Measuring glycolytic flux in single yeast cells with an orthogonal synthetic biosensor release_jcud7hbqrrebnj5nfrsyijhqai

by Francisca Monteiro, Georg Hubmann, Vakil Takhaveev, Silke R Vedelaar, Justin Norder, Johan Hekelaar, Joana Saldida, Athanasios Litsios, Hein J Wijma, Alexander Schmidt, Matthias Heinemann

Published in Molecular Systems Biology by EMBO.

2019   Volume 15, Issue 12, e9071

Abstract

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.
In text/plain format

Archived Files and Locations

application/pdf  2.0 MB
file_opkvqoirl5ey5jam5xsj2phbhu
www.rug.nl (web)
web.archive.org (webarchive)
application/pdf  1.9 MB
file_hrlgtgvnvnf5tgu3ij4njyglye
lirias2repo.kuleuven.be (web)
web.archive.org (webarchive)
Read Archived PDF
Preserved and Accessible
Type  article-journal
Stage   published
Year   2019
Language   en ?
Container Metadata
Open Access Publication
In DOAJ
In ISSN ROAD
In Keepers Registry
ISSN-L:  1744-4292
Work Entity
access all versions, variants, and formats of this works (eg, pre-prints)
Catalog Record
Revision: 24185e3b-98e5-463d-9996-75ee87a8e9d5
API URL: JSON