Expression of Recombinant Human Flavin Monooxygenase and application for N-oxidation of Moclobemide
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Hanlon, Steven, Camattari, Andrea, Abad, Sandra, Glieder, Anton, Kittelmann, Matthias, Luetz, Stephan, Wirz, Beat and Winkler, Margit (2012) Expression of Recombinant Human Flavin Monooxygenase and application for N-oxidation of Moclobemide. Chemical communications. ISSN 1359-7349
Abstract
Rec. h. flavine containing monooxygenases (FMOs), isoforms 1-5, were cloned and expressed in E. coli BL21. Highest activity (methimazole oxidation) of solubilized enzyme was obtained with the FMO3 construct in an NADPH depletion assay. Moclobemide, a MAO-inhibitor marketed by Roche, was applied as the substrate in whole cell biotransformation assays. Citrate and NADP+ were required for effective bioconversion with FMO3 containing E. coli cells. In a whole cell biotransformation on one L scale with rec h FMO3 and 40 mg of moclobemide 81 % conversion to the N-oxide was achieved. The product was isolated from the reaction mixture and its structure confirmed by MS and NMR spectroscopy (poduct isolation and structure elucidation still ongoing).
| Item Type: | Article |
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| Additional Information: | Collaboration with Roche AG, Basel and TU-Graz in the project "Non-CYP-Metabolizing Enzymes (NCME)" in the frame of the competence centre "ACIB - Austrian Research Centre of Industrial Biotechnology" |
| Keywords: | Flavine containing monooxygenase, heterologous expression, biocatalysis, biotransformation, N-oxidation, moclobemide |
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| Date Deposited: | 13 Oct 2015 13:14 |
| Last Modified: | 13 Oct 2015 13:14 |
| URI: | https://oak.novartis.com/id/eprint/6282 |