Cytochrome P450 enzymes from the metabolically diverse bacterium Rhodopseudomonas palustris.
Four (CYP195A2, CYP199A2, CYP203A1, and CYP153A5) of the seven P450 enzymes, and palustrisredoxin A, a ferredoxin associated with CYP199A2, from the metabolically diverse bacterium Rhodopseudomonas palustris have been expressed and purified. A range of substituted benzenes, phenols, benzaldehydes, and benzoic acids was shown to bind to the four P450 enzymes. Monooxygenase activity of CYP199A2 was reconstituted with palustrisredoxin A and putidaredoxin reductase of the P450cam system from Pseudomonas putida. We found that 4-ethylbenzoate and 4-methoxybenzoate were oxidized to single products, and 4-methoxybenzoate was demethylated to form 4-hydroxybenzoate. Crystals of substrate-free CYP199A2 which diffracted to approximately 2.0A have been obtained.
Department of Chemistry, University of Oxford, Inorganic Chemistry Laboratory, South Parks Road, Oxford, OX1 3QR, UK. stephen.bell@chem.ox.ac.uk
Stephen G
SG
Bell
Nicola
N
Hoskins
Feng
F
Xu
Domenico
D
Caprotti
Zihe
Z
Rao
Luet-Lok
LL
Wong
D002384:Catalysis; D003460:Crystallization; D003577:Cytochrome P-450 Enzyme System / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D015870:Gene Expression; D012241:Rhodopseudomonas / Q000145:classification / Q000201:enzymology* / Q000235:genetics / Q000378:metabolism; D013057:Spectrum Analysis; D013379:Substrate Specificity
D003577:Cytochrome P-450 Enzyme System