海参皂苷Echinoside A的酶解及其产物结构鉴定与溶血毒性

摘要

Sea cucumber saponin Echinoside A was hydrolyzed into low hemolytic secondary glycosides by commercial enzymes.In order to obtain less hemolysis from the rare secondary sea cucumber saponins,sea cucumber glycoside—Echinoside A was hydrolyzed by 5 glycosidases.The glycosidases with enzymatic hydrolysis effects for EA were screened out by TLC,HPLC -MS and ESI-MS/MS,and the enzymatic products structures of which were identified and hemolytic toxicity studies were performed.The results showed that EA could be hydrolyzed by Pectinase,Pectinex Ultra SP-L and Celluclast 1.5L enzymes.The enzymatic hydrolysates of the Pectinase were secondary sea cucumber saponin—Compound 1 with two saccharide groups and secondary sea cucumber saponin—compound 2 with one saccharide group;The enzymatic hydrolysates of the Pectinex Ultra SP-L and Celluclast 1.5 L were secondary sea cucumber saponin—Compound 1.The other two enzymes had no enzymatic hydrolysis effect.Hemolysis test showed that the hemolytic toxicity of the products was obviously reduced.The hemolysis of compound 2 was remarkably reduced to less than 30％.%利用商业酶将海参皂苷Echinoside A(EA)酶解成低溶血毒性的次级苷.为了得到溶血毒性更低的稀有次级海参皂苷,以海参皂苷Echinoside A为底物,选用5种商品糖苷酶,对其进行酶解.利用TLC、高效液相色谱-质谱法(HPLC-MS)和电喷雾串联质谱法(ESI-MS/MS)筛选出具有能将海参皂苷EA降解生成新物质的商品糖苷酶,并进行其酶解产物结构鉴定和溶血毒性研究.结果表明,果胶酶、果胶酶Ultra SP-L和纤维素酶有酶解效果,其中果胶酶酶解生成二糖基次级海参皂苷(化合物1)和一糖基次级海参皂苷(化合物2);果胶酶Ultra SP-L和纤维素酶酶解生成化合物1,其他两种酶对海参皂苷EA没有酶解作用.溶血实验证明酶解产物的溶血毒性相对于海参皂苷EA的溶血毒性有明显地降低,其中化合物2的溶血毒性降低到30％以下,效果显著.