Abstract
The ability of cytochrome P450 enzymes to catalyze highly regio- and stereospecific hydroxylations makes them attractive alternatives to approaches based on chemical synthesis but they require expensive cofactors, e.g. NAD(P)H, which limits their commercial potential. Ferredoxin (Fdx) is a multifunctional electron carrier that in plants accepts electrons from photosystem I (PSI) and facilitates photoreduction of NADP+ to NADPH mediated by ferredoxin-NAD(P)H oxidoreductase (FdR). In bacteria, the electron flow is reversed and Fdx accepts electrons from NADPH via FdR and serves as the direct electron donor to bacterial P450s. By combining the two systems, we demonstrate that irradiation of PSI can drive the activity of a bacterial P450, CYP124 from Mycobacterium tuberculosis. The substitution of the costly cofactor NADPH with sunlight illustrates the potential of the light-driven hydroxylation system for biotechnology applications.
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Acknowledgements
KJ was supported by a Ph.D. stipend from the Faculty of Life Sciences, University of Copenhagen and sponsored by a travel stipend from Bilateral network activities, Visiting Scientist Grants 2010: Improving plants for food, fuel and bioactive natural products (09-075501). JBJ was supported by a Heiser Postdoctoral Fellowship for Research in Leprosy and Tuberculosis sponsored by The New York Community Trust. This work was also funded by the Villum Foundation to the research centre “Pro-Active Plants” and from “Center for Synthetic Biology”, funded by the UNIK research initiative of the Danish Ministry of Science, Technology and Innovation (BLM) and NIH grants GM25515 and AI074824 (P.O.M.).
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Jensen, K., Johnston, J.B., de Montellano, P.R.O. et al. Photosystem I from plants as a bacterial cytochrome P450 surrogate electron donor: terminal hydroxylation of branched hydrocarbon chains. Biotechnol Lett 34, 239–245 (2012). https://doi.org/10.1007/s10529-011-0768-4
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DOI: https://doi.org/10.1007/s10529-011-0768-4