti-inflammatory drug or an anti-inflammatory cosmetic; derivatives of the bioactive peptides IAHPKLGKRIR are meant to have the same activity or better activity than the bioactive peptides IAHPKLGKRIR.

9. A product having an immunomodulatory function, comprising the biologically active peptide IAHPKLGKRIR of claim 1 or a derivative of the biologically active peptide IAHPKLGKRIR; the product with immunoregulation function comprises food with immunoregulation function or medicine with immunoregulation function; derivatives of the bioactive peptides IAHPKLGKRIR are meant to have the same activity or better activity than the bioactive peptides IAHPKLGKRIR.

10. An anti-inflammatory product according to claim 8 or a product with immunoregulatory function according to claim 9 wherein the derivative of bioactive peptide IAHPKLGKRIR is a derivative of bioactive peptide obtained by hydroxylation, carboxylation, carbonylation, methylation, acetylation, phosphorylation, esterification or glycosylation modification at the amino acid side chain group, amino terminus or carboxy terminus of bioactive peptide IAHPKLGKRIR.