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Cloning, Purification, and Partial Characterization of Bacillus subtilis Urate Oxidase Expressed in Escherichia coli
Joint Authors
Prates, Maura Vianna
Moraes, Lídia Maria Pepe de
Torres, Fernando Araripe Gonçalves
De Marco, Janice Lisboa
Bloch, Carlos
Pfrimer, Pollyana
Salles, Loise P.
Galdino, Alexsandro Sobreira
Reis, Viviane Castelo Branco
Source
Journal of Biomedicine and Biotechnology
Issue
Vol. 2010, Issue 2010 (31 Dec. 2010), pp.1-6, 6 p.
Publisher
Hindawi Publishing Corporation
Publication Date
2010-02-04
Country of Publication
Egypt
No. of Pages
6
Main Subjects
Abstract EN
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.
American Psychological Association (APA)
Pfrimer, Pollyana& Moraes, Lídia Maria Pepe de& Galdino, Alexsandro Sobreira& Salles, Loise P.& Reis, Viviane Castelo Branco& De Marco, Janice Lisboa…[et al.]. 2010. Cloning, Purification, and Partial Characterization of Bacillus subtilis Urate Oxidase Expressed in Escherichia coli. Journal of Biomedicine and Biotechnology،Vol. 2010, no. 2010, pp.1-6.
https://search.emarefa.net/detail/BIM-489541
Modern Language Association (MLA)
Pfrimer, Pollyana…[et al.]. Cloning, Purification, and Partial Characterization of Bacillus subtilis Urate Oxidase Expressed in Escherichia coli. Journal of Biomedicine and Biotechnology No. 2010 (2010), pp.1-6.
https://search.emarefa.net/detail/BIM-489541
American Medical Association (AMA)
Pfrimer, Pollyana& Moraes, Lídia Maria Pepe de& Galdino, Alexsandro Sobreira& Salles, Loise P.& Reis, Viviane Castelo Branco& De Marco, Janice Lisboa…[et al.]. Cloning, Purification, and Partial Characterization of Bacillus subtilis Urate Oxidase Expressed in Escherichia coli. Journal of Biomedicine and Biotechnology. 2010. Vol. 2010, no. 2010, pp.1-6.
https://search.emarefa.net/detail/BIM-489541
Data Type
Journal Articles
Language
English
Notes
Includes bibliographical references
Record ID
BIM-489541