Aktivitas urikase yang dihasilkan dari berbagai sel lactobacillus plantarum dan parameter kinetikanya

Dyah Iswantini, Novik Nurhidayat, . Trivadila, Eka Mardiah

Abstract


Uric acid concentration could be determined by spectrophotometry method. Uric acid was oxidized into allatonin in the presence of uricase and calculated by measuring the decrease of uric acid absorbance at 293 nm. These uricase were obtained from cells of Lactobacillus plantarum. L. plantarum K. Mar. E was isolated from passion fruit skin and L. plantarum Mgs. Psmb and Mgs. Bst from mangosteen. This research was conducted to observe the activity and kinetics of uricase from various cells of L. plantarum by spectrophotometric method. The plate assay method indicated that L. plantarum produced uricase, based on the clear zone about 0,2 mm on glucose yeast peptone medium contained 0,2% uric acid. The optimum condition of uricase activity from the three different sources occured in physiological condition. Uricase activity generated from cells of L. p/antarum K. Mar. E, Mgs. Psmb, and Mgs. Bst were 0,1073; 0,0867; and 0,0842 U/ml, respectively. The kinetic parameters for uricase, determined with uric acid as the substrate. Vmax produced by L. plantarum K. Mar. E, Mgs. Psmb, and Mgs. Bst were 1,3635; 0,0316; and 0,0418 U/ml of bacterial culture, respectively and KM 0,1541; 0,0061; and
0,0054 mM, respectively. Uricase activity in various bacterial cells of L. plantarum was stable until the second day.

Keywords


Activity; uricase; spectrophotometry; kinetics; Lactobacillus plantarum cells

Full Text:

PDF

References


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.

Iswantini, D,. Kato, K., Kano, K., Ikeda, T. 1998. Electrochemical measurements of glucose dehydrogenase activity exhibited by Escherichia coli cells; effects of the additions of pyrroloquinoline quinine, magnesium or calcium ions and ethylenediaminetetraacetic acid. Bioelectrochemistry and Bioenergetics 46:249-254.

Iswantini, D., Kano. K., Ikeda, T. 2000. Kinetics and thermodynamics of activation of quinoprotein glucose dehydrogenase apoenzyme in vivo and catalytic activity of the activated enzyme in Escherichia coli cells. Biochemistry Journal 350:917-923.

Liao et al., 2006. Evaluation of a kinetic uricase method for serum uric acid assay by predicting background serapance of uricase reaction solution with an integrated method. Journal of Zhejiang University SCIENCE B 7(6):497-502.

Luo, Y.C., Do, J.S., Liu, C.C. 2006. An amperometric uric acid biosensor based on modified Ir-C electrode. Biosensor and Bioelectronics 22:482-488.

Ogawa, J. 2006. Analysis of microbial purine metabolism and its application for hyperuricemia prevention. [Noda Institute for Scientific Research GRANT]. Outline of Research Result.

Trivedi, R.C., Rebar, L., Berta, E., Stong, L. 1978a. New ultraviolet (340 nm) methode for assay of uric acid in serum or plasma. Clin Chem 24(4):562-566.

Zhang et al., 2007. Development and analytical application of an uric acid biosensor using an urikase-immobilized eggshell membrane. Biosensor and Bioelectronics 22:1791-1797.

Zhou, X.L., Ma, X.H., Sun, Q.Q., Li, X., Guo, K.P. 2005. Isolation of a thermostable urikase-producing bacterium and study on its enzyme production conditions. Process Biochemistry 40:3749-3753.




View JIPI Stats

 

   Creative Commons License

This journal is published under the terms of the Creative Commons Attribution-NonCommercial 4.0 International License.

Editorial Office: Institute for Research and Community Services (LPPM), Andi Hakim Nasoetion Building, 5th Floor, Jl. Raya Darmaga, IPB Darmaga Campus, Bogor, West Java, Indonesia 16680, Telp/Fax: +62251-8622323, email: jipi-lppm@apps.ipb.ac.id; jipi-lppm@ipb.ac.id