Cofactor-Free Light-Driven Whole-Cell Cytochrome P450 Catalysis

• Published in: Angewandte Chemie International Edition Vol. 54; no. 3; pp. 969 - 973
• Main Authors: Park, Jong Hyun, Lee, Sahng Ha, Cha, Gun Su, Choi, Da Som, Nam, Dong Heon, Lee, Jae Hyung, Lee, Jung-Kul, Yun, Chul-Ho, Jeong, Ki Jun, Park, Chan Beum
• Format: Journal Article
• Language: English
• Published: Weinheim WILEY-VCH Verlag 12.01.2015; WILEY‐VCH Verlag; Wiley Subscription Services, Inc
• Edition: International ed. in English
• Subjects: Bacteria; Bacterial Proteins - chemistry; Bacterial Proteins - metabolism; Biocatalysis; Catalysis; Catalysts; cofactors; Cytochrome P-450 Enzyme System - chemistry; Cytochrome P-450 Enzyme System - metabolism; Cytochromes P450; Cytoplasm; Electron Transport; enzymatic catalysis; Escherichia coli; Escherichia coli - metabolism; Estradiol - chemistry; Estradiol - metabolism; Fluorescein - chemistry; Fluorescein - metabolism; Heme - chemistry; Heme - metabolism; Humans; Illumination; Light; Lovastatin - chemistry; Lovastatin - metabolism; Omeprazole - chemistry; Omeprazole - metabolism; Oxidation-Reduction; photocatalysis; photoinduced electron transfer; Photosensitizing Agents - chemistry; Photosensitizing Agents - metabolism; Platforms; Protein Structure, Tertiary; Simvastatin - chemistry; Simvastatin - metabolism; photoinduced electron transfer; cytochromes P450; cofactors; photocatalysis; enzymatic catalysis
• ISSN: 1433-7851; 1521-3773
• DOI: 10.1002/anie.201410059

Cytochromes P450 can catalyze various regioselective and stereospecific oxidation reactions of non‐functionalized hydrocarbons. Here, we have designed a novel light‐driven platform for cofactor‐free, whole‐cell P450 photo‐biocatalysis using eosin Y (EY) as a photosensitizer. EY can easily enter into the cytoplasm of Escherichia coli and bind specifically to the heme domain of P450. The catalytic turnover of P450 was mediated through the direct transfer of photoinduced electrons from the photosensitized EY to the P450 heme domain under visible light illumination. The photoactivation of the P450 catalytic cycle in the absence of cofactors and redox partners is successfully conducted using many bacterial P450s (variants of P450 BM3) and human P450s (CYPs 1A1, 1A2, 1B1, 2A6, 2E1, and 3A4) for the bioconversion of different substrates, including marketed drugs (simvastatin, lovastatin, and omeprazole) and a steroid (17β‐estradiol), to demonstrate the general applicability of the light‐driven, cofactor‐free system. Eosin Y (EY) can bind specifically to the heme domain of cytochromes P450 expressed in the cytoplasm of Escherichia coli. The catalytic turnover is mediated by direct transfer of photoinduced electrons from photosensitized EY to the heme domain of P450 under visible light illumination. The photoactivation of P450 catalytic cycle is a new platform for cost‐effective, cofactor‐free, light‐driven biocatalysis.