A kind of bioactive polypeptide skhssldcvl and its preparation method and application
A technology of biologically active peptides and drugs, applied in the field of protein, can solve problems such as human body risks, and achieve the effects of reducing injuries, enhancing proliferative ability, and improving human ability
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Embodiment 1
[0033] The artificial synthesis of embodiment 1 active peptide SKHSLDCVL
[0034] 1. Synthesis of Bioactive Peptides
[0035] 1. Weigh 3g of RINK resin (degree of substitution: 0.3mmol / g) into a 150ml reactor and soak with 50ml of dichloromethane (DCM).
[0036] 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.
[0037] 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.
[0038] 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.
[0039] 5. ...
Embodiment 2
[0082] Antioxidant activity experiment of embodiment 2 bioactive peptides
[0083] The antioxidant activity of the biologically active polypeptide SKHSLDCVL obtained in Example 1 was tested by free radical scavenging method (DPPH·method) and total antioxidant capacity method (Ferric Reducing AbilityPower FRAP method).
[0084] 1. Determination of in vitro antioxidant activity of bioactive peptide SKHSLDCVL by [DPPH·] method
[0085] 1) Experimental reagents and instruments
[0086] 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 SKHSLDCVL.
[0087] 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.
[0088] 2) Experimental method...
Embodiment 3
[0123] Example 3 Bioactive Peptide Promoting Body Immunity Activity Experiment
[0124] 1. MTT method to determine the in vitro lymphocyte proliferation ability of biologically active polypeptide SKHSLDCVL
[0125] 1) Experimental materials and instruments:
[0126] Reagents and materials: experimental animals balb / c mice (male 6-8 weeks old, Animal Experiment Center, School of Agriculture and Biology, Shanghai Jiao Tong University); milk-derived bioactive polypeptide SKHSLDCVL obtained in Example 1; mouse lymphocyte extract (purchased from Suo Laibao); RPMI1640 medium (purchased from GIBCO); 3-(4,5-dimethylthiazole-2)-2,5-diphenyltetrazolium bromide (MTT, purchased from from Amresco Company); Concanavalin (ConA, purchased from Sigma Company); bovine serum albumin (BSA, purchased from Genebase Company); pepsin (purchased from Sigma Company); pancreatin (Corolase PP, purchased from AB Company ).
[0127] Instruments: LRH-250F biochemical incubator, Shanghai Heng Technology C...
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