title: Identification of the specific electron transfer proteins, ferredoxin, and ferredoxin reductase, for CYP105D7 in Streptomyces avermitilis MA4680.
abstract: It was previously proposed that regiospecific hydroxylation of daidzein at 3'-position is mediated by cytochrome P450 hydroxylase (CYP105D7) in the presence of putidaredoxin (CamB) and putidaredoxin reductase (CamA) as electron transfer proteins from Pseudomonas putida. The genome sequence of Streptomyces avermitilis MA4680 revealed 33 P450 (CYPs) with 6 ferredoxin reductases (Fprs) and 9 ferredoxins (Fdxs) as their putative electron transfer partner proteins. To identify right endogenous electron transfer proteins for CYP105D7 activity, in vitro reconstitution, gene disruption, and quantitative reverse transcription polymerase chain reaction (qRT-PCR) mRNA expression profile analysis were examined. The most effective electron transfer proteins for CYP105D7 appear to be FdxH (SAV7470), which is located downstream to CYP105D7 as a cluster, and FprD (SAV5675). Throughout our overall analysis, we proposed that the primary electron transfer pathway for CYP105D7 follows as such NAD(P)H→FdxH→FprD→CYP105D7.
authors.affiliation: School of Chemical and Biological Engineering, Institute of Molecular Biology and Genetics, Institute of Bioengineering, Seoul National University, Seoul, South Korea.
authors.forename: Bishnu Prasad
authors.initials: BP
authors.lastname: Pandey
authors.forename: Nahum
authors.initials: N
authors.lastname: Lee
authors.forename: Kwon-Young
authors.initials: KY
authors.lastname: Choi
authors.forename: Ji-Nu
authors.initials: JN
authors.lastname: Kim
authors.forename: Eun-Jung
authors.initials: EJ
authors.lastname: Kim
authors.forename: Byung-Gee
authors.initials: BG
authors.lastname: Kim
mesh_terms: D003577:Cytochrome P-450 Enzyme System / Q000235:genetics*; D005288:Ferredoxins / Q000235:genetics* / Q000378:metabolism*; D017353:Gene Deletion; D020869:Gene Expression Profiling; D053858:Metabolic Networks and Pathways; D010088:Oxidoreductases / Q000235:genetics* / Q000378:metabolism*; D060888:Real-Time Polymerase Chain Reaction; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013302:Streptomyces / Q000201:enzymology* / Q000235:genetics
chemical_list: D005288:Ferredoxins; D003577:Cytochrome P-450 Enzyme System; D010088:Oxidoreductases