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dc.contributor.authorDoong, Stephanie
dc.contributor.authorGupta, Apoorv
dc.contributor.authorJones, Kristala L.
dc.date.accessioned2018-12-04T17:55:18Z
dc.date.available2018-12-04T17:55:18Z
dc.date.issued2018-03
dc.date.submitted2017-09
dc.identifier.issn0027-8424
dc.identifier.issn1091-6490
dc.identifier.urihttp://hdl.handle.net/1721.1/119416
dc.description.abstractMicrobial production of value-added chemicals from biomass is a sustainable alternative to chemical synthesis. To improve product titer, yield, and selectivity, the pathways engineered into microbes must be optimized. One strategy for optimization is dynamic pathway regulation, which modulates expression of pathwayrelevant enzymes over the course of fermentation. Metabolic engineers have used dynamic regulation to redirect endogenous flux toward product formation, balance the production and consumption rates of key intermediates, and suppress production of toxic intermediates until later in the fermentation. Most cases, however, have utilized a single strategy for dynamically regulating pathway fluxes. Here we layer two orthogonal, autonomous, and tunable dynamic regulation strategies to independently modulate expression of two different enzymes to improve production of D-glucaric acid from a heterologous pathway. The first strategy uses a previously described pathway-independent quorum sensing system to dynamically knock down glycolytic flux and redirect carbon into production of glucaric acid, thereby switching cells from "growth" to "production" mode. The second strategy, developed in this work, uses a biosensor for myo-inositol (MI), an intermediate in the glucaric acid production pathway, to induce expression of a downstream enzyme upon sufficient buildup of MI. The latter, pathway-dependent strategy leads to a 2.5-fold increase in titer when used in isolation and a fourfold increase when added to a strain employing the former, pathway-independent regulatory system. The dual-regulation strain produces nearly 2 g/L glucaric acid, representing the highest glucaric acid titer reported to date in Escherichia coli K-12 strains.en_US
dc.description.sponsorshipNational Science Foundation (U.S.). Graduate Research Fellowship Programen_US
dc.description.sponsorshipSynthetic Biology Engineering Research Center (Grant EEC-0540879)en_US
dc.description.sponsorshipNational Institute of General Medical Sciences (U.S.) Biotechnology Training Program (Grant T32GM008334)en_US
dc.description.sponsorshipNational Institute of General Medical Sciences (U.S.). Division of Molecular and Cellular Biosciences (Grant MCB-1517913)en_US
dc.publisherProceedings of the National Academy of Sciencesen_US
dc.relation.isversionofhttp://dx.doi.org/10.1073/PNAS.1716920115en_US
dc.rightsArticle is made available in accordance with the publisher's policy and may be subject to US copyright law. Please refer to the publisher's site for terms of use.en_US
dc.sourcePNASen_US
dc.titleLayered dynamic regulation for improving metabolic pathway productivity in Escherichia colien_US
dc.typeArticleen_US
dc.identifier.citationDoong, Stephanie J., Apoorv Gupta, and Kristala L. J. Prather. “Layered Dynamic Regulation for Improving Metabolic Pathway Productivity in Escherichia Coli.” Proceedings of the National Academy of Sciences 115, no. 12 (March 5, 2018): 2964–2969. © 2018 National Academy of Sciences.en_US
dc.contributor.departmentMassachusetts Institute of Technology. Department of Biological Engineeringen_US
dc.contributor.departmentMassachusetts Institute of Technology. Department of Chemical Engineeringen_US
dc.contributor.departmentMassachusetts Institute of Technology. Synthetic Biology Centeren_US
dc.contributor.mitauthorDoong, Stephanie
dc.contributor.mitauthorGupta, Apoorv
dc.contributor.mitauthorJones, Kristala L.
dc.relation.journalProceedings of the National Academy of Sciencesen_US
dc.eprint.versionFinal published versionen_US
dc.type.urihttp://purl.org/eprint/type/JournalArticleen_US
eprint.statushttp://purl.org/eprint/status/PeerRevieweden_US
dc.date.updated2018-12-03T21:11:01Z
dspace.orderedauthorsDoong, Stephanie J.; Gupta, Apoorv; Prather, Kristala L. J.en_US
dspace.embargo.termsNen_US
dc.identifier.orcidhttps://orcid.org/0000-0001-8089-2034
dc.identifier.orcidhttps://orcid.org/0000-0002-6548-9420
dc.identifier.orcidhttps://orcid.org/0000-0003-0437-3157
mit.licensePUBLISHER_POLICYen_US


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