Studies On Preparation And Bioactivities Of Oligo-peptides Of Collagen By Enzymatic Bonding Hydrolysis Technology From Trout Skin And Bone

The whole reseach study was aimed at preparative method of oligo-peptides of Mr<3000Da of collagen which can be easily absorbed by human body and easy to amplify by enzymatic bonding hydrolysis technology from skin and bone of cultivation Trout which live on cold spring.Using biochemistry and enzyme engineering technologys,process of decolorization,Decalcification,optimization by quadrature,process control,purification et al.have been done.The collagen peptides from Trout skin have been extracted by using papain and flavourzyme methods.Two kinds of enzyme were analyzed bonding with single factor test and orthogonal test method,at the same times,the way of bleaching and process improvement were tried.The results showed that the optimum addi-tion of papain was the ratio of material to liquid 1:22,the quantity of enzymethe 0.6% ,temprarure 60℃,pH=6.4,the time of enzymehysis was 3 hours,and the yield of extraction of the collagen peptides from Trout skin was 14.33%,DH 10.33%.The optimum addition of flavourzyme was the ratio of material to liquid 1:20,the quantity of enzymethe 0.35%,temprarure 55℃,pH=6.5,the time of enz-ymehysis was 2.5 hours,and the yield of extraction of the collagen peptides from Trout skin was 13.68%,DH 10.65%.Among three methods,using papain before and using flavourzyme after,which is the best method;Activated Carbon can be used as decolorant.Finished product was all right after the process of colation, microfiltration,ultrafiltration,concentration,drying and cribration were done and purity coefficient was 91.04%,collagen peptides less than 2000Da were 82.23 %.FARP value(3mg/mL) was 3.39mmol/L,efficiency of scavenging activity (3m g/mL) of hydroxy radical was 10.09% and efficiency of scavenging activity (3 mg/mL) of superoxide free radical was 3.95%. Pepsin and Flavourzyme were chosen from 9 protease and were used for mading bioactivity collagen polypeptides(Mr<3000Da) from decalcifying Trout bone,at the same time,antioxidative activity were detected.The results showed that the way of EDTA bonding with microwave was the best method,concentration of EDTA,times,power of microwave and so on were the comparatively large influe-ntial factors;The best process parameters of Pepsin was the ratio of material to liquid 1:30,Pepsin 1.2%,temperature 65℃,pH2.2,action time 6 hours and the extraction of collagen polypeptides was 8.68%,DH2.23%,Then changed the ratio of material to liquid 1:35,Flavourzyme 1.2%,temperature 57℃,pH6.4,action ti-me 4 hours,and the extraction of collagen polypeptides was 6.96%,DH 5.81%; Finished product was all right after the process of Purification were done and purity coefficient was 84.08%,Hpro 6.22%.the value of FRAP of finished prod-uct was 3.80mmol/L,clearance rate of OH was 11.24%.Apart from selecting protease,determining parameter of hydrolization,the way of combination of enzyme,Purification and Physicochemical Property of oligo-pep-tides of collagen,we have also studyed concerning pre-disposal treatment,impro-vement of reaction pattern etc.and to gain applied easily method and the process of preparatio.