Journal article
Assessing glycolytic flux alterations resulting from genetic perturbations in E. coli using a biosensor
Novo Nordisk Foundation Center for Biosustainability, Technical University of Denmark1
Bacterial Synthetic Biology, Novo Nordisk Foundation Center for Biosustainability, Technical University of Denmark2
Research Groups, Novo Nordisk Foundation Center for Biosustainability, Technical University of Denmark3
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.
Language: | English |
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Publisher: | Academic Press |
Year: | 2017 |
Pages: | 194-202 |
ISSN: | 10967176 and 10967184 |
Types: | Journal article |
DOI: | 10.1016/j.ymben.2017.07.002 |
ORCIDs: | Lehning, Christina Eva , Siedler, Solvej , Ellabaan, Mostafa M Hashim and Sommer, Morten Otto Alexander |