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

A technology of biologically active polypeptides and drugs, applied in the protein field, can solve problems such as human risks, and achieve the effects of reducing injury, enhancing proliferation ability, and regulating the body's immune activity.

Active Publication Date: 2020-04-14
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 atledspevi and its preparation method and application
  • A kind of bioactive polypeptide atledspevi and its preparation method and application
  • A kind of bioactive polypeptide atledspevi and its preparation method and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0036] The artificial synthesis of embodiment 1 active peptide ATLEDSPEVI

[0037] 1. Synthesis of Bioactive Peptides

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

[0039] 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.

[0040] 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.

[0041] 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.

[0042] 5....

Embodiment 2

[0078] Example 2 Antioxidant Activity Experiment of Bioactive Peptides

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

[0080] 1. Determination of in vitro antioxidant activity of biologically active peptide ATLEDSPEVI by [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 of Japan; Methanol, provided by Shanghai Sinopharm Company; obtained in Example 1 Milk-derived bioactive polypeptide ATLEDSPEVI.

[0083] 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.

[0084] 2) Experim...

Embodiment 3

[0118] Example 3 Experiment on the Immunity-Promoting Activity of Bioactive Peptides

[0119] 1. MTT method to determine the in vitro lymphocyte proliferation ability of biologically active polypeptide ATLEDSPEVI

[0120] 1) Experimental materials and instruments:

[0121] 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 ATLEDSPEVI 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 ).

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

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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 Patents(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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