Bioactive polypeptide LPLP, and preparation and application thereof

A technology of biologically active polypeptides and derivatives, applied in the protein field, can solve problems such as human risks, and achieve the effect of promoting the body's immune activity, improving the ability, and good antioxidant activity.

Active Publication Date: 2013-08-07
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) wer

Method used

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

[0126] Antioxidant activity experiment of embodiment 2 bioactive polypeptides

[0127] The antioxidant activity of the biologically active polypeptide LPLP obtained in Example 1 was tested by free radical scavenging method (DPPH·method) and total antioxidant capacity method (Ferric Reducing Ability Power FRAP method).

[0128] 1. Determination of in vitro antioxidant activity of biologically active polypeptide LPLP by [DPPH·] method

[0129] 1) Experimental reagents and instruments

[0130] 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 LPLP obtained by fermentation of Lactobacillus helveticus (collected peak B sample).

[0131] Main instruments: Sunrise microplate reader, product of Tecan Company of Austria; 96-well cell culture plate, manufactured by Millipore Company of the United States; ana...

Embodiment 3

[0168] Example 3 Bioactive Polypeptide LPLP Immunity-Promoting Activity Experiment

[0169] 1. MTT method to determine the in vitro macrophage proliferation ability of biologically active polypeptide LPLP

[0170] 1) Experimental reagents and instruments

[0171] Reagent: experimental animal balb / c mouse (male 6-8 weeks old) Animal Experiment Center, College of Agriculture and Biology, Shanghai Jiaotong University; milk-derived bioactive polypeptide LPLP fermented by Lactobacillus helveticus; 3-(4,5-di Methylthiazole-2)-2,5-diphenyltetrazolium bromide (MTT) Amresco; LPS (lipopolysaccharide), purchased from Sigma; bovine serum albumin (Bovine Serum Albumin, BSA) Genebase; Triple solution, containing 10% SDS, 5% isobutanol and 0.012mol / L HCl in water.

[0172] Equipment: LRH-250F biochemical incubator Shanghai Heng Technology Co., Ltd.; GL-22M high-speed refrigerated centrifuge Shanghai Luxiangyi Centrifuge Instrument Co., Ltd.; Hera cell 150CO 2 Incubator Heraeus; Dragon Wel...

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Abstract

The invention relates to the field of protein, and specifically relates to a milk-derived bioactive polypeptide LPLP with in-vitro antioxidant activity and immunity improving function. An amino acid sequence of the polypeptide is Leu-Pro-Leu-Pro. In-vitro anti-oxidation and in-vitro immune function promotion experiments prove that the polypeptide LPLP has good anti-oxidation bioactivity and cellular immunity promotion activity. With the polypeptide LPLP, free radicals in bodies can be removed, such that harm of the free radicals to human bodies can be reduced; and the bioactive polypeptide LPLP can enhance macrophage phagocytic function, can improve body capacity for resisting external pathogens, can reduce body incidence rate, and does not cause body immunological rejection. The polypeptide has important significance in developing dairy products, health products and medicines with anti-oxidation function and immunity enhancing function.

Description

technical field [0001] The invention relates to the field of proteins, in particular to a bioactive polypeptide LPLP 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 polypeptides, some have special physiological functions and are called "biologically active polypeptides". [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 e...

Claims

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Application Information

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IPC IPC(8): C07K5/103C12N15/12C12P21/06C07K1/36C07K1/34C07K1/20A23L1/305A61K38/07A61P39/06A61P37/04C12R1/225
CPCC12P21/06A61K38/07C07K5/101C07K1/20A23L1/305C07K1/36C12R1/225A61K38/018C07K1/12C07K1/34A23L1/3053A23L33/18A61P37/04A61P39/06C12P21/02
Inventor 张少辉卢姗姗孙冠华马鎏镠周婕慧李锡安占东升
Owner ZHEJIANG HUITAI LIFE HEALTH TECH CO LTD
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