Kinetic resolution of racemic styrene oxide at a high concentration by recombinant Aspergillus usamii epoxide hydrolase in an n -hexanol/buffer biphasic system

Enantiopure epoxides are versatile synthetic intermediates for producing optically active pharmaceuticals. In an effort to provide more options for the preparation of enantiopure epoxides, a variant of the epoxide hydrolase (vEH-Am) gene from a marine microorganism Agromyces mediolanus was synthesized and expressed in Escherichia coli. Recombiant vEH-Am displayed a molecular weight of 43 kDa and showed high stability with a half-life of 51.1 h at 30 °C. The purified vEH-Am exhibited high enantioselectivity towards styrene oxide (SO) and benzyl glycidyl ether (BGE). The vEH-Am preferentially converted (S)-SO, leaving (R)-SO with the enantiomeric excess (ee) >99%. However, (R)-BGE was preferentially hydrolyzed by vEH-Am, resulting in (S)-BGE with >99% ee. To investigate the origin of regioselectivity, the interactions between vEH-Am and enantiomers of SO and BGE were analyzed by molecular docking simulation. In addition, it was observed that the yields of (R)-SO and (S)-BGE decreased with the increase of substrate concentrations. The yield of (R)-SO was significantly increased by adding 2% (v/v) Tween-20 or intermittent supplementation of the substrate. To our knowledge, vEH-Am displayed the highest enantioselectivity for the kinetic resolution of racemic BGE among the known EHs, suggesting promising applications of vEH-Am in the preparation of optically active BGE.

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Production of (R)-styrene oxide by recombinant whole-cell biocatalyst in aqueous and biphasic system

Polish Journal of Chemical Technology ◽

10.2478/pjct-2018-0023 ◽

2018 ◽

Vol 20 (2) ◽

pp. 54-60 ◽

Cited By ~ 1

Author(s):

Feng Xue ◽

Jian Gao

Keyword(s):

Kinetic Resolution ◽

Styrene Oxide ◽

Enantiomeric Excess ◽

Tween 80 ◽

Aqueous System ◽

Biphasic System ◽

Whole Cells ◽

Enantioselective Resolution ◽

Cell Substrate ◽

Recombinant Cell

Abstract The enantioselective resolution of racemic styrene oxide (rac-SO) to (R)-SO by whole cells of a recombinant Escherichia coli expressing epoxide hydrolase (EH) activity in aqueous and biphasic system were studied. Some parameters that may alter this bio-resolution, such as the concentration of recombinant cell, substrate and product were evaluated. The effect of the addition of different additives on the course of rac-SO biotransformation was also investigated. The results showed that the yield and the enantiomeric excess (ee) of (R)-SO were dependent on these variables. When the kinetic resolution was conducted with 350 mM of rac-SO, enantiopure (R)-SO with high (≥99%) ee was obtained with a yield of 38.2% yield at 12.2 h in the presence of 10% (v/v) Tween 80. An isooctane/aqueous system was developed to overcome the adverse factors in the aqueous phase, resulting in an improvement of yield from 38.2% to 42.9%. The results will provide a useful guidance for further application of this enzyme in the biocatalytic production of chiral synthons.

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