Characteristics Of Collagenase From Intestine And Its Effect On Collagen Degradation From Body Wall In Sea Cucumber

Sea cucumber was an important marine invertebrate.Sea cucumber was susceptible to external stimulus.and would spit the intestine and undergo autolysis,which leaded to cause inconvenience in the storage and process of sea cucumber.In this study,the characteristics of endogenous enzymes were identified by gelatin zymography,and the effect of endogenous enzymes on the rheological behavior of crude collagen fiber(CCF)from body wall was analyzed by rheometer.Results showed that sea cucumber intestinal endogenous enzymes belong to gelatin enzymes,were mainly distributed in the molecular weight of 29.0?44.3 kDa and more than 116 kDa.Under the condition of pH9.0 at 40?,the enzymatic hydrolysis product of CCF showed G' ? G".the elastic characteristics was higher than the viscosity properties.and solid elastic behavior was predominant.Its liquidity was better than the other,and viscoelasticity was reduced and gel was enhanced.Inhibitors effects:the effect of erine protease inhibitors PMSF inhibition did not impact.These results suggested that PMSF inhibited the sea cucumber endogenous enzymes and maintained viscoelasticity of CCF,serine protease played a main role to degrade CCF.In addition,the characteristics of endogenous enzymes from sea cucumber intestine was investigated and its effect on collagen degradation from body wall was confirmed.Under the optimal conditions,the effects of protease inhibitors on degradation of pepsin-soluble collagen(PSC)by endogenous enzymes was analyzed by sodium dodecyl sulfate polyacrylamide gel electrophoresis.TCA-soluble oligopeptide and hydroxyproline release assays.The results showed that the collagenase displayed maximum activity at pH 9 and 40?.The endogenous enzymes showed strong degradation effect on PSC.PMSF inhibited the degradation of PSC,also 1.10-phenanthroline and E-64.These results suggested that endogenous enzymes involved in the degradation of the PSC.and serine protease played a main role.