海参皂苷Echinoside A的酶解及其产物结构鉴定与溶血毒性
Enzymatic hydrolysis of sea cucumber saponin Echinoside A and identification of its product structure and hemolytic activity
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摘要: 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。Abstract: 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%.
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