Study On The Preparation Of Functional Oligopeptides From Squid Viscera

Squid viscera as the research object, degree of hydrolysis and nitrogen recovery of the hydrolysis as measure indicators. Firstly, trypsin was selected the best one that favorable to the enzymolysis of squid viscera. Parameters including p H, enzymolysis temperature, enzymolysis time and enzyme concentration were optimized using Box-Behnken central composite design and response surface analysis methods, which degree of hydrolysis was used as response value, based on single factor investigation for choosing maximum the degree of hydrolysis. Flavourzyme were applied in enzymatic hydrolysis of squid visceral in this experiment. The superior protease(Trypsin) was selected by comparing hydrolysis degree and hydrolysis yield of different hydrolysate and measuring total nitrogen content and amino nitrogen content of the hydrolysate. Tryptic hydrolyzate was eluted with water, 20%, 60% and 80%methanol-water gradients through macroporous resin HP-20, and then analyzed by LC-MS. It showed that the molecular weight of short peptide in the hydrolysate was in the range of 200 to 2400 and mass distribution was more concentrated with 60%methanol-water gradient, According to the simulation model and combining with actual condition, the optimum conditions were as follows, p H 8.0, enzymolysis temperature43℃, enzymolysis time 4h, and enzyme concentration 0.27%, as a result, predict response value and the actual value was 72.62% and 73.58% respectively, which the relative deviation was 1.32%. The results indicated that response surface regression equation was highly significant and valid. In addition, through the model test of significance, main effect relationship of factors was: enzymolysis time > enzymolysis temperature > enzyme concentration >p H.The main results were as follows:1. The alkaline protease, Trypsin, Papain, Animal protease, Neutral protease,Flavourzyme were applied in enzymatic hydrolysis of squid visceral in this experiment.The superior protease(Trypsin) was selected by comparing hydrolysis degree and hydrolysis yield of different hydrolysate and measuring total nitrogen content and amino nitrogen content of the hydrolysate. Tryptic hydrolyzate was eluted with water,20%, 60% and 80% methanol-water gradients through macroporous resin HP-20, and then analyzed by LC-MS. It showed that the molecular weight of short peptide in the hydrolysate was in the range of 200 to 2400 and mass distribution was more concentrated with 60% methanol-water gradient, mainly on about 200 molecular weight compounds. DPPH radical scavenging capacity of each elution gradient hydrolyzate was analyzed, indicating that DPPH radical scavenging capacity of 60% methanol-water gradient hydrolyzate was 40.21%. Conclusion: The Superior Enzymatic hydrolysis protease is Trypsin, and 60% methanol-water is the best elution.concentration.2. According to the simulation model and combining with actual condition, the optimum conditions were as follows, p H 8.0, enzymolysis temperature 43℃,enzymolysis time 4h, and enzyme concentration 0.27%, as a result, predict response value and the actual value was 72.62% and 73.58% respectively, which the relative deviation was 1.32%. The alkaline protease, Trypsin, Papain, Animal protease, Neutral protease, Flavourzyme were applied in enzymatic hydrolysis of squid visceral in this experiment. The superior protease(Trypsin) was selected by comparing hydrolysis degree and hydrolysis yield of different hydrolysate and measuring total nitrogen content and amino nitrogen content of the hydrolysate.3. Squid viscera as the research object, degree of hydrolysis and nitrogen recovery of the hydrolysis as measure indicators. Firstly, trypsin was selected the best one that favorable to the enzymolysis of squid viscera. Parameters including p H, enzymolysis temperature, enzymolysis time and enzyme concentration were optimized using Box-Behnken central composite design and response surface analysis methods, which degree of hydrolysis was used as response value, based on single factor investigation for choosing maximum the degree of hydrolysis. According to the simulation model and combining with actual condition, the optimum conditions were as follows, p H 8.0,enzymolysis temperature 43℃, enzymolysis time 4h, and enzyme concentration 0.27%,as a result, predict response value and the actual value was 72.62% and 73.58% respectively, which the relative deviation was 1.32%. The results indicated that response surface regression equation was highly significant and valid. In addition, through the model test of significance, main effect relationship of factors was: enzymolysis time >enzymolysis temperature > enzyme concentration >p H.