earticle

영어

Cytochrome P450, the heme-thiolate proteins are the ubiquitous mono-oxygenase enzymes which plays a key role in almost all living forms; prokaryotes to eukaryotes. The P450 enzymes catalyze versatile oxidation reactions and have wide substrate specificity. Despite, the P450 enzyme plays a crucial role in fungi by its hydrophobic conversion of the primary and secondary metabolites and in the adaption of diverse ecological niches by degrading the ecological pollutants. Indeed, genome sequencing projects has revealed the presence of several thousands of putative P450 genes from various families in the fungal kingdom. Bioinformatic annotations showcase the presence of 169 putative P450 genes and 2 putative Cytochrome P450 reducatase (CPR) genes from the whole genome sequence of Fusarium oxysporum f. sp. lycopersici. We initiated the functional genomic research of this fungus by screening and selecting the target putative novel P450s based on the multiple alignment and the phylogenetic analysis with the reported P450 genes of other organisms with various novel functionalities. So far, the target P450s selected includes the Sterol 14α-Lanosterol demethylase, Hydroxylation of Flavonoids (flavine and naringenin), Hydroxylation of Benzoates, Oxidation of polycyclic aromatic hydrocarbons (PAH) and Toluene, etc. To characterize the functional FoCYPs, cDNAs are synthesized by optimizing the culture conditions and the transcriptional parameters. To date, about 20 P450 genes and CPR genes are been isolated and cloned, which will be expressed to analyze biochemically. Consequently, the FoCYP P450 gene library will be constructed, which drives us towards an intensive approach to determine the functionalities of novel P450s.