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Bioactive polypeptide ATLEDSPEVI as well as preparation method and application thereof

A technology of biologically active peptides and derivatives, applied in the field of protein, can solve problems such as human body risks, achieve the effects of reducing injuries, improving human ability, and enhancing proliferation ability

Active Publication Date: 2017-09-08
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 ATLEDSPEVI as well as preparation method and application thereof
  • Bioactive polypeptide ATLEDSPEVI as well as preparation method and application thereof
  • Bioactive polypeptide ATLEDSPEVI as well as preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0036] Example 1 Synthesis of Active Peptide ATLEDSPEVI

[0037] 1. Synthesis of Bioactive Peptides

[0038] 1. Weigh 3 g of RINK resin (degree of substitution: 0.3 mmol / g) into a 150 ml reactor, and soak it with 50 ml of dichloromethane (DCM).

[0039] After 2.2 hours, the resin was washed with nitrogen-dimethylformamide (DMF) 3 times the resin volume, and then drained. This was repeated four times, and the resin was drained for use.

[0040] 3. A certain amount of 20% piperidine (piperidine / DMF=1:4, v:v) was added to the reactor, and it was shaken on a decolorizing shaker for 20 minutes to remove the Fmoc protecting group on the resin. After deprotection, it was washed four times with 3 times the resin volume of DMF, and then drained.

[0041] 4. Take a small amount of resin and test it by ninhydrin (Ninhydrin hydrate) method (two drops each of test A and test B, react at 100°C for 1 min), the resin is colored, indicating that the deprotection is successful.

[0042] 5. W...

Embodiment 2

[0078] Example 2 Antioxidant activity test of bioactive peptides

[0079] The antioxidant activity of the biologically active polypeptide ATLEDSPEVI obtained in Example 1 was tested by using the free radical scavenging method (DPPH· method) and the total antioxidant capacity method (Ferric Reducing AbilityPower FRAP method).

[0080] 1. The in vitro antioxidant activity of the bioactive peptide ATLEDSPEVI was determined by the [DPPH·] method

[0081] 1) Experimental reagents and instruments

[0082] Reagent: 1,1-Diphenyl-2-trinitrophenylhydrazine (1,1-Diphenyl-2-picrylhydrazyl[DPPH·]), produced by Wako Company in Japan; methanol, provided by Shanghai Sinopharm Company; obtained in Example 1 Milk-derived biologically active polypeptide ATLEDSPEVI.

[0083] Main instruments: Sunrise microplate reader, product of Tecan Company, Austria; 96-well cell culture plate, manufactured by Millipore Company of the United States; analytical balance, product of Meitelei-tolido Company.

...

Embodiment 3

[0118] Example 3 Experiment on the promotion of immunity of biologically active peptides

[0119] 1. Determination of in vitro lymphocyte proliferation ability of biologically active polypeptide ATLEDSPEVI by MTT method

[0120] 1) Experimental materials and instruments:

[0121] 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 ATLEDSPEVI obtained in Example 1; mouse lymphocyte extract (purchased from Soleobor Company); RPMI1640 medium (purchased from GIBCO Company); 3-(4,5-Dimethylthiazole-2)-2,5-diphenyltetrazolium bromide (MTT, purchased from from Amresco Company); Concanavalin (ConA, from Sigma Company); Bovine Serum Albumin (BSA, from Genebase Company); Pepsin (from Sigma Company); Pancreatin (Corolase PP, from AB Company) ).

[0122] Instruments: LRH-250F biochemical incubator, Shanghai Heng Technology Co...

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Abstract

The invention relates to the field of proteins and particularly relates to bioactive polypeptide ATLEDSPEVI as well as a preparation method and application thereof. The amino acid sequence of the bioactive polypeptide ATLEDSPEVI is Ala-Thr-Leu-Glu-Asp-Ser-Pro-Glu-Val-Ile. In-vitro anti-oxidation experiments and in-vitro immune function promotion experiments verify that the polypeptide ATLEDSPEVI has better antioxidant biological activity and immune modulating function, so that on one hand, the polypeptide ATLEDSPEVI has better antioxidant activity, so that free radicals in a body can be removed, and the quality of life is improved; on the other hand, the polypeptide ATLEDSPEVI can enhance the in-vitro multiplication capacity of lymphocytes and macrophages, the macrophages are promoted to secrete cytokines, the capability of the body in defense for external pathogen infection is enhanced, and the incidence rate of the body is reduced. Therefore, the polypeptide ATLEDSPEVI has great significance in development of food, health-care products and drugs with immune modulating function and antioxidant function.

Description

technical field [0001] The invention relates to the field of protein, in particular to a biologically active polypeptide ATLEDSPEVI 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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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/47C12N15/12A23L33/18A61K38/17A61K8/64A61P39/06A61P37/02A61Q19/08
CPCA23L33/18A23V2002/00A61K8/64A61K38/00A61Q19/08C07K14/4732A23V2200/324
Inventor 张少辉李婉茹陈静汪超李阜烁袁芳豪范梦珠程志才
Owner ZHEJIANG HUITAI LIFE HEALTH TECH CO LTD
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