Journal
BIOTECHNOLOGY LETTERS
Volume 31, Issue 5, Pages 765-770Publisher
SPRINGER
DOI: 10.1007/s10529-009-9925-4
Keywords
Cofactor substitution; Conductive polymer; Cytochrome P450; Electrochemically driven biocatalysis; Immobilisation; P450 monooxygenase
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Funding
- German Federal Ministry of Economics and Technology (BMWi)
- Federation of Industrial Cooperative Research Associations [14781 N/1]
- German Federal Ministry for Food, Agriculture and Consumer Protection [22000504, 04NR005, 22030905]
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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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