猴头菌子实体粉的复合酶解及其功能特性

Multienzyme Complex Hydrolysis of Hericium erinaceus Fruiting Body Powder and in vitro Activity of the Released Polysaccharides

在筛选的复合酶(纤维素酶∶β-葡萄糖苷酶∶壳聚糖酶)配比(1∶1∶1)基础上,采用单因素实验研究料液比、加酶量、酶解时间、酶解温度、pH对猴头菌(Hericium erinaceus)子实体粉中多糖溶出量的影响,通过响应面法确定其最佳提取工艺参数,并对酶解后子实体粉溶出多糖的理化性质及保护胃黏膜上皮细胞效果进行分析。结果表明:猴头菌子实体粉复合酶解的最佳工艺条件为料液比1∶4(g∶mL),复合酶添加量150 U·g^(-1)、52℃、30 min、pH 5.5,酶解后猴头菌子实体粉多糖含量可达(8.41±0.31)%,与响应面预测值8.45%接近,与未酶解子实体粉多糖含量相比提高115%。酶解后子实体粉多糖重均分子量明显降低,单糖组成中葡萄糖胺、半乳糖比例增加,葡萄糖的比例降低。胃黏膜上皮细胞损伤修复研究表明,酶解猴头菌子实体粉处理组的胃黏膜上皮细胞(GES-1)存活率可达81.30%,与未酶解子实体粉的细胞存活率相比提高约20%。表明酶解后猴头菌子实体粉多糖溶出量和保护胃黏膜作用得到提升。

A multienzyme complex comprising cellulase,β-glucosidase and chitosanase at a molar ratio of 1∶1∶1 was used to hydrolyse Hericium erinaceus fruiting body powder to increase polysaccharide content in the powder.Single factor experiments were carried out to study the effect of material-liquid ratio,enzyme quantity,enzymatic hydrolysis time,enzymatic hydrolysis temperature and pH on polysaccharide release from the fruiting body powder.Then the optimal extraction conditions were determined by the response surface design.The resultant crude polysaccharide extract was then studied for physicochemical properties and its protective effect on gastric mucosa epithelial cells(GES-1)in vitro.The results showed that the optimal conditions were as follows:1∶4(g∶mL)material-liquid ratio,150 U·g^(-1) multienzyme complex,hydrolyzed at 52℃and pH 5.5 for 30 min.The polysaccharide content in H.erinaceus fruiting body powder after the enzymatic hydrolysis reached(8.4±0.31)%,which was close to the predicted value of the response surface(8.45%)and 115%greater than that of the fruiting body powder without enzymatic hydrolysis.After multienzyme complex hydrolysis,the weight average molecular weight of the released polysaccharides decreased significantly,the proportion of glucosamine and galactose both increased,and the proportion of glucose decreased.The multienzyme complex treated powder effectively protected gastric mucosal epithelial cells from H_(2)O_(2) induced damage and the cell viability reached 81.3%,a 20%increase compared with that of the fruiting body powder without enzymatic hydrolysis.