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Heterologous expression and purification of a multiheme cytochrome from a Gram-positive bacterium capable of performing extracellular respiration.
Costa, N L; Carlson, H K; Coates, J D; Louro, R O; Paquete, C M.
Afiliación
  • Costa NL; Instituto de Tecnologia Química e Biológica António Xavier, Universidade Nova de Lisboa, Av. da República - EAN, 2780-157 Oeiras, Portugal.
  • Carlson HK; Energy Bioscience Institute, University of California, Berkeley, CA 94720, United States.
  • Coates JD; Energy Bioscience Institute, University of California, Berkeley, CA 94720, United States; Department of Plant and Microbial Biology, University of California, Berkeley, CA 94720, United States.
  • Louro RO; Instituto de Tecnologia Química e Biológica António Xavier, Universidade Nova de Lisboa, Av. da República - EAN, 2780-157 Oeiras, Portugal.
  • Paquete CM; Instituto de Tecnologia Química e Biológica António Xavier, Universidade Nova de Lisboa, Av. da República - EAN, 2780-157 Oeiras, Portugal. Electronic address: cpaquete@itqb.unl.pt.
Protein Expr Purif ; 111: 48-52, 2015 Jul.
Article en En | MEDLINE | ID: mdl-25797208
Microbial electrochemical technologies are emerging as environmentally friendly biotechnological processes. Recently, a thermophilic Gram-positive bacterium capable of electricity production in a microbial fuel cell was isolated. Thermincola potens JR contains several multiheme c-type cytochromes that were implicated in the process of electricity production. In order to understand the molecular basis by which Gram-positive bacteria perform extracellular electron transfer, the relevant proteins need to be characterized in detail. Towards this end, a chimeric gene containing the signal peptide from Shewanella oneidensis MR-1 small tetraheme cytochrome c (STC) and the gene sequence of the target protein TherJR_0333 was constructed. This manuscript reports the successful expression of this chimeric gene in the Gram-negative bacterium Escherichia coli and its subsequent purification and characterization. This methodology opens the possibility to study other multiheme cytochromes from Gram-positive bacteria, allowing the extracellular electron transfer mechanisms of this class of organisms to be unraveled.
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Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Consumo de Oxígeno / Proteínas Bacterianas / Citocromos c / Escherichia coli / Bacterias Grampositivas Idioma: En Revista: Protein Expr Purif Asunto de la revista: BIOLOGIA MOLECULAR Año: 2015 Tipo del documento: Article País de afiliación: Portugal Pais de publicación: Estados Unidos

Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Consumo de Oxígeno / Proteínas Bacterianas / Citocromos c / Escherichia coli / Bacterias Grampositivas Idioma: En Revista: Protein Expr Purif Asunto de la revista: BIOLOGIA MOLECULAR Año: 2015 Tipo del documento: Article País de afiliación: Portugal Pais de publicación: Estados Unidos