A Sense of Balance: Experimental Investigation and Modeling of a Malonyl-CoA Sensor in Escherichia coli

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• dc.contributor.author Fehér, Tamásca
• dc.contributor.author Libis, Vincentca
• dc.contributor.author Carbonell, Pabloca
• dc.contributor.author Faulon, Jean-Loupca
• dc.date.accessioned 2015-06-11T08:19:07Z
• dc.date.available 2015-06-11T08:19:07Z
• dc.date.issued 2015
• dc.description.abstract Production of value-added chemicals in microorganisms is regarded as a viable alternative to chemical synthesis. In the past decade, several engineered pathways producing such chemicals, including plant secondary metabolites in microorganisms have been reported; upscaling their production yields, however, was often challenging. Here, we analyze a modular device designed for sensing malonyl-CoA, a common precursor for both fatty acid and flavonoid biosynthesis. The sensor can be used either for high-throughput pathway screening in synthetic biology applications or for introducing a feedback circuit to regulate production of the desired chemical. Here, we used the sensor to compare the performance of several predicted malonyl-CoA-producing pathways, and validated the utility of malonyl-CoA reductase and malonate-CoA transferase for malonyl-CoA biosynthesis. We generated a second-order dynamic linear model describing the relation of the fluorescence generated by the sensor to the biomass of the host cell representing a filter/amplifier with a gain that correlates with the level of induction. We found the time constants describing filter dynamics to be independent of the level of induction but distinctively clustered for each of the production pathways, indicating the robustness of the sensor. Moreover, by monitoring the effect of the copy-number of the production plasmid on the dose-response curve of the sensor, we managed to coarse-tune the level of pathway expression to maximize malonyl-CoA synthesis. In addition, we provide an example of the sensor's use in analyzing the effect of inducer or substrate concentrations on production levels. The rational development of models describing sensors, supplemented with the power of high-throughput optimization provide a promising potential for engineering feedback loops regulating enzyme levels to maximize productivity yields of synthetic metabolic pathways.ca
• dc.description.sponsorship PC is supported by Genopole through an ATIGE Grant by PRES UniverSud Paris, by Agence Nationale de la Recherche, and by UPFellows program with the support of the Marie Curie COFUND program. VL is supported by a DGA (French Ministry of Defense) graduate fellowship. The authors acknowledge funding provided by Toulouse White Biotech (TWB) and the GIP GENOPOLE.
• dc.format.mimetype application/pdfca
• dc.identifier.citation Fehér T, Libis V, Carbonell P, Faulon JL. A Sense of Balance: Experimental Investigation and Modeling of a Malonyl-CoA Sensor in Escherichia coli. Front Bioeng Biotechnol. 2015 Apr 8;(2)3:46. DOI: 10.3389/fbioe.2015.00046ca
• dc.identifier.doi http://dx.doi.org/10.3389/fbioe.2015.00046
• dc.identifier.issn 2296-4185
• dc.identifier.uri http://hdl.handle.net/10230/23785
• dc.language.iso engca
• dc.publisher Frontiersca
• dc.relation.ispartof Frontiers in Bioengineering and Biotechnology. 2015 Apr 8;(2)3:46
• dc.rights Copyright © 2015 Fehér, Libis, Carbonell and Faulon. This is an open-access article/ndistributed under the terms of the Creative Commons Attribution License (CC BY). https://creativecommons.org/licenses/by/4.0//nThe use, distribution or reproduction in other forums is permitted, provided the original/nauthor(s) or licensor are credited andthatthe original publication inthis journal is cited,/nin accordance with accepted academic practice. No use, distribution or reproduction is/npermitted which does not comply with these terms.ca
• dc.rights.accessRights info:eu-repo/semantics/openAccessca
• dc.rights.uri https://creativecommons.org/licenses/by/4.0/ca
• dc.subject.other Biologia de sistemesca
• dc.subject.other Sistemes biològicsca
• dc.title A Sense of Balance: Experimental Investigation and Modeling of a Malonyl-CoA Sensor in Escherichia colica
• dc.type info:eu-repo/semantics/articleca
• dc.type.version info:eu-repo/semantics/publishedVersionca