Biological active polypeptide DDVTEVM, preparation method and applications thereof
A technology of biologically active peptides and derivatives, applied in the field of protein, can solve problems such as human body risks, achieve the effects of reducing body aging, improving quality, and scavenging free radicals
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Embodiment 1
[0039] The artificial synthesis of embodiment 1 active peptide DDVTEVM
[0040] 1. Synthesis of Bioactive Peptides
[0041] 1. Weigh 3g of RINK resin (degree of substitution: 0.3mmol / g) into a 150ml reactor and soak with 50ml of dichloromethane (DCM).
[0042] 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.
[0043] 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.
[0044] 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.
[0045] 5. We...
Embodiment 2
[0081] Antioxidant activity experiment of embodiment 2 bioactive peptides
[0082] 1. Determination of in vitro antioxidant activity of bioactive peptide DDVTEVM by [DPPH·] method
[0083] 1. Experimental reagents and instruments:
[0084]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 Biologically active polypeptide DDVTEVM.
[0085] 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.
[0086] 2. Experimental method:
[0087] (1) 1mmol / L [DPPH·]methanol solution
[0088] 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. The effect of biologically active polypeptide DDVTEVM on the structure of spleen tissue in vivo
[0122] 1. Experimental reagents and instruments:
[0123] 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. Ltd.; the biologically active polypeptide DDVTEVM obtained in Example 1.
[0124] 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.
[0125] 2. Experimental method:
[0126] (1) Modeling of animal aging models
[0127] After one week of adaptive feeding, the ICR mice were divide...
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