Conservation and cloning of CYP51: a sterol 14 alpha-demethylase from Mycobacterium smegmatis.
The genetic locus encoding cytochrome P450 51 (CYP51; P450(14DM)) in Mycobacterium smegmatis is described here together with confirmation of activity in lanosterol 14 alpha-demethylation. The protein bound azole antifungals with high affinity and the rank order based on affinity matched the ranked order for microbiological sensitivity of the organism, thus supporting a possible role for CYP51 as a target in the antimycobacterial activity of these compounds. Non-saponifiable lipids were extracted from the bacteria grown on minimal medium. Unlike a previous report using growth on complex medium, no cholesterol was detected in two strains of M. smegmatis, but a novel lipid was detected. The genetic locus of CYP51 is discussed in relation to function; it is conserved as part of a putative operon in M. smegmatis, Mycobacterium tuberculosis, Mycobacterium avium, and Mycobacterium bovis and consists of six open-reading frames including two CYPs and a ferredoxin under a putative Tet-R regulated promoter.
Jackson
Colin J
CJ
Wolfson Laboratory of P450 Biodiversity, Institute of Biological Sciences, University of Wales, Aberystwyth, Wales, SY23 3DA, UK.
Lamb
David C
DC
Marczylo
Timothy H
TH
Parker
Josie E
JE
Manning
Nigel L
NL
Kelly
Diane E
DE
Kelly
Steven L
SL
D000595:Amino Acid Sequence; D000818:Animals; D003001:Cloning, Molecular; D003577:Cytochrome P-450 Enzyme System / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D005798:Genes, Bacterial; D006801:Humans; D007810:Lanosterol / Q000378:metabolism; D008055:Lipids / Q000737:chemistry; D008969:Molecular Sequence Data; D020102:Mycobacterium smegmatis / Q000201:enzymology* / Q000235:genetics; D010088:Oxidoreductases / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D010802:Phylogeny; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D016415:Sequence Alignment; D058886:Sterol 14-Demethylase
C548418:CYP51A1 protein, human; D008055:Lipids; D011994:Recombinant Proteins; D007810:Lanosterol; D003577:Cytochrome P-450 Enzyme System; D010088:Oxidoreductases; D058886:Sterol 14-Demethylase