Bioactive polypeptide SKHSSLDCVL, and preparation method and application thereof

A technology of biologically active polypeptides and derivatives, applied in the field of proteins, can solve problems such as human risks, and achieve the effects of reducing injury, regulating immunity, and enhancing proliferation ability.

Active Publication Date: 2017-10-03
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) wer

Method used

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  • Bioactive polypeptide SKHSSLDCVL, and preparation method and application thereof
  • Bioactive polypeptide SKHSSLDCVL, and preparation method and application thereof
  • Bioactive polypeptide SKHSSLDCVL, and preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Example Embodiment

[0033] Example 1 Artificial synthesis of active peptide SKHSSLDCVL

[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 it with 50ml of dichloromethane (DCM).

[0036] After 2.2 hours, the resin was washed with 3 times the resin volume of nitrogen-dimethylformamide (DMF), and then drained. Repeat this four times to drain the resin for use.

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

[0038] 4. Take a small amount of resin and test it with ninhydrin (Ninewell Ninhydrin) method (two drops each for test A and test B, reaction at 100°C for 1 min), the resin is colored, indicating successful deprotection.

[0039] 5. Weigh appropriate amount of am...

Example Embodiment

[0082] Example 2 Antioxidant activity experiment of biologically active peptide

[0083] The free radical scavenging method (DPPH·method) and the total antioxidant capacity method (Ferric Reducing AbilityPower FRAP method) were used to test the antioxidant activity of the biologically active polypeptide SKHSSLDCVL obtained in Example 1.

[0084] 1. [DPPH·] method to determine the in vitro antioxidant activity of biologically active peptide SKHSSLDCVL

[0085] 1) Experimental reagents and instruments

[0086] Reagents: 1,1-Diphenyl-2-picrylhydrazyl (1,1-Diphenyl-2-picrylhydrazyl[DPPH·]), produced by Wako, Japan; methanol, provided by Shanghai Sinopharm; obtained in Example 1 Milk-derived biologically active polypeptide SKHSSLDCVL.

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

[0088] 2) Experimental method

[0089] (1) 1mmol / L[DPPH·] methanol solu...

Example Embodiment

[0123] Example 3 Experiments on the activity of biologically active peptides to promote immunity

[0124] 1. In vitro lymphocyte proliferation ability test of biologically active peptide SKHSSLDCVL by MTT method

[0125] 1) Experimental materials and instruments:

[0126] Reagents and materials: experimental animal balb / c mice (male 6-8 weeks old, Animal Experiment Center, School of Agriculture and Biology, Shanghai Jiaotong University); milk-derived biologically active polypeptide SKHSSLDCVL obtained in Example 1; mouse lymphocyte extract (Purchased from Soleibao); RPMI1640 medium (purchased from GIBCO); 3-(4,5-dimethylthiazole-2)-2,5-diphenyltetrazolium bromide (MTT, purchased From Amresco); Concanavalin (ConA, purchased from Sigma); Bovine Serum Albumin (BSA, purchased from Genebase); Pepsin (purchased from Sigma); Pancreatin (Corolase PP, purchased from AB) ).

[0127] Instruments: LRH-250F biochemical incubator, Shanghai Heng Technology Co., Ltd.; GL-22M high-speed refrigerated...

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Abstract

The invention relates to the field of protein, and in particular to a bioactive polypeptide SKHSSLDCVL, and a preparation method and application thereof. An amino acid sequence of the bioactive polypeptide SKHSSLDCVL is Ser-Lys-His-Ser-Ser-Leu-Asp-Cys-Val-Leu. An in-vitro antioxidant experiment and an in-vitro immunological function promoting experiment verify that the bioactive polypeptide SKHSSLDCVL has better antioxidant biological activity and a better immune modulating function; on one hand, the bioactive polypeptide SKHSSLDCVL has better antioxidant activity and is capable of removing free radicals in a body and increasing the quality of life; on the other hand, the bioactive polypeptide SKHSSLDCVL is capable of enhancing in-vitro proliferation ability of lymphocytes and macrophages, promoting the ability of swallowing neutral red by the macrophages, increasing the ability of defending external pathogen infection by the body and reducing the morbidity of the body, and has a very important significance in developing food, healthcare products and drugs having an immune modulating function and an antioxidant function.

Description

technical field [0001] The invention relates to the field of proteins, in particular to a biologically active polypeptide SKHSLDCVL 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 protecti...

Claims

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Application Information

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IPC IPC(8): C07K14/79C12N15/12A23L33/18A61K38/17A61K8/64A61P39/06A61P37/02A61Q19/08
CPCA23L33/18A23V2002/00A61K8/64A61K38/00A61Q19/08C07K14/79A23V2200/324
Inventor 张少辉李阜烁汪超袁芳豪范梦珠陈静李婉茹
Owner ZHEJIANG HUITAI LIFE HEALTH TECH CO LTD
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