Homeostasis of metabolites in Escherichia coli on transition from anaerobic to aerobic conditions and the transient secretion of pyruvate
We have developed a method for rapid quenching of samples taken from chemostat cultures of Escherichia coli that gives reproducible and reliable measurements of extracellular and intracellular metabolites by 1H NMR and have applied it to study the major central metabolites during the transition from...
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| Format: | Article |
| Language: | English |
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The Royal Society Publishing
2016
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| Online Access: | http://psasir.upm.edu.my/id/eprint/54446/ http://psasir.upm.edu.my/id/eprint/54446/1/Homeostasis%20of%20metabolites%20in%20Escherichia%20coli.pdf |
| _version_ | 1848852546901245952 |
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| author | Yasid, Nur Adeela Rolfe, Matthew D. Green, Jeffrey Williamson, Mike P. |
| author_facet | Yasid, Nur Adeela Rolfe, Matthew D. Green, Jeffrey Williamson, Mike P. |
| author_sort | Yasid, Nur Adeela |
| building | UPM Institutional Repository |
| collection | Online Access |
| description | We have developed a method for rapid quenching of samples taken from chemostat cultures of Escherichia coli that gives reproducible and reliable measurements of extracellular and intracellular metabolites by 1H NMR and have applied it to study the major central metabolites during the transition from anaerobic to aerobic growth. Almost all metabolites showed a gradual change after perturbation with air, consistent with immediate inhibition of pyruvate formate-lyase, dilution of overflow metabolites and induction of aerobic enzymes. Surprisingly, although pyruvate showed almost no change in intracellular concentration, the extracellular concentration transiently increased. The absence of intracellular accumulation of pyruvate suggested that one or more glycolytic enzymes might relocate to the cell membrane. To test this hypothesis, chromosomal pyruvate kinase (pykF) was modified to express either PykF-green fluorescent protein or PykF-FLAG fusion proteins. Measurements showed that PykF-FLAG relocates to the cell membrane within 5 min of aeration and then slowly returns to the cytoplasm, suggesting that on aeration, PykF associates with the membrane to facilitate secretion of pyruvate to maintain constant intracellular levels. |
| first_indexed | 2025-11-15T10:39:48Z |
| format | Article |
| id | upm-54446 |
| institution | Universiti Putra Malaysia |
| institution_category | Local University |
| language | English |
| last_indexed | 2025-11-15T10:39:48Z |
| publishDate | 2016 |
| publisher | The Royal Society Publishing |
| recordtype | eprints |
| repository_type | Digital Repository |
| spelling | upm-544462018-03-16T09:00:10Z http://psasir.upm.edu.my/id/eprint/54446/ Homeostasis of metabolites in Escherichia coli on transition from anaerobic to aerobic conditions and the transient secretion of pyruvate Yasid, Nur Adeela Rolfe, Matthew D. Green, Jeffrey Williamson, Mike P. We have developed a method for rapid quenching of samples taken from chemostat cultures of Escherichia coli that gives reproducible and reliable measurements of extracellular and intracellular metabolites by 1H NMR and have applied it to study the major central metabolites during the transition from anaerobic to aerobic growth. Almost all metabolites showed a gradual change after perturbation with air, consistent with immediate inhibition of pyruvate formate-lyase, dilution of overflow metabolites and induction of aerobic enzymes. Surprisingly, although pyruvate showed almost no change in intracellular concentration, the extracellular concentration transiently increased. The absence of intracellular accumulation of pyruvate suggested that one or more glycolytic enzymes might relocate to the cell membrane. To test this hypothesis, chromosomal pyruvate kinase (pykF) was modified to express either PykF-green fluorescent protein or PykF-FLAG fusion proteins. Measurements showed that PykF-FLAG relocates to the cell membrane within 5 min of aeration and then slowly returns to the cytoplasm, suggesting that on aeration, PykF associates with the membrane to facilitate secretion of pyruvate to maintain constant intracellular levels. The Royal Society Publishing 2016-08 Article PeerReviewed text en http://psasir.upm.edu.my/id/eprint/54446/1/Homeostasis%20of%20metabolites%20in%20Escherichia%20coli.pdf Yasid, Nur Adeela and Rolfe, Matthew D. and Green, Jeffrey and Williamson, Mike P. (2016) Homeostasis of metabolites in Escherichia coli on transition from anaerobic to aerobic conditions and the transient secretion of pyruvate. Royal Society Open Science, 3 (8). pp. 1-12. ISSN 2054-5703 http://rsos.royalsocietypublishing.org/content/3/8/160187 Escherichia coli; Metabolomics; Metabolon; Glycolysis; Nuclear magnetic resonance 10.1098/rsos.160187 |
| spellingShingle | Escherichia coli; Metabolomics; Metabolon; Glycolysis; Nuclear magnetic resonance Yasid, Nur Adeela Rolfe, Matthew D. Green, Jeffrey Williamson, Mike P. Homeostasis of metabolites in Escherichia coli on transition from anaerobic to aerobic conditions and the transient secretion of pyruvate |
| title | Homeostasis of metabolites in Escherichia coli on transition from anaerobic to aerobic conditions and the transient secretion of pyruvate |
| title_full | Homeostasis of metabolites in Escherichia coli on transition from anaerobic to aerobic conditions and the transient secretion of pyruvate |
| title_fullStr | Homeostasis of metabolites in Escherichia coli on transition from anaerobic to aerobic conditions and the transient secretion of pyruvate |
| title_full_unstemmed | Homeostasis of metabolites in Escherichia coli on transition from anaerobic to aerobic conditions and the transient secretion of pyruvate |
| title_short | Homeostasis of metabolites in Escherichia coli on transition from anaerobic to aerobic conditions and the transient secretion of pyruvate |
| title_sort | homeostasis of metabolites in escherichia coli on transition from anaerobic to aerobic conditions and the transient secretion of pyruvate |
| topic | Escherichia coli; Metabolomics; Metabolon; Glycolysis; Nuclear magnetic resonance |
| url | http://psasir.upm.edu.my/id/eprint/54446/ http://psasir.upm.edu.my/id/eprint/54446/ http://psasir.upm.edu.my/id/eprint/54446/ http://psasir.upm.edu.my/id/eprint/54446/1/Homeostasis%20of%20metabolites%20in%20Escherichia%20coli.pdf |