Enhanced Semi-Preparative Biotransformation of Cumene Dioxygenase: From Analytical Scale to Product Isolation

Scale-up of oxygenase catalyzed reactions is often challenging due to the limited oxygen mass transfer in aqueous solutions. To overcome such limitation, we studied different scale-up conditions using recombinant resting cells of E. coli JM109(DE3), harboring the cumene dioxygenase of Pseudomonas fluorescens IP01, for the dihydroxylation of naphthalene to (1R,2S)-cis-1,2-dihydro-1,2-naphthalenediol. Thereby, vigorous stirring of the biotransformation in a 2 L round bottom flask in combination with an oxygen-enriched headspace exhibited outstanding product formation after 1 h. Furthermore, the enhanced setup was used for the cumene dioxygenase catalyzed biosynthesis of 240 mg of valuable (+)-trans-carveol from (R)-(+)-limonene, demonstrating the application of our workflow for volatile compounds.