Defluorination of 4-fluorophenol by cytochrome P450(BM₃)-F87G: activation by long chain fatty aldehydes.
Cytochrome P450(BM3)-F87G catalyzed the oxidative defluorination of 4-fluorophenol, followed by reduction of the resulting benzoquinone to hydroquinone via the NADPH P450-reductase activity of the enzyme. The k (cat) and K (m) for this reaction were 71 ± 5 min(-1) and 9.5 ± 1.3 mM, respectively. Co-incubation of the reaction mixture with long chain aldehydes stimulated the defluorination reaction, with the 2,3-unsaturated aldehyde, 2-decenal producing a 12-fold increase in catalytic efficiency. At 150 μM aldehyde, k (cat) increased to 158 ± 4, while K (m) decreased to 1.8 ± 0.2. The effects of catalase, glutathione and ascorbate on the reaction were all consistent with a direct oxygen insertion mechanism, as opposed to a radical mechanism. The study demonstrates the potential use of P450(BM3) mutants in oxidative defluorination reactions, and characterizes the novel stimulatory action of straight chain aldehydes on this activity.
The Department of Chemistry, The University of North Carolina at Greensboro, 416 Sullivan Science Building, Greensboro, NC 27402, USA.
Alexandria
A
Harkey
Hye-Jin
HJ
Kim
Suneel
S
Kandagatla
Gregory M
GM
Raner
D000447:Aldehydes / Q000378:metabolism*; D000475:Alkenes / Q000378:metabolism*; D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism*; D003577:Cytochrome P-450 Enzyme System / Q000235:genetics / Q000378:metabolism*; D020536:Enzyme Activators / Q000378:metabolism*; D007700:Kinetics; D050505:Mutant Proteins / Q000235:genetics / Q000378:metabolism; D010084:Oxidation-Reduction; D010636:Phenols / Q000378:metabolism*
C476489:(2E)-decenal; D000447:Aldehydes; D000475:Alkenes; D001426:Bacterial Proteins; D020536:Enzyme Activators; D050505:Mutant Proteins; D010636:Phenols; C106880:4-fluorophenol; D003577:Cytochrome P-450 Enzyme System