Activity and Stability of Uricase from Lactobacillus plantarum Immobilizated on Natural Zeolite for Uric Acid Biosensor

Abstract

Determination of uric acid concentration in human urine and blood is needed to diagnose several diseases, especially the occurrence of kidney disease in gout patients. Therefore, it is needed to develop a simple and inexpensive method for uric acid detection. The purpose of the research was to observe the use of Indonesian microbe that was immobilized on natural zeolite as a source of uricase for uric acid biosensor. Selection of mediators and determination of optimum condition measurement, the stability and kinetic properties of L. plantarum uricase were performed using carbon paste electrode. Cyclic voltammetry was employed to investigate the catalytic behavior of the biosensor. The result indicated that the best mediator for measurement of L. plantarum uricase activity was Qo (2,3-dimethoxy-5-methyl-1,4 benzoquinone). Optimum conditions for immobilization of L. plantarum uricase on zeolite were obtained at pH 7.6, with temperature of 28 degrees C, using uric acid concentration of 0.015 mM and zeolite mass at 135 mg K(M) and V(Max) of L. plantarum uricase obtained from Lineweaver-burk equation for the immobilization uricase on zeolite were 8.6728 x 10(-4) mM and 6.3052 mM, respectively. K(M) value of L. plantarum uricase directly immobilized onto the electrode surface was smaller than K(M) value of L. plantarum uricase immobilized on zeolite. The smaller K(M) value shows the higher affinity toward the substrate. The Electrode when kept at 10 degrees C was stable until 6 days, however the immobilized electrode on zeolite was stable until 18 days. Therefore, Indonesian L. plantarum could be used as a uric acid biosensor.

Uric Acid Biosensor Using Immobilised Lactobacillus plantarum Mar8 on Zeolite/K-Carrageenan Membrane

“…Mar8 is a lactic acid bacterial isolate producing uricase that has good catalytic activity on uric acid (Iswantini et al, 2014). Therefore immobilisation of this isolate on a membrane that facilitates the bacteria to extend its life time and produce catalytic uricase, would be an advantage in an uric acid biosensor development.…”

Isolation of Bile Salt Hydrolase and Uricase Producing Lactobacillus brevis SF121 from Pak Sian Dong (Fermented Spider Plant) for using as Probiotics

“…Therefore, catalytic for uric acid activity, uricase is interested to be used for hyperuricemia and gout. Variety genus of bacteria have been screened and isolated with uricase activity, such as Bacillus 23,24 , Pseudomonas 25 and Lactobacillus 26 .…”

Selection of uric acid oxidizing-Lactobacillus plantarum isolates based on their genetic determinant and uricase kinetics

“…In most mammalian species uric acid is further metabolized by the enzyme uricase to the more soluble metabolite which is subsequently excreted in the urine (Benn et al, 2018). Human and certain primates, however, lost their uric acid oxidizing enzyme, the uricase, consequently uric acid is the inal product of purine catabolism and can not be metabolized to the soluble allantoin, urea, or ammonia for excretion from the body (Kratzer et al, 2014;Iswantini et al, 2014;Tan et al, 2016).…”

“…Notably, an isolate of probiotic bacteria of Lactobacillus plantarum has been reported to show an uric acid oxidizing activity (Iswantini et al, 2014). L. plantarum is commonly found in plantderived foods such as fruits, pickles, wine, and bean sauces (Li et al, 2014).…”