Penentuan kinetika urikase dari sel bacillus subtilis, B. megaterium, dan B. cereus
Abstract
Uric acid concentration could be determined based on the oxidation of uric acid into alantoin in the presence of uricase. Determination of uric acid concentration is needed to diagnose the occurrence of kidney disease in gout patients. The aim of the research was to study the kinetics of uricase in Bacillus subtilis, B. megaterium, and B. cereus cells using spectrophotometry method by measurement the decrease of uric acid absorbance at 293 nm. The optimum conditions of uricase activity from the three bacterial cells occured in physiological conditions and uricase activity was stable until the second day. The values of maximum velocity (Vmax) for B. subtilis, B. megaterium, and B. cereuswere 0.1642, 0.0824, and 0.0412 mM/min, respectively. The values of Michaelis-Menten constant (Km) for B. subtilis, B. megaterium and B. cereus were 0.0003, 0.0036, and 0.0036 mM, respectively. The value of catalytic constant (kkat) for B. subtilis, B. megaterium, and B. cereus were 410.5000, 171.6667, and 257.5000 mM/min ml 00=1, respectively. Based on these results, among all bacteria tested, the highest of uricase binding with substrate in B. substilis cells was observed because of the smallest of Km value and greatest of Vmax and kkat .
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References
Arslan F. 2008. An amperometric biosensor for uric acid determination prepared from uricase immobilized in polyaniline-polypyrrole. Sensor 8:5492-5500.
Azab E.A., Ali M.M., Farred M.F. 2005. Studies on uricase induction in certain bacteria. Egyptian Journal of Biology 7:44-54.
Bergmeyer H.U., Gawehn K., Grassl M. 1974. In Methods on Enzymatic Analysis (Bergmeyer HU). Ed ke-2. Volume ke-1 (518). New York: Academic Pr.
Bongaerst G.P.A., Uitzetter J., Brouns R. and Vogels G.D. 1978. Uricase of Bacillus fastidiosus properties and regulation of syntesis. Biochimica et Biophysica Acta 527:348-358.
Bomalaski et al., 2002. Uricase with polyethylene glycol (uricase PEG 20): biochemical and preclinical studies. J Reumatol 29:1942-1949.
Fattah Y., Saeed H., Gohar M., El-Baz M. 2005. Improved production of Pseudomonas aeruginosa uricase by optimization of process parameters through statistical experimental designs. Process Biochemistry 40:1707-1714.
Hall L.M., Macvicar R. 1954. Ammonia as an intermediate in nitrate redijction by Bacillus subtilis. J. Bio Chem 197:305-310.
Huang S.H., Wu T.K. 2004. Modified colorimetric assay for uricase activity and a screen for mutan Bacillus subtillis gene following StEP mutagenesis. The Journal of Reumatol 271:517-523.
Lehninger A.L., Nelson D.L., Cox M.M. 2004. The Princiles of Biochemistry. Ed ke-4. New York: Worth Publisher.
Lotfy W. 2008. Production of a thermostable uricase by a novel Bacillus thermocatenulatus strain. Bioresource Technology 99:699-702.
Priest F.G., Goodfellow M., Todd C. 1988. Bacillus heterogenety, Journal oF Genetics Microbiology 134:1847-1882.
Trivedi et al., 1978. New Ultraviolet (340 nm) Method for assay of uric acid in serum or plasma. Clinical Chemistry, 24(4):562-566.
Zang Y et al., 2007. Development and analytical application of an uric acid biosensor using an uricase-immobilized eggshell membrane. Biosensor and Bioelectronic. 22:1791-1797.
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