title: Electron donation to an archaeal cytochrome P450 is enhanced by PCNA-mediated selective complex formation with foreign redox proteins.
abstract: Cytochrome P450 monooxygenases (P450s) are environmentally friendly biocatalysts that catalyze diverse chemical reactions using molecular oxygen under mild reaction conditions. P450s are activated upon receiving electrons from specific redox partner proteins, although the redox partners for most bacterial/archaeal P450s are not yet identified. Thus, it is important to establish a variety of efficient and versatile electron transfer systems from NAD(P)H to P450s for the design of biocatalysts. Sulfolobus solfataricus possesses a heterotrimeric proliferating cell nuclear antigen (PCNA). Fusion of the PCNA subunits to S. acidocaldarius P450 (CYP119) and the Pseudomonas putida redox proteins, putidaredoxin (PdX) and putidaredoxin reductase (PdR), yielded a selective protein complex containing one molecule each of the three proteins. The PCNA-mediated heterotrimerization of CYP119, PdX, and PdR enhanced the CYP119 activity, likely as a result of high local concentrations of the two redox proteins toward CYP119. Therefore, the PCNA-mediated formation of the complex containing PdX and PdR might be applicable for harnessing the utility of P450s whose redox partners are not yet identified.
authors.affiliation: Department of Bioengineering, School of Engineering, The University of Tokyo, Tokyo, Japan.
authors.forename: Risa
authors.initials: R
authors.lastname: Suzuki
authors.forename: Hidehiko
authors.initials: H
authors.lastname: Hirakawa
authors.forename: Teruyuki
authors.initials: T
authors.lastname: Nagamune
mesh_terms: D019843:Archaeal Proteins / Q000737:chemistry / Q000378:metabolism*; D003577:Cytochrome P-450 Enzyme System / Q000737:chemistry / Q000378:metabolism*; D004579:Electron Transport; D005288:Ferredoxins / Q000737:chemistry / Q000378:metabolism*; D006900:Hydroxylation; D007850:Lauric Acids / Q000737:chemistry / Q000378:metabolism; D009247:NADH, NADPH Oxidoreductases / Q000737:chemistry / Q000378:metabolism*; D018809:Proliferating Cell Nuclear Antigen / Q000737:chemistry / Q000378:metabolism*; D055550:Protein Stability
chemical_list: D019843:Archaeal Proteins; D005288:Ferredoxins; D007850:Lauric Acids; D018809:Proliferating Cell Nuclear Antigen; C030358:lauric acid; C100137:putidaredoxin; D003577:Cytochrome P-450 Enzyme System; C408874:CYP119 protein, Sulfolobus solfataricus; D009247:NADH, NADPH Oxidoreductases; C022486:putidaredoxin reductase
keywords: Artificial protein complex; CYP119; Proliferating cell nuclear antigen; Putidaredoxin; Sulfolobus solfataricus