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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
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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.
作者 林禹欣 刘惠麟 韩葳葳 詹冬玲 刘景圣
机构地区 吉林农业大学食品科学与工程学院 吉林大学分子酶学工程教育部重点实验室
出处 《吉林大学学报（理学版）》 CAS CSCD 北大核心 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)
关键词 嗜热菌Thermaerobacter subterraneus DSM13965 磷酸三酯酶 原核表达 性质 Thermaerobacter subterraneus DSM13965 phosphotriesterase prokaryotic expression property
分类号 Q786 [生物学—分子生物学]
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参考文献12

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吉林大学学报（理学版）
2013年 第6期

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