北冬虫夏草活性多肽的制备工艺研究

Study on the preparation method of enzymatic hydrolysis process of Cordyceps militaris active peptides

目的通过酶解法制备北冬虫夏草活性多肽并筛选出最佳酶解条件。方法分别用胃蛋白酶和胰蛋白酶对北冬虫夏草进行单酶酶解,然后先加胃蛋白酶后加胰蛋白酶进行双酶酶解。按时间,加酶量和底物浓度不同条件采用正交试验,通过茚三酮比色法检测水解度优化最佳酶解工艺,采用MTT法检测单酶酶解液和双酶酶解液对PC12细胞活性的影响。结果胃蛋白酶和胰蛋白酶水解度测定吸光度最大值分别为0.899和0.788,对应酶解时间、加酶量和底物浓度都是5h、2.5%和12%;双酶酶解在此条件下的吸光度值为0.875。酶解液对PC12细胞修复率影响最大值为139.59%,对应样品为胃蛋白酶酶解原液稀释10^(-6)倍。结论胃蛋白酶酶解效果高于胰蛋白酶和双酶。

Objective Cordyceps militaris active peptides are prepared through enzyme hydrolysis and to screen out the best conditions of enzymatic hydrolysis. Methods Respectively with pepsin and trypsin to single enzyme digestion of north aweto,then and pepsin and trypsin after double enzyme digestion. According to time,add amount of enzyme and substrate concentration in different conditions by orthogonal test,by ninhydrin colorimetry to detect hydrolysis to optimize the best enzyme hydrolysis,determined by MTT method is used to detect the single enzyme digestion and double enzyme enzymolysis liquid effects on PC12 cell activity.Results The determination of pepsin and trypsin hydrolysis degree of a maximum absorbance were 0. 899 and 0. 788,corresponding to the digestion time,enzyme and substrate concentration is 5 h,2. 5% and 12%; Under the condition of double enzyme digestion in the absorbance value of 0. 875. Enzymolysis liquid on PC12 cell repair rate affects a maximum of 139. 59%,corresponding to concentrate samples for pepsin enzyme solution diluted 10^-6times. Conclusion Trypsin and pepsin enzyme effect is higher than the double enzyme.