FimH-based display of functional eukaryotic proteins on bacteria surfaces

Abstract The demand for recombinant proteins for analytic and therapeutic purposes is increasing; however, most currently used bacterial production systems accumulate the recombinant proteins in the intracellular space, which requires denaturating procedures for harvesting and functional testing. We...

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Autores principales:	Markus Chmielewski, Johannes Kuehle, Danuta Chrobok, Nicole Riet, Michael Hallek, Hinrich Abken
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2019
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/6f5191feaea74029b69a14bd4e59208a
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spelling	oai:doaj.org-article:6f5191feaea74029b69a14bd4e59208a2021-12-02T15:09:31ZFimH-based display of functional eukaryotic proteins on bacteria surfaces10.1038/s41598-019-44883-z2045-2322https://doaj.org/article/6f5191feaea74029b69a14bd4e59208a2019-06-01T00:00:00Zhttps://doi.org/10.1038/s41598-019-44883-zhttps://doaj.org/toc/2045-2322Abstract The demand for recombinant proteins for analytic and therapeutic purposes is increasing; however, most currently used bacterial production systems accumulate the recombinant proteins in the intracellular space, which requires denaturating procedures for harvesting and functional testing. We here present a novel FimH-based expression system that enables display of fully functional eukaryotic proteins while preventing technical difficulties in translocating, folding, stabilizing and isolating the displayed proteins. As examples, Gaussia Luciferase (GLuc), epidermal growth factor (EGF), transforming growth factor-α (TGF-α) and epiregulin (EPRG) were expressed as FimH fusion proteins on the surface of E. coli bacteria. The fusion proteins were functionally active and could be released from the bacterial surface by specific proteolytic cleavage into the culture supernatant allowing harvesting of the produced proteins. EGFR ligands, produced as FimH fusion proteins and released by proteolytic cleavage, bound to the EGF receptor (EGFR) on cancer cells inducing EGFR phosphorylation. In another application of the technology, GLuc-FimH expressed on the surface of bacteria was used to track tumor-infiltrating bacteria by bioluminescence imaging upon application to mice, thereby visualizing the colonization of transplanted tumors. The examples indicate that the FimH-fusion protein technology can be used in various applications that require functionally active proteins to be displayed on bacterial surfaces or released into the culture supernatant.Markus ChmielewskiJohannes KuehleDanuta ChrobokNicole RietMichael HallekHinrich AbkenNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 9, Iss 1, Pp 1-10 (2019)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Markus Chmielewski Johannes Kuehle Danuta Chrobok Nicole Riet Michael Hallek Hinrich Abken FimH-based display of functional eukaryotic proteins on bacteria surfaces
description	Abstract The demand for recombinant proteins for analytic and therapeutic purposes is increasing; however, most currently used bacterial production systems accumulate the recombinant proteins in the intracellular space, which requires denaturating procedures for harvesting and functional testing. We here present a novel FimH-based expression system that enables display of fully functional eukaryotic proteins while preventing technical difficulties in translocating, folding, stabilizing and isolating the displayed proteins. As examples, Gaussia Luciferase (GLuc), epidermal growth factor (EGF), transforming growth factor-α (TGF-α) and epiregulin (EPRG) were expressed as FimH fusion proteins on the surface of E. coli bacteria. The fusion proteins were functionally active and could be released from the bacterial surface by specific proteolytic cleavage into the culture supernatant allowing harvesting of the produced proteins. EGFR ligands, produced as FimH fusion proteins and released by proteolytic cleavage, bound to the EGF receptor (EGFR) on cancer cells inducing EGFR phosphorylation. In another application of the technology, GLuc-FimH expressed on the surface of bacteria was used to track tumor-infiltrating bacteria by bioluminescence imaging upon application to mice, thereby visualizing the colonization of transplanted tumors. The examples indicate that the FimH-fusion protein technology can be used in various applications that require functionally active proteins to be displayed on bacterial surfaces or released into the culture supernatant.
format	article
author	Markus Chmielewski Johannes Kuehle Danuta Chrobok Nicole Riet Michael Hallek Hinrich Abken
author_facet	Markus Chmielewski Johannes Kuehle Danuta Chrobok Nicole Riet Michael Hallek Hinrich Abken
author_sort	Markus Chmielewski
title	FimH-based display of functional eukaryotic proteins on bacteria surfaces
title_short	FimH-based display of functional eukaryotic proteins on bacteria surfaces
title_full	FimH-based display of functional eukaryotic proteins on bacteria surfaces
title_fullStr	FimH-based display of functional eukaryotic proteins on bacteria surfaces
title_full_unstemmed	FimH-based display of functional eukaryotic proteins on bacteria surfaces
title_sort	fimh-based display of functional eukaryotic proteins on bacteria surfaces
publisher	Nature Portfolio
publishDate	2019
url	https://doaj.org/article/6f5191feaea74029b69a14bd4e59208a
work_keys_str_mv	AT markuschmielewski fimhbaseddisplayoffunctionaleukaryoticproteinsonbacteriasurfaces AT johanneskuehle fimhbaseddisplayoffunctionaleukaryoticproteinsonbacteriasurfaces AT danutachrobok fimhbaseddisplayoffunctionaleukaryoticproteinsonbacteriasurfaces AT nicoleriet fimhbaseddisplayoffunctionaleukaryoticproteinsonbacteriasurfaces AT michaelhallek fimhbaseddisplayoffunctionaleukaryoticproteinsonbacteriasurfaces AT hinrichabken fimhbaseddisplayoffunctionaleukaryoticproteinsonbacteriasurfaces
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FimH-based display of functional eukaryotic proteins on bacteria surfaces

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