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

Protein Expression and Purification
N L CostaCatarina M Paquete

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.

References

Dec 22, 1987·Proceedings of the Royal Society of London. Series B, Containing Papers of a Biological Character·J D Pettigrew, B Dreher
Jun 1, 1997·Protein Science : a Publication of the Protein Society·R E DalbeyJ M van Dijl
Aug 28, 1998·Molecular Microbiology·R KranzS Merchant
Feb 19, 2000·Biochemical and Biophysical Research Communications·P N da CostaL M Saraiva
Aug 29, 2001·FEMS Microbiology Reviews·K H van WelyA J Driessen
Apr 5, 2002·Chembiochem : a European Journal of Chemical Biology·Lucia BanciAndrew R Thompsett
Dec 15, 2006·Journal of Biochemistry·Yuki TakayamaHideo Akutsu
Aug 11, 2007·Protein Expression and Purification·Yuki Takayama, Hideo Akutsu
Mar 18, 2008·Protein Expression and Purification·Ana P FernandesCarlos A Salgueiro
Sep 5, 2008·The ISME Journal·Kelly C WrightonJohn D Coates
Sep 2, 2009·Microbiology and Molecular Biology Reviews : MMBR·Robert G KranzElaine R Frawley
Oct 20, 2009·Applied and Environmental Microbiology·Bjoern SchuetzJohannes Gescher
Jun 8, 2010·Journal of Bacteriology·Kathryne G Byrne-BaileyJohn D Coates
Sep 13, 2011·Applied and Environmental Microbiology·K C WrightonJ D Coates
Feb 7, 2012·Proceedings of the National Academy of Sciences of the United States of America·Hans K CarlsonJohn D Coates
Mar 1, 2012·Frontiers in Microbiology·Liang ShiJames K Fredrickson
Mar 3, 2012·Journal of Biological Inorganic Chemistry : JBIC : a Publication of the Society of Biological Inorganic Chemistry·Matthew YoungblutA Andrew Pacheco
Jun 5, 2012·The Journal of Biological Chemistry·Ivo H SaraivaRicardo O Louro
Jul 4, 2012·Annual Review of Microbiology·Derek R Lovley
Nov 28, 2012·Biochemical Society Transactions·Ricardo O Louro, Catarina M Paquete
Mar 27, 2013·Applied Microbiology and Biotechnology·Kheng Oon LowRosli Md Illias
Mar 19, 2014·Biochimica Et Biophysica Acta·Andreia F Verissimo, Fevzi Daldal
Jul 16, 2014·Frontiers in Microbiology·Catarina M PaqueteRicardo O Louro
Aug 21, 2014·Applied Microbiology and Biotechnology·Anne SydowDirk Holtmann

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