Purification and Properties of Uricase from Candida Sp. and Its Application in Uric Acid Analysis in Serum

Liu, Jianguo; Li, Gao Xiang; Liu, Hong; Zhou, Xiying

doi:10.1111/j.1749-6632.1995.tb20000.x

Cited by 15 publications

(19 citation statements)

References 8 publications

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“…The loss of uricase activity might be caused by the degradation in acidic environment by proteases secreted by the host cells or released from host cell lyses. On the other hand, low pH may cause instability of uricase (Liu et al 1994).…”

Section: Recombinant Uricase Production In Bioreactor Culturesmentioning

confidence: 99%

Uricase production by a recombinant Hansenula polymorpha strain harboring Candida utilis uricase gene

Chen

Wang

et al. 2008

Appl Microbiol Biotechnol

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Uricase is an important medical enzyme which can be used to determine urate in clinical analysis, to therapy gout, hyperuricemia, and tumor lysis syndrome. Uricase of Candida utilis was successfully expressed in Hansenula polymorpha under the control of methanol oxidase promoter using Saccharomyces cerevisiae α-factor signal peptide as the secretory sequence. Recombinant H. polymorpha MU200 with the highest extracellular uricase production was characterized with three copies of expression cassette and selected for process optimization for the production of recombinant enzyme. Among the parameters investigated in shaking flask cultures, the pH value of medium and inoculum size had great influence on the recombinant uricase production. The maximum extracellular uricase yield of 2.6 U/ml was obtained in shaking flask culture. The yield of recombinant uricase was significantly improved by the combined use of a high cell-density cultivation technique and a pH control strategy of switching culture pH from 5.5 to 6.5 in the induction phase. After induction for 58 h, the production of recombinant uricase reached 52.3 U/ml (about 2.1 g/l of protein) extracellularly and 60.3 U/ml (about 2.4 g/l) intracellularly in fed-batch fermentation, which are much higher than those expressed in other expression systems. To our knowledge, this is the first report about the heterologous expression of uricase in H. polymorpha.

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Section: Recombinant Uricase Production In Bioreactor Culturesmentioning

confidence: 99%

Uricase production by a recombinant Hansenula polymorpha strain harboring Candida utilis uricase gene

Chen

Wang

et al. 2008

Appl Microbiol Biotechnol

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“…Known urate oxidase subunits range from 30 to 80 kDa. Different enzyme structures have been described: monomer [Liu et al, 1994], dimer [Lucas et al, 1983], homotetramer [Giffard et al, 2011;Nishimura et al, 1982] and heterotetramer [Bongaerts et al, 1978].…”

Section: Discussionmentioning

confidence: 99%

“…[Liu et al, 1994] using enzyme activity assays. They found that directly after the addition of purines, such as adenine and uric acid, the respective genes are activated.…”

Section: Discussionmentioning

confidence: 99%

“…Urate oxidase is an inducible enzyme that is found in a variety of organisms such as bacteria [Arima and Nose, 1968;Lotfy, 2008], fungi [Franke and Krieg, 1952], yeast [Liu et al, 1994], algae [Alamillo et al, 1991], plants [Montalbini et al, 1997] and is expressed in mammalian peroxisomes in specific tissues [Antonenkov and Panchenko, 1978;Rajoka et al, 2006]. It is however absent in higher primates (hominoids and some New World monkeys), birds, terrestrial reptiles and most insects [Keebaugh and Thomas, 2010;Keilin, 1959;Oda et al, 2002].…”

Section: Introductionmentioning

confidence: 99%

“…The product, allantoin, is 5-10 times more soluble than uric acid and does not precipitate at physiological concentrations. Urate oxidase can also be used to analytically to determine uric acid content of serum [Liu et al, 1994;Zhao et al, 2009] or therapeutically for uric acid reduction in biological fluids for resolving hyperuricemic disorders such as tumor lysis syndrome, which can occur during chemotherapy [Wang et al, 2006].…”

Section: Introductionmentioning

confidence: 99%

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Arxula adeninivorans Recombinant Urate Oxidase and Its Application in the Production of Food with Low Uric Acid Content

Trautwein-Schult

Jankowska

Cordes³

et al. 2013

Microb Physiol

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Hyperuricemia and its symptoms are becoming increasingly common worldwide. Elevated serum uric acid levels are caused by increased uric acid synthesis from food constituents and reduced renal excretion. Treatment in most cases involves reducing alcohol intake and consumption of meat and fish or treatment with pharmaceuticals. Another approach could be to reduce uric acid level in food, either during production or consumption. This work reports the production of recombinant urate oxidase by Arxula adeninivorans and its application to reduce uric acid in a food product. The A. adeninivorans urate oxidase amino acid sequence was found to be similar to urate oxidases from other fungi (61-65% identity). In media supplemented with adenine, hypoxanthine or uric acid, induction of the urate oxidase (AUOX) gene and intracellular accumulation of urate oxidase (Auoxp) was observed. The enzyme characteristics were analyzed from isolates of the wild-type strain A. adeninivorans LS3, as well as from those of transgenic strains expressing the AUOX gene under control of the strong constitutive TEF1 promoter or the inducible AYNI1 promoter. The enzyme showed high substrate specificity for uric acid, a broad temperature and pH range, high thermostability and the ability to reduce uric acid content in food.

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Purification and characterization of caprine kidney uricase, possessing novel kinetic and thermodynamic properties

Rajoka

Rehman

Tabish

et al. 2005

World J Microbiol Biotechnol

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Purified uricase from a caprine kidney, possessed K m and V max values of 1.1 mg ml )1 and 3512 IU (mg protein) )1 for uric acid hydrolysis, respectively. The optimum temperature and pH for catalytic activity were 40°C and 8.5, respectively. The activation energy for formation of ES complex was 13.6 kJ mol )1 . Enthalpy (DH*), entropy of activation (DS*) and Gibbs free energy demand of uricase inactivation were 62.8 kJ mol )1 , )102 J mol )1 K )1 and 104.3 kJ mol )1 , respectively. Gibbs free enrgy demand for substrate binding and transition state stabilization were also determined which were comparable with those for themostable enzymes.

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Purification and Properties of Uricase from Candida Sp. and Its Application in Uric Acid Analysis in Serum

Cited by 15 publications

References 8 publications

Uricase production by a recombinant Hansenula polymorpha strain harboring Candida utilis uricase gene

Uricase production by a recombinant Hansenula polymorpha strain harboring Candida utilis uricase gene

<b><i>Arxula adeninivorans</i></b> Recombinant Urate Oxidase and Its Application in the Production of Food with Low Uric Acid Content

Purification and characterization of caprine kidney uricase, possessing novel kinetic and thermodynamic properties

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