Application of Uricase Isolated from Bacillus subtilis SP6 in Uric Acid Assay Diagnostic Kit

Abstract

Uricase enzyme is a major component of the diagnostic kit for the estimation of uric acid for diagnosis, monitoring and treatment of gout and joint inflammations. Uric Acid/Uricase assay kit is a simple assay for measuring uric acid concentrations in biological samples such as serum, plasma, and urine without any need for pretreatment. The level of uricase enzyme activity was detected in the crude extract of some animal liver tissues, plant leaves and a microbial source (Bacillus subtilis SP6 bacteria) and expressed as specific activity (unit/mg protein). It was found difficult to produce high yield of uricase in short time from animal or plant tissues and stability of uricase from both sources is still unclear; therefore, it was isolated from Bacillus subtilis SP6. The extraction procedure of uricase from Bacillus subtilis SP6 involved, isolation and extraction of bacterial cells, determining the uricase activity in both intracellular and extracellular fractions, pooling for both fractions and ammonium sulfate precipitation which seemed to be convenient since 74.7 % of uricase activity was recovered. The isolated uricase was applied in the preparation of uric acid diagnostic kit that found sensitive and comparable with commercially available ones.