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

A technology of biologically active polypeptides and drugs, applied in the field of protein, can solve problems such as human body risks, and achieve the effect of improving the quality of life, improving vitality, and good anti-aging activity

Active Publication Date: 2021-03-23
SHANGHAI JIAOTONG UNIV +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 dklaq and its preparation method and application
  • A kind of bioactive polypeptide dklaq and its preparation method and application
  • A kind of bioactive polypeptide dklaq and its preparation method and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

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

[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] 5. Weig...

Embodiment 2

[0078] Antioxidant activity experiment of embodiment 2 bioactive peptides

[0079] 1. Determination of in vitro antioxidant activity of bioactive peptide DKLAQ 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 Bioactive polypeptide DKLAQ.

[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 [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 immediately after mi...

Embodiment 3

[0118] Anti-aging activity experiment of embodiment 3 bioactive peptides

[0119] 1. Experiment on the effect of biologically active polypeptide DKLAQ on the lifespan of Caenorhabditis elegans

[0120] 1. Experimental reagents and instruments:

[0121] 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; 5-fluorouracil, Sigma Corporation, USA; the biologically active polypeptide DKLAQ obtained in Example 1.

[0122] 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 Preci...

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Abstract

The invention relates to the protein field, in particular to bioactive polypeptide DKLAQ as well as a preparation method and an application thereof. An amino acid sequence of the bioactive polypeptideDKLAQ is Asp-Lys-Leu-Ala-Gln. In vitro anti-oxidation experiments and in vivo anti-aging experiments prove that the polypeptide DKLAQ has the better anti-oxidation bioactivity and anti-aging activity, on one hand, the bioactive polypeptide DKLAQ has better anti-oxidation activity and can scavenge free radicals in organisms and improve the quality of life; on the other hand, the bioactive polypeptide DKLAQ can improve the vitality of in vivo anti-peroxidase systems and enhance functions of organisms for resisting exogenous stimulation to reduce the probability of organism aging, senescence anddisease and has very important significance in development of food, healthcare products and drugs with anti-oxidation and anti-aging functions.

Description

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

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K14/335C12N15/31A23L33/18A61K38/16A61K8/64A61P39/00A61P39/06A61Q19/08
CPCA23V2002/00A61K8/64A61K38/00A61Q19/08A23L33/18A61P39/00A61P39/06C07K14/335A23V2250/55A23V2200/302A23V2200/30
Inventor 张少辉李阜烁占文静陈静李婉如汪超范梦珠张伯宇
Owner SHANGHAI JIAOTONG UNIV
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