Thermal Adaptation of Enzymes: Impacts of Conformational Shifts on Catalytic Activation Energy and Optimum Temperature

Thermal adaptation of enzymes is essential for both living organism development in extreme conditions and efficient biocatalytic applications. However, the molecular mechanisms leading to a shift in catalytic activity optimum temperatures remain unclear, and there is increasing experimental evidence that thermal adaptation involves complex changes in both structural and reactive properties. Here, a combination of enhanced protein conformational sampling with an explicit chemical reaction description was applied to mesophilic and thermophilic homologues of the dihydrofolate reductase enzyme, and a quantitative description of the stability and catalytic activity shifts between homologues was obtained. The key role played by temperature-induced shifts in protein conformational distributions is revealed. In contrast with pictures focusing on protein flexibility and dynamics, it is shown that while the homologues’ reaction free energies are similar, the striking discrepancy between their activation energies is caused by their different conformational changes with temperature. An analytic model is proposed that combines catalytic activity and structural stability, and which quantitatively predicts the shift in homologue optimum temperatures. It is shown that this general model provides a molecular explanation of changes in optimum temperatures for several other enzymes.