Identification and characterization of novel cytochrome P450 genes from the white-rot basidiomycete, Coriolus versicolor

• Published in: Applied microbiology and biotechnology 2002, Vol.58 (1), p.97-105
• Main Authors: Ichinose, H, Wariishi, H, Tanaka, H
• Format: Article
• Language: English
• Subjects: Amino Acid Sequence; amino acid sequences; Amino acids; Aspergillus terreus; Base Sequence; Basidiomycota - enzymology; Basidiomycota - genetics; Basidiomycota - growth & development; Biological and medical sciences; Biology of microorganisms of confirmed or potential industrial interest; Biotechnology; Cloning; complementary DNA; Coriolus versicolor; CYP512A1 protein; Cytochrome; cytochrome P-450; Cytochrome P-450 Enzyme System - chemistry; Cytochrome P-450 Enzyme System - genetics; Cytochrome P-450 Enzyme System - metabolism; cytochrome P450; Fundamental and applied biological sciences. Psychology; Fusarium; gene expression regulation; Gene Expression Regulation, Fungal; genes; Genes, Fungal; Genetics; Gibberella fujikuroi; Heat-Shock Response; microorganisms; Mission oriented research; Molecular Sequence Data; Neurospora crassa; new family; nucleotides; Proteins; rapid amplification of cDNA ends; Reverse Transcriptase Polymerase Chain Reaction; Sequence Analysis, DNA; Thiophenes - chemistry; Thiophenes - pharmacology
• ISSN: 0175-7598; 1432-0614
• DOI: 10.1007/s00253-001-0868-5

Using a reverse-transcription polymerase chain reaction (RT-PCR) technique, cytochrome P450 genes were cloned from the lignin-degrading basidiomycete, Coriolus versicolor. One possible P450 gene was identified, which consisted of 1,672 nucleotides and a poly(A) tail and encoded a deduced protein containing 449 amino acids. The deduced amino acid sequence revealed the presence of the P450 heme-binding motif, strongly suggesting that this protein belongs to the P450 superfamily, then designated CYP512A1. The deduced protein showed sequential similarity to other known P450s from several micro-organisms, such as Aspergillus terreus, Gibberella fujikuroi, and Neurospora crassa, with 30-35% identity. Since the identity of the amino acid sequence was less than 40% with any other P450s, this protein was suggested to be the first member of a new family of cytochrome P450. In addition, a differential display RT-PCR analysis showed the expression of the other P450 genes, which were up-regulated by the addition of dibenzothiophene and 4-methyldibenzothiophene-5-oxide. Using the 5' rapid amplification of cDNA ends method, a 520-nucleotide sequence, including the P450 motif-coding region, was determined for one clone. The deduced protein showed high similarity to CYP512A1 but less than 40% identity with P450s from other organisms. A chemical stress-responsive expression of P450 is suggested for the first time in basidiomycetes.