caa a22 4500 605501017 CHVBK 20210128100558.0 cr unu---uuuuu 210128e20151101xx s 000 0 eng 10.1007/s00253-015-6716-9 doi (NATIONALLICENCE)springer-10.1007/s00253-015-6716-9 Efficient kinetic resolution of phenyl glycidyl ether by a novel epoxide hydrolase from Tsukamurella paurometabola [Elektronische Daten] [Kai Wu, Hualei Wang, Huihui Sun, Dongzhi Wei] Enantioselective hydrolysis of racemic epoxides mediated by epoxide hydrolases (EHs) is one of the most promising approaches to obtain enantiopure epoxides. In this study, we identified and characterized a novel EH (TpEH1) from Tsukamurella paurometabola by analyzing the conserved catalytic residues of EH. TpEH1 was overexpressed and purified, and its catalytic properties were studied using racemic phenyl glycidyl ether (PGE) and its derivatives as substrates. TpEH1 showed excellent enantioselectivity to the substrates PGE, 3-methylPGE, and 3-nitroPGE. The highest enantioselectivity (E>100) was achieved when 3-nitroPGE was used as the substrate. The recombinant Escherichia coli TpEH1 demonstrated high substrate tolerance toward PGE and could hydrolyze PGE at concentrations of up to 400mM (60g/L) with high enantioselectivity (E=65), giving (R)-PGE with enantiomeric excess of more than 99% ee and 45% yield within 1h. This concentration of PGE is the highest reported concentration catalyzed by native EHs to date. Thus, the easily available and highly active E. coli TpEH1 showed great potential for the practical preparation of optically pure (R)-PGE. Springer-Verlag Berlin Heidelberg, 2015 Epoxide hydrolase nationallicence Enantioselectivity nationallicence Phenyl glycidyl ether nationallicence Kinetic resolution nationallicence High substrate tolerance nationallicence Wu Kai State Key Laboratory of Bioreactor Engineering, New World Institute of Biotechnology, East China University of Science and Technology, 200237, Shanghai, China aut Wang Hualei State Key Laboratory of Bioreactor Engineering, New World Institute of Biotechnology, East China University of Science and Technology, 200237, Shanghai, China aut Sun Huihui State Key Laboratory of Bioreactor Engineering, New World Institute of Biotechnology, East China University of Science and Technology, 200237, Shanghai, China aut Wei Dongzhi State Key Laboratory of Bioreactor Engineering, New World Institute of Biotechnology, East China University of Science and Technology, 200237, Shanghai, China aut Applied Microbiology and Biotechnology Springer Berlin Heidelberg 99/22(2015-11-01), 9511-9521 0175-7598 99:22<9511 2015 99 253 https://doi.org/10.1007/s00253-015-6716-9 text/html Onlinezugriff via DOI BK010053 XK010053 XK010000 Metadata rights reserved Springer special CC-BY-NC licence nationallicence 1 research-article jats NATIONALLICENCE NATIONALLICENCE NL-springer NATIONALLICENCE 856 40 https://doi.org/10.1007/s00253-015-6716-9 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Wu Kai State Key Laboratory of Bioreactor Engineering, New World Institute of Biotechnology, East China University of Science and Technology, 200237, Shanghai, China aut NATIONALLICENCE 700 1- Wang Hualei State Key Laboratory of Bioreactor Engineering, New World Institute of Biotechnology, East China University of Science and Technology, 200237, Shanghai, China aut NATIONALLICENCE 700 1- Sun Huihui State Key Laboratory of Bioreactor Engineering, New World Institute of Biotechnology, East China University of Science and Technology, 200237, Shanghai, China aut NATIONALLICENCE 700 1- Wei Dongzhi State Key Laboratory of Bioreactor Engineering, New World Institute of Biotechnology, East China University of Science and Technology, 200237, Shanghai, China aut NATIONALLICENCE 773 0- Applied Microbiology and Biotechnology Springer Berlin Heidelberg 99/22(2015-11-01), 9511-9521 0175-7598 99:22<9511 2015 99 253