Production, Purification and Characterization of Uricase Produced by Pseudomonas aeruginosa

Abstract

In this study, detection of uricase production from Pseudomonas aeruginosa
isolates was done by applying colorimetric method, Uricase was purified from the
most potent isolate by precipitation using ammonium sulphate (80% saturation) then
purification was achieved using DEAE –Cellulose ion exchange and Sepharose 6B
gel filtration chromatography column, 16.4% of total enzyme was recovered with
specific activity 2337.5U/mg and 22.21folds of purification. Characterization of
uricase involved detection of optimal conditions for uricase activity, the maximal
activity was obtained at temperature 45ºC,while uricase appeared to be stable at
40ºC. Uricase showed optimal activity at pH 9 while pH stability was in the
range of 7.5-9. Molecular weight of uricase obtained in this study determined by
sodium dedosyl sulfate electrophoresis (SDS-PAGE) technique was approximately
35kDa.