NADH-dependent formate dehydrogenase mutants for efficient carbon dioxide fixation

Bioconversion of CO2 to high-valuable products is a globally pursued sustainable technology for carbon neutrality. However, low CO2 activation with formate dehydrogenase (FDH) remains a major challenge for further upcycling due to the poor CO2 affinity, reduction activity and stability of currently used FDHs. Here, we present two recombined mutants, AFDHPa48 and AFDHPa4814, which exhibit high CO2 reduction activity and antioxidative activity. Compared to FDHPa, the reduction activity of AFDHPa48 was increased up to 743 % and the yield in the reduction of CO2 to methanol was increased by 3.16-fold. Molecular dynamics identified that increasing the width of the substrate pocket of AFDHPa48 could improve the enzyme reduction activity. Meanwhile, the enhanced rigidity of C-terminal residues effectively protected the active center. These results fundamentally advanced our understanding of the CO2 activation process and efficient FDH for enzymatic CO2 activation and conversion.

Automated summary

This study developed two recombinant mutants with enhanced CO2 reduction activity, leading to improved efficiency in converting CO2 to methanol. Molecular dynamics analysis revealed that increasing the width of the substrate pocket and enhancing the rigidity of C-terminal residues were key factors in improving the enzyme's reduction activity.