Universität zu Köln

Chmielewski, Markus, Kuehle, Johannes, Chrobok, Danuta, Riet, Nicole, Hallek, Michael and Abken, Hinrich (2019). FimH-based display of functional eukaryotic proteins on bacteria surfaces. Sci Rep, 9. LONDON: NATURE PUBLISHING GROUP. ISSN 2045-2322

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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-alpha (TGF-alpha) 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.

Item Type:	Journal Article
Creators:	Creators Email ORCID ORCID Put Code Chmielewski, Markus UNSPECIFIED UNSPECIFIED UNSPECIFIED Kuehle, Johannes UNSPECIFIED UNSPECIFIED UNSPECIFIED Chrobok, Danuta UNSPECIFIED UNSPECIFIED UNSPECIFIED Riet, Nicole UNSPECIFIED UNSPECIFIED UNSPECIFIED Hallek, Michael UNSPECIFIED UNSPECIFIED UNSPECIFIED Abken, Hinrich UNSPECIFIED UNSPECIFIED UNSPECIFIED
URN:	urn:nbn:de:hbz:38-137958
DOI:	10.1038/s41598-019-44883-z
Journal or Publication Title:	Sci Rep
Volume:	9
Date:	2019
Publisher:	NATURE PUBLISHING GROUP
Place of Publication:	LONDON
ISSN:	2045-2322
Language:	English
Faculty:	Unspecified
Divisions:	Unspecified
Subjects:	no entry
Uncontrolled Keywords:	Keywords Language OUTER-MEMBRANE; ESCHERICHIA-COLI; CELL-SURFACE; BETA-LACTAMASE; AUTODISPLAY; MECHANISM; FIMBRIAE; ADHESIN; YIELDS; PILUS Multiple languages Multidisciplinary Sciences Multiple languages
Refereed:	Yes
URI:	http://kups.ub.uni-koeln.de/id/eprint/13795