Physiological regulation, xenobiotic induction, and heterologous expression of P450 monooxygenase gene pc-3 (CYP63A3), a new member of the CYP63 gene cluster in the white-rot fungus Phanerochaete chrysosporium.

TitlePhysiological regulation, xenobiotic induction, and heterologous expression of P450 monooxygenase gene pc-3 (CYP63A3), a new member of the CYP63 gene cluster in the white-rot fungus Phanerochaete chrysosporium.
Publication TypeJournal Article
Year of Publication2005
AuthorsDoddapaneni, HV, Subramanian, V, Yadav, JS
JournalCurr Microbiol
Volume50
Issue6
Pagination292-8
Date Published2005 Jun
ISSN0343-8651
KeywordsAdaptation, Physiological, Amino Acid Sequence, Culture Media, Cytochrome P-450 Enzyme System, Enzyme Induction, Escherichia coli, Gene Expression Regulation, Enzymologic, Gene Expression Regulation, Fungal, Hydrocarbons, Aromatic, Lignin, Molecular Sequence Data, Multigene Family, Phanerochaete, Xenobiotics
Abstract

In order to characterize the functional diversity in CYP63 cluster of tandemly linked P450 genes (pc-1, pc-2, and pc-3) in Phanerochaete chrysosporium, here we report the functional characterization of pc-3 (CYP63A3), a newly cloned member of this group. pc-3 expression was favored in nutrient-limited versus nutrient-rich media in 3-6-day-old cultures and was upregulated by starch as a carbon source or by oxygenation of cultures. pc-3 was induced by various xenobiotics in defined nutrient-limited (3-9-fold) and nutrient-rich (2-5-fold) cultures. Particularly, a range of unsubstituted and substituted aliphatic hydrocarbons (alkanes and fatty acids) induced the expression under the two nutrient conditions albeit in a differential manner. Interestingly, pc-3 was also inducible by certain oxygenated mono aromatics (nitrophenol, benzoate, and resorcinol), lower molecular weight (2 to 4 ring size) polycyclic aromatic hydrocarbons (PAHs) and alkali-treated lignin derivatives in nutrient-rich malt extract cultures. The study further establishes that the three CYP63 genes (CYP63A1, A2, and A3) are independently regulated despite being members of the tandem gene cluster with high gene structural similarity (13-14 introns) and protein sequence homology (59-85%). The pc-3 cDNA (1,812 bp) was expressed in E. coli as a His-tagged protein (approximately 74 kDa). This constitutes the first report on heterologous expression of a P450 monooxygenase enzyme from this model white-rot fungus.

DOI10.1007/s00284-005-4480-2
Alternate JournalCurr. Microbiol.
PubMed ID15968506
Grant ListR01-ES10210 / ES / NIEHS NIH HHS / United States