P450(camr), a cytochrome P450 catalysing the stereospecific 6- endo-hydroxylation of (1 R)-(+)-camphor.
Rhodococcus sp. NCIMB 9784 accumulated 6- endo-hydroxycamphor 3 when grown on (1 R)-(+)-camphor 1 as sole carbon source. The structure of 3 has been unambiguously assigned for the first time using X-ray crystallography. A soluble cytochrome P450 hydroxylase, induced by growth on (1 R)-(+)-camphor and designated P450(camr), has been isolated from the bacterium Rhodococcus sp. NCIMB 9784. Using authentic 6- endo hydroxycamphor as standard, a cell-free system consisting of pure P450(camr) and putidaredoxin and putidaredoxin reductase from Pseudomonas putida confirmed that the enzyme hydroxylates (1 R)-(+)-camphor specifically in the 6- endoposition, in contrast to the 5- exo hydroxylation catalysed by the well-studied P450(cam) from P. putida. P450(camr) has a molecular mass of approximately 44 kDa, and a pI of 4.8.
Grogan
G
G
Edinburgh Centre for Protein Technology, Department of Chemistry, University of Edinburgh, The King's Buildings, Edingburg, UK.
Roberts
G A
GA
Parsons
S
S
Turner
N J
NJ
Flitsch
S L
SL
D002164:Camphor / Q000737:chemistry* / Q000378:metabolism*; D018360:Crystallography, X-Ray; D003577:Cytochrome P-450 Enzyme System / Q000302:isolation & purification / Q000378:metabolism*; D004591:Electrophoresis, Polyacrylamide Gel; D006900:Hydroxylation; D008958:Models, Molecular; D012240:Rhodococcus / Q000201:enzymology* / Q000254:growth & development; D013237:Stereoisomerism
D002164:Camphor; D003577:Cytochrome P-450 Enzyme System