title: 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.
abstract: 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.
authors.affiliation: B.D. Summers, M. Omar, T. Ronson, Dr G. Grogan, York Structural Biology Laboratory, Department of Chemistry University of York, York YO10 5DD, UK. gideon.grogan@york.ac.uk.
authors.forename: Benjamin D
authors.initials: BD
authors.lastname: Summers
authors.forename: Muhiadin
authors.initials: M
authors.lastname: Omar
authors.forename: Thomas O
authors.initials: TO
authors.lastname: Ronson
authors.forename: Jared
authors.initials: J
authors.lastname: Cartwright
authors.forename: Michael
authors.initials: M
authors.lastname: Lloyd
authors.forename: Gideon
authors.initials: G
authors.lastname: Grogan
mesh_terms: D055162:Biocatalysis; D019149:Bioreactors; D004926:Escherichia coli / Q000166:cytology / Q000235:genetics* / Q000378:metabolism; D015971:Gene Expression Regulation, Enzymologic / Q000235:genetics; D007659:Ketones / Q000737:chemistry / Q000378:metabolism*; D008968:Molecular Conformation; D010105:Oxygenases / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D012240:Rhodococcus / Q000201:enzymology* / Q000378:metabolism; D013379:Substrate Specificity
chemical_list: D007659:Ketones; D010105:Oxygenases; C019961:cyclohexanone oxygenase