Bioactive polypeptide QPEVMGVSKVKEAMAPKQKEMPFPKY as well as preparation and application thereof
A technology of biologically active peptides and derivatives, applied in the field of protein, can solve problems such as human body risks, and achieve the effect of enhancing function, improving vitality, and good anti-aging activity
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Embodiment 1
[0037] The artificial synthesis of embodiment 1 active peptide QPEVMGVSKVKEAMAPKQKEMPFPKY
[0038] 1. Synthesis of Bioactive Peptides
[0039] 1. Weigh 3g of RINK resin (degree of substitution: 0.3mmol / g) into a 150ml reactor and soak in 50ml of dichloromethane (DCM).
[0040] 2. After 2 hours, wash the resin with nitrogen-dimethylformamide (DMF) that is 3 times the volume of the resin, and then drain it. Repeat this four times, and drain the resin for use.
[0041] 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.
[0042] 4. Take a small amount of resin and use ninhydrin (Nine Wells Ninhydrin) method to test it (Two drops each of Test A and Test B, react at 100°C for 1 min), if the resin is colored, it means that the deprotection is successful.
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Embodiment 2
[0080] Antioxidant activity experiment of embodiment 2 bioactive peptides
[0081] 1. Determination of in vitro antioxidant activity of bioactive peptide QPEVMGVSKVKEAMAPKQKEMPFPKY 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 peptide QPEVMGVSKVKEAMAPKQKEMPFPKY.
[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...
Embodiment 3
[0120] Anti-aging activity experiment of embodiment 3 bioactive peptides
[0121] 1. Experiment on the effect of biologically active polypeptide QPEVMGVSKVKEAMAPKQKEMPFPKY on the exercise ability of Caenorhabditis elegans
[0122] 1. Experimental reagents and instruments:
[0123] 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 QPEVMGVSKVKEAMAPKQKEMPFPKY obtained in Example 1.
[0124]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 osci...
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