scholarly journals Conformational diversity and enantioconvergence in potato epoxide hydrolase 1

2016 ◽  
Vol 14 (24) ◽  
pp. 5639-5651 ◽  
Author(s):  
P. Bauer ◽  
Å. Janfalk Carlsson ◽  
B. A. Amrein ◽  
D. Dobritzsch ◽  
M. Widersten ◽  
...  
Keyword(s):  
Epoxide Hydrolase ◽  
Styrene Oxide ◽  
Computational Studies ◽  
Conformational Diversity ◽  
Enantioconvergent Hydrolysis ◽  
Hydrolysis Of

Computational studies highlight the importance of conformational diversity in the enantioconvergent hydrolysis of styrene oxide by potato epoxide hydrolase 1.

Enantioselective hydrolysis of racemic styrene oxide by epoxide hydrolase ofRhodosporidium kratochvilovae SYU-08

2003 ◽  
Vol 8 (5) ◽  
pp. 306-308 ◽  
Author(s):  
Ji Won Lee ◽  
Eun Jung Lee ◽  
Seung Sik Yoo ◽  
Sung Hoon Park ◽  
Hee Sook Kim ◽  
...  
Keyword(s):  
Epoxide Hydrolase ◽  
Styrene Oxide ◽  
Enantioselective Hydrolysis ◽  
Hydrolysis Of

scholarly journals Substantially improving the enantioconvergence of PvEH1, a Phaseolus vulgaris epoxide hydrolase, towards m-chlorostyrene oxide by laboratory evolution

2019 ◽  
Vol 18 (1) ◽  
Author(s):  
Xun-Cheng Zong ◽  
Chuang Li ◽  
Yao-Hui Xu ◽  
Die Hu ◽  
Bo-Chun Hu ◽  
...  
Keyword(s):  
Phaseolus Vulgaris ◽  
Epoxide Hydrolase ◽  
Substrate Binding ◽  
Binding Pocket ◽  
Saturation Mutagenesis ◽  
Substrate Binding Pocket ◽  
Laboratory Evolution ◽  
E Coli ◽  
Enantioconvergent Hydrolysis ◽  
Hydrolysis Of

Abstract Background Epoxide hydrolase can regioselectively catalyze the oxirane ring-opening hydrolysis of rac-epoxides producing the corresponding chiral diols. In our laboratory, a gene named pveh1 encoding an EH from Phaseolus vulgaris was cloned. Although the directed modification of PvEH1 was carried out, the mutant PvEH1Y3 showed a limited degree of enantioconvergence towards racemic (rac-) m-chlorostyrene oxide (mCSO). Results PvEH1 and PvEH1Y3 were combinatively subjected to laboratory evolution to further enhance the enantioconvergence of PvEH1Y3 towards rac-mCSO. Firstly, the substrate-binding pocket of PvEH1 was identified using a CAVER 3.0 software, and divided into three zones. After all residues in zones 1 and 3 were subjected to leucine scanning, two E. coli transformants, E. coli/pveh1Y149L and /pveh1P184L, were selected, by which rac-mCSO was transformed into (R)-m-chlorophenyl-1,2-ethanediol (mCPED) having 55.1% and 27.2% eep. Secondly, two saturation mutagenesis libraries, E. coli/pveh1Y149X and /pveh1P184X (X: any one of 20 residues) were created at sites Y149 and P184 of PvEH1. Among all transformants, both E. coli/pveh1Y149L (65.8% αS and 55.1% eep) and /pveh1P184W (66.6% αS and 59.8% eep) possessed the highest enantioconvergences. Finally, the combinatorial mutagenesis was conducted by replacements of both Y149L and P184W in PvEH1Y3, constructing E. coli/pveh1Y3Z2, whose αS reached 97.5%, higher than that (75.3%) of E. coli/pveh1Y3. In addition, the enantioconvergent hydrolysis of 20 mM rac-mCSO was performed by E. coli/pveh1Y3Z2, giving (R)-mCPED with 95.2% eep and 97.2% yield. Conclusions In summary, the enantioconvergence of PvEH1Y3Z2 was successfully improved by laboratory evolution, which was based on the study of substrate-binding pocket by leucine scanning. Our present work introduced an effective strategy for the directed modification of enantioconvergence of PvEH1.

Reprogramming Epoxide Hydrolase to Improve Enantioconvergence in Hydrolysis of Styrene Oxide Scaffolds

2020 ◽  
Vol 362 (21) ◽  
pp. 4699-4706
Author(s):  
Fu‐Long Li ◽  
Yan‐Yan Qiu ◽  
Yu‐Cong Zheng ◽  
Fei‐Fei Chen ◽  
Xu–Dong Kong ◽  
...  
Keyword(s):  
Epoxide Hydrolase ◽  
Styrene Oxide ◽  
Hydrolysis Of
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