PURIFICATION AND CHARACTERIZATION OF URICASE FOR RENEWABLE ENERGY

Abstract

This research focused on harnessing uricase, a crucial enzyme in purine metabolism, from poultry waste for potential applications. Despite its absence in humans due to genetic mutations, uricase was successfully extracted from poultry waste with an initial activity of 3721.04 U/mL and a specific activity of 135.681 U/mg. Notably, the enzyme's activity increased to 9581.843 U/mL and specific activity to 534.641 U/mg after ammonium sulfate precipitation, resulting in decreased protein content. Further purification through ion exchange chromatography using DEAE cellulose and gel filtration with Sephadex G-150 demonstrated an enhanced activity of 3768.114 U/mL and specific activity of 1689.73 U/mg. The enzyme was purified 63-fold, with protein content reduced to 2.23 mg/mL. Optimal conditions for enzyme activity were observed at pH 8 and 37°C. The study delved into substrate and enzyme concentration effects, revealing maximum activity at 2.5 mg substrate (poultry waste) and 750μL enzyme concentrations. Additionally, the research explored the thermal stability of uricase, noting a maximum activity of 4095.18 U/mL after 20 minutes of incubation at elevated temperatures. The irreversible thermal denaturation decreased half-life from 660 to 149.4 min at 60°C compared to 37°C, with the enzyme showing positive ∆S*, ∆H*, and ∆G*. This work contributes valuable insights into utilizing uricase from poultry waste.