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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) 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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  • 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

Embodiment 1

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

[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 with 50ml of dichloromethane (DCM).

[0036] After 2.2 hours, the resin was washed with nitrogen-dimethylformamide (DMF) 3 times the volume of the resin, and then drained. This was repeated four times, and the resin was drained 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 test it with ninhydrin (Ninhydrin Nine Wells) method (two drops each of test A and test B, react at 100°C for 1 min), if the resin has color, it means that the deprotection is successful.

[0039] 5. ...

Embodiment 2

[0082] Antioxidant activity experiment of embodiment 2 bioactive peptides

[0083] The antioxidant activity of the biologically active polypeptide SKHSLDCVL 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 SKHSLDCVL 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 SKHSLDCVL.

[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 method...

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 SKHSLDCVL

[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 Jiao Tong University); milk-derived bioactive polypeptide SKHSLDCVL 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 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 C...

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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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