Bioactive polypeptide QEPV, and preparation and application thereof

A bioactive polypeptide and polypeptide technology, applied in the field of protein, can solve the problems of unreachable and loss of physiological activity, etc., and achieve the effect of increasing the amount of induction, reducing damage, and improving the ability

Active Publication Date: 2013-04-03
ZHEJIANG HUITAI LIFE HEALTH TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In the complex environment of the human gastrointestinal tract, peptides are easily degraded by a series of enzymes such

Method used

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  • Bioactive polypeptide QEPV, and preparation and application thereof
  • Bioactive polypeptide QEPV, and preparation and application thereof
  • Bioactive polypeptide QEPV, and preparation and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0059] The preparation of embodiment 1 active peptide QEPVL

[0060] 1. Preparation of fermented milk

[0061] 1) Lactobacillus helveticus fermented milk

[0062] Use skim milk powder (New Zealand NZMP brand skim milk powder) and water to prepare 12wt% skim milk (12wt% skim milk is prepared by adding 12g skim milk powder to 88g water, the same below). Under aseptic conditions, the tricyclic colony of Lactobacillus helveticus (CICC6024) was picked, added to sterilized 12wt% skim milk, and stirred evenly. After inoculation, seal with aluminum foil to prevent contamination. Place in an incubator at 37°C for 19 hours. After the cultivation is over, the curd is stirred evenly under aseptic conditions to complete the activation of the Lactobacillus helveticus and prepare the starter for preparing the Lactobacillus helveticus fermented milk.

[0063] Inoculate 10mL of the prepared Lactobacillus helveticus starter into 500mL of sterilized 12wt% skim milk (inoculation rate 2v / v%), ...

Embodiment 2

[0136] Preparation and Confirmation of Example 2 Bioactive Peptide QEPV

[0137] 1. Enzymatic hydrolysis of bioactive peptide QEPVL by simulating gastrointestinal digestion in vitro

[0138] The simulated gastrointestinal digestion experiment is mainly divided into two steps. First, use sterilized deionized water to prepare a bioactive peptide QEPVL solution with a concentration of 500 μg / mL, and add pepsin (purchased from Sigma Company) to the QEPVL solution with a concentration of 500 μg / mL at a ratio of 20 mg of pepsin per gram of QEPVL. The pH value of the reaction solution was brought to 2.0, and incubated in a constant temperature water bath at 37°C for 90 minutes; then the pH value of the reaction solution was adjusted to 7.5, and trypsin (Corolase PP, purchased from AB Company in Germany) was added at the ratio of adding trypsin per gram of QEPVL 40 mg, kept in a constant temperature water bath at 37°C for 150 minutes; finally placed in a water bath at 95°C for 5 minu...

Embodiment 3

[0152] Antioxidant activity experiment of embodiment 3 bioactive peptide QEPV

[0153] The antioxidant activity of the biologically active polypeptide QEPV obtained in Example 1 was tested by free radical scavenging method (DPPH·method) and total antioxidant capacity method (Ferric Reducing Ability Power FRAP method).

[0154] 1. Determination of in vitro antioxidant activity of bioactive peptide QEPV by [DPPH·] method

[0155] 1) Experimental reagents and instruments

[0156] Reagent: 1,1-diphenyl-2-trinitrophenylhydrazine (1,1-Diphenyl-2-picrylhydrazyl[DPPH·]), produced by Wako Company, Japan; Methanol, provided by Shanghai Sinopharm Company; obtained in Example 2 Milk-derived bioactive polypeptide QEPV.

[0157] 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.

[0158] 2) Experimental method

[0159]...

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Abstract

The invention relates to the field of proteins and specifically relates to a milk-derived bioactive polypeptide with in-vitro antioxidant activity and capability of enhancing immunity. The amino acid sequence is Gln-Glu-Pro-Val. In-vitro anti-oxidation experiments and in-vitro immune function promotion experiments verify that the polypeptide QEPV has better antioxidant bioactivity and activity of promoting cellular immunity; on the one hand, free radicals in an organism can be cleared, and the harm of the free radicals to a human body can be reduced; and on the other hand, the bioactive polypeptide QEPV can enhance the immunity, enhance the multiplication capacity of lymphocytes and macrophages, increase the induction quantity of nitrogen oxide from the macrophages, promote the secretion of cell factors by the macrophages, improve the capability of resisting external pathogen infection of the organism, reduce the disease incidence of the organism and realize the great significance for developing milk products, health care products and medicaments with anti-oxidation function and immunity-enhancing function.

Description

technical field [0001] The invention relates to the field of proteins, in particular to a biologically active polypeptide QEPV and its preparation 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] Immunoactive peptides are a class of bioactive peptides obtained from milk for the first time after the discovery of opioid peptides and their physiological activity has been proved. In 1981, Jolles et al. discovered for the first time that a hexap...

Claims

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Application Information

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IPC IPC(8): C07K5/103C07K1/20C12N15/11C12P21/02C12P21/06A23L1/29A61K38/07A61P39/06A61P37/04C12R1/225A23L33/00
CPCC12P21/06C07K1/36C07K1/12A23L1/305C07K1/20C12R1/225A23L1/3053C07K1/34A61K38/00C07K5/1021A61K38/07A23L33/18A61P37/04A61P39/06C07K14/4732
Inventor 张少辉卢姗姗马鎏镠孙冠华崔磊金赢凯徐海红
Owner ZHEJIANG HUITAI LIFE HEALTH TECH CO LTD
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