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Dynamic in vivo (31)P nuclear magnetic resonance study of Saccharomyces cerevisiae in glucose-limited chemostat culture during the aerobic-anaerobic shift.
Yeast. 2000 Apr; 16(6):483-97.Y

Abstract

The purpose of this work was to analyse in vivo the influence of sudden oxygen depletion on Saccharomyces cerevisiae, grown in glucose-limited chemostat culture, using a recently developed cyclone reactor coupled with (31)P NMR spectroscopy. Before, during and after the transition, intracellular and extracellular phosphorylated metabolites as well as the pHs in the different cellular compartments were monitored with a time resolution of 2.5 min. The employed integrated NMR bioreactor system allowed the defined glucose-limited continuous cultivation of yeast at a density of 75 g DW/l and a p(O(2)) of 30% air saturation. A purely oxidative metabolism was maintained at all times. In vivo (31)P NMR spectra obtained were of excellent quality and even allowed the detection of phosphoenolpyruvate (PEP). During the switch from aerobic to anaerobic conditions, a rapid, significant decrease of intracellular ATP and PEP levels was observed and the cytoplasmic pH decreased from 7.5 to 6.8. This change, which was accompanied by a transient influx of extracellular inorganic phosphate (P(i)), appeared to correlate linearly with the decrease of the ATP concentration, suggesting that the cause of the partial collapse of the plasma membrane pH gradient was a reduced availability of ATP. The complete phosphorous balance established from our measurement data showed that polyphosphate was not the source of the increased intracellular P(i). The derived intracellular P(i), ATP and ADP concentration data confirmed that the glycolytic flux at the level of glyceraldehyde-3-phosphate dehydrogenase, 3-phosphoglycerate kinase and enolase enzymes is mainly controlled by thermodynamic constraints.

Authors+Show Affiliations

Laboratoire de Technologie de la Nutrition et de l'Alimentation, Centre de Recherche en Nutrition Humaine de Clermont-Ferrand, France.No affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article

Language

eng

PubMed ID

10790685

Citation

Gonzalez, B, et al. "Dynamic in Vivo (31)P Nuclear Magnetic Resonance Study of Saccharomyces Cerevisiae in Glucose-limited Chemostat Culture During the Aerobic-anaerobic Shift." Yeast (Chichester, England), vol. 16, no. 6, 2000, pp. 483-97.
Gonzalez B, de Graaf A, Renaud M, et al. Dynamic in vivo (31)P nuclear magnetic resonance study of Saccharomyces cerevisiae in glucose-limited chemostat culture during the aerobic-anaerobic shift. Yeast. 2000;16(6):483-97.
Gonzalez, B., de Graaf, A., Renaud, M., & Sahm, H. (2000). Dynamic in vivo (31)P nuclear magnetic resonance study of Saccharomyces cerevisiae in glucose-limited chemostat culture during the aerobic-anaerobic shift. Yeast (Chichester, England), 16(6), 483-97.
Gonzalez B, et al. Dynamic in Vivo (31)P Nuclear Magnetic Resonance Study of Saccharomyces Cerevisiae in Glucose-limited Chemostat Culture During the Aerobic-anaerobic Shift. Yeast. 2000;16(6):483-97. PubMed PMID: 10790685.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Dynamic in vivo (31)P nuclear magnetic resonance study of Saccharomyces cerevisiae in glucose-limited chemostat culture during the aerobic-anaerobic shift. AU - Gonzalez,B, AU - de Graaf,A, AU - Renaud,M, AU - Sahm,H, PY - 2000/5/3/pubmed PY - 2000/6/10/medline PY - 2000/5/3/entrez SP - 483 EP - 97 JF - Yeast (Chichester, England) JO - Yeast VL - 16 IS - 6 N2 - The purpose of this work was to analyse in vivo the influence of sudden oxygen depletion on Saccharomyces cerevisiae, grown in glucose-limited chemostat culture, using a recently developed cyclone reactor coupled with (31)P NMR spectroscopy. Before, during and after the transition, intracellular and extracellular phosphorylated metabolites as well as the pHs in the different cellular compartments were monitored with a time resolution of 2.5 min. The employed integrated NMR bioreactor system allowed the defined glucose-limited continuous cultivation of yeast at a density of 75 g DW/l and a p(O(2)) of 30% air saturation. A purely oxidative metabolism was maintained at all times. In vivo (31)P NMR spectra obtained were of excellent quality and even allowed the detection of phosphoenolpyruvate (PEP). During the switch from aerobic to anaerobic conditions, a rapid, significant decrease of intracellular ATP and PEP levels was observed and the cytoplasmic pH decreased from 7.5 to 6.8. This change, which was accompanied by a transient influx of extracellular inorganic phosphate (P(i)), appeared to correlate linearly with the decrease of the ATP concentration, suggesting that the cause of the partial collapse of the plasma membrane pH gradient was a reduced availability of ATP. The complete phosphorous balance established from our measurement data showed that polyphosphate was not the source of the increased intracellular P(i). The derived intracellular P(i), ATP and ADP concentration data confirmed that the glycolytic flux at the level of glyceraldehyde-3-phosphate dehydrogenase, 3-phosphoglycerate kinase and enolase enzymes is mainly controlled by thermodynamic constraints. SN - 0749-503X UR - https://www.unboundmedicine.com/medline/citation/10790685/Dynamic_in_vivo__31_P_nuclear_magnetic_resonance_study_of_Saccharomyces_cerevisiae_in_glucose_limited_chemostat_culture_during_the_aerobic_anaerobic_shift_ L2 - https://doi.org/10.1002/(SICI)1097-0061(200004)16:6<483::AID-YEA542>3.0.CO;2-E DB - PRIME DP - Unbound Medicine ER -