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

作　　者：郭艳琼[1,2] 吴海花[1] 宣涛[1] 张建珍[1] 郭亚平[1] 马恩波[1] 

机构地区：[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

出　　处：《应用昆虫学报》2011年第4期826-830,共5页Chinese Journal of Applied Entomology

基　　金：国家自然科学基金重大国际合作项目(30810103907);国家自然科学基金(30870302);中国博士后科学基金(201003656和20090451359)

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0.3046μmol/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8.5185μmol/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25.4ku。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。Glutathione S-transferase （GST） in the fifth-instar nymphs of Oxya chinensis（Thunberg）was purified by ammonium sulfate and GSH-agrose 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 1.82-fold.Further GSH-agrose affinity chromatography indicated that purification reached 50.88 with specific activity of 8.5185 μmol/min/mg protein.SDS-PAGE 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.

关 键 词：谷胱甘肽硫转移酶 中华稻蝗 纯化 亲和层析 

分 类 号：S433.2[农业科学—农业昆虫与害虫防治] Q966[农业科学—植物保护]