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Electrochemically Activated Catalytic Pathways of Human Metabolic Cytochrome P450s in Ultrathin Films

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Electrochemistry of N4 Macrocyclic Metal Complexes

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Abstract

Electrochemical understanding of metalloenzyme biocatalysts can guide in the development of green bioreactors for stereoselective syntheses and tailoring of efficient bio-inspired catalysts for large-scale applications. In addition, if the enzymes themselves can be stabilized into viable catalytic materials such as films or nanoparticles, valuable and unique stereo- and regiospecific reactions can be catalyzed in simple aqueous medium. In particular, biological electrocatalysis is an emerging area that is expected to play a significant role in fine chemical and drug synthesis, niche electronic devices, sustainable energy, and biomedical fields. Our research has specifically been inspired by the broad stereoselective biocatalytic properties of human cytochrome P450 (cyt P450) enzymes. Cyt P450s are a large family of heme iron monooxygenases with high expression in liver and other human organs that serve as the major oxidative catalysts in human metabolism. Electrochemical bioreactors featuring cyt P450s have the potential to complement and accelerate drug development processes by facilitating candidate identification and toxicity evaluation of metabolites. Challenging aspects associated with electrochemical studies of cyt P450s in thin films include enzyme stability, electronic connectivity between cyt P450-heme cofactor and electrodes, density of immobilized electrocatalytically active enzyme molecules, and bioactive cyt P450 conformations in films on electrodes. To achieve these features, various electrode surface environments have been explored. We designed ultrathin bioactive films of cyt P450 enzymes with polyions or insoluble surfactants that demonstrated the first direct electron transfer and biocatalytic applications of this class of enzymes on electrodes. Subsequently, we constructed films of genetically engineered microsomes, rat and human liver microsomes, and cyt P450s assembled with microsomal cyt P450 reductase (CPR) that enabled a bioelectrocatalytic pathway that closely mimicked the in vivo cyt P450 biocatalytic mechanism. These systems allowed us to develop a clearer understanding of cyt P450 electron transfer and enabled uses in microfluidic toxicity screening arrays. In this review, we provide an overview of different catalytic pathways that are accessed and driven electrochemically using pure human cyt P450 enzymes and those present in membrane-bound forms along with CPR.

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Acknowledgments

The authors thank colleagues named in joint publications who collaborated on research in this area, and without whom progress would not have been possible. JFR thanks the National Institute of Environmental Health Sciences (NIEHS), NIH, USA, Grant No. ES03154 for financial support. SK is grateful for financial support by the National Institute of Diabetes and Digestive and Kidney Diseases of the National Institutes of Health under Award Number R15DK103386.

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Authors and Affiliations

  1. Department of Chemistry, Oklahoma State University, Stillwater, 74078, USA

    Sadagopan Krishnan

  2. Department of Chemistry, University of Connecticut, Storrs, CT, 06269, USA

    James F. Rusling

  3. Department of Surgery and Neag Cancer Center, University of Connecticut Health Center, Farmington, 06032, USA

    James F. Rusling

  4. Institute of Materials Science, University of Connecticut, Storrs, CT, 06269, USA

    James F. Rusling

  5. School of Chemistry, National University of Ireland at Galway, Galway, Ireland

    James F. Rusling

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  1. Sadagopan Krishnan
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Correspondence to Sadagopan Krishnan .

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  1. Universidad de Santiago de Chile, Santiago, Chile

    Jose H. Zagal

  2. CNRS-Chimie ParisTech, Paris, France

    Fethi Bedioui

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Krishnan, S., Rusling, J.F. (2016). Electrochemically Activated Catalytic Pathways of Human Metabolic Cytochrome P450s in Ultrathin Films. In: Zagal, J., Bedioui, F. (eds) Electrochemistry of N4 Macrocyclic Metal Complexes. Springer, Cham. https://doi.org/10.1007/978-3-319-31332-0_2

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