{"database":"biostudies-literature","file_versions":[],"scores":null,"additional":{"submitter":["Giang PD"],"funding":["Basic Energy Sciences","Australian Research Council"],"pagination":["6035-6049"],"full_dataset_link":["https://www.ebi.ac.uk/biostudies/studies/S-EPMC11891784"],"repository":["biostudies-literature"],"omics_type":["Unknown"],"volume":["16(14)"],"pubmed_abstract":["Formate dehydrogenase (FdsDABG) from <i>Cupriavidus necator</i> is a Mo-containing enzyme capable of catalysing both formate oxidation to CO<sub>2</sub> and the reverse CO<sub>2</sub> reduction to formate by utilising NAD<sup>+</sup> or NADH, respectively. This enzyme is part of the NADH dehydrogenase superfamily. Its subcomplex, FdsBG, lacking the formate oxidizing/CO<sub>2</sub>-reducing Mo-cofactor, but harbouring an FMN as well as [2Fe-2S] and [4Fe-4S] clusters, reversibly interconverts the NAD<sup>+</sup>/NADH redox pair. UV-vis spectroelectrochemistry across the range 6 < pH < 8 determined the redox potentials of these three cofactors. Cyclic voltammetry was used to explore mechanistic and kinetic properties of each oxidation- and reduction-half reaction. Through mediated enzyme electrochemistry experiments, the Michaelis constant for NADH oxidation (<i>K</i> <sub>M,NADH</sub> = 1.7 × 10<sup>2</sup> μM) was determined using methylene blue as a redox mediator. For the reverse NAD<sup>+</sup> reduction reaction using methyl viologen as electron donor a similar analysis yielded the value of <i>K</i> <sub>M,NAD<sup>+</sup></sub> = 1.2 mM. All experimental voltammetry data were reproduced by electrochemical simulations furnishing a set of self-consistent rate constants for the catalytic FdsBG system for both NAD<sup>+</sup> reduction and NADH oxidation. This comprises the first electrochemical kinetic analysis of its kind for a reversible NADH dehydrogenase enzyme and provides new insight to the function of the FdsDABG formate dehydrogenase holoenzyme."],"journal":["Chemical science"],"pubmed_title":["Reversible enzyme-catalysed NAD&lt;sup&gt;+&lt;/sup&gt;/NADH electrochemistry."],"pmcid":["PMC11891784"],"funding_grant_id":["DE-SC0010666","DP220103268"],"pubmed_authors":["Niks D","Hille R","Bernhardt PV","Giang PD","Hakopian S"],"additional_accession":[]},"is_claimable":false,"name":"Reversible enzyme-catalysed NAD&lt;sup&gt;+&lt;/sup&gt;/NADH electrochemistry.","description":"Formate dehydrogenase (FdsDABG) from <i>Cupriavidus necator</i> is a Mo-containing enzyme capable of catalysing both formate oxidation to CO<sub>2</sub> and the reverse CO<sub>2</sub> reduction to formate by utilising NAD<sup>+</sup> or NADH, respectively. This enzyme is part of the NADH dehydrogenase superfamily. Its subcomplex, FdsBG, lacking the formate oxidizing/CO<sub>2</sub>-reducing Mo-cofactor, but harbouring an FMN as well as [2Fe-2S] and [4Fe-4S] clusters, reversibly interconverts the NAD<sup>+</sup>/NADH redox pair. UV-vis spectroelectrochemistry across the range 6 < pH < 8 determined the redox potentials of these three cofactors. Cyclic voltammetry was used to explore mechanistic and kinetic properties of each oxidation- and reduction-half reaction. Through mediated enzyme electrochemistry experiments, the Michaelis constant for NADH oxidation (<i>K</i> <sub>M,NADH</sub> = 1.7 × 10<sup>2</sup> μM) was determined using methylene blue as a redox mediator. For the reverse NAD<sup>+</sup> reduction reaction using methyl viologen as electron donor a similar analysis yielded the value of <i>K</i> <sub>M,NAD<sup>+</sup></sub> = 1.2 mM. All experimental voltammetry data were reproduced by electrochemical simulations furnishing a set of self-consistent rate constants for the catalytic FdsBG system for both NAD<sup>+</sup> reduction and NADH oxidation. This comprises the first electrochemical kinetic analysis of its kind for a reversible NADH dehydrogenase enzyme and provides new insight to the function of the FdsDABG formate dehydrogenase holoenzyme.","dates":{"release":"2025-01-01T00:00:00Z","publication":"2025 Apr","modification":"2025-06-27T03:05:39.256Z","creation":"2025-04-07T07:49:47.005Z"},"accession":"S-EPMC11891784","cross_references":{"pubmed":["40070472"],"doi":["10.1039/d5sc00570a"]}}