Abstract
In vitro biocatalysis with cytochrome P450 BM-3 was investigated aiming for the substitution of the expensive natural cofactor NADPH by electrochemistry. The monooxygenase was immobilized on electrodes by entrapment in polypyrrole as a conductive polymer for electrochemically wiring the enzyme. Electropolymerization of pyrrole proved to be a useful means of immobilising an active cytochrome P450 BM-3 mutein on platinum and glassy carbon electrodes without denaturation. Repeatedly sweeping the electric potential between −600 and +600 mV versus Ag/AgCl led to enzymatically-catalysed product formation while in the absence of the enzyme no product formed under otherwise identical conditions.
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Acknowledgement
We thank R.D. Schmid and V. Urlacher, Stuttgart University, for kindly providing the P450 BM-3 mutant and for helpful advices regarding the enzyme and Ulrich Schwaneberg, Jacobs University Bremen, for kindly providing the pNCA. Parts of the research were supported by the German Federal Ministry of Economics and Technology (BMWi) via the Federation of Industrial Cooperative Research Associations “Otto von Guericke” e.V. (14781 N/1) and by the German Federal Ministry for Food, Agriculture and Consumer Protection (22000504 (04NR005) and 22030905).
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Holtmann, D., Mangold, KM. & Schrader, J. Entrapment of cytochrome P450 BM-3 in polypyrrole for electrochemically-driven biocatalysis. Biotechnol Lett 31, 765–770 (2009). https://doi.org/10.1007/s10529-009-9925-4
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DOI: https://doi.org/10.1007/s10529-009-9925-4