The effect of external oxygen conditions on the metabolic flux distribution of Saccharomyces cerevisiae

Paula Jouhten, Eija Rintala, Anne Huuskonen, Anu Tamminen, Merja Toivari, Marilyn Wiebe, A. Rantanen, Laura Ruohonen, Merja Penttilä, Hannu Maaheimo

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

Abstract

The yeast Saccharomyces cerevisiae is a facultative anaerobic organism able to switch the metabolism to fermentation when the aerobic respiration is limited or completely prevented. The availablity of oxygen is a major determinant of the metabolism of S. cerevisiae mainly because when the oxygen availability is restricted, cells need other acceptors to the electrons of NADH and FADH2. Furthermore, when the function of the respiratory system is limited additional means for ATP generation and cross-membrane transport of metabolites and ions are required. Growth under limited respiration thus requires major redistribution of metabolic fluxes compared to the aerobic metabolism. S. cerevisiae CEN.PK113-1A was grown in glucose-limited chemostat culture with 0%, 0.5%, 1.0%, 2.8% and 20.9% O2 in the inlet gas (D = 0.10 h-1, pH 5, 30 ?C) and labeled with 10% of [U-13C]-glucose to study the metabolic flux distribution response to oxygen. The labeled biomass was sampled for METAFoR (Metabolic Flux Ratio) analysis using 13C HSQC NMR spectroscopic detection (Szyperski et al., 1999). A stoichiometric model of the central carbon metabolism of S. cerevisiae was set up comprising of 29 reactions and transport fluxes and 20 metabolites in two intracellular compartments. Additional constraint equations for the stoichiometric model were obtained from the six metabolic flux ratios from METAFoR analysis (Fischer et al., 2004). The uptake and production rates were experimentally measured and the precursor requirements for biomass synthesis were taken from literature. The metabolic net fluxes were solved using an in-house program for weighted optimization based on the matlab function fmincon. A slight decrease in the oxygen uptake rate resulted in low ethanol production already in the 2.8% oxygen conditions. However, major changes in the intracellular metabolic flux distribution compared to aerobic conditions, in particular in the pyruvate branching point, were observed in the 1% oxygen conditions when the oxygen availability was severely reduced.
Original languageEnglish
Title of host publication3rd European Federation of Biotechnology Conference
Subtitle of host publicationPhysiology of Yeasts and Filamentous Fungi PYFF3
Place of PublicationEspoo
PublisherVTT Technical Research Centre of Finland
Pages117
ISBN (Electronic)978-951-38-6314-2
ISBN (Print)978-951-38-6313-5
Publication statusPublished - 2007
Event3rd European Federation of Biotechnology Conference : Physiology of Yeasts and Filamentous Fungi - Helsinki, Finland
Duration: 13 Jun 200716 Jun 2007

Publication series

NameVTT Symposium
PublisherVTT
Number245
ISSN (Print)0357-9387
ISSN (Electronic)1455-0873

Conference

Conference3rd European Federation of Biotechnology Conference
Abbreviated titlePYFF3
CountryFinland
CityHelsinki
Period13/06/0716/06/07

Fingerprint

Saccharomyces cerevisiae
oxygen
metabolism
metabolites
uptake mechanisms
glucose
metabolic studies
aerobiosis
biomass
aerobic conditions
ethanol production
cell respiration
respiratory system
branching
fermentation
electrons
gases
ions
yeasts
synthesis

Cite this

Jouhten, P., Rintala, E., Huuskonen, A., Tamminen, A., Toivari, M., Wiebe, M., ... Maaheimo, H. (2007). The effect of external oxygen conditions on the metabolic flux distribution of Saccharomyces cerevisiae. In 3rd European Federation of Biotechnology Conference : Physiology of Yeasts and Filamentous Fungi PYFF3 (pp. 117). [P63] Espoo: VTT Technical Research Centre of Finland. VTT Symposium, No. 245
Jouhten, Paula ; Rintala, Eija ; Huuskonen, Anne ; Tamminen, Anu ; Toivari, Merja ; Wiebe, Marilyn ; Rantanen, A. ; Ruohonen, Laura ; Penttilä, Merja ; Maaheimo, Hannu. / The effect of external oxygen conditions on the metabolic flux distribution of Saccharomyces cerevisiae. 3rd European Federation of Biotechnology Conference : Physiology of Yeasts and Filamentous Fungi PYFF3. Espoo : VTT Technical Research Centre of Finland, 2007. pp. 117 (VTT Symposium; No. 245).
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abstract = "The yeast Saccharomyces cerevisiae is a facultative anaerobic organism able to switch the metabolism to fermentation when the aerobic respiration is limited or completely prevented. The availablity of oxygen is a major determinant of the metabolism of S. cerevisiae mainly because when the oxygen availability is restricted, cells need other acceptors to the electrons of NADH and FADH2. Furthermore, when the function of the respiratory system is limited additional means for ATP generation and cross-membrane transport of metabolites and ions are required. Growth under limited respiration thus requires major redistribution of metabolic fluxes compared to the aerobic metabolism. S. cerevisiae CEN.PK113-1A was grown in glucose-limited chemostat culture with 0{\%}, 0.5{\%}, 1.0{\%}, 2.8{\%} and 20.9{\%} O2 in the inlet gas (D = 0.10 h-1, pH 5, 30 ?C) and labeled with 10{\%} of [U-13C]-glucose to study the metabolic flux distribution response to oxygen. The labeled biomass was sampled for METAFoR (Metabolic Flux Ratio) analysis using 13C HSQC NMR spectroscopic detection (Szyperski et al., 1999). A stoichiometric model of the central carbon metabolism of S. cerevisiae was set up comprising of 29 reactions and transport fluxes and 20 metabolites in two intracellular compartments. Additional constraint equations for the stoichiometric model were obtained from the six metabolic flux ratios from METAFoR analysis (Fischer et al., 2004). The uptake and production rates were experimentally measured and the precursor requirements for biomass synthesis were taken from literature. The metabolic net fluxes were solved using an in-house program for weighted optimization based on the matlab function fmincon. A slight decrease in the oxygen uptake rate resulted in low ethanol production already in the 2.8{\%} oxygen conditions. However, major changes in the intracellular metabolic flux distribution compared to aerobic conditions, in particular in the pyruvate branching point, were observed in the 1{\%} oxygen conditions when the oxygen availability was severely reduced.",
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Jouhten, P, Rintala, E, Huuskonen, A, Tamminen, A, Toivari, M, Wiebe, M, Rantanen, A, Ruohonen, L, Penttilä, M & Maaheimo, H 2007, The effect of external oxygen conditions on the metabolic flux distribution of Saccharomyces cerevisiae. in 3rd European Federation of Biotechnology Conference : Physiology of Yeasts and Filamentous Fungi PYFF3., P63, VTT Technical Research Centre of Finland, Espoo, VTT Symposium, no. 245, pp. 117, 3rd European Federation of Biotechnology Conference , Helsinki, Finland, 13/06/07.

The effect of external oxygen conditions on the metabolic flux distribution of Saccharomyces cerevisiae. / Jouhten, Paula; Rintala, Eija; Huuskonen, Anne; Tamminen, Anu; Toivari, Merja; Wiebe, Marilyn; Rantanen, A.; Ruohonen, Laura; Penttilä, Merja; Maaheimo, Hannu.

3rd European Federation of Biotechnology Conference : Physiology of Yeasts and Filamentous Fungi PYFF3. Espoo : VTT Technical Research Centre of Finland, 2007. p. 117 P63 (VTT Symposium; No. 245).

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

TY - CHAP

T1 - The effect of external oxygen conditions on the metabolic flux distribution of Saccharomyces cerevisiae

AU - Jouhten, Paula

AU - Rintala, Eija

AU - Huuskonen, Anne

AU - Tamminen, Anu

AU - Toivari, Merja

AU - Wiebe, Marilyn

AU - Rantanen, A.

AU - Ruohonen, Laura

AU - Penttilä, Merja

AU - Maaheimo, Hannu

PY - 2007

Y1 - 2007

N2 - The yeast Saccharomyces cerevisiae is a facultative anaerobic organism able to switch the metabolism to fermentation when the aerobic respiration is limited or completely prevented. The availablity of oxygen is a major determinant of the metabolism of S. cerevisiae mainly because when the oxygen availability is restricted, cells need other acceptors to the electrons of NADH and FADH2. Furthermore, when the function of the respiratory system is limited additional means for ATP generation and cross-membrane transport of metabolites and ions are required. Growth under limited respiration thus requires major redistribution of metabolic fluxes compared to the aerobic metabolism. S. cerevisiae CEN.PK113-1A was grown in glucose-limited chemostat culture with 0%, 0.5%, 1.0%, 2.8% and 20.9% O2 in the inlet gas (D = 0.10 h-1, pH 5, 30 ?C) and labeled with 10% of [U-13C]-glucose to study the metabolic flux distribution response to oxygen. The labeled biomass was sampled for METAFoR (Metabolic Flux Ratio) analysis using 13C HSQC NMR spectroscopic detection (Szyperski et al., 1999). A stoichiometric model of the central carbon metabolism of S. cerevisiae was set up comprising of 29 reactions and transport fluxes and 20 metabolites in two intracellular compartments. Additional constraint equations for the stoichiometric model were obtained from the six metabolic flux ratios from METAFoR analysis (Fischer et al., 2004). The uptake and production rates were experimentally measured and the precursor requirements for biomass synthesis were taken from literature. The metabolic net fluxes were solved using an in-house program for weighted optimization based on the matlab function fmincon. A slight decrease in the oxygen uptake rate resulted in low ethanol production already in the 2.8% oxygen conditions. However, major changes in the intracellular metabolic flux distribution compared to aerobic conditions, in particular in the pyruvate branching point, were observed in the 1% oxygen conditions when the oxygen availability was severely reduced.

AB - The yeast Saccharomyces cerevisiae is a facultative anaerobic organism able to switch the metabolism to fermentation when the aerobic respiration is limited or completely prevented. The availablity of oxygen is a major determinant of the metabolism of S. cerevisiae mainly because when the oxygen availability is restricted, cells need other acceptors to the electrons of NADH and FADH2. Furthermore, when the function of the respiratory system is limited additional means for ATP generation and cross-membrane transport of metabolites and ions are required. Growth under limited respiration thus requires major redistribution of metabolic fluxes compared to the aerobic metabolism. S. cerevisiae CEN.PK113-1A was grown in glucose-limited chemostat culture with 0%, 0.5%, 1.0%, 2.8% and 20.9% O2 in the inlet gas (D = 0.10 h-1, pH 5, 30 ?C) and labeled with 10% of [U-13C]-glucose to study the metabolic flux distribution response to oxygen. The labeled biomass was sampled for METAFoR (Metabolic Flux Ratio) analysis using 13C HSQC NMR spectroscopic detection (Szyperski et al., 1999). A stoichiometric model of the central carbon metabolism of S. cerevisiae was set up comprising of 29 reactions and transport fluxes and 20 metabolites in two intracellular compartments. Additional constraint equations for the stoichiometric model were obtained from the six metabolic flux ratios from METAFoR analysis (Fischer et al., 2004). The uptake and production rates were experimentally measured and the precursor requirements for biomass synthesis were taken from literature. The metabolic net fluxes were solved using an in-house program for weighted optimization based on the matlab function fmincon. A slight decrease in the oxygen uptake rate resulted in low ethanol production already in the 2.8% oxygen conditions. However, major changes in the intracellular metabolic flux distribution compared to aerobic conditions, in particular in the pyruvate branching point, were observed in the 1% oxygen conditions when the oxygen availability was severely reduced.

M3 - Conference abstract in proceedings

SN - 978-951-38-6313-5

T3 - VTT Symposium

SP - 117

BT - 3rd European Federation of Biotechnology Conference

PB - VTT Technical Research Centre of Finland

CY - Espoo

ER -

Jouhten P, Rintala E, Huuskonen A, Tamminen A, Toivari M, Wiebe M et al. The effect of external oxygen conditions on the metabolic flux distribution of Saccharomyces cerevisiae. In 3rd European Federation of Biotechnology Conference : Physiology of Yeasts and Filamentous Fungi PYFF3. Espoo: VTT Technical Research Centre of Finland. 2007. p. 117. P63. (VTT Symposium; No. 245).