Study of Uricase Properties Partially Purified from Pisum sativum Seeds and its Effect on Uric Acid Level in Mice
JOURNAL OF EDUCATION AND SCIENCE,
2013, Volume 26, Issue 4, Pages 11-21
Uricase (urate oxidase) was partially purified from Pisum sativum seeds by ammonium sulfate precipitation ,dialysis and anion exchange chromatography techniques. One proteinous peak was obtained containing two isomers of uricase I and II with specific activity (5.92×10-3 and 4.9955×10-3) unit/mg protein and with purification fold(31.32and 26.43) respectively compared to crude enzyme.
The optimum conditions for the purified enzyme was determined using uric acid as a substrate. The results showed that maximum activity with glycine-NaOH buffer at pH=7, 40°C, substrate concentration 100mM Vmax=6.25×10-2 U/mg protein and the Km=50mM, 100µl enzyme volume and the reaction velocity was increased with copper ion.
The active dose of uricase for reducing uric acid was determined in healthy mice 400µg/kg body weight, and the best treatment was obtained by the injection of enzyme with allopurinal intraperitonally.
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