title: Immobilization of a Bacterial Cytochrome P450 Monooxygenase System on a Solid Support.
abstract: Bacterial cytochrome P450s (P450s), which catalyze regio- and stereoselective oxidations of hydrocarbons with high turnover rates, are attractive biocatalysts for fine chemical production. Enzyme immobilization is needed for cost-effective industrial manufacturing. However, immobilization of P450s is difficult because electron-transfer proteins are involved in catalysis and anchoring these can prevent them from functioning as shuttle molecules for carrying electrons. We studied a heterotrimeric protein-mediated co-immobilization of a bacterial P450, and its electron-transfer protein and reductase. Fusion with subunits of a heterotrimeric Sulfolobus solfataricus proliferating cell nuclear antigen (PCNA) enabled immobilization of the three proteins on a solid support. The co-immobilized enzymes catalyzed monooxygenation because the electron-transfer protein fused to PCNA via a single peptide linker retained its electron-transport function.
authors: Department of Bioengineering, School of Engineering, The University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo, 113-8656, Japan.
authors: Cheau Yuaan
authors: CY
authors: Tan
authors: Department of Chemistry and Biotechnology, School of Engineering, The University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo, 113-8656, Japan.
authors: Hidehiko
authors: 0000-0003-0623-8573
authors: H
authors: Hirakawa
authors: Department of Bioengineering, School of Engineering, The University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo, 113-8656, Japan.|Department of Biotechnology, Graduate School of Engineering, Nagoya University, Furo-cho, Chikusa-ku, Nagoya, Aichi, 464-8603, Japan.
authors: Risa
authors: 0000-0002-1565-5339
authors: R
authors: Suzuki
authors: Department of Chemistry and Biotechnology, School of Engineering, The University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo, 113-8656, Japan.
authors: Tomoaki
authors: T
authors: Haga
authors: Department of Chemistry and Biotechnology, School of Engineering, The University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo, 113-8656, Japan.
authors: Fumiya
authors: F
authors: Iwata
authors: Department of Bioengineering, School of Engineering, The University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo, 113-8656, Japan.|Department of Chemistry and Biotechnology, School of Engineering, The University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo, 113-8656, Japan.
authors: Teruyuki
authors: 0000-0002-2435-0270
authors: T
authors: Nagamune
mesh_terms: D003577:Cytochrome P-450 Enzyme System / Q000378:metabolism*; D004800:Enzymes, Immobilized / Q000378:metabolism*; D008958:Models, Molecular; D016958:Pseudomonas putida / Q000201:enzymology*
chemical_list: D004800:Enzymes, Immobilized; D003577:Cytochrome P-450 Enzyme System
keywords: biocatalysis; cytochromes; enzymes; hydroxylation; immobilization