Geier, Martina, Bachler, Thorsten, Hanlon, Steven P., Eggimann, Fabian, Kittelmann, Matthias, Weber, Hansjörg, Luetz, Stephan, Wirz, Beat and Winkler, Margit (2015) Human FMO2-based microbial whole-cell catalysts for drug metabolite synthesis. Journal Microbial Cell Factories, 14 (82).

Abstract

Background: Getting access to authentic human drug metabolites is an important issue during the drug discovery and development process. Employing recombinant microorganisms as whole-cell biocatalysts constitutes an elegant alternative to organic synthesis to produce these compounds. The present work aimed for the generation of an efficient whole-cell catalyst based on the flavin monooxygenase isoform 2 (FMO2), which is part of the human phase I metabolism.
Results: We show for the first time the functional expression of human FMO2 in E. coli. Truncations of the C-terminal membrane anchor region did not result in soluble FMO2 protein, but had a significant effect on levels of recombinant protein. The FMO2 biocatalysts were employed for substrate screening purposes, revealing trifluoperazine and propranolol as FMO2 substrates. Biomass cultivation on the 100 L scale afforded active catalyst for biotransformations on preparative scale. The whole-cell conversion of trifluoperazine resulted in perfectly selective oxidation to 48 mg (46% yield) of the corresponding N1-oxide with a purity >98%.
Conclusions: The generated FMO2 whole-cell catalysts are not only useful as screening tool for human metabolites of drug molecules but more importantly also for their chemo- and regioselective preparation on the multi-milligram scale.

Item Type:	Article
Keywords:	Flavin monooxygenase isoform 2, Escherichia coli, Whole-cell biocatalysis, Drug metabolites, Trifluoperazine, Propranolol, FMO2
Date Deposited:	26 Apr 2016 23:45
Last Modified:	26 Apr 2016 23:45
URI:	https://oak.novartis.com/id/eprint/24859