Late Microaerobic Growth for Efficient Production of Human Cytochrome P450 3A4 in E. coli

Detailed preclinical characterization of metabolites formed in vivo from candidate drug substances is mandatory prior to the initiation clinical trials. Therefore, inexpensive and efficient methods for metabolite synthesis are high importance rapid advancement development process. A large fraction small molecule drugs modified by monooxygenase cytochrome P450 3A4 produced human liver intestine. this enzyme frequently employed catalyze vitro, making availability a critical requirement early development. Unfortunately, recombinant production microbial hosts notoriously difficult. Maintaining low oxygen transfer rates use rich media host cultivation required production. However, detailed studies on relationship between supply space-time yields missing. We describe an improved biotechnological process heterologous expression together with its redox partner, reductase, Escherichia coli. Enzyme was most under so-called “late microaerobic” growth conditions, which cells have just not yet made switch anaerobic metabolism, characterized limited leading concentrations liquid phase that far below detection limit standard electrodes. Furthermore, feeding carbon source glycerol as well controlling cellular acetate formation productivity. The presented protocol resulted functional at up 680 nmol L-1.