Spectroscopic studies of the oxidation of ferric CYP153A6 by peracids: Insights into P450 higher oxidation states.
Our previous rapid-scanning stopped-flow studies of the reaction of substrate-free cytochrome P450cam with peracids [T. Spolitak, J.H. Dawson, D.P. Ballou, J. Biol. Chem. 280 (2005) 20300-20309; J. Inorg. Biochem. 100 (2006) 2034-2044; J. Biol. Inorg. Chem. 13 (2008) 599-611] spectrally characterized compound I (ferryl iron plus a porphyrin pi-cation radical (Fe(IV)O/Por(.+))), Cpd ES, and Cpd II (Fe(IV)O/Tyr() or Fe(IV)O). We now report that reactions of CYP153A6 with peracids yield all these intermediates, with kinetic profiles allowing better resolution of all forms at pH 8.0 compared to similar reactions with WT P450cam. Properties of the reactions of these higher oxidation state intermediates were determined in double-mixing experiments in which intermediates are pre-formed and ascorbate is then added. Reactions of heptane-bound CYP153A6 (pH 7.4) with mCPBA resulted in conversion of P450 to the low-spin ferric form, presumably as heptanol was formed, suggesting that CYP 153A6 is a potential biocatalyst that can use peracids with no added NAD(P)H or reducing systems for bioremediation and other industrial applications.
Spolitak
Tatyana
T
Department of Biological Chemistry, Medical School, University of Michigan, Ann Arbor, MI 48109-5606, USA.
Funhoff
Enrico G
EG
Ballou
David P
DP
D002723:Chlorobenzoates / Q000737:chemistry* / Q000494:pharmacology; D003577:Cytochrome P-450 Enzyme System / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D005290:Ferric Compounds / Q000737:chemistry / Q000378:metabolism; D006863:Hydrogen-Ion Concentration; D009161:Mycobacterium / Q000201:enzymology* / Q000235:genetics; D016877:Oxidants / Q000737:chemistry* / Q000494:pharmacology; D010084:Oxidation-Reduction / Q000187:drug effects
D002723:Chlorobenzoates; D005290:Ferric Compounds; D016877:Oxidants; D003577:Cytochrome P-450 Enzyme System; C000433:3-chloroperbenzoic acid