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

• Published in: Chemie ingenieur technik 2023-04, Vol.95 (4), p.607-611
• Main Authors: Schelle, Jona T., Lepoittevin, William, Hauer, Bernhard
• Format: Article
• Language: English
• Subjects: Biocatalysis; Cumene dioxygenase; Hydroxylation; Rieske non‐heme iron oxygenases; Scale‐up
• ISSN: 0009-286X; 1522-2640
• DOI: 10.1002/cite.202200162

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. In this article, we enhanced the semi‐preparative biotransformation of Rieske non‐heme iron oxygenases at the example of the cumene dioxygenase, by improving oxygen transfer in a sealed system suitable for volatile products. The usefulness of the setup was demonstrated at the biosynthesis of (1R, 2S)‐cis‐1,2‐dihydro‐1,2‐naphthalenediol and (+)‐trans‐carveol.