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A kind of bioactive polypeptide enpraf and its preparation method and application

A bioactive peptide and drug technology, applied in the protein field, can solve problems such as human body risks, and achieve the effects of reducing body aging, improving vitality, and improving anti-aging activity.

Active Publication Date: 2020-06-09
SHANGHAI BOHUI BIOLOGICAL 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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  • A kind of bioactive polypeptide enpraf and its preparation method and application
  • A kind of bioactive polypeptide enpraf and its preparation method and application
  • A kind of bioactive polypeptide enpraf and its preparation method and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

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

[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. Wei...

Embodiment 2

[0080] Antioxidant activity experiment of embodiment 2 bioactive peptides

[0081] 1. Determination of in vitro antioxidant activity of biologically active peptide ENPRAF 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 Biologically active polypeptide ENPRAF.

[0084] 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.

[0085] 2. Experimental method:

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

[0087] Weigh 0.349 mg [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 use ...

Embodiment 3

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

[0123] 1. Experiment on the effect of biologically active polypeptide ENPRAF 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 biologically active polypeptide ENPRAF 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 Experimental Equipmen...

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Abstract

The invention relates to the field of proteins, in particular to a bioactive polypeptide ENPRAF, a preparation method thereof and an application of the bioactive polypeptide. The amino acid sequence of the bioactive polypeptide ENPRAF is Glu-Asn-Pro-Arg-Ala-Phe. An in-vitro antioxidant experiment and an in-vivo anti-aging experiment verify that the polypeptide ENPRAF has good antioxidant biological activity and anti-aging activity. On one hand, the bioactive polypeptide ENPRAF has good antioxidant activity, free radicals in a body can be removed, and life quality can be improved. On the otherhand, in-vivo anti-peroxidase system activity can be improved, the exogenous stimulation resisting function of the body is enhanced, so that the probability of aging, senescence and sickness of the body is reduced, and the bioactive polypeptide ENPRAF is of great significance for developing food, health care products and medicines with antioxidant functions and anti-aging functions.

Description

technical field [0001] The present invention relates to the protein field, in particular to a biologically active polypeptide ENPRAF 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. Lactic acid bacteria cells also have some self-synthesized protein polypeptide fragments for bacterial growth and utilization. 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...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K14/335C12N15/31A61K38/16A61P29/00A61P39/06A23L33/18A61K8/64A61Q19/00A61Q19/08
CPCA23L33/18A23V2002/00A61K8/64A61K38/00A61P29/00A61P39/06A61Q19/00A61Q19/08C07K14/335A23V2200/302A23V2200/30A23V2250/55
Inventor 张少辉占文静张伯宇汪超李阜烁范梦珠陈静李婉茹
Owner SHANGHAI BOHUI BIOLOGICAL TECH CO LTD
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