Controlled functional expression of the bacteriocins pediocin PA-1 and bactofencin A in Escherichia coli

Abstract The bacteriocins bactofencin A (class IId) and pediocin PA-1 (class IIa) are encoded by operons with a similarly clustered gene organization including a structural peptide, an immunity protein, an ABC transporter and accessory bacteriocin transporter protein. Cloning of these operons in E....

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Autores principales:	Beatriz Mesa-Pereira, Paula M. O’Connor, Mary C. Rea, Paul D. Cotter, Colin Hill, R. Paul Ross
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2017
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/70ee0db8db5645a7b55daa74ed25d185
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spelling	oai:doaj.org-article:70ee0db8db5645a7b55daa74ed25d1852021-12-02T12:32:31ZControlled functional expression of the bacteriocins pediocin PA-1 and bactofencin A in Escherichia coli10.1038/s41598-017-02868-w2045-2322https://doaj.org/article/70ee0db8db5645a7b55daa74ed25d1852017-06-01T00:00:00Zhttps://doi.org/10.1038/s41598-017-02868-whttps://doaj.org/toc/2045-2322Abstract The bacteriocins bactofencin A (class IId) and pediocin PA-1 (class IIa) are encoded by operons with a similarly clustered gene organization including a structural peptide, an immunity protein, an ABC transporter and accessory bacteriocin transporter protein. Cloning of these operons in E. coli TunerTM (DE3) on a pETcoco-2 derived vector resulted in successful secretion of both bacteriocins. A corresponding approach, involving the construction of vectors containing different combinations of these genes, revealed that the structural and the transporter genes alone are sufficient to permit heterologous production and secretion in this host. Even though the accessory protein, usually associated with optimal disulfide bond formation, was not required for bacteriocin synthesis, its presence did result in greater pediocin PA-1 production. The simplicity of the system and the fact that the associated bacteriocins could be recovered from the extracellular medium provides an opportunity to facilitate protein engineering and the overproduction of biologically-active bacteriocins at industrial scale. Additionally, this system could enable the characterization of new bacteriocin operons where genetic tools are not available for the native producers.Beatriz Mesa-PereiraPaula M. O’ConnorMary C. ReaPaul D. CotterColin HillR. Paul RossNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 7, Iss 1, Pp 1-11 (2017)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Beatriz Mesa-Pereira Paula M. O’Connor Mary C. Rea Paul D. Cotter Colin Hill R. Paul Ross Controlled functional expression of the bacteriocins pediocin PA-1 and bactofencin A in Escherichia coli
description	Abstract The bacteriocins bactofencin A (class IId) and pediocin PA-1 (class IIa) are encoded by operons with a similarly clustered gene organization including a structural peptide, an immunity protein, an ABC transporter and accessory bacteriocin transporter protein. Cloning of these operons in E. coli TunerTM (DE3) on a pETcoco-2 derived vector resulted in successful secretion of both bacteriocins. A corresponding approach, involving the construction of vectors containing different combinations of these genes, revealed that the structural and the transporter genes alone are sufficient to permit heterologous production and secretion in this host. Even though the accessory protein, usually associated with optimal disulfide bond formation, was not required for bacteriocin synthesis, its presence did result in greater pediocin PA-1 production. The simplicity of the system and the fact that the associated bacteriocins could be recovered from the extracellular medium provides an opportunity to facilitate protein engineering and the overproduction of biologically-active bacteriocins at industrial scale. Additionally, this system could enable the characterization of new bacteriocin operons where genetic tools are not available for the native producers.
format	article
author	Beatriz Mesa-Pereira Paula M. O’Connor Mary C. Rea Paul D. Cotter Colin Hill R. Paul Ross
author_facet	Beatriz Mesa-Pereira Paula M. O’Connor Mary C. Rea Paul D. Cotter Colin Hill R. Paul Ross
author_sort	Beatriz Mesa-Pereira
title	Controlled functional expression of the bacteriocins pediocin PA-1 and bactofencin A in Escherichia coli
title_short	Controlled functional expression of the bacteriocins pediocin PA-1 and bactofencin A in Escherichia coli
title_full	Controlled functional expression of the bacteriocins pediocin PA-1 and bactofencin A in Escherichia coli
title_fullStr	Controlled functional expression of the bacteriocins pediocin PA-1 and bactofencin A in Escherichia coli
title_full_unstemmed	Controlled functional expression of the bacteriocins pediocin PA-1 and bactofencin A in Escherichia coli
title_sort	controlled functional expression of the bacteriocins pediocin pa-1 and bactofencin a in escherichia coli
publisher	Nature Portfolio
publishDate	2017
url	https://doaj.org/article/70ee0db8db5645a7b55daa74ed25d185
work_keys_str_mv	AT beatrizmesapereira controlledfunctionalexpressionofthebacteriocinspediocinpa1andbactofencinainescherichiacoli AT paulamoconnor controlledfunctionalexpressionofthebacteriocinspediocinpa1andbactofencinainescherichiacoli AT marycrea controlledfunctionalexpressionofthebacteriocinspediocinpa1andbactofencinainescherichiacoli AT pauldcotter controlledfunctionalexpressionofthebacteriocinspediocinpa1andbactofencinainescherichiacoli AT colinhill controlledfunctionalexpressionofthebacteriocinspediocinpa1andbactofencinainescherichiacoli AT rpaulross controlledfunctionalexpressionofthebacteriocinspediocinpa1andbactofencinainescherichiacoli
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Controlled functional expression of the bacteriocins pediocin PA-1 and bactofencin A in Escherichia coli

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