Double substituted variant of Bacillus amyloliquefaciens esterase with enhanced enantioselectivity and high activity towards 1-(3-4-methylenedioxyphenyl)ethyl acetate
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- 作者：Jia-Yan Liu ; Han-Ping Bian ; Yun Tang…
- 关键词：Bacillus amyloliquefaciens ; Esterase ; Enantioselectivity ; Iterative saturation mutagenesis ; 1 ; (3-4-Methylenedioxyphenyl)ethanol
- 刊名：Applied Microbiology and Biotechnology
- 出版年：2015
- 出版时间：February 2015
- 年：2015
- 卷：99
- 期：4
- 页码：1701-1708
- 全文大小：642 KB
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15. - 作者单位：Jia-Yan Liu (1)
Han-Ping Bian (2)
Yun Tang (2)
Yun-Peng Bai (1)
Jian-He Xu (1)
1. State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, 130 Meilong Road, Shanghai, 200237, China
2. Shanghai Key Laboratory of New Drug Design, School of Pharmacy, East China University of Science and Technology, 130 Meilong Road, Shanghai, 200237, China - 刊物类别：Chemistry and Materials Science
- 刊物主题：Chemistry
Biotechnology
Microbiology
Microbial Genetics and Genomics - 出版者：Springer Berlin / Heidelberg
- ISSN：1432-0614
文摘
Bacillus amyloliquefaciens esterase (BAE) was applied to produce (R)-1-(3-4-methylenedioxyphenyl)ethanol, a chiral drug intermediate. In this study, we improved the enantioselectivity of BAE by protein engineering instead of process engineering as used in our previous work. Saturation mutagenesis was carried out on eight positions of BAE based on structure modeling and substrate docking. A double substituted variant V10 (K358D/A396C) showed an excellent enantioselectivity without decreasing the activity. The functions of these two mutations (K358D and A396C) were investigated, revealing a synergic effect on the BAE enantioselectivity. Using the variant V10, enantiopure (R)-1-(3-4-methylenedioxyphenyl)ethanol could be readily prepared in >97?% ee, affording a high space-time yield (123?g?L??day?) and a high ratio of substrate/catalyst (40?g?g?) in 1-L reaction.