E. coli cells expressing the Baeyer-Villiger monooxygenase 'MO14' (ro03437) from Rhodococcus jostii RHA1 catalyse the gram-scale resolution of a bicyclic ketone in a fermentor

• Published in: Organic & biomolecular chemistry Vol. 13; no. 6; pp. 1897 - 1903
• Main Authors: Summers, Benjamin D, Omar, Muhiadin, Ronson, Thomas O, Cartwright, Jared, Lloyd, Michael, Grogan, Gideon
• Format: Journal Article
• Language: English
• Published: England 14.02.2015
• Subjects: Biocatalysis; Bioreactors; Escherichia coli - cytology; Escherichia coli - genetics; Escherichia coli - metabolism; Gene Expression Regulation, Enzymologic - genetics; Ketones - chemistry; Ketones - metabolism; Molecular Conformation; Oxygenases - chemistry; Oxygenases - genetics; Oxygenases - metabolism; Rhodococcus - enzymology; Rhodococcus - metabolism; Substrate Specificity
• ISSN: 1477-0520; 1477-0539
• DOI: 10.1039/c4ob01441c

The Baeyer-Villiger monooxygenase (BVMO) 'MO14' from Rhodococcus jostii RHA1, is an enantioselective BVMO that catalyses the resolution of the model ketone substrate bicyclo[3.2.0]hept-2-en-6-one to the (1S,5R)-2-oxa lactone and the residual (1S,5R)-substrate enantiomer. This regio-plus enantioselective behaviour is highly unusual for BVMOs, which often perform enantiodivergent biotransformations of this substrate. The scaleability of the transformation was investigated using fermentor-based experiments, in which variables including gene codon optimisation, temperature and substrate concentration were investigated. E. coli cells expressing MO14 catalysed the resolution of bicyclo[3.2.0]hept-2-en-6-one to yield (1S,5R)-2-oxa lactone of >99% ee and (1S,5R)-ketone of 96% ee after 14 h at a temperature of 16 °C and a substrate concentration of 0.5 g L(-1) (4.5 mM). MO14 is thus a promising biocatalyst for the production of enantio-enriched ketones and lactones derived from the [3.2.0] platform.