A kind of bioactive polypeptide vpitptlnr and its preparation method and application
A bioactive polypeptide and drug technology, applied in the protein field, can solve problems such as human body risks, and achieve the effects of reducing body aging, good anti-aging activity, and good antioxidant activity
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Embodiment 1
[0035] The artificial synthesis of embodiment 1 active peptide VPITPTLNR
[0036] 1. Synthesis of Bioactive Peptides
[0037] 1. Weigh 3g of RINK resin (degree of substitution: 0.3mmol / g) into a 150ml reactor and soak with 50ml of dichloromethane (DCM).
[0038] 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.
[0039] 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.
[0040] 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.
[0041] 5. Weigh an...
Embodiment 2
[0077] Antioxidant activity experiment of embodiment 2 bioactive peptides
[0078] 1. Determination of in vitro antioxidant activity of biologically active peptide VPITPTLNR by [DPPH·] method
[0079] 1. Experimental reagents and instruments:
[0080] 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 VPITPTLNR.
[0081] 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.
[0082] 2. Experimental method:
[0083] (1) 1mmol / L [DPPH·]methanol solution
[0084] 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 foi...
Embodiment 3
[0118] Anti-aging activity experiment of embodiment 3 bioactive peptides
[0119] 1. The experiment of biologically active polypeptide VPITPTLNR improving the viability of Drosophila
[0120] 1. Experimental reagents and instruments:
[0121] Reagent: Oregon K wild-type Drosophila melanogaster, Genetics Laboratory, College of Agriculture, Shanghai Jiao Tong University; agar powder, Sinopharm Chemical Reagent Co., Ltd.; milk-derived biologically active polypeptide VPITPTLNR obtained in Example 1.
[0122] Equipment: CM-230 Mole ultra-clean water, Shanghai Mole Scientific Instrument Co., Ltd.; G136T Zealway intelligent high-temperature sterilization pot, Xiamen Zhiwei Instrument Technology Co., Ltd.; BJ-CD SERIES biological incubator, Shanghai Boxun Industrial Company; GRX-9073 hot air disinfection box, Shanghai Yiheng Technology Co., Ltd.
[0123] 2. Experimental method:
[0124] Take Drosophila as the experimental model: collect newly eclosioned Drosophila adults within 8 h...
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