Entrapment of cytochrome P450 BM-3 in polypyrrole for electrochemically-driven biocatalysis.
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.
Holtmann
Dirk
D
Biochemical Engineering Group, Karl-Winnacker-Institut, DECHEMA e.V., 60486, Frankfurt am Main, Germany.
Mangold
Klaus-Michael
KM
Schrader
Jens
J
D001426:Bacterial Proteins / Q000302:isolation & purification / Q000378:metabolism*; D055162:Biocatalysis; D002244:Carbon; D003577:Cytochrome P-450 Enzyme System / Q000302:isolation & purification / Q000378:metabolism*; D055664:Electrochemical Techniques / Q000379:methods*; D004566:Electrodes; D004800:Enzymes, Immobilized / Q000378:metabolism*; D009251:NADPH-Ferrihemoprotein Reductase / Q000302:isolation & purification / Q000378:metabolism*; D010984:Platinum; D011108:Polymers / Q000378:metabolism*; D011485:Protein Binding; D011758:Pyrroles / Q000378:metabolism*
D001426:Bacterial Proteins; D004800:Enzymes, Immobilized; D011108:Polymers; D011758:Pyrroles; C067635:polypyrrole; D010984:Platinum; D002244:Carbon; D003577:Cytochrome P-450 Enzyme System; D009251:NADPH-Ferrihemoprotein Reductase; C052423:flavocytochrome P450 BM3 monoxygenases