Bioactive polypeptide VVPPFL, and preparation method and application thereof
A technology of biologically active polypeptides and derivatives, applied in the field of protein, can solve the problems of poor digestibility and absorption, and can achieve the effect of reducing the aging of the body, improving the vitality, and improving the quality of life.
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Embodiment 1
[0038] The artificial synthesis of embodiment 1 active peptide VVPPFL
[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 bioactive peptide VVPPFL 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 polypeptide VVPPFL.
[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 immedia...
Embodiment 3
[0120] Anti-aging activity experiment of embodiment 3 bioactive peptides
[0121] 1. Experiment on the effect of biologically active polypeptide VVPPFL on the lifespan 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; 5-fluorouracil, Sigma Corporation of the United States; the milk-derived biologically active polypeptide VVPPFL 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 tempera...
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