CN112679598A - Bioactive peptide SGVSLAALKKALAAAGYDVEK, and preparation method and application thereof - Google Patents
Bioactive peptide SGVSLAALKKALAAAGYDVEK, and preparation method and application thereof Download PDFInfo
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Abstract
The invention relates to the field of protein, and in particular relates to a bioactive peptide SGVSLAALKKALAAAGYDVEK, a preparation method and application thereof, wherein the amino acid sequence of the bioactive peptide SGVSLAALKKALAAAGYDVEK is Ser-Gly-Val-Ser-Leu-Ala-Ala-Ala-Leu-Lys-Lys-Ala-Leu-Ala-Ala-Gly-Tyr-Asp-Val-Glu-Lys. In vitro immunoregulation function experiments prove that the bioactive peptide SGVSLAALKKALAAAGYDVEK has a good immunoregulation function. The bioactive peptide SGVSLAALKKALAAAGYDVEK can promote macrophage to secrete cell factor, can improve the capability of an organism to resist infection of external pathogens, enhances the in vitro proliferation capability of lymphocytes, reduces the morbidity of the organism, improves the quality of life, and has very important significance for developing foods, health-care products and medicines with the immunoregulation function.
Description
Technical Field
The invention relates to the field of protein, in particular to a bioactive peptide SGVSLAALKKALAAAGYDVEK, and a preparation method and application thereof.
Background
In recent years, bioactive peptides have become a word of great energy in the ear. Immunomodulatory peptides generally refer to small, relatively small molecular weight peptides with immunomodulatory activity. 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.
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 Histone H1.3 protein is shown as SEQ ID NO: 2, respectively. At present, the related functions of the polypeptide fragments of Histone H1.3 protein are not researched in the prior art.
Disclosure of Invention
The invention aims to provide a bioactive peptide SGVSLAALKKALAAAGYDVEK, 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 SGVSLAALKKALAAAGYDVEK is provided, which has an amino acid sequence shown in SEQ ID NO: 1, specifically:
Ser-Gly-Val-Ser-Leu-Ala-Ala-Leu-Lys-Lys-Ala-Leu-Ala-Ala-Ala-Gly-Tyr-Asp-Val-Glu-Lys。
preferably, the bioactive peptide is mouse spleen derived lymphocyte peptide. Specifically, the protein is derived from Histone H1.3 protein and is the amino acid residue at the 56 th to 76 th positions of the Histone H1.3 protein. The amino acid sequence of Histone H1.3 protein is shown as SEQ ID NO: 2, respectively.
The amino acid sequence and the corresponding nucleotide sequence of the Histone H1.3 protein are the prior art, and the nucleotide fragment for coding the 56 th-76 th amino acid residues of the Histone H1.3 protein can code mature bioactive peptide SGVSLAALKKALAAAGYDVEK.
Preferably, the bioactive peptide has anti-inflammatory and immunoregulatory functions.
The present invention also provides polynucleotides encoding the biologically active peptide SGVSLAALKKALAAAGYDVEK.
In the second aspect of the present invention, there is provided a method for preparing the bioactive peptide SGVSLAALKKALAAAGYDVEK, 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 SGVSLAALKKALAAAGYDVEK 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 SGVSLAALKKALAAAGYDVEK, the bioactive peptide SGVSLAALKKALAAAGYDVEK 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 SGVSLAALKKALAAAGYDVEK in the preparation of a medicament or a cosmetic having an anti-inflammatory function.
In particular, the bioactive peptide SGVSLAALKKALAAAGYDVEK of the present invention may be used in the preparation of medicaments with anti-inflammatory and/or anti-oxidant properties.
Further, the use of the bioactive peptide SGVSLAALKKALAAAGYDVEK in the manufacture of a medicament for inhibiting inflammation due to oxidation.
In a fourth aspect of the present invention, there is provided a use of the bioactive peptide SGVSLAALKKALAAAGYDVEK in the preparation of food or medicine with immunoregulatory function.
Further, the use of the bioactive peptide SGVSLAALKKALAAAGYDVEK in the preparation of a food or medicament for promoting the in vitro proliferation of lymphocytes.
Further, the application of the bioactive peptide SGVSLAALKKALAAAGYDVEK in preparing foods or medicines for promoting macrophage to secrete cytokines is provided.
In a fifth aspect of the invention, there is provided an anti-inflammatory product comprising said biologically active peptide SGVSLAALKKALAAAGYDVEK or a derivative of said biologically active peptide SGVSLAALKKALAAAGYDVEK; the anti-inflammatory product comprises an anti-inflammatory drug or an anti-inflammatory cosmetic.
In a sixth aspect of the present invention, there is provided a product having an immunoregulatory function, comprising said biologically active peptide SGVSLAALKKALAAAGYDVEK or a derivative of said biologically active peptide SGVSLAALKKALAAAGYDVEK; the product with immunoregulatory function comprises food with immunoregulatory function or medicine with immunoregulatory function.
Derivatives of the bioactive peptides SGVSLAALKKALAAAGYDVEK are meant to have the same activity or better activity than the bioactive peptides SGVSLAALKKALAAAGYDVEK.
The derivative of the bioactive peptide SGVSLAALKKALAAAGYDVEK refers to a polypeptide derivative obtained by modifying the amino acid side chain group, amino terminal or carboxyl terminal of the bioactive peptide SGVSLAALKKALAAAGYDVEK by hydroxylation, carboxylation, carbonylation, methylation, acetylation, phosphorylation, esterification or glycosylation.
The bioactive peptide SGVSLAALKKALAAAGYDVEK has the beneficial effects that: the bioactive peptide SGVSLAALKKALAAAGYDVEK has good anti-inflammatory activity; the bioactive peptide SGVSLAALKKALAAAGYDVEK can promote macrophage to secrete cell factor, improve the capability of an organism to resist infection of external pathogens, promote the in vitro proliferation of lymphocytes, reduce the morbidity of the organism, improve the quality of life and have 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 688.0604 (m/z 688.0604);
FIG. 2: a secondary mass spectrum of a fragment with a mass-to-charge ratio of 688.0604 and the breakage conditions of the polypeptides 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: ALABORATORY 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 invention is described in detail below with reference to the figures and specific embodiments.
Example 1 Artificial Synthesis of active peptide SGVSLAALKKALAAAGYDVEK
Synthesis of bioactive peptide
1.3 g 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 a proper amount of amino acid Ser and a proper amount of 1-hydroxy-benzotriazole (HOBT) into a 50ml centrifuge tube, adding 20ml of DMF to dissolve the amino acid Ser 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. The amino acids Ser, Gly, Val, Ser, Leu, Ala, Leu, Lys, Ala, Leu, Ala, Gly, Tyr, Asp, Val, Glu and Lys are grafted in sequence 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 SGVSLAALKKALAAAGYDVEK 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 SGVSLAALKKALAAAGYDVEK 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 SGVSLAALKKALAAAGYDVEK 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 688.0604, and the retention time is 51.1 min.
3) Results
As can be seen from fig. 2, the fragment sequence of mass-to-charge ratio 688.0604 was calculated by Mascot software analysis based on the cases of az and by cleavage, and found to be Ser, Gly, Val, Ser, Leu, Ala, Leu, Lys, Ala, Leu, Ala, Gly, Tyr, Asp, Val, Glu, Lys (SGVSLAALKKALAAAGYDVEK), and is represented as SEQ ID NO: 1. the fragment corresponds to the residue sequence of 55-75 th sites of the Histone H1.3 protein, the GenBank number of the amino acid sequence of the Histone H1.3 protein is BAC35711.1, and the sequence is shown in SEQ ID NO: 2.
example 2 immunological Activity assay of bioactive peptides
First, experiment (ELISA method) of promoting macrophage secretion cell factor of biological active peptide SGVSLAALKKALAAAGYDVEK
1. Experimental reagents and instruments:
reagent: experimental animals balb/c mice (male 6, 8 weeks old), Shanghai Slek Experimental animals, Inc.; mouse lymphocyte extract, shanghai solibao biotechnology limited; RPMI1640 medium, GIBCO; bovine Serum Albumin (BSA), Genebase; the mouse spleen lymphocyte-derived bioactive peptide SGVSLAALKKALAAAGYDVEK obtained in example 1; ELISA cytokine Rapid kits (TNF-. alpha., IL-1. beta. and IL-6), Wuhan Dr bioengineering, Inc.
The instrument equipment comprises: LRH, 250F biochemical incubator shanghai constant technology ltd; GL, 22M high speed refrigerated centrifuge Shanghai Luxiang apparatus centrifuge instruments 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 μ l/well of cell suspension/ml, 200 μ l/well of peptide-containing RPMI1640 complete medium (10% FBS) after adherent purification, LPS to a final concentration of 10 μ g/ml at 24 hours in the inflammation group, continuous culture for 48 hours, and LPS to a final concentration of 100ng/ml at 24 hours before termination of the culture in the inflammation group. After the termination of the culture, the cell culture supernatant was collected by centrifugation. Adding 100 μ l of supernatant to an ELISA plate coated with a cytokine antibody, reacting at 37 ℃ for 90 minutes, adding a biotin-labeled antibody, reacting at 37 ℃ for 60 minutes, washing with PBS, adding avidin-peroxidase complex, and reacting for 30 minutes. After washing with PBS, a developing solution was added thereto, and the reaction was carried out for 20 minutes. After addition of the chromogenic stop solution, the absorbance value (OD450) was measured at a wavelength of 450nm using a microplate reader.
3. Experimental results and analysis:
TABLE 1 determination of the Effect of bioactive peptide SGVSLAALKKALAAAGYDVEK on macrophage cytokine levels
Note: significant difference compared to negative control (P < 0.05); the difference in the negative control group was very significant (P <0.01)
As can be seen from Table 1, in the experimental results of three cytokines, namely TNF-alpha, IL-1 beta and IL-6, the significant difference (P <0.01) occurs between TNF-alpha and IL-1 beta at 0.2mg/ml and above, and the significant difference (P <0.01) occurs between IL-6 at 0.5mg/ml, so that SGVSLAALKKALAAAGYDVEK at a certain concentration can promote the activation of mouse abdominal cavity macrophages and release TNF-alpha, IL-1 beta and IL-6, and the TNF-alpha, IL-1 beta and IL-6 can induce the differentiation and antibody production of B cells, induce the activation, proliferation and differentiation of T cells and participate in the immune response of organisms. Therefore, SGVSLAALKKALAAAGYDVEK at a certain concentration can improve the action of these cytokines in the resting state of normal macrophages, thereby regulating the immunity of the organism.
Second, in vitro lymphocyte proliferation potency assay (MTT method) for bioactive peptide SGVSLAALKKALAAAGYDVEK
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 SGVSLAALKKALAAAGYDVEK 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.; ALPHA1-2-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 68h, adding 20 μ L MTT into each well under aseptic condition, culturing for 4h, carefully removing supernatant, adding 100 μ L dimethyl sulfoxide into each well, incubating at 37 deg.C for 10min, shaking, and measuring absorbance at 570nm with microplate reader.
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 SGVSLAALKKALAAAGYDVEK on lymphocyte proliferation in vitro
| Experiment grouping | Stimulation index SI |
| BSA | 1 |
| SGVSLAALKKALAAAGYDVEK | 1.165±0.022** |
Note: the number marked as significant difference (P < 0.05) compared to the negative control.
The results are shown in Table 2. As shown in Table 2, under the condition that the mass concentration of the bioactive peptide SGVSLAALKKALAAAGYDVEK is 100 mug/mL, the stimulation index of the bioactive peptide SGVSLAALKKALAAAGYDVEK is greater than that of BSA, which indicates that SGVSLAALKKALAAAGYDVEK can stimulate the proliferation of mouse lymphocytes in vitro to a certain extent. And SGVSLAALKKALAAAGYDVEK reached a stimulation index of 1.165, which was very significantly different from the negative control group (P < 0.01). Therefore, the bioactive peptide SGVSLAALKKALAAAGYDVEK 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> Zhejiang ghui peptide Life health science and technology Limited
<120> bioactive peptide SGVSLAALKKALAAAGYDVEK, and preparation method and application thereof
<160> 2
<170> SIPOSequenceListing 1.0
<210> 1
<211> 21
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 1
Ser Gly Val Ser Leu Ala Ala Leu Lys Lys Ala Leu Ala Ala Ala Gly
1 5 10 15
Tyr Asp Val Glu Lys
20
<210> 2
<211> 221
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 2
Met Ser Glu Thr Ala Pro Ala Ala Pro Ala Ala Pro Ala Pro Val Glu
1 5 10 15
Lys Thr Pro Val Lys Lys Lys Ala Lys Lys Thr Gly Ala Ala Ala Gly
20 25 30
Lys Arg Lys Ala Ser Gly Pro Pro Val Ser Glu Leu Ile Thr Lys Ala
35 40 45
Val Ala Ala Ser Lys Glu Arg Ser Gly Val Ser Leu Ala Ala Leu Lys
50 55 60
Lys Ala Leu Ala Ala Ala Gly Tyr Asp Val Glu Lys Asn Asn Ser Arg
65 70 75 80
Ile Lys Leu Gly Leu Lys Ser Leu Val Ser Lys Gly Thr Leu Val Gln
85 90 95
Thr Lys Gly Thr Gly Ala Ser Gly Ser Phe Lys Leu Asn Lys Lys Ala
100 105 110
Ala Ser Gly Glu Ala Lys Pro Lys Ala Lys Lys Ala Gly Ala Ala Lys
115 120 125
Ala Lys Lys Pro Ala Gly Ala Ala Lys Lys Pro Lys Lys Ala Thr Gly
130 135 140
Ala Ala Thr Pro Lys Lys Thr Ala Lys Lys Thr Pro Lys Lys Ala Lys
145 150 155 160
Lys Pro Ala Ala Ala Ala Gly Ala Lys Lys Val Ser Lys Ser Pro Lys
165 170 175
Lys Val Lys Ala Ala Lys Pro Lys Lys Ala Ala Lys Ser Pro Ala Lys
180 185 190
Ala Lys Ala Pro Lys Ala Lys Ala Ser Lys Pro Lys Ala Ser Lys Pro
195 200 205
Lys Ala Thr Lys Ala Lys Lys Ala Ala Pro Arg Lys Lys
210 215 220
Claims (10)
1. A bioactive peptide SGVSLAALKKALAAAGYDVEK, characterized by the amino acid sequence:
Ser-Gly-Val-Ser-Leu-Ala-Ala-Leu-Lys-Lys-Ala-Leu-Ala-Ala-Ala-Gly-Tyr-Asp-Val-Glu-Lys。
2. a polynucleotide encoding the biologically active peptide SGVSLAALKKALAAAGYDVEK of claim 1.
3. The method of claim 1, wherein the bioactive peptide SGVSLAALKKALAAAGYDVEK is synthesized by genetic engineering, isolated from cells, purified, or chemically synthesized.
4. The use of bioactive peptide SGVSLAALKKALAAAGYDVEK as claimed in claim 1, wherein the use of bioactive peptide SGVSLAALKKALAAAGYDVEK in the manufacture of a medicament or cosmetic product with anti-inflammatory properties.
5. The use of biologically active peptide SGVSLAALKKALAAAGYDVEK of claim 1, wherein the use of biologically active peptide SGVSLAALKKALAAAGYDVEK in the preparation of a food or a medicament having immunomodulatory properties.
6. The use of biologically active peptide SGVSLAALKKALAAAGYDVEK of claim 5, wherein said biologically active peptide SGVSLAALKKALAAAGYDVEK is used in the manufacture of a food or medicament for promoting the in vitro proliferation of lymphocytes.
7. The use of biologically active peptide SGVSLAALKKALAAAGYDVEK of claim 5, wherein said biologically active peptide SGVSLAALKKALAAAGYDVEK is used in the manufacture of a food or a pharmaceutical product for promoting the secretion of cytokines by macrophages.
8. An anti-inflammatory product comprising the biologically active peptide SGVSLAALKKALAAAGYDVEK of claim 1 or a derivative of the biologically active peptide SGVSLAALKKALAAAGYDVEK; the anti-inflammatory product comprises an anti-inflammatory drug or an anti-inflammatory cosmetic; derivatives of the bioactive peptides SGVSLAALKKALAAAGYDVEK are meant to have the same activity or better activity than the bioactive peptides SGVSLAALKKALAAAGYDVEK.
9. A product having an immunomodulatory function, comprising the biologically active peptide SGVSLAALKKALAAAGYDVEK of claim 1 or a derivative of the biologically active peptide SGVSLAALKKALAAAGYDVEK; the product with immunoregulation function comprises food with immunoregulation function or medicine with immunoregulation function; derivatives of the bioactive peptides SGVSLAALKKALAAAGYDVEK are meant to have the same activity or better activity than the bioactive peptides SGVSLAALKKALAAAGYDVEK.
10. An anti-inflammatory product according to claim 8 or a product with immunomodulatory activity according to claim 9, wherein said derivative of bioactive peptide SGVSLAALKKALAAAGYDVEK is a polypeptide derivative obtained by modification of amino acid side chain groups, amino-terminal or carboxy-terminal of bioactive peptide SGVSLAALKKALAAAGYDVEK by hydroxylation, carboxylation, carbonylation, methylation, acetylation, phosphorylation, esterification or glycosylation.
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