中华稻蝗谷胱甘肽S-转移酶的分离纯化
Affinity chromatography for the purification of glutathione S-transferase from Oxya chinensis
郭艳琼1,2,吴海花1,宣涛1,张建珍1,郭亚平1,马恩波1**
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作者单位:1山西大学应用生物学研究所太原030006; 2山西农业大学农学院太谷030801
中文关键词:谷胱甘肽硫转移酶,中华稻蝗,纯化,亲和层析
英文关键词:glutathione Stransferases (GSTs), Oxya chinensis, purification, affinity chromatography
中文摘要: 采用硫酸铵沉淀法和GSH-agarose亲和层析法,对中华稻蝗Oxya chinensis(Thunberg)5龄若虫谷胱甘肽S-转移酶(glutathione S-transferases,GSTs)进行了分离纯化。结果表明:经硫酸铵沉淀,饱和度在60%~80%下沉淀中GSTs比活力较高,饱和度90%时比活力达到最高,为0.3046 μmol/min/mg protein,纯化倍数为1.82。进一步经GSH-agarose亲和层析纯化后,比活力最高达到8.5185 μmol/min/mg protein,纯化倍数为50.88。经SDS-PAGE电泳检测,呈现单一的条带,亚基的分子量约为25.4 ku。此组分的纯化成功为进一步研究中华稻蝗谷胱甘肽S-转移酶的酶学性质、结构特点和作用原理提供了基础资料。
英文摘要: Glutathione Stransferase (GST) in the fifthinstar nymphs of Oxya chinensis(Thunberg)was purified by ammonium sulfate and GSHagrose affinity chromatography. Higher specific activity of GSTs could be detected at 60%-80% purity, and the highest specific activity was found at 90%. The specific activity was 0.3046 μmol/min/mg protein, and the purification factor was 1.82fold. Further GSHagrose affinity chromatography indicated that purification reached 50.88 with specific activity of 8.5185 μmol/min/mg protein. SDSPAGE analysis indicated that the purified GSTs preparation had a single band with a relative molecular mass of 25.4 ku. These results provide the basis for further study of the enzymatic properties, structural characteristics and functions of the GSTs of O. chinensis.