| 嗜热菌Thermaerobacter subterraneus DSM13965磷酸三酯酶的原核表达、 ]纯化及性质表征 |
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| 引用本文： | 林禹欣,刘惠麟,韩葳葳,詹冬玲,刘景圣.嗜热菌Thermaerobacter subterraneus DSM13965磷酸三酯酶的原核表达、 ]纯化及性质表征[J].吉林大学学报(理学版),2013,51(6):1182-1186. |
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| 作者姓名： | 林禹欣 刘惠麟 韩葳葳 詹冬玲 刘景圣 |
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| 作者单位： | 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 |
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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.
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| 关 键 词： | 嗜热菌Thermaerobacter subterraneus DSM13965 磷酸三酯酶 原核表达 性质 |
| 收稿时间： | 2013-02-18 |
Prokaryotic Expression,Purification and Properties ofPhosphotriesterase from Thermophilic Archaeon Thermaerobacter subterraneusDSM13965 |
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| LIN Yu xin,LIU Hui lin,HAN Wei wei,ZHAN Dong ling,LIU Jing sheng.Prokaryotic Expression,Purification and Properties ofPhosphotriesterase from Thermophilic Archaeon Thermaerobacter subterraneusDSM13965[J].Journal of Jilin University: Sci Ed,2013,51(6):1182-1186. |
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| Authors: | LIN Yu xin LIU Hui lin HAN Wei wei ZHAN Dong ling LIU Jing sheng |
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| Institution: | 1. College of Food Science and Engineering, Jilin Agricultural University, Changchun 130118, China; 2. Key Laboratoryfor Molecular Enzymology and Engineering of the Ministry of Education, Jilin University, Changchun 130012, China |
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| Abstract: | Gene phosphotriesterase from archeaon Thermaerobactersubterraneus DSM13965 was cloned and expressed at high levels in E.coli BL21. The recombinant enzyme was purified to homogeneity via ultrasonic, heating andhis 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. |
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| Keywords: | Thermaerobacter subterraneus DSM13965 phosphotriesterase prokaryotic expression property |
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