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Bioactive polypeptide DELQ and preparation method and application thereof

A bioactive polypeptide and drug technology, applied in the protein field, can solve problems such as human risk, achieve the effect of promoting the body's immune activity, improving the ability to resist external pathogen infection, and reducing damage

Active Publication Date: 2014-12-24
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 DELQ and preparation method and application thereof
  • Bioactive polypeptide DELQ and preparation method and application thereof
  • Bioactive polypeptide DELQ and preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0050] The preparation of embodiment 1 active peptide

[0051] 1. Preparation of fermented milk

[0052] 1) Lactobacillus helveticus fermented milk

[0053] Use skimmed milk powder (New Zealand NZMP brand skimmed milk powder) and water to prepare 12wt% skimmed milk (12g skimmed milk powder is added to 88g water, the same below). Under aseptic conditions, the tricyclic colony of Lactobacillus helveticus (CICC6024) was picked, added into sterilized 12wt% skim milk, and stirred evenly under aseptic conditions. 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.

[0054] Inoculate 10mL of the prepared Lactobacillus helveticus starter into 500mL of sterilized 12wt% skim milk (inoculation rate 2v / v...

Embodiment 2

[0127] Antioxidant activity experiment of embodiment 2 bioactive peptides

[0128] The antioxidant activity of the biologically active polypeptide DELQ obtained in Example 1 was tested by free radical scavenging method (DPPH·method) and total antioxidant capacity method (Ferric Reducing Ability Power FRAP method).

[0129] 1. Determination of in vitro antioxidant activity of bioactive peptide DELQ by [DPPH·] method

[0130] 1) Experimental reagents and instruments

[0131] 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 DELQ (collected peak C sample) obtained from fermentation of Lactobacillus helveticus.

[0132] 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,...

Embodiment 3

[0167] Example 3 Bioactive Peptide Promoting Body Immunity Activity Experiment

[0168] In vitro Lymphocyte Proliferation Ability Test of Bioactive Polypeptide DELQ by MTT Method

[0169] 1) Experimental materials and instruments:

[0170] 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 DELQ fermented by Lactobacillus helveticus; mouse lymphocyte extraction RPMI1640 medium (purchased from GIBCO); 3-(4,5-dimethylthiazole-2)-2,5-diphenyltetrazolium bromide (MTT, purchased from Amresco Company); Concanavalin (ConA, purchased from Sigma Company); bovine serum albumin (BSA, purchased from Genebase Company); pepsin (purchased from Sigma Company); pancreatin (CorolasePP, purchased from AB Company ).

[0171] Instruments: LRH-250F biochemical incubator, Shanghai Heng Technology Co., Ltd.; GL-22M high-speed refrigerated centrif...

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Abstract

Provided is a milk-derived biologically active polypeptide having an amino acid sequence of DELQ, which has in-vitro antioxidation activity and promotes immune activity of human body. In-vitro antioxidation and in-vitro immunity promotion experiments prove that the peptide has good biological antioxidation activity, and has the function of enhancing cellular immunity.

Description

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

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K5/113C12N15/12C12P21/06C07K1/36C07K1/34C07K1/20A23L1/305A61K38/07A61P39/06A61P37/04C12R1/225
CPCC07K5/1021A23V2200/00C12P21/06A23L1/3053C12P21/02A61K38/00A23L1/3056A23L33/18A23L33/19A23V2002/00A61P37/04A61P39/06A23V2250/5424A23V2200/324
Inventor 张少辉卢姗姗孙冠华马鎏镠周婕慧李锡安占东升高扬
Owner ZHEJIANG HUITAI LIFE HEALTH TECH CO LTD
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