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

Research output: Contribution to journalJournal articleResearchpeer-review

145 Downloads (Pure)

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

Cite this

@article{b3fa12443799477e8c82aff5ce21c5af,
title = "Assessing glycolytic flux alterations resulting from genetic perturbations in E. coli using a biosensor",
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.",
author = "Lehning, {Christina Eva} and Solvej Siedler and Ellabaan, {Mostafa M Hashim} and Sommer, {Morten Otto Alexander}",
note = "Open Access funded by European Research Council Under a Creative Commons license",
year = "2017",
doi = "10.1016/j.ymben.2017.07.002",
language = "English",
volume = "42",
pages = "194--202",
journal = "Metabolic Engineering",
issn = "1096-7176",
publisher = "Academic Press",

}

Assessing glycolytic flux alterations resulting from genetic perturbations in E. coli using a biosensor. / Lehning, Christina Eva; Siedler, Solvej; Ellabaan, Mostafa M Hashim; Sommer, Morten Otto Alexander.

In: Metabolic Engineering, Vol. 42, 2017, p. 194-202.

Research output: Contribution to journalJournal articleResearchpeer-review

TY - JOUR

T1 - Assessing glycolytic flux alterations resulting from genetic perturbations in E. coli using a biosensor

AU - Lehning, Christina Eva

AU - Siedler, Solvej

AU - Ellabaan, Mostafa M Hashim

AU - Sommer, Morten Otto Alexander

N1 - Open Access funded by European Research Council Under a Creative Commons license

PY - 2017

Y1 - 2017

N2 - 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.

AB - 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.

U2 - 10.1016/j.ymben.2017.07.002

DO - 10.1016/j.ymben.2017.07.002

M3 - Journal article

VL - 42

SP - 194

EP - 202

JO - Metabolic Engineering

JF - Metabolic Engineering

SN - 1096-7176

ER -