Using a Baeyer-Villiger Monooxygenase

6 Enantioselective Kinetic Resolution of Racemic 3-Phenylbutan-2-one Using a Baeyer-Villiger Monooxygenase 

1 Procedure 1 Recombinant Expression of the BVMO from P. fluorescens DSM 50106 in E. coli 

Tryptone (5 g), yeast extract (2.5 g) and NaCl (5 g) were dissolved in distilled water, the volume was adjusted to 500 mL and then autoclaved (20 min, 120 °C). A small portion of this Luria-Bertani (LB) medium (10 mL) was placed into a sterile 100 mL shake flask and ampicillin and chloramphenicol solutions were added (LBamp+cm) to final concentrations of 100 p,g mL and 20 p.g mL respectively. The solution was inoculated with E. coli JM109 pGro7 pJOE4072.6 and shaken overnight at 37 °C and 200 rpm. This overnight culture (2 mL) was used to inoculate 200 mL LBamp+cm in a 

Cells were then harvested by centrifugation for 20 min at 4400g and 4 °C. The medium was removed and the cell pellet was washed once with 50 mL phosphate buffer solution and centrifuged again. The cells can be stored in the fridge for a few days or used directly for biotransformation. 

Phosphate buffer solution (50 mM, pH 7.5) racemic 3-phenylbutan-2-one (0.15 g, 1 mmol) /3-cyclodextrin (0.07 g, 0.5 mmol) glucose solution (1 m, 4 mL) ethyl acetate 

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