嗜热菌Thermaerobacter subterraneus DSM13965磷酸三酯酶的原核表达、纯化及性质表征  

Prokaryotic Expression,Purification and Properties of Phosphotriesterase from Thermophilic Archaeon Thermaerobacter subterraneus DSM13965

作　　者：林禹欣[1] 刘惠麟[1] 韩葳葳[2] 詹冬玲[1] 刘景圣[1] 

机构地区：[1]吉林农业大学食品科学与工程学院,长春130118 [2]吉林大学分子酶学工程教育部重点实验室,长春130012

出　　处：《吉林大学学报（理学版）》2013年第6期1182-1186,共5页Journal of Jilin University:Science Edition

基　　金：国家自然科学基金(批准号:31070638);吉林省自然科学基金(批准号:20105109);吉林农业大学博士科研启动基金(批准号:201223)

摘　　要：将来源于嗜热菌Thermaerobacter subterraneus DSM13965的磷酸三酯酶基因在大肠杆菌BL21中高效表达,并采用超声、热失活和组氨酸标签镍柱对产物进行分离纯化,通过SDS-PAGE和Western-blot对表达产物进行鉴定.结果表明:该重组酶以二聚体的寡聚体形式存在;其最适温度为70℃,最适pH=10.0,具有良好的热稳定性.Gene phosphotriesterase from archeaon Thermaerobacter subterraneus DSM13965 was cloned and expressed at high levels in E.coli BL21.The recombinant enzyme was purified to homogeneity via ultrasonic,heating and his-tag in Ni-sepharose chromatography.With the help of SDS-PAGE and Western-blotting,the phosphotriesterase was identified as a homo-oligomer existed in dimer form.The enzyme was characterized with ethyl-paraoxon,its optimum temperature and pH were 70 ℃ and 10.0,respectively.The enzyme has a favourable heat resistance.

关 键 词：嗜热菌Thermaerobacter subterraneus DSM13965 磷酸三酯酶 原核表达 性质 

分 类 号：Q786[生物学—分子生物学]