Purification and Optimization of Uricase Enzyme Produced by Pseudomonas aeruginosa
Keywords:
Uricase, Pseudomonas aeruginosa, purification, optimizationAbstract
The uricase (urate oxidase) enzyme was extracted from Pseudomonas aeruginosa and purified by ammonium sulphate precipitation. The molecular weight of purified uricase was determined by SDS-PAGE electrophoresis using marker proteins of known molecular weight. The optimum temperature, pH and the effect of various metal ions on uricase activity were evaluated. The results showed that 70% ammonium sulphate concentration proved high uricase activity by 40.0 U/ml than other concentration and cell free supernatant. The molecular weight of purified uricase enzyme was estimated to be 33 kDa by SDS-PAGE profile. The optimum temperature and pH was detected at 35°C and 8.5 for maximum uricase activity. Metal ions such as Co2+, Mn2+, Mg2+, Fe2+, Zn2+ and Cu2+ were reduced the enzyme activity to 33.17%, 92.18%, 72.47%, 32.73%, 64.32% and 90.10%, whereas Ca2+ enhanced uricase activity to 126%.    ÂÂ