Enhancing Uricase Stability Using a Urea-Glycerol-Choline Chloride Deep Eutectic Solvent: Optimization of DES Concentration through Enzymatic Assays

Urate oxidase is an oxidoreductase enzyme that catalyzes the conversion of uric acid to ۵-hydroxyisourate. This enzyme is employed in the management of various pathological conditions, including gout, hyperuricemia, renal failure, tumor lysis syndrome, and cardiovascular complications arising from hyperuricemic. Nevertheless, the clinical application of this enzyme faces significant challenges due to its structural instability and susceptibility to conformational changes under physiological conditions. One effective strategy to enhance the stability and functional performance of urate oxidase involves the use of deep eutectic solvents (DESS), which have been shown to improve enzyme stability and catalytic efficiency. In the present study, Escherichia coli BL۲۱ (DE۳) cells harboring the pET۲۸a(+)-uricase plasmid were cultured overnight and induced with isopropyl B-D-۱-thiogalactopyranoside (IPTG) and lactose. The recombinant enzyme was purified using Ni-NTA affinity chromatography. The purified urate oxidase was then treated with a DES formulation composed of urea, glycerol, and choline chloride at varying concentrations. The optimal DES concentration was determined to be ۲.۵% (v/v).

کلیدواژه ها:

Urate oxidase ، Gout ، Hyperuricemia ، Deep eutectic solvents (DESS) ، Optimal concentration