Enzyme Activity Measurement for Formate Dehydrogenase (NAD+)

Creative Enzymes has been serving the enzyme industry with reliable spectrophotometric activity assays for formate dehydrogenase. We are a leading company in the field of enzyme services and specialized in customized enzyme activity measurement. In order to ensure the high quality of our service, our staff have always been devoting themselves into discovering and developing innovative enzyme activity tests.

Formate dehydrogenase (EC 1.2.1.2; FDH) is a NAD⁺-dependent enzyme which catalyzes the formate ion to carbon dioxide with the coupled reduction of NAD⁺ to NADH. This is the final reaction in the linear oxidation pathway for methanol dissimilation in methlotrophic yeasts. Formate dehydrogenase consists of two identical subunits, containing no metal ions or prosthetic groups, and cannot use one-electron carriers as oxidizers. The enzyme is highly specific to both NAD⁺ and formate. NAD⁺-dependent formate dehydrogenase has been found in all methanol-utilizing yeasts of Candida, Pichia, and Hansenula genera and isolated and characterized from various strains. However, this enzyme cannot be found so widely in bacteria. This enzyme is a member of the superfamily of D-specific 2-hydroxy acid dehydrogenases. The catalytic mechanism of this enzyme is specified by a transfer of hydride ion from the substrate onto the C-4 position of the nicotinamide moiety of NAD⁺ without stages of acid-base catalysis, which are present in reactions catalyzed by other related dehydrogenases. Thus, formate dehydrogenase is widely accepted as a model enzyme for studies on the mechanism of hydride ion transferring in the active center of dehydrogenases. Moreover, this enzyme has many alternative names including the followings:

• formate-NAD⁺ oxidoreductase;
• FDH I;
• FDH II;
• N-FDH;
• formic hydrogen-lyase;
• formate hydrogenlyase;
• hydrogenlyase;
• NAD⁺-linked formate dehydrogenase;
• NAD⁺-dependent formate dehydrogenase;
• formate dehydrogenase (NAD⁺);
• NAD⁺-formate dehydrogenase;
• formate benzyl-viologen oxidoreductase;
• formic acid dehydrogenase.

Furthermore, formate dehydrogenase is also used for regenerating NADH in enzymatic syntheses of optically active compounds with dehydrogenases. Due to the irreversibility and the wide pH optimum of the reaction catalyzed by formate dehydrogenase, this enzyme becomes a versatile biocatalyst that is efficient for many processes. Owing to the great potential of formate dehydrogenase in biotechnology and pharmaceutical chemistry, the enzyme activity assay of this enzyme has been desired for a long time. Nonetheless, a standardized and efficient activity assay has not become available. Thus, Creative Enzymes developed the reliable activity assays of formate dehydrogenase. Even for complicated research purposes, we can still help to determine the enzymatic activity of formate dehydrogenase rapidly and accurately.

Figure: The crystal structure of NAD+-dependent formate dehydrogenase from Pseudomonas sp.101 in complex with formate.
PDB: 2GUG