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

2019-12Journal article Open access
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dc.contributor.authorMonteiro, Francisca
dc.contributor.authorHubmann, Georg
dc.contributor.authorTakhaveev, Vakil
dc.contributor.authorVedelaar, Silke R
dc.contributor.authorNorder, Justin
dc.contributor.authorHekelaar, Johan
dc.contributor.authorSaldida, Joana
dc.contributor.authorLitsios, Athanasios
dc.contributor.authorWijma, Hein J
dc.contributor.authorSchmidt, Alexander
dc.contributor.authorHeinemann, Matthias
dc.date.accessioned2024-01-26T14:27:00Z
dc.date.available2024-01-26T14:27:00Z
dc.date.issued2019-12
dc.description.abstractMetabolic 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.pt_PT
dc.description.versioninfo:eu-repo/semantics/publishedVersionpt_PT
dc.identifier.doi10.15252/msb.20199071pt_PT
dc.identifier.urihttp://hdl.handle.net/10451/62255
dc.language.isoengpt_PT
dc.peerreviewedyespt_PT
dc.publisherWileypt_PT
dc.relationEuropean Commission (EC) under grant agreement no. 613745, PROMYSpt_PT
dc.relationEuropean Commission (EC) under grant agreement no. 642738, MetaRNApt_PT
dc.relationEuropean Commission (EC) under grant agreement no. no 289995, ISOLATEpt_PT
dc.rights.urihttp://creativecommons.org/licenses/by/4.0/pt_PT
dc.titleMeasuring glycolytic flux in single yeast cells with an orthogonal synthetic biosensorpt_PT
dc.typejournal article
dspace.entity.typePublication
oaire.citation.issue12pt_PT
oaire.citation.titleMolecular Systems Biologypt_PT
oaire.citation.volume15pt_PT
person.familyNameSarreira Simões Horta Monteiro
person.givenNameFrancisca
person.identifier.ciencia-idA815-CE3A-53C9
person.identifier.orcid0000-0002-5275-714X
rcaap.rightsopenAccesspt_PT
rcaap.typearticlept_PT
relation.isAuthorOfPublication8063a7ff-bf07-4b7d-83a4-61786b528709
relation.isAuthorOfPublication.latestForDiscovery8063a7ff-bf07-4b7d-83a4-61786b528709

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