Study of Uricase Properties Partially Purified from Pisum sativum Seeds and its Effect on Uric Acid Level in Mice
JOURNAL OF EDUCATION AND SCIENCE,
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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