Open AccessMeasuring glycolytic flux in single yeast cells with an orthogonal synthetic biosensor
Author(s) -
Monteiro Francisca,
Hubmann Georg,
Takhaveev Vakil,
Vedelaar Silke R,
Norder Justin,
Hekelaar Johan,
Saldida Joana,
Litsios Athanasios,
Wijma Hein J,
Schmidt Alexander,
Heinemann Matthias
Publication year - 2019
Publication title -
molecular systems biology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 8.523
H-Index - 148
ISSN - 1744-4292
DOI - 10.15252/msb.20199071
Subject(s) - biological sciences , library science , biology , computational biology , computer science
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.