Biologically active polypeptide WNIPMGLIVNQ 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, enhancing proliferation ability, and improving ability

Active Publication Date: 2017-09-15
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) ...

Method used

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  • Biologically active polypeptide WNIPMGLIVNQ and preparation method and application thereof
  • Biologically active polypeptide WNIPMGLIVNQ and preparation method and application thereof
  • Biologically active polypeptide WNIPMGLIVNQ and preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

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

[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] 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 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] ...

Embodiment 2

[0082] Antioxidant activity experiment of embodiment 2 bioactive peptides

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

[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) Ex...

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 WNIPMGLIVNQ

[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 WNIPMGLIVNQ 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 Technolog...

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Abstract

The present invention relates to the field of protein, and in particular relates to biologically active polypeptide WNIPMGLIVNQ and a preparation method and the application thereof, and the amino acid sequence of the biologically active polypeptide WNIPMGLIVNQ is Trp-Asn-Ile-Pro-Met-Gly-Leu-Ile-Val-Asn-Gln. In vitro antioxidant experiments and in vitro immune function promotion experiments verify the biologically active polypeptide WNIPMGLIVNQ has good antioxidant biological activity and immunity regulating function, on the one hand, the biologically active polypeptide WNIPMGLIVNQ has good antioxidant activity to remove free radicals in the body and improve the quality of life; on the other hand, the biologically active polypeptide WNIPMGLIVNQ can enhance the in-vitro proliferation ability of lymphocytes and macrophages to promote the increase of macrophage induced nitric oxide, the biologically active polypeptide WNIPMGLIVNQ can improve the external pathogen infection resistance of the body and reduce the disease incidence of the body, and has very important significance for development of food, health care products and drugs with immune function regulating and antioxidant function.

Description

technical field [0001] The invention relates to the field of protein, in particular to a biologically active polypeptide WNIPMGLIVNQ 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 protect...

Claims

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Application Information

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IPC IPC(8): C07K14/79C12N15/12A23L33/18A61K38/17A61K8/64A61P39/06A61P37/02A61Q19/08
CPCA23L33/18A23V2002/00A61K8/64A61K38/00C07K14/79A23V2200/324
Inventor 张少辉王乾宇李文瑾吴静王凯袁芳豪范梦珠汪超李阜烁
Owner ZHEJIANG HUITAI LIFE HEALTH TECH CO LTD
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