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Biological active polypeptide LPPLL, preparation method and application thereof

A technology of biologically active polypeptides and derivatives, applied in the field of proteins, can solve problems such as human body risks, and achieve the effects of reducing body aging, improving antioxidant activity and improving quality.

Inactive Publication Date: 2018-04-10
ZHEJIANG HUITAI LIFE HEALTH TECH CO LTD +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] 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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  • Biological active polypeptide LPPLL, preparation method and application thereof
  • Biological active polypeptide LPPLL, preparation method and application thereof
  • Biological active polypeptide LPPLL, preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0038] The artificial synthesis of embodiment 1 active peptide LPPLL

[0039] 1. Synthesis of Bioactive Peptides

[0040] 1. Weigh 3g of RINK resin (degree of substitution: 0.3mmol / g) into a 150ml reactor and soak with 50ml of dichloromethane (DCM).

[0041] 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.

[0042] 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.

[0043] 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.

[0044] 5. Weig...

Embodiment 2

[0080] Antioxidant activity experiment of embodiment 2 bioactive peptides

[0081] 1. Determination of in vitro antioxidant activity of biologically active peptide LPPLL by [DPPH·] method

[0082] 1. Experimental reagents and instruments:

[0083] 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 LPPLL.

[0084] Main instruments: Sunrise microplate reader, product of Tecan Company, Austria; 96-well cell culture plate, manufactured by Millipore Company of the United States; analytical balance, product of Meitelei-tolido Company.

[0085] 2. Experimental method:

[0086] (1) 1mmol / L [DPPH·]methanol solution

[0087] Weigh 0.349 mg of [DPPH·] with an analytical balance and dissolve it in 1 mL of methanol solution to prepare a 1 mmol / L [DPPH·] methanol solution, store it in the dark on tin foil, and ...

Embodiment 3

[0122] Anti-aging activity experiment of embodiment 3 bioactive peptides

[0123] 1. Experiment on the effect of biologically active polypeptide LPPLL on the reproductive ability of Caenorhabditis elegans

[0124] 1. Experimental reagents and instruments:

[0125] Reagents: Caenorhabditis elegans, Institute of Integrating Chinese and Western Medicine affiliated to Fudan University; Escherichia coli strain E.coli OP50, Institute of Integrating Chinese and Western Medicine affiliated to Fudan University; Agar powder, Sinopharm Chemical Reagent Co., Ltd.; Yeast powder, Sinopharm Chemical Reagent Co., Ltd. Company; the milk-derived biologically active polypeptide LPPLL obtained in Example 1.

[0126] Instruments and equipment: Likang RO15 pure water system, Likang Biomedical Technology Co., Ltd.; G136T Zealway intelligent high-temperature sterilization pot, Xiamen Zhiwei Instrument Technology Co., Ltd.; THZ-32 desktop constant temperature oscillator, Shanghai Precision Experiment...

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Abstract

The invention relates to the field of proteins, and in particular relates to a biological active polypeptide LPPLL as well as a preparation method and application thereof. The amino acid sequence of the biological active polypeptide LPPLL is Leu-Pro-Pro-Leu-Leu. In-vitro anti-oxidation experiments and in-vivo anti-aging experiments show that the polypeptide LPPLL has relatively good antioxidant biological activity and anti-aging activity, and on one hand, the biological active polypeptide LPPLL disclosed by the invention has relatively good antioxidant activity, free radicals in the body can be removed, and the quality of life can be improved; on the other hand, the activity of the antioxidant enzyme system in the body can be improved, so that the function of resisting exogenous stimulation of the organism is enhanced, so that the probability of aging and disease of the organism is reduced, and the invention has very important significance for developing foods, health products and medicines with anti-oxidation functions and anti-aging functions.

Description

technical field [0001] The invention relates to the field of proteins, in particular to a biologically active polypeptide LPPLL 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] It is especially important to find safe bioactive peptides in natural food sources. In recent years, it has been found that some food-derived polypeptides have good biological activity, such as corn short peptides, soybean peptides, and mil...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/47C12N15/12A23L33/18A61K38/17A61K8/64A61P39/06A61Q19/08
Inventor 张少辉张伯宇林学海汪超李云飞李阜烁范梦珠李婉如
Owner ZHEJIANG HUITAI LIFE HEALTH TECH CO LTD
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