| High-throughput Screening of the Enantioselectivity of Hyperthermophilic Mutant Esterases from Archaeon Aeropyrum pernix K1 for Resolution of （ R, S ） -2-Octanol Acetate |
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| 作者姓名： | ZHANG Gui-rong GAO Ren-jun ZHANG Ai-jun RAO Lang CAO Shu-gui |
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| 作者单位： | ZHANG Gui-rong，GAO Ren-jun，ZHANG Ai-jun，RAO Lang and CAO Shu-guiKey Laboratory for Molecular Enzymology and Engineering of Ministry of Education，Jilin University，Changchun 130021，P. R. China |
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| 基金项目： | 国家自然科学基金，国家高技术研究发展计划(863计划) |
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| 摘 要： | To identify the desired hyperthermophilic variants within a mutant esterase library for the resolution of (R,S)-2-octanol acetate, a simple, reliable, and versatile method was developed in this study. We built a screening strategy including two steps, first we selected agar plate with substrate to screen the enzymatic activity; secondly we used a pH indicator to screen the enantioselectivity. This method could rapidly detect favorable mutants with high activity and enantioselectivity. A total of 96.2% of tedious screening work can be precluded using this screening strategy. It is an effective screening for alkyl ester and can be applied to relative screening researches. The four improved mutants were screened from the mutant esterase library. Their enantioselectivities, activities, and structures were investigated at different temperatures.
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| 关 键 词： | 超嗜热需氧古生菌 立体选择性 超嗜热突变体 高通量筛选 酯酶 定向进化 （R S）-2-醋酸辛酯 拆分 |
| 收稿时间： | 17 April 2006. |
| 修稿时间： | 2006-04-17 |
High-throughput Screening of the Enantioselectivity of Hyperthermophilic Mutant Esterases from Archaeon Aeropyrum pernix K1 for Resolution of (R,S) -2-Octanol Acetate |
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| ZHANG Gui-rong GAO Ren-jun ZHANG Ai-jun RAO Lang CAO Shu-gui.High-throughput Screening of the Enantioselectivity of Hyperthermophilic Mutant Esterases from Archaeon Aeropyrum pernix K1 for Resolution of (R,S) -2-Octanol Acetate[J].Chemical Research in Chinese University,2007,23(3):319-324. |
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| Authors: | Gui-rong ZHANG Ren-jun GAO Ai-jun ZHANG Lang RAO Shu-gui CAO |
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| Institution: | aKey Laboratory for Molecular Enzymology and Engineering of Ministry of Education, Jilin University, Changchun 130021, P. R. China |
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| Abstract: | To identify the desired hyperthermophilic variants within a mutant esterase library for the resolution of (R,S)-2-octanol acetate, a simple, reliable, and versatile method was developed in this study. We built a screening strategy including two steps, first we selected agar plate with substrate to screen the enzymatic activity; secondly we used a pH indicator to screen the enantioselectivity. This method could rapidly detect favorable mutants with high activity and enantioselectivity. A total of 96.2% of tedious screening work can be precluded using this screening strategy. It is an effective screening for alkyl ester and can be applied to relative screening researches. The four improved mutants were screened from the mutant esterase library. Their enantioselectivities, activities, and structures were investigated at different temperatures. |
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| Keywords: | High-throughput screening Enantioselectivity Hyperthermophilic esterase Directed evolution |
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