P450camr, a cytochrome P450 catalysing the stereospecific 6-endo-hydroxylation of (1R)-(+)-camphor

• Published in: Applied microbiology and biotechnology Vol. 59; no. 4-5; pp. 449 - 454
• Main Authors: GROGAN, G, ROBERTS, G. A, PARSONS, S, TURNER, N. J, FLITSCH, S. L
• Format: Journal Article
• Language: English
• Published: Berlin Springer 2002; Springer Nature B.V
• Subjects: Bacteria; Bioconversions. Hemisynthesis; Biological and medical sciences; Biotechnology; Carbon sources; Crystallography; Cytochrome; Enzymes; Fundamental and applied biological sciences. Psychology; Methods. Procedures. Technologies; Microbiology; Monoterpene; Stereospecificity; Hydroxylation; Camphor; Cytochrome P450
• ISSN: 0175-7598; 1432-0614
• DOI: 10.1007/s00253-002-1054-0

Rhodococcus sp. NCIMB 9784 accumulated 6-endo-hydroxycamphor 3 when grown on (1R)-(+)-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 (1R)-(+)-camphor and designated P450^sub 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^sub camr^ and putidaredoxin and putidaredoxin reductase from Pseudomonas putida confirmed that the enzyme hydroxylates (1R)-(+)-camphor specifically in the 6-endo position, in contrast to the 5-exo hydroxylation catalysed by the well-studied P450^sub cam^ from P. putida. P450^sub camr^ has a molecular mass of approximately 44 kDa, and a pI of 4.8.[PUBLICATION ABSTRACT]