Study On Preparation Technology Of Enzymolysis Technology Derived Collagen Peptides From Cyanea And Polysaccharide From Mactra Veneriformis

This thesis consists of two parts:1.Study on bioactive enzymatic peptides of CyaneaTrypsin were used to prepare enzymatic hydrolysate from Cyanea and the pharmacodynamics index indicated that enzymatic hydrolysate from anea have a definite ACE-inhibitory activity.Then furtherly used ACE-inhibitory activity as an index,Response surface method was subjected to optimize the enzymatic technology and different molecular mass segments of Cyanea enzymatic hydrolysatesthe(<1k Da,1-3 k Da,>3 k Da)were obtained by ultratiltration.The mass segment that molecular weight less than 1k Da had highest ACE inhibitory activity.Following parts were mainly included:(1).Literature studyResearch papers about Cyanea were referred,and the progress in studies on Cyanea was understood comprehensively,including biological characteristics,status of resources,medical history,pharmacology and related ingredients contained,etc.The summary was to provide research ideas for further development.(2).Preparation of bioactive activity peptide of CyaneaStudy on the content of water and ash and amino acide compositions of Cyanea.Trypsin were used to prepare enzymatic hydrolysate from Cyanea,enzymatic hydrolysate from Cyanea have a definite ACE-inhibitory activity,Tyrosinase inhibitory activity and protective effects of oxidative stress in endothelial cells and myocardial hypoxia/reoxygenation death.ACE inhibitory rate as an index,different one-factor experimental design(including enzyme-to-substrate ratio,temperature,pH and reaction time)was accomplished with the enzymatic and Centural Composite model response surface design was furtherly used to optimize the enzymatic technology of Cyanea enzymatic hydrolysate.The Cyanea enzymatic hydrolysate which possessed best ACE inhibitory activity was prepared under following condition:trypsin was selected to hydrolyze at pH 7.8,49.6 ℃,2.0 h reaction time and enzyme-to-substrate ratio 0.69%.At this condition,the ACE inhibitory rate could be 56.79%.The ultratiltration membrane was subjected to obtain different molecular mass segments of Cyanea enzymatic hydrolysate.The cut-off molecular weight of ultratiltration membrane was 1k,3k,the mass segment that molecular weight less than 1k Da had highest ACE inhibitory activity,which is 89.70%.2.Study on preparation process of Mactra veneriformis polysaccharideUsed polysaccharide yield and polysaccharide transfer rate as indexes,the applicability about preparation technology of I(once alcohol precipitation)and II(secondary alcohol precipitation)polysaccharide extracted from Mactra veneriformis was studied.And the the applicability of different preparation technology was also evaluated by using the same indexes.Try to improve the preparation technology through leading enzymatic hydrolysis into it just before the alcohol precipitation.Model of oral glucose tolerance in normal mice,normal mice gluconeogenesis model and model of diabetic mice were established.Pharmacodynamics difference of polysaccharide preparation Ⅰ,Ⅱ and Ⅲ was compared.(1).Study on polysaccharide content of different month in order to choose the best harvest time.(2).Study on polysaccharide content and transfer rate of original Mactra veneriformis polysaccharide preparation I,II.The results showed that the transfer rate and polysaccharide content were low after alcohol precipitation,process should be further optimized.(3).Effects on yield and transfer rate of different extraction methods were investigated,including boiling water extraction,acid solution extraction,alkali solution extraction,water immersion extraction and enzymatic hydrolysis extraction.The results showed that boiling water extraction method was the best method to extract Mactra veneriformis polysaccharides.(4).Enzymatic hydrolysis was subjected to deal with Mactra veneriformis water extract concentrate to remove macromolecular protein substances before alcohol precipitation.Compared the affects of transfer rate and polysaccharide content of polysaccharide in precipitation,which was prepared by trypsin,pepsin,papain and neutral protease.Neutral protease is more suitable for water extraction and the preparation was furtherly optimized by orthogonal test.(5).Model of oral glucose tolerance in normal mice,normal mice gluconeogenesis model and model of diabetic mice created by Alloxan were established.Compared the pharmacodynamic difference between the Mactra veneriformis polysaccharides of two processes.