A kind of bioactive polypeptide vpitptln and its preparation method and application
A bioactive peptide and drug technology, applied in the protein field, can solve problems such as human body risks, and achieve the effects of reducing body aging, improving vitality, and improving anti-aging activity.
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Embodiment 1
[0038] The artificial synthesis of embodiment 1 active peptide VPITPTLN
[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 in 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 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.
[0044] 5. ...
Embodiment 2
[0080] Antioxidant activity experiment of embodiment 2 bioactive peptides
[0081] 1. Determination of in vitro antioxidant activity of biologically active peptide VPITPTLN 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 VPITPTLN.
[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,...
Embodiment 3
[0119] Anti-aging activity experiment of embodiment 3 bioactive peptides
[0120] 1. Experiments on the effect of biologically active polypeptide VPITPTLN on the structure of spleen tissue in vivo
[0121] 1. Experimental reagents and instruments:
[0122] Reagents: experimental animal ICR mice (male 5 weeks old), Shanghai Experimental Animal Center; D-gal, Sinopharm Chemical Reagent Co., Ltd.; paraformaldehyde, Sinopharm Chemical Reagent Co., Ltd.; sodium chloride, Sinopharm Chemical Reagent Co., Ltd. Co., Ltd.; the milk-derived bioactive polypeptide VPITPTLN obtained in Example 1.
[0123] Instruments and equipment: CM-230 Mole ultra-clean water, Shanghai Moller Scientific Instrument Co., Ltd.; MilliporeMILLEX GP0.22μm filter membrane, Millipore Corporation; GL-22M high-speed refrigerated centrifuge, Shanghai Luxiangyi Centrifuge Instrument Co., Ltd.
[0124] 2. Experimental method:
[0125] (1) Modeling of animal aging models
[0126] After one week of adaptive feeding,...
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