Assessing glycolytic flux alterations resulting from genetic perturbations in E. coli using a biosensor

Christina Eva Lehning, Solvej Siedler, Mostafa M Hashim Ellabaan, Morten Otto Alexander Sommer

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Abstract

We describe the development of an optimized glycolytic flux biosensor and its application in detecting altered flux in a production strain and in a mutant library. The glycolytic flux biosensor is based on the Cra-regulated ppsA promoter of E. coli controlling fluorescent protein synthesis. We validated the glycolytic flux dependency of the biosensor in a range of different carbon sources in six different E. coli strains and during mevalonate production. Furthermore, we studied the flux-altering effects of genome-wide single gene knock-outs in E. coli in a multiplex FlowSeq experiment. From a library consisting of 2126 knock-out mutants, we identified 3 mutants with high-flux and 95 mutants with low-flux phenotypes that did not have severe growth defects. This approach can improve our understanding of glycolytic flux regulation improving metabolic models and engineering efforts.
Original languageEnglish
JournalMetabolic Engineering
Volume42
Pages (from-to)194-202
ISSN1096-7176
DOIs
Publication statusPublished - 2017

Bibliographical note

Open Access funded by European Research Council
Under a Creative Commons license

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