Sahng Ha Lee and Yong-Chan Kwon contributed equally to this work.
Cytochrome P450-catalyzed O-dealkylation coupled with photochemical NADPH regeneration†
Article first published online: 5 OCT 2012
Copyright © 2012 Wiley Periodicals, Inc.
Biotechnology and Bioengineering
Volume 110, Issue 2, pages 383–390, February 2013
How to Cite
Lee, S. H., Kwon, Y.-C., Kim, D.-M. and Park, C. B. (2013), Cytochrome P450-catalyzed O-dealkylation coupled with photochemical NADPH regeneration. Biotechnol. Bioeng., 110: 383–390. doi: 10.1002/bit.24729
- Issue published online: 20 DEC 2012
- Article first published online: 5 OCT 2012
- Accepted manuscript online: 24 SEP 2012 07:26AM EST
- Manuscript Accepted: 10 SEP 2012
- Manuscript Revised: 2 SEP 2012
- Manuscript Received: 15 JUL 2012
- National Research Foundation (NRF) via National Research Laboratory. Grant Number: R0A-2008-000-20041-0
- Intelligent Synthetic Biology Center of Global Frontier Project. Grant Numbers: 2011-0031957, 2011-0031946
- Converging Research Center, Republic of Korea. Grant Numbers: 2012k001388, 2011K000841
- redox enzymes;
- cytochrome P450;
- cofactor regeneration;
- cell-free protein synthesis
Cytochrome P450 monooxygenases are multifunctional enzymes with potential applications in chemoenzymatic synthesis of complex chemicals as well as in studies of metabolism and xenobiotics. Widespread application of cytochrome P450s, however, is encumbered by the critical need for redox equivalents in their catalytic function. To overcome this limitation, we studied visible light-driven regeneration of NADPH for P450-catalyzed O-dealkylation reaction; we used eosin Y as a photosensitizing dye, triethanolamine as an electron donor, and [Cp*Rh(bpy)H2O] as an electron mediator. We analyzed catalytic activity of cell-free synthesized P450 BM3 monooxygenase variant (Y51F/F87A, BM3m2) in the presence of key components for NADPH photoregeneration. The P450-catalyzed O-dealkylation reaction sustainably maintained its turnover with the continuous supply of photoregenerated NADPH. Visible light-driven, non-enzymatic NADPH regeneration provides a new route for efficient, sustainable utilization of P450 monooxygenases. Biotechnol. Bioeng. 2013; 110: 383–390. © 2012 Wiley Periodicals, Inc.