Biologically active polypeptide QQPVLGPVRGP and preparation method and application thereof
A technology of biologically active polypeptides and derivatives, applied in the protein field, can solve problems such as human body risks, and achieve the effects of reducing body aging, improving anti-aging activity and improving quality.
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Embodiment 1
[0038] The artificial synthesis of embodiment 1 active peptide QQPVLGPVRGP
[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...
Embodiment 2
[0080] Antioxidant activity experiment of embodiment 2 bioactive peptides
[0081] 1. Determination of in vitro antioxidant activity of bioactive peptide QQPVLGPVRGP 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 QQPVLGPVRGP.
[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 [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 i...
Embodiment 3
[0122] Anti-aging activity experiment of embodiment 3 bioactive peptides
[0123] 1. Experiment on the effect of biologically active polypeptide QQPVLGPVRGP 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 QQPVLGPVRGP 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 Precisio...
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