Study of the stability of uricase from Aspergillus flavus and its stabilization by Glucose

Uricase or urate oxidase is an enzyme that converts uric acid (with low solubility) to 5-hydroxyisourate and finally to allantoin. The possibility of developing some diseases like gout and kidney stones will be increased in high levels of uric acid. Thus, uricase can be used as drug enzyme to reduce uric acid levels in the blood. The low stability of proteins (such as drug enzymes) is a challenge in the use of them. There are several approaches such as use of additives for protein stabilization. In this study, E. Coli BL21 (DE3) was transformed by pET28a () vector carrying Aspergillus flavus uricase gene. The recombinant protein was expressed and then purified by a Ni-NTA agarose chromatography column. After purification, the thermal stability of the purified enzyme was evaluated and then it stabilized by additives. The results showed that enzyme is active and purified very well. Thermal stability results indicated that uricase maintains its stability up to 20°C and then loses its stability. The half-life of enzyme was 30minutes at 40 °. The results of enzyme stabilization by 20% (v/w) concentration of glucose and sorbitol as well as by 20 % (v/v) of glycerol showed that glucose had the most stabilization effect on the uricase among the additives. The stability (half life) of enzyme was increased more than two times in the presence of glucose. Finally, we conclude that additives like glucose which increase surface tension have the most stabilization effect on the uricase enzyme stability.