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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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Main Authors: Yasid, Nur Adeela, Rolfe, Matthew D., Green, Jeffrey, Williamson, Mike P.
Format: Article
Language:English
Published: The Royal Society Publishing 2016
Online Access:http://psasir.upm.edu.my/id/eprint/54446/1/Homeostasis%20of%20metabolites%20in%20Escherichia%20coli.pdf
http://psasir.upm.edu.my/id/eprint/54446/
http://rsos.royalsocietypublishing.org/content/3/8/160187
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spelling my.upm.eprints.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 10.1098/rsos.160187
institution Universiti Putra Malaysia
building UPM Library
collection Institutional Repository
continent Asia
country Malaysia
content_provider Universiti Putra Malaysia
content_source UPM Institutional Repository
url_provider http://psasir.upm.edu.my/
language English
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.
format Article
author Yasid, Nur Adeela
Rolfe, Matthew D.
Green, Jeffrey
Williamson, Mike P.
spellingShingle 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
author_facet Yasid, Nur Adeela
Rolfe, Matthew D.
Green, Jeffrey
Williamson, Mike P.
author_sort Yasid, Nur Adeela
title 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_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_sort homeostasis of metabolites in escherichia coli on transition from anaerobic to aerobic conditions and the transient secretion of pyruvate
publisher The Royal Society Publishing
publishDate 2016
url http://psasir.upm.edu.my/id/eprint/54446/1/Homeostasis%20of%20metabolites%20in%20Escherichia%20coli.pdf
http://psasir.upm.edu.my/id/eprint/54446/
http://rsos.royalsocietypublishing.org/content/3/8/160187
_version_ 1643835638571597824
score 13.211869

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