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Bioactive peptide SKVLPVPEKAVPYPQ as well as preparation method and application thereof

A technology of biologically active polypeptides and derivatives, applied in the field of protein, can solve problems such as human body risks, achieve the effects of reducing morbidity, enhancing proliferation ability, good antioxidant activity and regulating body immunity activity

Active Publication Date: 2017-09-19
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 peptide SKVLPVPEKAVPYPQ as well as preparation method and application thereof
  • Bioactive peptide SKVLPVPEKAVPYPQ as well as preparation method and application thereof
  • Bioactive peptide SKVLPVPEKAVPYPQ 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 SKVLPVPEKAVPYPQ

[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.

[003...

Embodiment 2

[0082] Antioxidant activity experiment of embodiment 2 bioactive peptides

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

[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) ...

Embodiment 3

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

[0123] 1. MTT method to determine the in vitro lymphocyte proliferation ability of biologically active polypeptide SKVLPVPEKAVPYPQ

[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 SKVLPVPEKAVPYPQ 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 T...

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Abstract

The invention relates to the field of protein, in particular to a bioactive peptide SKVLPVPEKAVPYPQ as well as a preparation method and an application thereof. An amino acid sequence of the bioactive peptide SKVLPVPEKAVPYPQ is Ser-Lys-Val-Leu-Pro-Val-Pro-Glu-Lys-Ala-Val-Pro-Tyr-Pro-Gln. An in-vitro anti-oxidation experiment and an in-vitro immunity function promoting experiment prove that the bioactive peptide SKVLPVPEKAVPYPQ has better anti-oxidation biological activity and immunity regulation function, on one hand, the bioactive peptide SKVLPVPEKAVPYPQ has better anti-oxidation activity, can clear free radicals in an organism and improves life quality; on the other hand, by the aid of the bioactive peptide SKVLPVPEKAVPYPQ, in-vitro proliferation capacity of lymphocytes and macrophages can be enhanced, the phagocytosis capacity of the macrophages for neutral red is improved, the organism's capacity of resisting outside pathogen infection is improved, morbidity of the organism is reduced, and therefore, the bioactive peptide SKVLPVPEKAVPYPQ has quite great significance in development of food, healthcare products and drugs with an immune regulation function and an anti-oxidation function.

Description

technical field [0001] The present invention relates to the protein field, in particular to a biologically active polypeptide SKVLPVPEKAVPYPQ 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 th...

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
CPCA23L33/18A23V2002/00A61K8/64A61K38/00C07K14/4732A23V2200/324
Inventor 张少辉袁芳豪范梦珠汪超李阜烁陈静李婉茹
Owner ZHEJIANG HUITAI LIFE HEALTH TECH CO LTD
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