Characterization of a second alkane-inducible cytochrome P450-encoding gene, CYP52A2, from Candida tropicalis.
A second alkane-inducible cytochrome P450-encoding gene (CYP52A2) from the yeast Candida tropicalis was sequenced and characterized. CYP52A2 is located 1 kb upstream from CYP52A1, the previously characterized P450 gene [Sanglard and Loper, Gene 76 (1989) 121-136] and shows the same orientation. Like CYP52A1, CYP52A2 is induced by growth on alkane. Both promoter regions share repeats of the sequence CATGTGAA that could be of importance for the induction of the two genes. At the amino acid level, alk2 shows an overall identity of 68.2% and an overall similarity of 81.6% to alk1. Regions of high homology between the two proteins are found in the distal and proximal heme binding sites which contain the highly conserved cysteine residue as the fifth ligand to the heme iron. However, marked differences between the two proteins exist at their N-terminal end, which includes the transmembrane domain, and at the putative substrate-binding domain. Upon expression of CYP52A2 in Saccharomyces cerevisiae, alk2 was shown to hydroxylate hexadecane, but had no hydroxylation activity towards lauric acid, whereas alk1 showed both activities. Comparative immunoblots demonstrate that neither alk1 nor alk2 expressed in S. cerevisiae corresponds to the main cytochrome P450 present in C. tropicalis when grown on alkane.
Institute of Biotechnology, Swiss Federal Institute of Technology, ETH Hönggerberg, Zürich.
W
W
Seghezzi
D
D
Sanglard
A
A
Fiechter
D000473:Alkanes / Q000378:metabolism*; D000595:Amino Acid Sequence; D001483:Base Sequence; D015152:Blotting, Northern; D015153:Blotting, Western; D002175:Candida / Q000235:genetics*; D003577:Cytochrome P-450 Enzyme System / Q000235:genetics*; D004271:DNA, Fungal; D005656:Fungal Proteins; D015870:Gene Expression; D005800:Genes, Fungal; D006900:Hydroxylation; D008861:Microsomes / Q000378:metabolism; D008969:Molecular Sequence Data; D012331:RNA, Fungal / Q000235:genetics; D015183:Restriction Mapping; D012441:Saccharomyces cerevisiae / Q000235:genetics; D016415:Sequence Alignment; D012689:Sequence Homology, Nucleic Acid; D013379:Substrate Specificity
D000473:Alkanes; D004271:DNA, Fungal; D005656:Fungal Proteins; D012331:RNA, Fungal; D003577:Cytochrome P-450 Enzyme System; C499277:Alk2 protein, Candida