Heterologous expression and purification of a multiheme cytochrome from a Gram-positive bacterium capable of performing extracellular respiration

N. L. Costa, H. K. Carlson, J. D. Coates, R. O. Louro, C. M. Paquete

Research output: Contribution to journalArticle

12 Citations (Scopus)

Abstract

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.

Original languageEnglish
Pages (from-to)48-52
Number of pages5
JournalProtein Expression And Purification
Volume111
DOIs
Publication statusPublished - 1 Jul 2015

Keywords

  • Extracellular electron transfer
  • Gram-positive bacteria
  • Microbial fuel cells
  • Multiheme-cytochrome
  • Thermincola

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