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NAD-dependent formate dehydrogenase (FDH) from  Candida boidinii  (CbFDH) has been widely used in various CO 2 -reduction systems but its practical applications are often impeded due to low CO 2 -reducing activity. In this study, we demonstrated superior CO 2 -reducing properties of FDH from  Thiobacillus  sp. KNK65MA (TsFDH) for production of formate from CO 2  gas. To discover more efficient CO 2 -reducing FDHs than a reference enzyme, i.e. CbFDH, five FDHs were selected with biochemical properties and then, their CO 2 -reducing activities were evaluated. All FDHs including CbFDH showed better CO 2 -reducing activities at acidic pHs than at neutral pHs and four FDHs were more active than CbFDH in the CO 2  reduction reaction. In particular, the FDH from  Thiobacillus  sp. KNK65MA (TsFDH) exhibited the highest CO 2 -reducing activity and had a dramatic preference for the reduction reaction, i.e., a 84.2-fold higher ratio of CO 2  reduction to formate oxidation in catalytic efficiency ( k   cat / K   B ) compared to CbFDH. Formate was produced from CO 2  gas using TsFDH and CbFDH, and TsFDH showed a 5.8-fold higher formate production rate than CbFDH. A sequence and structural comparison showed that FDHs with relatively high CO 2 -reducing activities had elongated N- and C-terminal loops. The experimental results demonstrate that TsFDH can be an alternative to CbFDH as a biocatalyst in CO 2  reduction systems.

Efficient CO2-Reducing Activity of NAD-Dependent Formate Dehydrogenase from Thiobacillus sp. KNK65MA for Formate Production from CO2 Gas