Exploration of the cofactor specificity of wild-type phosphite dehydrogenase and its mutant using molecular dynamics simulations

Phosphite dehydrogenase (Pdh) catalyzes the NAD-dependent oxidation of phosphite to phosphate with the formation of NADH. It can be used in several bioorthogonal systems for metabolic control and related applications, for example, bioelectricity. At present, NAD has poor stability at high concentrations and costs are expensive. Implementation of a non-natural cofactor alternative to the ubiquitous redox cofactor nicotinamide adenosine dinucleotide (NAD) is of great scientific and biotechnological interest. Several Pdhs have been engineered to favor a smaller-sized NAD analogue with a cheaper price and better thermal stability, namely, nicotinamide cytosine dinucleotide (NCD). However, the conformational changes of two cofactors binding to Pdh remain unknown. In this study, five molecular dynamics (MD) simulations were performed to exploit the different cofactors binding to wild-type (WT) Pdh and mutant-type (MT) Pdh (I151R/P176E/M207A). The results were as follows: First, compared with WT Pdh, the cofactor-binding pocket of mutant Pdh became smaller, which may favor a smaller-sized NCD. Second, secondary structure analysis showed that the alpha helices in residues 151-207 partly disappeared in mutant Pdh binding to NAD or NCD. Our theoretical results may provide a basis for further studies on the Pdh family.

Automated summary

The phosphite dehydrogenase (Pdh) catalyzes the oxidation of phosphite to phosphate with the formation of NADH. Different cofactors binding to Pdh, including a smaller-sized NAD analogue nicotinamide cytosine dinucleotide (NCD), can lead to changes in cofactor-binding pockets and secondary structures. These findings can provide a basis for further studies on the Pdh family.