Impact of Deep Eutectic Solvents on Kinetics and Folding Stability of Formate Dehydrogenase

Specifically designed co-solvent mixtures are an efficient way to enhance the kinetics of enzyme-catalyzed reactions without compromising enzyme stability; among them, several deep eutectic solvents have emerged as exciting for biocatalytic reactions. DESs nature allows one tailor enzyme-co-solvent interactions by using constituents diverse functional groups. In this work, influence co-solvents (betaine, glycerol, and sorbitol) two (betaine:glycerol betaine:sorbitol) on candida boidinii Formate dehydrogenase was evaluated. The results showed a 30% increase in catalytic efficiency adding 15 wt.-% betaine buffered aqueous reaction media. Further, cbFDH folded-state stability evaluated differential scanning fluorimetry finally obtain binding affinity, unfolding curves, thermodynamic parameters. addition sorbitol, increased protection against thermal stress, effect could be improved increasing concentrations. Moreover, ability reduce irreversibility process. Betaine only that had negative effect, which offset betaine-based DESs. latter result additivity certain individual effects stability. Non-monotonous were obtained sorbitol buffer solutions, probably because hydrogen bond dynamics between cbFDH/co-solvent/water change dramatically with amount water present. Finally, NAD+ affinity interestingly direct correlation kinetics.