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Bioactive polypeptide LVYPFPG as well as preparation method and application thereof

A technology of biologically active peptides and derivatives, applied in the field of proteins, can solve problems such as human risks, and achieve the effects of reducing injuries, regulating the body's immune activity, and enhancing antioxidant activity

Inactive Publication Date: 2017-09-26
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 LVYPFPG as well as preparation method and application thereof
  • Bioactive polypeptide LVYPFPG as well as preparation method and application thereof
  • Bioactive polypeptide LVYPFPG as well as preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

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

[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] 5. W...

Embodiment 2

[0082] Antioxidant activity experiment of embodiment 2 bioactive peptides

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

[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

[0122] Example 3 Bioactive Peptide Promoting Body Immunity Activity Experiment

[0123] 1. In vitro lymphocyte proliferation ability test of biologically active polypeptide LVYPFPG determined by MTT method

[0124] 1) Experimental materials and instruments:

[0125] 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 LVYPFPG 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 ).

[0126] Instruments: LRH-250F biochemical incubator, Shanghai Heng Technology Co.,...

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Abstract

The invention relates to the field of proteins, in particular to bioactive polypeptide LVYPFPG as well as a preparation method and application thereof. An amino acid sequence of the bioactive polypeptide LVYPFPG is Leu-Val-Tyr-Pro-Phe-Pro-Gly. In-vitro antioxidant experiments and in-vitro immune function promotion experiments prove that the polypeptide LVYPFPG has better antioxidant biological activity and an immune regulation function; on the one hand, the polypeptide LVYPFPG has the better antioxidant activity, thus being capable of removing free radicals in a human body and further improving the quality of life; on the other hand, the bioactive polypeptide LVYPFPG can enhance the in vitro proliferative capacity of lymphocytes and macrophages and promote the neutral red phagocytosis of the macrophages, thus improving the external pathogen infection resistance of the organism and reducing the morbidity of the organism; therefore, the bioactive polypeptide LVYPFPG has a very important significance for developing food, health care products and medicines which have the immune regulation function and the antioxidant function.

Description

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

Claims

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Application Information

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IPC IPC(8): C07K14/47C12N15/12A23L33/18A61K38/17A61P39/06A61P37/02A61K8/64A61Q19/08
CPCC07K14/4732A23L33/18A23V2002/00A61K8/64A61K38/00A61Q19/08A23V2200/324
Inventor 张少辉陈静李婉茹范梦珠袁芳豪李阜烁汪超沈鹏
Owner ZHEJIANG HUITAI LIFE HEALTH TECH CO LTD
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