초록
<P>The bacterial molybdenum (Mo)-containing formate dehydrogenase (FdsDABG) from <I>Cupriavidus necator</I> is a soluble NAD<SUP>+</SUP>-dependent enzyme belonging to the DMSO reductase family. The holoenzyme is complex and possesses nine redox-active cofactors including a bis(molybdopterin guanine dinucleotide) (bis-MGD) active site, seven iron-sulfur clusters, and 1 equiv of flavin mononucleotide (FMN). FdsDABG catalyzes the two-electron oxidation of HCOO<SUP>-</SUP> (formate) to CO<SUB>2</SUB> and reversibly reduces CO<SUB>2</SUB> to HCOO<SUP>-</SUP> under physiological conditions close to its thermodynamic redox potential. Here we develop an electrocatalytically active formate oxidation/CO<SUB>2</SUB> reduction system by immobilizing FdsDABG on a glassy carbon electrode in the presence of coadsorbents such as chitosan and glutaraldehyde. The reversible enzymatic interconversion between HCOO<SUP>-</SUP> and CO<SUB>2</SUB> by FdsDABG has been realized with cyclic voltammetry using a range of artificial electron transfer mediators, with methylene blue (MB) and phenazine methosulfate (PMS) being particularly effective as electron acceptors for FdsDABG in formate oxidation. Methyl viologen (MV) acts as both an electron acceptor (MV<SUP>2+</SUP>) in formate oxidation and an electron donor (MV<SUP>+•</SUP>) for CO<SUB>2</SUB> reduction. The catalytic voltammetry was reproduced by electrochemical simulation across a range of sweep rates and concentrations of formate and mediators to provide new insights into the kinetics of the FdsDABG catalytic mechanism.</P><BR>[FIG OMISSION]</BR>