Application of Uricase Isolated from Bacillus subtilis SP6 in Uric Acid Assay Diagnostic Kit
Mohamed S. Helmy
Molecular Biology Department, National Research Centre, El-Tahrir st., Dokki, Giza, Egypt and Proteome Research Laboratory, Central Laboratories Network and Centers of Excellence, National Research Centre, El-Tahrir st., Dokki, Giza, Egypt.
Hassan M. M. Masoud *
Molecular Biology Department, National Research Centre, El-Tahrir st., Dokki, Giza, Egypt and Proteome Research Laboratory, Central Laboratories Network and Centers of Excellence, National Research Centre, El-Tahrir st., Dokki, Giza, Egypt.
Doaa A. Darwish
Molecular Biology Department, National Research Centre, El-Tahrir st., Dokki, Giza, Egypt and Proteome Research Laboratory, Central Laboratories Network and Centers of Excellence, National Research Centre, El-Tahrir st., Dokki, Giza, Egypt.
Mohamed M. Abdel-Monsef
Molecular Biology Department, National Research Centre, El-Tahrir st., Dokki, Giza, Egypt and Proteome Research Laboratory, Central Laboratories Network and Centers of Excellence, National Research Centre, El-Tahrir st., Dokki, Giza, Egypt.
Sayed S. Esa
Molecular Biology Department, National Research Centre, El-Tahrir st., Dokki, Giza, Egypt and Proteome Research Laboratory, Central Laboratories Network and Centers of Excellence, National Research Centre, El-Tahrir st., Dokki, Giza, Egypt.
Mahmoud A. Ibrahim
Molecular Biology Department, National Research Centre, El-Tahrir st., Dokki, Giza, Egypt and Proteome Research Laboratory, Central Laboratories Network and Centers of Excellence, National Research Centre, El-Tahrir st., Dokki, Giza, Egypt.
*Author to whom correspondence should be addressed.
Abstract
Uricase is a major component enzyme of uric acid estimation kit which used for diagnosis, monitoring and treatment of gout and joint inflammations. Uricase/Uric Acid diagnostic kit used for determination of uric acid concentrations in plasma, serum, and urine. 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 diagnostic kit of uric acid preparation that found comparable and sensitive with available commercially ones.
Keywords: Uricase, mammalian tissues, plant tissues, Bacillus subtilis SP6, diagnostic kit