Abstract
Gout is a metabolic disorder due to the deposition of uric acid crystals within articular or periarticular tissues. Uricase (urate oxidase) catalyzes the oxidation of less water soluble uric acid (7 mg/dl) to a compound allantoin which is more water soluble(11g/L at 40°C) resulting into the ease of excretion of uric acid. The objective of the work was to develop a new method for screening of microbes for uricase production and estimation of uricase thereof. This was achieved by utilizing the fact that uric acid dissolves on being acted upon by uricase. The proposed method is a novel, inexpensive, simple and sensitive technique for screening and estimation of uricase. Biomass and uricase production at different stages of microbial growth curve for the uricase producing microbe was studied. Effects of different medium components affecting uricase production by microbes were studied using Placket Burman statistical design. Addition of uric acid in the nutrient medium was found to be effective in increasing the uricase production by microbes growing in the medium.
Highlights
Enzymes are biocatalysts, thermo labile in nature and highly specific molecules for their action on substrate
Uric acid is degraded by urate oxidase to allantoin as described previously[3,4,5]
An attempt was made to correlate the fact that uric acid has a solubility of 7 mg/dL in water while allantoin is more soluble than uric acid having a solubility of 400 mg/dL and that allantoin is formed when uricase acts upon uric acid[3]
Summary
Thermo labile in nature and highly specific molecules for their action on substrate. These enzymes are synthesized by living cells only[1]. Identification of natural microbes which produce uricase is a difficult task, because it requires each microbial colony to be tested for the presence of uricase, by using estimation kits available in the market. These have a high cost and require specific critical environmental conditions for conduction of the tests. Allantoin has a much higher solubility which is 400 mg/dl in plasma and water at 250C
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