title: Comparative functional characterization of a novel benzoate hydroxylase cytochrome P450 of Fusarium oxysporum.
abstract: FoCYP53A19, a novel cytochrome P450 capable of performing benzoate hydroxylation, was identified and characterized from the ascomycete Fusarium oxysporum f.sp. lycopersici. Comparative functional analysis of FoCYP53A19 with the heterologous and homologous cytochrome P450 reductases (CPR) such as Saccharomyces cerevisiae (ScCPR), Candida albicans (CaCPR) and F. oxysporum (FoCPR) revealed novel catalytic properties. The catalytic efficiency and substrate specificity of FoCYP53A19 were significantly influenced and altered by the source of the reductase employed. The yeast reconstitution system of FoCYP53A19 with ScCPR performed the hydroxylation of benzoic acid (BA) and demethylation of 3-methoxybenzoic acid (3-MBA); but when reconstituted with CaCPR, FoCYP53A19 performed only the essential hydroxylation of fungal benzoate catabolism. Remarkably, FoCYP53A19 with its homologous reductase FoCPR, not only demonstrated the improved conversion rates of BA and 3-MBA, but also exhibited activity toward the hydroxylation of 3-hydroxybenzoic acid. The electron transfer compatibility and the coupling efficiency between the homologous FoCYP-FoCPR system are significant and it favored enhanced monooxygenase activity with broader substrate specificity.
authors: School of Biotechnology, Yeungnam University, Gyeongsan, South Korea.
authors: Pradeepraj
authors: P
authors: Durairaj
authors: School of Chemical and Biological Engineering, Seoul National University, Seoul, South Korea.
authors: Eunok
authors: E
authors: Jung
authors: School of Biotechnology, Yeungnam University, Gyeongsan, South Korea.
authors: Hyun Ho
authors: HH
authors: Park
authors: School of Chemical and Biological Engineering, Seoul National University, Seoul, South Korea.
authors: Byung-Gee
authors: BG
authors: Kim
authors: Department of Bioscience and Biotechnology, Konkuk University, Seoul, South Korea. Electronic address: hyungdon@konkuk.ac.kr.
authors: Hyungdon
authors: H
authors: Yun
mesh_terms: D000595:Amino Acid Sequence; D001709:Biotechnology; D002176:Candida albicans / Q000201:enzymology / Q000235:genetics; D003577:Cytochrome P-450 Enzyme System / Q000235:genetics / Q000378:metabolism*; D005656:Fungal Proteins / Q000235:genetics / Q000378:metabolism*; D005670:Fusarium / Q000201:enzymology* / Q000235:genetics; D005800:Genes, Fungal; D008969:Molecular Sequence Data; D010084:Oxidation-Reduction; D010105:Oxygenases / Q000235:genetics / Q000378:metabolism*; D010802:Phylogeny; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000201:enzymology / Q000235:genetics; D017386:Sequence Homology, Amino Acid
chemical_list: D005656:Fungal Proteins; D011994:Recombinant Proteins; D003577:Cytochrome P-450 Enzyme System; D010105:Oxygenases; C019766:benzoate 1,2-dioxygenase
keywords: Benzoate hydroxylase; Cytochrome P450; Cytochrome P450 reductase; Heterologous expression; Saccharomyces cerevisiae; cDNA gene cloning