Bioactive polypeptide ATLEDSPEVI as well as preparation method and application 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 injuries, improving human ability, and enhancing proliferation ability
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Embodiment 1
[0036] Example 1 Synthesis of Active Peptide ATLEDSPEVI
[0037] 1. Synthesis of Bioactive Peptides
[0038] 1. Weigh 3 g of RINK resin (degree of substitution: 0.3 mmol / g) into a 150 ml reactor, and soak it with 50 ml of dichloromethane (DCM).
[0039] After 2.2 hours, the resin was washed with nitrogen-dimethylformamide (DMF) 3 times the resin volume, and then drained. This was repeated four times, and the resin was drained for use.
[0040] 3. A certain amount of 20% piperidine (piperidine / DMF=1:4, v:v) was added to the reactor, and it was shaken on a decolorizing shaker for 20 minutes to remove the Fmoc protecting group on the resin. After deprotection, it was washed four times with 3 times the resin volume of DMF, and then drained.
[0041] 4. Take a small amount of resin and test it by ninhydrin (Ninhydrin hydrate) method (two drops each of test A and test B, react at 100°C for 1 min), the resin is colored, indicating that the deprotection is successful.
[0042] 5. W...
Embodiment 2
[0078] Example 2 Antioxidant activity test of bioactive peptides
[0079] The antioxidant activity of the biologically active polypeptide ATLEDSPEVI obtained in Example 1 was tested by using the free radical scavenging method (DPPH· method) and the total antioxidant capacity method (Ferric Reducing AbilityPower FRAP method).
[0080] 1. The in vitro antioxidant activity of the bioactive peptide ATLEDSPEVI was determined by the [DPPH·] method
[0081] 1) Experimental reagents and instruments
[0082] Reagent: 1,1-Diphenyl-2-trinitrophenylhydrazine (1,1-Diphenyl-2-picrylhydrazyl[DPPH·]), produced by Wako Company in Japan; methanol, provided by Shanghai Sinopharm Company; obtained in Example 1 Milk-derived biologically active polypeptide ATLEDSPEVI.
[0083] Main instruments: Sunrise microplate reader, product of Tecan Company, Austria; 96-well cell culture plate, manufactured by Millipore Company of the United States; analytical balance, product of Meitelei-tolido Company.
...
Embodiment 3
[0118] Example 3 Experiment on the promotion of immunity of biologically active peptides
[0119] 1. Determination of in vitro lymphocyte proliferation ability of biologically active polypeptide ATLEDSPEVI by MTT method
[0120] 1) Experimental materials and instruments:
[0121] Reagents and materials: experimental animal balb / c mice (male, 6-8 weeks old, Animal Experiment Center, School of Agriculture and Biology, Shanghai Jiaotong University); milk-derived biologically active polypeptide ATLEDSPEVI obtained in Example 1; mouse lymphocyte extract (purchased from Soleobor Company); RPMI1640 medium (purchased from GIBCO Company); 3-(4,5-Dimethylthiazole-2)-2,5-diphenyltetrazolium bromide (MTT, purchased from from Amresco Company); Concanavalin (ConA, from Sigma Company); Bovine Serum Albumin (BSA, from Genebase Company); Pepsin (from Sigma Company); Pancreatin (Corolase PP, from AB Company) ).
[0122] Instruments: LRH-250F biochemical incubator, Shanghai Heng Technology Co...
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