Abstract
The reduction potentials of two relatively high-molecular-mass enzymes were determined directly at an edge pyrolytic graphite electrode by using square-wave voltammetry. The equilibrium reduction potential versus standard hydrogen electrode was determined for Clostridium pasteurianum hydrogenase I (E'0 = -377 +/- 10 mV; molecular mass 60 kDa) and Rhodospirillum rubrum carbon monoxide dehydrogenase (E'0 = -418 +/- 7 mV; molecular mass 62 kDa). The reduction potential of each enzyme was pH-independent, and one electron was transferred per redox centre. The reduction potential was determined to be identical for the CO dehydrogenase, semi-apo-(CO dehydrogenase), and CO dehydrogenase with carbonyl sulphide (COS) or cyanide bound. The electron-transferring efficiency of CO dehydrogenase was affected by two inhibitors, COS and cyanide, as indicated by a diminished analytic current.