A kind of bioactive polypeptide vavvkkgsnfq and its preparation method and application

A technology of biologically active peptides and drugs, applied in the field of protein, can solve problems such as human body risks, and achieve the effects of reducing damage, improving antioxidant activity, and improving the ability of human body

Active Publication Date: 2020-04-10
ZHEJIANG HUITAI LIFE HEALTH TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] Some early synthetic antioxidants such as butyl hydroxyanisole (BHA) and 2,6-di-tert-butyl-4-methylphenol (BHT) were used in food as lipid antioxidants, but these artificial Synthetic additives are potentially dangerous to the human body

Method used

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  • A kind of bioactive polypeptide vavvkkgsnfq and its preparation method and application
  • A kind of bioactive polypeptide vavvkkgsnfq and its preparation method and application
  • A kind of bioactive polypeptide vavvkkgsnfq and its preparation method and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] The artificial synthesis of embodiment 1 active peptide VAVVKKGSNFQ

[0034] 1. Synthesis of Bioactive Peptides

[0035] 1. Weigh 3g of RINK resin (degree of substitution: 0.3mmol / g) into a 150ml reactor and soak in 50ml of dichloromethane (DCM).

[0036] 2. After 2 hours, wash the resin with nitrogen-dimethylformamide (DMF) that is 3 times the volume of the resin, and then drain it. Repeat this four times, and drain the resin for use.

[0037] 3. Add a certain amount of 20% piperidine (piperidine / DMF=1:4, v:v) into the reactor, and shake it on a decolorizing shaker for 20 minutes to remove the Fmoc protecting group on the resin. After deprotection, wash four times with 3 times the resin volume of DMF, and then drain.

[0038] 4. Take a small amount of resin and use ninhydrin (Nine Wells Ninhydrin) method to test it (Two drops each of Test A and Test B, react at 100°C for 1 min), if the resin is colored, it means that the deprotection is successful.

[0039] 5. Weigh...

Embodiment 2

[0082] Antioxidant activity experiment of embodiment 2 bioactive peptides

[0083]The antioxidant activity of the biologically active polypeptide VAVVKKGSNFQ obtained in Example 1 was tested by free radical scavenging method (DPPH·method) and total antioxidant capacity method (Ferric Reducing AbilityPower FRAP method).

[0084] 1. Determination of in vitro antioxidant activity of bioactive peptide VAVVKKGSNFQ by [DPPH·] method

[0085] 1) Experimental reagents and instruments

[0086] Reagent: 1,1-diphenyl-2-trinitrophenylhydrazine (1,1-Diphenyl-2-picrylhydrazyl[DPPH·]), produced by Wako Company of Japan; Methanol, provided by Shanghai Sinopharm Company; obtained in Example 1 Milk-derived bioactive polypeptide VAVVKKGSNFQ.

[0087] Main instruments: Sunrise microplate reader, product of Tecan Company of Austria; 96-well cell culture plate, manufactured by Millipore Company of the United States; analytical balance, product of Meitelei-tolido Company.

[0088] 2) Experimental...

Embodiment 3

[0123] Example 3 Bioactive Peptide Promoting Body Immunity Activity Experiment

[0124] 1. MTT method to determine the in vitro lymphocyte proliferation ability of biologically active polypeptide VAVVKKGSNFQ

[0125] 1) Experimental materials and instruments:

[0126] Reagents and materials: experimental animals balb / c mice (male 6-8 weeks old, Animal Experiment Center, School of Agriculture and Biology, Shanghai Jiaotong University); milk-derived bioactive polypeptide VAVVKKGSNFQ obtained in Example 1; mouse lymphocyte extract (purchased from Suo Laibao); RPMI1640 medium (purchased from GIBCO); 3-(4,5-dimethylthiazole-2)-2,5-diphenyltetrazolium bromide (MTT, purchased from Amresco Company); Concanavalin (ConA, purchased from Sigma Company); bovine serum albumin (BSA, purchased from Genebase Company); pepsin (purchased from Sigma Company); pancreatin (Corolase PP, purchased from AB Company ).

[0127] Instruments: LRH-250F biochemical incubator, Shanghai Heng Technology Co....

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Abstract

The invention relates to the field of proteins, in particular to bioactive polypeptide VAVVKKGSNFQ as well as a preparation method and application thereof. An amino acid sequence of the bioactive polypeptide VAVVKKGSNFQ is Val-Ala-Val-Val-Lys-Lys-Gly-Ser-Asn-Phe-Gln. In-vitro antioxidant experiments and in-vitro immune function promotion experiments prove that the polypeptide VAVVKKGSNFQ has better antioxidant biological activity and an immune regulation function; on the one hand, the polypeptide VAVVKKGSNFQ has the better antioxidant activity, thus being capable of removing free radicals in a human body and further improving the quality of life; on the other hand, the bioactive polypeptide VAVVKKGSNFQ can enhance the in vitro proliferative capacity of lymphocytes and macrophages and promote the increment of nitrogen monoxide induction amount of the macrophages, thus improving the external pathogen infection resistance of the organism and reducing the morbidity of the organism; therefore, the bioactive polypeptide VAVVKKGSNFQ has a very important significance for developing food, health care products and medicines which have the immune regulation function and the antioxidant function.

Description

technical field [0001] The present invention relates to the protein field, in particular to a biologically active polypeptide VAVVKKGSNFQ and its preparation method and application. Background technique [0002] During the fermentation process of milk by lactic acid bacteria, a part of protein in milk is metabolized and utilized by lactic acid bacteria, and a series of physiological and biochemical reactions occur, making the protein into polypeptides or free amino acids, which are digested and absorbed by the human body or absorbed and transported by small intestinal epithelial cells directly into the blood circulation of the human body. Among these peptides, some have special physiological functions and are called "bioactive peptides". [0003] Oxidation reaction and oxidative metabolism are crucial for food and human body, and free radicals and reactive oxygen species cause a series of oxidation reactions. When excessive free radicals are formed, they will exceed the pr...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K14/79C12N15/12A23L33/18A61K38/17A61K8/64A61P39/06A61P37/02A61Q19/08
CPCA23L33/18A23V2002/00A61K8/64A61K38/00C07K14/79A23V2200/324
Inventor 张少辉黄犇余佳琪王凯吴静范梦珠袁芳豪李阜烁汪超
Owner ZHEJIANG HUITAI LIFE HEALTH TECH CO LTD
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