The effect of oxygen level on metabolic flux distribution of Saccharomyces cerevisoae

Research output: Chapter in Book/Report/Conference proceedingConference abstract in proceedingsScientific

Abstract

The metabolic flux responses of the yeast Saccharomyces cerevisiae to different oxygen levels were studied by growing the yeast in chemostat conditions under five different oxygen levels, corresponding to 21 (fully aerobic), 2.5, 1.0, 0.5 and 0% of the influx gas of the fermentor. The growth rate was maintained at 0.10 h-1. Intra-cellular metabolic flux ratios of the central carbon metabolism were determined by 13C tracer experiments. Approximately 10% of the carbon source glucose was replaced by its uniformly 13C labelled variant for 1.5 residence times and the flux ratios were determined from the fine structure of the 13C NMR signals of the proteinogenic amino acids in [1H, 13C] HSQC spectra. The distribution of metabolic net fluxes was estimated by 13C MFA, i.e. metabolic flux analysis with the flux ratios of the 13C tracer experiments as additional constraints. The metabolic reaction network used in the analysis consists of 25 metabolites and 28 reactions. In addition to the fluxes to the biomass, four extra-cellular fluxes and six intra-cellular flux ratios from the 13C tracer experiments were used in the flux balancing. Due to the additional constraints from the flux ratio analysis, it was not necessary to include the co-factors in the flux balancing. The result from both the flux ratio analysis and from the 13C-MFA showed that the most dramatic changes in the flux distribution take place at low oxygen levels, while the flux ratios of the 2.5% oxygen cultivation resembled the fully aerobic case. However, the fraction of glucose processed through the pentose phosphate pathway responded to the oxygen level already at high oxygen levels, while the relative anaplerotic flux to the TCA cycle was virtually unaffected until very low oxygen levels are reached.
Original languageEnglish
Title of host publicationInternational Specialised Symposium on Yeasts ISSY25
Subtitle of host publicationSystems Biology of Yeasts - from Models to Applications
PublisherVTT Technical Research Centre of Finland
Pages79
ISBN (Electronic) 951-38-6308-5
ISBN (Print)951-38-6307-7
Publication statusPublished - 2006
EventInternational Specialised Symposium on Yeasts, ISSY 25 - Espoo, Finland
Duration: 18 Jun 200621 Jun 2006

Publication series

SeriesVTT Symposium
Number242
ISSN0357-9387

Conference

ConferenceInternational Specialised Symposium on Yeasts, ISSY 25
Abbreviated titleISSY 25
CountryFinland
CityEspoo
Period18/06/0621/06/06

Fingerprint

oxygen
tracer
yeast
glucose
chemostat
experiment
carbon
distribution
effect
nuclear magnetic resonance
residence time
metabolite
amino acid
metabolism
phosphate
analysis
biomass
gas

Cite this

Jouhten, P., Rintala, E., Tamminen, A., Huuskonen, A., Wiebe, M., Salusjärvi, L., ... Maaheimo, H. (2006). The effect of oxygen level on metabolic flux distribution of Saccharomyces cerevisoae. In International Specialised Symposium on Yeasts ISSY25: Systems Biology of Yeasts - from Models to Applications (pp. 79). [P27] VTT Technical Research Centre of Finland. VTT Symposium, No. 242
Jouhten, Paula ; Rintala, Eija ; Tamminen, Anu ; Huuskonen, Anne ; Wiebe, Marilyn ; Salusjärvi, Laura ; Toivari, Mervi ; Ruohonen, Laura ; Penttilä, Merja ; Maaheimo, Hannu. / The effect of oxygen level on metabolic flux distribution of Saccharomyces cerevisoae. International Specialised Symposium on Yeasts ISSY25: Systems Biology of Yeasts - from Models to Applications. VTT Technical Research Centre of Finland, 2006. pp. 79 (VTT Symposium; No. 242).
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title = "The effect of oxygen level on metabolic flux distribution of Saccharomyces cerevisoae",
abstract = "The metabolic flux responses of the yeast Saccharomyces cerevisiae to different oxygen levels were studied by growing the yeast in chemostat conditions under five different oxygen levels, corresponding to 21 (fully aerobic), 2.5, 1.0, 0.5 and 0{\%} of the influx gas of the fermentor. The growth rate was maintained at 0.10 h-1. Intra-cellular metabolic flux ratios of the central carbon metabolism were determined by 13C tracer experiments. Approximately 10{\%} of the carbon source glucose was replaced by its uniformly 13C labelled variant for 1.5 residence times and the flux ratios were determined from the fine structure of the 13C NMR signals of the proteinogenic amino acids in [1H, 13C] HSQC spectra. The distribution of metabolic net fluxes was estimated by 13C MFA, i.e. metabolic flux analysis with the flux ratios of the 13C tracer experiments as additional constraints. The metabolic reaction network used in the analysis consists of 25 metabolites and 28 reactions. In addition to the fluxes to the biomass, four extra-cellular fluxes and six intra-cellular flux ratios from the 13C tracer experiments were used in the flux balancing. Due to the additional constraints from the flux ratio analysis, it was not necessary to include the co-factors in the flux balancing. The result from both the flux ratio analysis and from the 13C-MFA showed that the most dramatic changes in the flux distribution take place at low oxygen levels, while the flux ratios of the 2.5{\%} oxygen cultivation resembled the fully aerobic case. However, the fraction of glucose processed through the pentose phosphate pathway responded to the oxygen level already at high oxygen levels, while the relative anaplerotic flux to the TCA cycle was virtually unaffected until very low oxygen levels are reached.",
author = "Paula Jouhten and Eija Rintala and Anu Tamminen and Anne Huuskonen and Marilyn Wiebe and Laura Salusj{\"a}rvi and Mervi Toivari and Laura Ruohonen and Merja Penttil{\"a} and Hannu Maaheimo",
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Jouhten, P, Rintala, E, Tamminen, A, Huuskonen, A, Wiebe, M, Salusjärvi, L, Toivari, M, Ruohonen, L, Penttilä, M & Maaheimo, H 2006, The effect of oxygen level on metabolic flux distribution of Saccharomyces cerevisoae. in International Specialised Symposium on Yeasts ISSY25: Systems Biology of Yeasts - from Models to Applications., P27, VTT Technical Research Centre of Finland, VTT Symposium, no. 242, pp. 79, International Specialised Symposium on Yeasts, ISSY 25 , Espoo, Finland, 18/06/06.

The effect of oxygen level on metabolic flux distribution of Saccharomyces cerevisoae. / Jouhten, Paula; Rintala, Eija; Tamminen, Anu; Huuskonen, Anne; Wiebe, Marilyn; Salusjärvi, Laura; Toivari, Mervi; Ruohonen, Laura; Penttilä, Merja; Maaheimo, Hannu.

International Specialised Symposium on Yeasts ISSY25: Systems Biology of Yeasts - from Models to Applications. VTT Technical Research Centre of Finland, 2006. p. 79 P27 (VTT Symposium; No. 242).

Research output: Chapter in Book/Report/Conference proceedingConference abstract in proceedingsScientific

TY - CHAP

T1 - The effect of oxygen level on metabolic flux distribution of Saccharomyces cerevisoae

AU - Jouhten, Paula

AU - Rintala, Eija

AU - Tamminen, Anu

AU - Huuskonen, Anne

AU - Wiebe, Marilyn

AU - Salusjärvi, Laura

AU - Toivari, Mervi

AU - Ruohonen, Laura

AU - Penttilä, Merja

AU - Maaheimo, Hannu

PY - 2006

Y1 - 2006

N2 - The metabolic flux responses of the yeast Saccharomyces cerevisiae to different oxygen levels were studied by growing the yeast in chemostat conditions under five different oxygen levels, corresponding to 21 (fully aerobic), 2.5, 1.0, 0.5 and 0% of the influx gas of the fermentor. The growth rate was maintained at 0.10 h-1. Intra-cellular metabolic flux ratios of the central carbon metabolism were determined by 13C tracer experiments. Approximately 10% of the carbon source glucose was replaced by its uniformly 13C labelled variant for 1.5 residence times and the flux ratios were determined from the fine structure of the 13C NMR signals of the proteinogenic amino acids in [1H, 13C] HSQC spectra. The distribution of metabolic net fluxes was estimated by 13C MFA, i.e. metabolic flux analysis with the flux ratios of the 13C tracer experiments as additional constraints. The metabolic reaction network used in the analysis consists of 25 metabolites and 28 reactions. In addition to the fluxes to the biomass, four extra-cellular fluxes and six intra-cellular flux ratios from the 13C tracer experiments were used in the flux balancing. Due to the additional constraints from the flux ratio analysis, it was not necessary to include the co-factors in the flux balancing. The result from both the flux ratio analysis and from the 13C-MFA showed that the most dramatic changes in the flux distribution take place at low oxygen levels, while the flux ratios of the 2.5% oxygen cultivation resembled the fully aerobic case. However, the fraction of glucose processed through the pentose phosphate pathway responded to the oxygen level already at high oxygen levels, while the relative anaplerotic flux to the TCA cycle was virtually unaffected until very low oxygen levels are reached.

AB - The metabolic flux responses of the yeast Saccharomyces cerevisiae to different oxygen levels were studied by growing the yeast in chemostat conditions under five different oxygen levels, corresponding to 21 (fully aerobic), 2.5, 1.0, 0.5 and 0% of the influx gas of the fermentor. The growth rate was maintained at 0.10 h-1. Intra-cellular metabolic flux ratios of the central carbon metabolism were determined by 13C tracer experiments. Approximately 10% of the carbon source glucose was replaced by its uniformly 13C labelled variant for 1.5 residence times and the flux ratios were determined from the fine structure of the 13C NMR signals of the proteinogenic amino acids in [1H, 13C] HSQC spectra. The distribution of metabolic net fluxes was estimated by 13C MFA, i.e. metabolic flux analysis with the flux ratios of the 13C tracer experiments as additional constraints. The metabolic reaction network used in the analysis consists of 25 metabolites and 28 reactions. In addition to the fluxes to the biomass, four extra-cellular fluxes and six intra-cellular flux ratios from the 13C tracer experiments were used in the flux balancing. Due to the additional constraints from the flux ratio analysis, it was not necessary to include the co-factors in the flux balancing. The result from both the flux ratio analysis and from the 13C-MFA showed that the most dramatic changes in the flux distribution take place at low oxygen levels, while the flux ratios of the 2.5% oxygen cultivation resembled the fully aerobic case. However, the fraction of glucose processed through the pentose phosphate pathway responded to the oxygen level already at high oxygen levels, while the relative anaplerotic flux to the TCA cycle was virtually unaffected until very low oxygen levels are reached.

M3 - Conference abstract in proceedings

SN - 951-38-6307-7

T3 - VTT Symposium

SP - 79

BT - International Specialised Symposium on Yeasts ISSY25

PB - VTT Technical Research Centre of Finland

ER -

Jouhten P, Rintala E, Tamminen A, Huuskonen A, Wiebe M, Salusjärvi L et al. The effect of oxygen level on metabolic flux distribution of Saccharomyces cerevisoae. In International Specialised Symposium on Yeasts ISSY25: Systems Biology of Yeasts - from Models to Applications. VTT Technical Research Centre of Finland. 2006. p. 79. P27. (VTT Symposium; No. 242).