Identification of a novel cytochrome P-450 gene from the white rot fungus Phanerochaete chrysosporium.
A gene fragment belonging to the cytochrome P-450 superfamily has been cloned and identified from stationary cultures of the filamentous fungus Phanerochaete chrysosporium by reverse transcriptase (RT)-PCR. A set of degenerate primers homologous to highly conserved regions of known cytochrome P-450 sequences were used for initial RT-PCRs. Individual PCR products were cloned, sequenced, and identified as those belonging to the cytochrome P-450 superfamily based on amino acid sequence homologies and the presence of the highly conserved heme binding region. The nucleotide sequence of a single cDNA clone indicated the presence of an open reading frame encoding a partial cytochrome P-450 protein of 208 amino acids. Comparisons of the deduced amino acid sequence of the partial protein to other known cytochrome P-450 sequences indicate that it is the first member of a new family of cytochrome P-450s, designated CYP63-1A. Northern blot analysis suggests that CYP63-1A is expressed under both nitrogen-rich and nitrogen-deficient culture conditions and thus not under the same regulatory constraints as the well-studied lignin and manganese peroxidases. Western blot analyses using antibodies raised to the heme binding region of CYP63-1A indicate that the protein has a molecular mass of approximately 44,000 Da.
Kullman
S W
SW
Department of Environmental Toxicology, University of California, Davis 95616, USA.
Matsumura
F
F
D000595:Amino Acid Sequence; D001483:Base Sequence; D001487:Basidiomycota / Q000235:genetics*; D015152:Blotting, Northern; D015153:Blotting, Western; D003001:Cloning, Molecular; D003577:Cytochrome P-450 Enzyme System / Q000235:genetics*; D015966:Gene Expression Regulation, Fungal; D006418:Heme / Q000235:genetics / Q000276:immunology; D008969:Molecular Sequence Data; D009584:Nitrogen / Q000378:metabolism; D016366:Open Reading Frames; D016133:Polymerase Chain Reaction; D012331:RNA, Fungal / Q000032:analysis; D017386:Sequence Homology, Amino Acid
D012331:RNA, Fungal; D006418:Heme; D003577:Cytochrome P-450 Enzyme System; D009584:Nitrogen