Improving Thermal Stability of Uricase Using Deep Eutectic Solvent Containing Fructose, Sucrose, and Glycerol

Uricase is an enzyme classified as an oxidoreductase, responsible for the catalytic conversion of uric acid to allantoin. This enzyme has significant therapeutic applications in the management of conditions such as tumor lysis syndrome, gout, and hyperuricemia. However, its clinical utility is often limited by its pronounced sensitivity to environmental factors, including temperature and pH. Therefore, the development of methodologies for enhancing the thermal stability of uricase has become a focal point of research. One promising approach involves the utilization of deep eutectic solvents (DES). In the present study, a competent cell was prepared, and a plasmid encoding the uricase gene was successfully transferred. Bacterial cultures were subsequently established, and expression of the enzyme was induced using IPTG. Purification was conducted via nickel affinity chromatography. The effects of DES formulations containing sucrose, fructose, and glycerol on enzyme activity were evaluated and compared with that of the free enzyme. The results demonstrated a significant reduction in the inactivation constant (Kin) alongside an increase in both activation energy (Eact) and melting temperature (Tm) of the enzyme.

کلیدواژه ها:

Uricase ، Thermal stability ، Deep eutectic solvents ، Inactivation constant ، Activation energy ، Melting temperature