蜈蚣粉酶解工艺最佳用酶的初步筛选  

Preliminary screening of optimal enzymes for the enzymatic hydrolysis process of centipede powder

作　　者：刘春雨[1] 张盼盼[1] 王颖[1] 王立娜[1] 王彦多[1] 刘玉慧[1] 李晓辉[1] 郑海珠[1] 孙景瑞[1] 蔺叶桐 王集会[2] 高世杰[1] 

机构地区：[1]山东中医药大学药学院,济南250355 [2]山东中医药大学实验中心,济南250355

出　　处：《中南药学》2017年第6期759-762,共4页Central South Pharmacy

基　　金：山东中医药大学大学生研究训练(SRT)计划资助项目(编号:2016032)

摘　　要：目的分别利用胃蛋白酶、胰蛋白酶、胰凝乳蛋白酶、弹性蛋白酶对蜈蚣粉进行酶解并选取最佳用酶来制备抗肿瘤多肽。方法利用4种酶对蜈蚣粉进行酶解,采用MTT体外抗肿瘤实验方法,以蛋白质的水解度及酶解提取液对乳腺癌MCF-7细胞的抑制率为指标,同时以蜈蚣粉的水溶性蛋白质含量为标准,评价4种酶对蜈蚣粉的酶解效果并筛选出最佳用酶。结果胃蛋白酶对蜈蚣粉的水解度为10.44%;胰蛋白酶对蜈蚣粉的水解度为28.29%,在0.125 mg·m L^(-1)及0.25 mg·m L^(-1)的较小浓度下对乳腺癌MCF-7细胞的抑制率低于对照组,但随浓度的增大抑制率逐渐增大,在0.5~2 mg·m L^(-1)浓度时,胰蛋白酶在4种酶中达到最大抑瘤效果;胰凝乳蛋白酶和弹性蛋白酶的水解度分别为50.45%、44.16%,当浓度>0.5 mg·m L^(-1)时其对乳腺癌MCF-7细胞的抑制率高于对照组,在浓度为0.5mg·m L^(-1)时抑制率分别为2.86%、36.37%,且随浓度增大抑制率增大。结论蜈蚣粉酶解工艺最佳用酶为胰蛋白酶,其次是胰凝乳蛋白酶和弹性蛋白酶,而胃蛋白酶活性较低可以去除。Objective To conduct enzymolysis of centipede powder and to select the best enzyme to prepare antitu- mor peptide by pepsin, trypsin, chymotrypsin, and elastase. Methods The centipede powder was hydrolyzed by 4 enzymes, with MTT antitumor experiment method in vitro. We used the hydrolysis degree of the protein and breast cancer MCF-7 cells cell inhibition rate as the index to screen the best enzyme. The enzymolysis effect of 4 enzyme was evaluated and the benefit enzymes were optimized. Results The hydrolysis degree of centipede powder on pep- sin was 10.44%. Trypsin hydrolysis degree was 28.29%. The inhibition of breast cancer MCF-7 cell line was lower than the control group at low concentration of 0.125 mg mL - 1 to 0.25 mg ~ mL - 1, but with increasing concentra- tion, the inhibition rate was increasing gradually. At 0.5 to 2 mg mL - 1, the inhibition rate reached the maximum effect in the 4 enzymes. Hydrolysis degree for ehymotrypsin and elastase was 50.45% and 44.16% respectively, and the inhibition of breast cancer MCF-7 cells was higher than that of the control group. The inhibition rates were 2.86% and 36.37% at the concentration of 0.5 mg mL - 1 and they increased with concentration. Conclusion Trypsin is the best enzyme for centipede powder enzymolysis, followed by chymotrypsin and elastase.

关 键 词：蜈蚣粉 蛋白酶 水解度 乳腺癌MCF-7细胞株 细胞抑制率 

分 类 号：R283.6[医药卫生—中药学]