蚯蚓谷胱甘肽转硫酶分离纯化的初步研究  

Extraction and Purification of Glutathione S-Transferase from Earthworm Eisenia fetida

作　　者：杜智勇[1] 荣永海[1] 荣龙[1] 

机构地区：[1]北京航空航天大学生物与医学工程学院,北京100191

出　　处：《天然产物研究与开发》2016年第6期898-903,共6页Natural Product Research and Development

摘　　要：本研究利用闪提技术,通过连续两次硫酸铵沉淀与柱层析的方法从蚯蚓中提取了谷胱甘肽转硫酶（GST）,对提取条件和GST性质进行了初步探究。结果表明,GST的提取条件为：p H值7.0,闪提时间30 s,料液比1∶8,连续两次硫酸铵沉淀选取最佳饱和度分别为40%和80%。提取的GST比活力达到了1.236 U/mg Pr,纯化倍数达到了14倍,收率17.7%。利用SDS-PAGE电泳,得到GST亚基分子量约为25 k Da,GST分子量约为50 k Da。GST保持稳定的p H和温度的范围分别为p H 5.0~9.0和温度0~40℃。Glutathione S-Transferase （GST） is a kind of multifunctional enzyme in organism. In this study, GST was ex- tracted and purified from Earthworm Eisenia fetida, and the extraction condition was optimized as follows ： pH of 7.0, ex- traction time of 30 s, solid/liquid ratio of 1 ： 8, the saturation degrees for two times ammonium sulfate precipitations were 40% and 80%, respectively. Under these conditions, GST was purified 14-fold ;the specific activity of 1. 236 U/mg Pr. and the yield of 17.7% were obtained. GST is composed of two subunits, the molecular weight of one subunit is 25 KDa determined by SDS-PAGE, and therefore the molecular weight of GST is 50 KDa. GST was stable in the range of pH 5. 0-9.0 and temperature 0-40℃.

关 键 词：蚯蚓 谷胱甘肽转硫酶 闪提 纯化 

分 类 号：Q814.1[生物学—生物工程]