Measuring glycolytic flux in single yeast cells with an orthogonal synthetic biosensor

Monteiro, Francisca and Hubmann, Georg and Takhaveev, Vakil and Vedelaar, Silke R. and Norder, Justin and Hekelaar, Johan and Saldida, Joana and Litsios, Athanasios and Wijma, Hein J. and Schmidt, Alexander and Heinemann, Matthias. (2019) Measuring glycolytic flux in single yeast cells with an orthogonal synthetic biosensor. Molecular systems biology, 15 (12). e9071.

Full text not available from this repository.

Official URL: https://edoc.unibas.ch/75071/

Downloads: Statistics Overview


Metabolic heterogeneity between individual cells of a population harbors significant challenges for fundamental and applied research. Identifying metabolic heterogeneity and investigating its emergence require tools to zoom into metabolism of individual cells. While methods exist to measure metabolite levels in single cells, we lack capability to measure metabolic flux, i.e., the ultimate functional output of metabolic activity, on the single-cell level. Here, combining promoter engineering, computational protein design, biochemical methods, proteomics, and metabolomics, we developed a biosensor to measure glycolytic flux in single yeast cells. Therefore, drawing on the robust cell-intrinsic correlation between glycolytic flux and levels of fructose-1,6-bisphosphate (FBP), we transplanted the B. subtilis FBP-binding transcription factor CggR into yeast. With the developed biosensor, we robustly identified cell subpopulations with different FBP levels in mixed cultures, when subjected to flow cytometry and microscopy. Employing microfluidics, we were also able to assess the temporal FBP/glycolytic flux dynamics during the cell cycle. We anticipate that our biosensor will become a valuable tool to identify and study metabolic heterogeneity in cell populations.
Faculties and Departments:05 Faculty of Science > Departement Biozentrum > Services Biozentrum > Proteomics (Schmidt)
UniBasel Contributors:Schmidt, Alexander
Item Type:Article, refereed
Article Subtype:Research Article
Note:Publication type according to Uni Basel Research Database: Journal article
Identification Number:
Last Modified:05 Apr 2020 17:14
Deposited On:05 Apr 2020 17:14

Repository Staff Only: item control page