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Bioactive polypeptide KEPMIGVNQELA as well as 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 quality, and improving antioxidant activity.

Active Publication Date: 2018-05-11
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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  • Bioactive polypeptide KEPMIGVNQELA as well as preparation method and application thereof
  • Bioactive polypeptide KEPMIGVNQELA as well as preparation method and application thereof
  • Bioactive polypeptide KEPMIGVNQELA as well as preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0036] The artificial synthesis of embodiment 1 active peptide KEPMIGVNQELA

[0037] 1. Synthesis of Bioactive Peptides

[0038] 1. Weigh 3g of RINK resin (degree of substitution: 0.3mmol / g) into a 150ml reactor and soak with 50ml of dichloromethane (DCM).

[0039] 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.

[0040] 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.

[0041] 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.

[0042] ...

Embodiment 2

[0078] Antioxidant activity experiment of embodiment 2 bioactive peptides

[0079] 1. Determination of in vitro antioxidant activity of bioactive peptide KEPMIGVNQELA by [DPPH·] method

[0080] 1. Experimental reagents and instruments:

[0081] 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 KEPMIGVNQELA.

[0082] 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.

[0083] 2. Experimental method:

[0084] (1) 1mmol / L [DPPH·]methanol solution

[0085] 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...

Embodiment 3

[0119] Anti-aging activity experiment of embodiment 3 bioactive peptides

[0120] 1. Acute heat stress survival experiment of biologically active polypeptide KEPMIGVNQELA

[0121] 1. Experimental reagents and instruments:

[0122] 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 KEPMIGVNQELA obtained in Example 1.

[0123] 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 Experimental Equipment Co., Ltd. ; T...

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Abstract

The invention relates to the field of proteins, in particular to bioactive polypeptide KEPMIGVNQELA as well as a preparation method and an application thereof. The amino acid sequence of the bioactivepolypeptide KEPMIGVNQELA is LysGlu-Pro-Met-Ile-Gly-Val-Asn-Gln-Glu-LeuAla. In-vitro antioxidant experiments and in-vivo antiaging experiments prove that the polypeptide KEPMIGVNQELA has better antioxidant bioactivity and antiaging activity; on one hand, the bioactive polypeptide KEPMIGVNQELA has better antioxidant activity and can scavenge free radicals in the organism, and quality of life is improved; on the other hand, activity of an in-vivo peroxidase-resistant system can be improved, resistance of the organism to exogenous stimulation is enhanced, so that probabilities of aging and sickness of the organism are reduced, and the polypeptide has great significance in developing food, health care products and drugs with antioxidant and antiaging functions.

Description

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

Claims

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Application Information

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IPC IPC(8): C07K14/47C12N15/12A61K38/17A61P37/04A61P39/06A23L33/18A61K8/64A61Q19/08
CPCA23L33/18A23V2002/00A61K8/64A61K38/00A61Q19/08C07K14/4732A23V2250/55A23V2200/302
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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