Gaseous alkene biotransformation and enantioselective epoxyalkane formation by Nocardioides sp. strain JS614

Abstract

Enantiopure epoxides are valuable intermediates in the synthesis of optically pure biologically active fine chemicals (e.g., pharmaceuticals) that are often difficult to produce by chemical approaches. An attractive alternative is biological synthesis by microorganisms expressing stereoselective enzymes. In this study, we investigated the ability of ethene-grown Nocardioides sp. strain JS614 to produce highly enantio-enriched epoxyalkanes via stereoselective monooxygenase-mediated alkene epoxidation. Ethene-grown JS614 cells transformed propene, 1-butene, and trans-2-butene to their corresponding epoxyalkanes at rates ranging from 27.1 to 44.0 nmol/min mg protein. Chiral gas chromatography analysis revealed that R-1,2-epoxypropane, R-1,2-epoxybutane, and trans-2R,3R-epoxybutane were produced in enantiomeric excess (e.e.) of 98%, 74%, and 82%, respectively. Ethene-grown JS614 cells also preferentially transformed trans-2S,3S-epoxybutane from a racemic mixture, but could not resolve racemic 1,2-epoxypropane. Glucose facilitated increased epoxyalkane production by ethene-grown JS614 cells. However, after 22 h of propene biotransformation with 20 mM glucose, 84% of ethene-grown JS614 cells lost membrane integrity and the remaining live cells were not viable. Propene biotransformation by JS614 was extended beyond 22 h and 54% more epoxypropane was produced when cells were resuspended in fresh buffer + glucose at 8-h intervals. We conclude that JS614 is a promising new biocatalyst for applications that involve enantiopure epoxide production.