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Title: Cloning, purification, and partial characterization of Bacillus subtilis urate oxidase expressed in Escherichia coli
Authors: Pfrimer, Pollyanna
Moraes, Lídia Maria Pepe de
Galdino, Alexsandro Sobreira
Salles, Loise Pedrosa
Reis, Viviane Castelo Branco
De Marco, Janice Lisboa
Prates, Maura Vianna
Bloch Júnior, Carlos
Torres, Fernando Araripe Gonçalves
Assunto:: Enzimas - análise
Escherichia coli
Bactérias
Issue Date: 2010
Publisher: Hindawi Publishing Corporation
Citation: PFRIMER, Pollyanna et al. Cloning, purification, and partial characterization of Bacillus subtilis urate oxidase expressed in Escherichia coli. BioMed Research International, v. 2010, 2010. Disponível em: <http://www.hindawi.com/journals/bmri/2010/674908/>. Acesso em: 27 ago. 2013. DOI: http://dx.doi.org/10.1155/2010/674908.
Abstract: Urate oxidase (EC 1.7.3.3) is an enzyme involved in purine metabolism which is used in the treatment of gout and as diagnostic reagent for detection of uric acid. In order to produce this enzyme in large quantities for biotechnological purposes, the gene coding for the Bacillus subtilis urate oxidase was cloned and heterologously expressed in Escherichia coli. Time course induction in E. coli showed an induced protein with an apparent molecular mass of ∼60 kDa. Soluble recombinant enzyme was purified in a single-step procedure using Ni-NTA column. The enzyme was purified 2.1-fold with a yield of 56% compared to the crude extract. MALDI-TOF analysis revealed an ion with a mass of 58675 Da which is in agreement with the expected mass of the recombinant protein. The purified enzyme showed an optimal pH and temperature of 8.0 and 37◦C, respectively, and retained 90% of its activity after 72 hours of incubation at −20◦C and 4◦C.
Licença:: Copyright © 2010 Pollyana Pfrimer et al. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. Fonte: http://www.hindawi.com/journals/bmri/2010/674908/. Acesso em 27 ago. 2013.
DOI: http://dx.doi.org/10.1155/2010/674908
Appears in Collections:CEL - Artigos publicados em periódicos
FAR - Artigos publicados em periódicos

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