Geethani Bandara and Christopher R. Otey contributed equally to this study.
Preparation of human metabolites of propranolol using laboratory-evolved bacterial cytochromes P450†
Article first published online: 13 OCT 2005
Copyright © 2005 Wiley Periodicals, Inc.
Biotechnology and Bioengineering
Volume 93, Issue 3, pages 494–499, 20 February 2006
How to Cite
Otey, C. R., Bandara, G., Lalonde, J., Takahashi, K. and Arnold, F. H. (2006), Preparation of human metabolites of propranolol using laboratory-evolved bacterial cytochromes P450. Biotechnol. Bioeng., 93: 494–499. doi: 10.1002/bit.20744
- Issue published online: 5 JAN 2006
- Article first published online: 13 OCT 2005
- Manuscript Accepted: 31 AUG 2005
- Manuscript Received: 27 JUN 2005
- National Institutes of Health. Grant Number: R01 GM068664-01
- cytochrome P450 BM3;
- drug metabolites;
- drug metabolism;
- peroxide shunt;
- directed evolution
Testing the toxicities and biological activities of the human metabolites of drugs is important for development of safe and effective pharmaceuticals. Producing these metabolites using human cytochrome P450s is difficult, however, because the human enzymes are costly, poorly stable, and slow. We have used directed evolution to generate variants of P450 BM3 from Bacillus megaterium that function via the “peroxide shunt” pathway, using hydrogen peroxide in place of the reductase domain, oxygen and NADPH. Here, we report further evolution of the P450 BM3 heme domain peroxygenase to enhance production of the authentic human metabolites of propranolol by this biocatalytic route. This system offers a versatile, cost-effective, and scaleable route to the synthesis of drug metabolites. © 2005 Wiley Periodicals, Inc.