[HTML][HTML] Direct electrochemistry of nitrate reductase from the fungus Neurospora crassa

P Kalimuthu, P Ringel, T Kruse, PV Bernhardt - Biochimica et Biophysica …, 2016 - Elsevier
P Kalimuthu, P Ringel, T Kruse, PV Bernhardt
Biochimica et Biophysica Acta (BBA)-Bioenergetics, 2016Elsevier
We report the first direct (unmediated) catalytic electrochemistry of a eukaryotic nitrate
reductase (NR). NR from the filamentous fungus Neurospora crassa, is a member of the
mononuclear molybdenum enzyme family and contains a Mo, heme and FAD cofactor which
are involved in electron transfer from NAD (P) H to the (Mo) active site where reduction of
nitrate to nitrite takes place. NR was adsorbed on an edge plane pyrolytic graphite (EPG)
working electrode. Non-turnover redox responses were observed in the absence of nitrate …
Abstract
We report the first direct (unmediated) catalytic electrochemistry of a eukaryotic nitrate reductase (NR). NR from the filamentous fungus Neurospora crassa, is a member of the mononuclear molybdenum enzyme family and contains a Mo, heme and FAD cofactor which are involved in electron transfer from NAD(P)H to the (Mo) active site where reduction of nitrate to nitrite takes place. NR was adsorbed on an edge plane pyrolytic graphite (EPG) working electrode. Non-turnover redox responses were observed in the absence of nitrate from holo NR and three variants lacking the FAD, heme or Mo cofactor. The FAD response is due to dissociated cofactor in all cases. In the presence of nitrate, NR shows a pronounced cathodic catalytic wave with an apparent Michaelis constant (KM) of 39 μM (pH 7). The catalytic cathodic current increases with temperature from 5 to 35 °C and an activation enthalpy of 26 kJ mol− 1 was determined. In spite of dissociation of the FAD cofactor, catalytically activity is maintained.
Elsevier
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