Magda ALY, Sanaa TORK, Saleh AL-GARNI, Reda ALLAM

Production and characterization of uricase from Streptomyces exfoliatus UR10 isolated from farm wastes

Uricase plays an important role in nitrogen metabolism and can be used medically as a diagnostic reagent. From soil, wastewater, and poultry waste samples collected in Jeddah, 49 bacterial isolates were obtained on either nutrient agar or starch nitrate agar. All the obtained isolates were screened on minimal medium containing 0.5% uric acid for uricase production. The most active bacterium (isolate UR10) produced about 0.5 U/mL of intracellular uricase and was identified as a species belonging to the genus Streptomyces using morphological, physiological, and biochemical characters. By 16S rDNA, it was identified as Streptomyces exfoliatus UR10. Maximum uricase production was obtained using medium 2 with 0.2% uric acid as an inducer, an initial pH of 6.5, and an incubation temperature of 37 °C at 100 rpm. At the end of the incubation period, the cells were collected and disturbed, and the uricase enzyme was precipitated by ammonium sulfate. The enzyme was purified using different column chromatography methods, and the molecular weight of the purified uricase was determined by SDS-PAGE electrophoresis. The optimum temperature for maximum uricase activity was 45 °C; the optimum pH was 8. Co2+, Ni2+, Zn2+, Cu2+, and Pb2+ decreased the enzyme activity, whereas Ca2+, Mn2+, Mg2+, and Fe2+ stimulated it. In conclusion, uricase was produced by Streptomyces in a medium containing uric acid as inducer, and this enzyme can be used to detect and quantify uric acid in urine and/or blood.

Anahtar Kelimeler:

Key words: Uricase, Streptomyces, molecular weight, uric acid, enzyme activity, 16S rDNA

Production and characterization of uricase from Streptomyces exfoliatus UR10 isolated from farm wastes

Keywords:

Key words: Uricase, Streptomyces, molecular weight, uric acid, enzyme activity, 16S rDNA,

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Nishiya Y, Hibi T, Oda J. A purification method of the diagnostic enzyme Bacillus uricase using magnetic beads and non-specific protease. Protein Expres Purif 25: 426–429, 2002.
Colloc’h N, El Hajji M, Bachet B et al. Crystal structure of the protein drug urate oxidase-inhibitor complex at 2.05 Å resolution. Nat Struct Biol 4: 947–952, 1997.
Cheristians S, Kaltwasser H. Nickel-content of urease from Bacillus pasteurii. Arch Microbiol 145: 51–55, 1986.
Rando D, Steglitz U, Morsdorf G et al. Nickel availability and urease expression in Proteus mirabilis. Arch Microbiol 154: 428–432, 1990.
Nakagawa S, Ishino S, Teshiba S. Construction of catalase deficient Escherichia coli strains of the production of uricase. Biosci Biotech Bioch 60: 415–420, 1996.
Ammar MS, Elwan SH, El-Shahed AS. A uricolytic Streptomyces albogriseolus from an Egyptian soil. Taxonomy and uricase production and properties. J Microbiol 22: 261–279, 1987.
Zhou XL, Ma XH, Sun GQ et al. Isolation of a thermostable uricase producing bacterium and study on its enzyme production conditions. Process Biochem 40: 3749–3753, 2005.
Lofty WA. Production of a thermostable uricase by a novel Bacillus thermocatenulatus strain. Bioresource Technol 99: 699–702, 2008.
Tanaka A, Yamamura M, Kawamoto S et al. Production of uricase by Candida tropicalis using n-alkane as substrate. Appl Environ Microb 34: 342–346, 1977.
Abd El Fattah YR, Saeed HM, Gohar Y et al. Improved production of Pseudomonas aeruginosa uricase by optimization of process parameters through statistical experimental designs. Process Biochem 40: 1707–1714, 2005.
Shirling EB, Gottlieb D. Methods for characterization of Streptomyces species. Int J Syst Bacteriol 16: 313–340, 1966.
Lehejckova R, Demnerova K, Kralova B. Screening of microorganisms with uricase activity. Biotechnology Letters 8: 341–342, 1986.
Liu JG, Li GX. Culture conditions for uricase formation of Candida utilis. Wei Sheng Wu Xue Bao 29: 45–50, 1989.
Adamek Y, Kralova B, Suchova M et al. Purification of microbial uricase. J Chrom 497: 268–275, 1989.
Aly MM, El-Sabbagh S, El-Shouny W et al. Physiological response of Zea mays to NaCl stress with respect to Azotobacter chroococcum and Streptomyces niveus. Pak J Biol Sci 6: 2073– 2080, 2003.
Hasegawa T, Takizawa M, Tanida S. A rapid analysis for chemical grouping of aerobic actinomycetes. J Gen Appl Microbiol 29: 319–322, 1983.
Butte W. Rapid method for the determination of fatty acid profiles from fats and oils using trimethyl sulphonium hydroxide for transesterification. J Chrom 261: 142–145, 1983. Hoischen C, Gura K, Luge C et al. Lipid and fatty acid composition of cytoplasmic membranes from Streptomyces hygroscopicus and its stable protoplast-type L form. J Bacteriol 179: 3430–3436, 1997.
Weisberg WG, Barns SM, Pelletier DA et al. 16S ribosomal DNA amplification for phylogenetic study. J Bacteriol 173: 697–703, 1991.
Demnerova K, Kralova B, Lehejokova R et al. Purification of uricase by different strains of Streptomycetes. Biotech Lett 8: 577–578, 1986.
Rouf MA, Lomprey TF. Degradation of uric acid by certain aerobic bacteria. J Bacteriol 96: 617–622, 1968.
Agate A, Bhat JY. Microflora associated with the rhizosphere of Calotropis gigantea. J Indian Inst Sci 46: 1–10, 1964.
Abd El Fattah MG, Abo-Hamed NA. Bioconversion of poultry waste I-factors influencing the assay and productivity of crude uricase by three uricolytic filamentous fungi. Acta Microbiol Imm H 49: 445–454, 2002.
Aly MM, Tork S, Al-Garni SM et al. Chitinolytic enzyme production and genetic improvement of a new isolate belonging to Streptomyces anulatus. Ann Microbiol 61: 453–461, 2011.
Aly MM, Tork S, Al-Garni S et al. Production of lipase from genetically improved Streptomyces exfoliates LP10 isolated from oil contaminated soil. Afr J Microbiol Res 5: 1125–1137, 20
Azab EA, Aly MM, Fareed MF. Studies on uricase induction in certain bacteria. Egypt J Biol 7: 44–54, 2005.
Lookwood GF, Garrison RG. The possible role of uric acid in the ecology of Histoplasma capsuatum. Mycol Appl 35: 377– 379, 1968.
Mead GC. Anaerobic utilization of uric acid by some group D streptococci. J Gen Microbiol 82: 421–423, 1974.
Nour El-Dein M, El-Fallal A. Screening of some fungi for uricolytic activity. Qatar Univ Sci J 16: 71–76, 1996.
Khucharoenphaisan K, Sinma K. Production and partial characterization of uric acid degrading enzyme from new source Saccharopolyspora sp. PNR11. Pak J Biol Sci 14: 226– 231, 2011.
Williams ST, Sharpe ME, Molt JC. In: Krieg NR, Ludwig W, Whitman WB. eds. Bergey’s Manual of Systematic Bacteriology, Vol. Williams and Wilkins; 1989: pp. 2300–2648.
Ölmezoğlu E, Herand BK, Öncel MS, Tunç K, Özkan M. Copper bioremoval by novel bacterial isolates and their identification by 16S rRNA gene sequence analysis. Turk J Biol 36: 469–476, 2012.
Gupta N, Mishra S, Basak UC. Occurrence of different morphotypes of Streptomyces exfoliatus in mangrove ecosystems of Bhitarkanika, Orissa. Indian J Cult Collec 6: 21–27, 2009.
Atalla MM, Farag MM, Eman RH et al. Optimum conditions for uricase enzyme production by Gliomastix gueg. Malaysian J Microbiol 5: 45–50, 2009.
Ammar MS, Elwan SH, El-Desouky EM. Purification and some properties of uricase from Aspergillus flavus S-97. Egypt J Microbiol 23: 83–87, 1988.
Yazdi MT, Zarrini G, Mohit E et al. Mucor hiemalis: a new source for uricase production. World J Microb Biot 22: 325– 330, 2006.
Tohamy EY, Shindia AA. Partial purification and some properties of uricase produced by strain of Aspergillus terreus. Egypt J Microbiol 36: 77–87, 2001.
Kai L, Ma X, Zhou X et al. Purification and characterization of a thermostable uricase from Microbacterium sp. strain ZZJ4-1. World J Microb Biot 24: 401–406, 2008.
Yokoyama S, Ogawa A, Obayashi A. Rapid extraction of uricase from Candida utilis cells by use of reducing agent plus surfactant. Enzyme Microb Tech 10: 52–55, 1988.
Bongaerts GPA, Uitzetter J, Brouns R et al. Uricase of Bacillus fastidiosus properties and regulation of synthesis. Biochem Biophys Acta 527: 348–358, 1978.
Saeed HM, Abdel-Fattah YR, Gohar YM et al. Purification and characterization of extracellular Pseudomonas aeruginosa urate oxidase enzyme. Pol J Microbiol 53: 45–52, 2004.
Koyama Y, Ichikawa T, Nakano E. Cloning, sequence analysis, and expression in Escherichia coli of the gene encoding the Candida utilis urate oxidase (uricase). J Biochem (Tokyo) 120: 969–973, 1996.
Arrizubieta MJ, Polaina J. Increased thermal resistance and modification of the catalytic properties of α-glucosidase by random mutagenesis and in vitro recombination. J Biol Chem 275: 28843–28848, 2000.
Schiavon O, Caliceti P, Ferruti P et al. Therapeutic proteins: a comparison of chemical and biological properties of uricase conjugated to linear or branched poly ethylene glycol and poly N acryloyl morpholine. Il Farmaco 55: 264–269, 2000.
Wu XW, Lee CC, Muzny DM. Urate oxidase: primary structure and evolutionary implications. P Natl Acad Sci USA 86: 9412– 9416, 1989.
Suzuki K, Sakasegawa S, Misaki H et al. Molecular cloning and expression of uricase gene from Arthrobacter globiformis in Escherichia coli and characterization of the gene product. J Biosci Bioeng 98: 153–158, 2004.