A kind of bioactive polypeptide vpitptlnre and its preparation method and application
A bioactive polypeptide and drug technology, applied in the field of protein, can solve problems such as human body risks, achieve the effects of reducing body aging, good anti-aging activity, and improving quality
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Embodiment 1
[0036] The artificial synthesis of embodiment 1 active peptide VPITPTLNRE
[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....
Embodiment 2
[0078] Antioxidant activity experiment of embodiment 2 bioactive peptides
[0079] 1. Determination of in vitro antioxidant activity of bioactive peptide VPITPTLNRE 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 Milk-derived bioactive polypeptide VPITPTLNRE.
[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 u...
Embodiment 3
[0118] Anti-aging activity experiment of embodiment 3 bioactive peptides
[0119] 1. Experiment on the effect of biologically active polypeptide VPITPTLNRE 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 milk-derived biologically active polypeptide VPITPTLNRE 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 osc...
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