Integrative approaches for studying pentose metabolism in Saccharomyces cerevisiae

Laura Ruohonen, Laura Salusjärvi, Matej Oresic, Hannu Maaheimo, Eija Rintala, Helena Simolin, Merja Penttilä

Research output: Contribution to conferenceConference articleScientific

Abstract

We are interested in broadening the substrate utilisation range of the yeast Saccharomyces cerevisiae and in connection to this to study nutritional responses and signalling at a global level. S.cerevisiae does not naturally utilise pentose sugars unlike most other fungi, and recombinant S.cerevisiae strains have been constructed that contain the xylose utilisation pathway from other yeasts. The recombinant xylose pathway is believed to create a redox imbalance in the cells, due to different cofactor requirements of the enzymes of the pathway, and limit xylose fermentation. In order to understand redox and nutrient regulation in general, and the physiology of xylose utilising S.cerevisiae, we carried out chemostat and batch cultures at various oxygenation levels on xylose as a carbon source, and compared those with glucose cultures. Transcriptional profiling, total proteomics and metabolite analyses were carried out, as well as metabolix flux analysis (MFA) and measurements of metabolic fluxes with 13C-NMR. New responses towards xylose as a carbon source were discovered such as the upregulation of pathways for alternative carbon source utilisation and responses for nutritional control and starvation. The physiology of the recombinant yeast appears to be neither fully repressed (fermentative) nor derepressed (gluconeogenic). Own references [1] H. Maaheimo, J. Fiaux, Z. P. Cakar, J. E. Bailey, U. Sauer, T. Szyperski, Eur J Biochem. 268, 2464-2479 (2001). [2] J. P. Pitkänen, A. Aristidou, L. Salusjärvi, L. Ruohonen, M. Penttilä, Metab Eng. 5, 16-31 (2003). [3] L. Salusjärvi, M. Poutanen, J. P. Pitkänen, H. Koivistoinen, A. Aristidou, N. Kalkkinen, L. Ruohonen, M. Penttilä, Yeast 20, 295-314 (2003). [4] Salusjärvi, L., Pitkänen, J.-P., Aristidou, A., Ruohonen, L. and Penttilä, M. (2003a) (in preparation). [5] Toivari, M.H., Aristidou, A., Ruohonen, L. & Penttilä, M. 2001. Metab. Eng. 3, 236-249.
Original languageEnglish
Publication statusPublished - 2004
EventMetabolic Engineering V: Genome to Product - Lake Tahoe, United States
Duration: 19 Sep 200423 Sep 2004

Conference

ConferenceMetabolic Engineering V
CountryUnited States
CityLake Tahoe
Period19/09/0423/09/04

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pentoses
xylose
Saccharomyces cerevisiae
metabolism
yeasts
carbon
physiology
proteomics
starvation
fermentation
metabolites
sugars
glucose
fungi
nutrients
enzymes
cells

Cite this

Ruohonen, L., Salusjärvi, L., Oresic, M., Maaheimo, H., Rintala, E., Simolin, H., & Penttilä, M. (2004). Integrative approaches for studying pentose metabolism in Saccharomyces cerevisiae. Paper presented at Metabolic Engineering V, Lake Tahoe, United States.
Ruohonen, Laura ; Salusjärvi, Laura ; Oresic, Matej ; Maaheimo, Hannu ; Rintala, Eija ; Simolin, Helena ; Penttilä, Merja. / Integrative approaches for studying pentose metabolism in Saccharomyces cerevisiae. Paper presented at Metabolic Engineering V, Lake Tahoe, United States.
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abstract = "We are interested in broadening the substrate utilisation range of the yeast Saccharomyces cerevisiae and in connection to this to study nutritional responses and signalling at a global level. S.cerevisiae does not naturally utilise pentose sugars unlike most other fungi, and recombinant S.cerevisiae strains have been constructed that contain the xylose utilisation pathway from other yeasts. The recombinant xylose pathway is believed to create a redox imbalance in the cells, due to different cofactor requirements of the enzymes of the pathway, and limit xylose fermentation. In order to understand redox and nutrient regulation in general, and the physiology of xylose utilising S.cerevisiae, we carried out chemostat and batch cultures at various oxygenation levels on xylose as a carbon source, and compared those with glucose cultures. Transcriptional profiling, total proteomics and metabolite analyses were carried out, as well as metabolix flux analysis (MFA) and measurements of metabolic fluxes with 13C-NMR. New responses towards xylose as a carbon source were discovered such as the upregulation of pathways for alternative carbon source utilisation and responses for nutritional control and starvation. The physiology of the recombinant yeast appears to be neither fully repressed (fermentative) nor derepressed (gluconeogenic). Own references [1] H. Maaheimo, J. Fiaux, Z. P. Cakar, J. E. Bailey, U. Sauer, T. Szyperski, Eur J Biochem. 268, 2464-2479 (2001). [2] J. P. Pitk{\"a}nen, A. Aristidou, L. Salusj{\"a}rvi, L. Ruohonen, M. Penttil{\"a}, Metab Eng. 5, 16-31 (2003). [3] L. Salusj{\"a}rvi, M. Poutanen, J. P. Pitk{\"a}nen, H. Koivistoinen, A. Aristidou, N. Kalkkinen, L. Ruohonen, M. Penttil{\"a}, Yeast 20, 295-314 (2003). [4] Salusj{\"a}rvi, L., Pitk{\"a}nen, J.-P., Aristidou, A., Ruohonen, L. and Penttil{\"a}, M. (2003a) (in preparation). [5] Toivari, M.H., Aristidou, A., Ruohonen, L. & Penttil{\"a}, M. 2001. Metab. Eng. 3, 236-249.",
author = "Laura Ruohonen and Laura Salusj{\"a}rvi and Matej Oresic and Hannu Maaheimo and Eija Rintala and Helena Simolin and Merja Penttil{\"a}",
year = "2004",
language = "English",
note = "Metabolic Engineering V : Genome to Product ; Conference date: 19-09-2004 Through 23-09-2004",

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Ruohonen, L, Salusjärvi, L, Oresic, M, Maaheimo, H, Rintala, E, Simolin, H & Penttilä, M 2004, 'Integrative approaches for studying pentose metabolism in Saccharomyces cerevisiae' Paper presented at Metabolic Engineering V, Lake Tahoe, United States, 19/09/04 - 23/09/04, .

Integrative approaches for studying pentose metabolism in Saccharomyces cerevisiae. / Ruohonen, Laura; Salusjärvi, Laura; Oresic, Matej; Maaheimo, Hannu; Rintala, Eija; Simolin, Helena; Penttilä, Merja.

2004. Paper presented at Metabolic Engineering V, Lake Tahoe, United States.

Research output: Contribution to conferenceConference articleScientific

TY - CONF

T1 - Integrative approaches for studying pentose metabolism in Saccharomyces cerevisiae

AU - Ruohonen, Laura

AU - Salusjärvi, Laura

AU - Oresic, Matej

AU - Maaheimo, Hannu

AU - Rintala, Eija

AU - Simolin, Helena

AU - Penttilä, Merja

PY - 2004

Y1 - 2004

N2 - We are interested in broadening the substrate utilisation range of the yeast Saccharomyces cerevisiae and in connection to this to study nutritional responses and signalling at a global level. S.cerevisiae does not naturally utilise pentose sugars unlike most other fungi, and recombinant S.cerevisiae strains have been constructed that contain the xylose utilisation pathway from other yeasts. The recombinant xylose pathway is believed to create a redox imbalance in the cells, due to different cofactor requirements of the enzymes of the pathway, and limit xylose fermentation. In order to understand redox and nutrient regulation in general, and the physiology of xylose utilising S.cerevisiae, we carried out chemostat and batch cultures at various oxygenation levels on xylose as a carbon source, and compared those with glucose cultures. Transcriptional profiling, total proteomics and metabolite analyses were carried out, as well as metabolix flux analysis (MFA) and measurements of metabolic fluxes with 13C-NMR. New responses towards xylose as a carbon source were discovered such as the upregulation of pathways for alternative carbon source utilisation and responses for nutritional control and starvation. The physiology of the recombinant yeast appears to be neither fully repressed (fermentative) nor derepressed (gluconeogenic). Own references [1] H. Maaheimo, J. Fiaux, Z. P. Cakar, J. E. Bailey, U. Sauer, T. Szyperski, Eur J Biochem. 268, 2464-2479 (2001). [2] J. P. Pitkänen, A. Aristidou, L. Salusjärvi, L. Ruohonen, M. Penttilä, Metab Eng. 5, 16-31 (2003). [3] L. Salusjärvi, M. Poutanen, J. P. Pitkänen, H. Koivistoinen, A. Aristidou, N. Kalkkinen, L. Ruohonen, M. Penttilä, Yeast 20, 295-314 (2003). [4] Salusjärvi, L., Pitkänen, J.-P., Aristidou, A., Ruohonen, L. and Penttilä, M. (2003a) (in preparation). [5] Toivari, M.H., Aristidou, A., Ruohonen, L. & Penttilä, M. 2001. Metab. Eng. 3, 236-249.

AB - We are interested in broadening the substrate utilisation range of the yeast Saccharomyces cerevisiae and in connection to this to study nutritional responses and signalling at a global level. S.cerevisiae does not naturally utilise pentose sugars unlike most other fungi, and recombinant S.cerevisiae strains have been constructed that contain the xylose utilisation pathway from other yeasts. The recombinant xylose pathway is believed to create a redox imbalance in the cells, due to different cofactor requirements of the enzymes of the pathway, and limit xylose fermentation. In order to understand redox and nutrient regulation in general, and the physiology of xylose utilising S.cerevisiae, we carried out chemostat and batch cultures at various oxygenation levels on xylose as a carbon source, and compared those with glucose cultures. Transcriptional profiling, total proteomics and metabolite analyses were carried out, as well as metabolix flux analysis (MFA) and measurements of metabolic fluxes with 13C-NMR. New responses towards xylose as a carbon source were discovered such as the upregulation of pathways for alternative carbon source utilisation and responses for nutritional control and starvation. The physiology of the recombinant yeast appears to be neither fully repressed (fermentative) nor derepressed (gluconeogenic). Own references [1] H. Maaheimo, J. Fiaux, Z. P. Cakar, J. E. Bailey, U. Sauer, T. Szyperski, Eur J Biochem. 268, 2464-2479 (2001). [2] J. P. Pitkänen, A. Aristidou, L. Salusjärvi, L. Ruohonen, M. Penttilä, Metab Eng. 5, 16-31 (2003). [3] L. Salusjärvi, M. Poutanen, J. P. Pitkänen, H. Koivistoinen, A. Aristidou, N. Kalkkinen, L. Ruohonen, M. Penttilä, Yeast 20, 295-314 (2003). [4] Salusjärvi, L., Pitkänen, J.-P., Aristidou, A., Ruohonen, L. and Penttilä, M. (2003a) (in preparation). [5] Toivari, M.H., Aristidou, A., Ruohonen, L. & Penttilä, M. 2001. Metab. Eng. 3, 236-249.

M3 - Conference article

ER -

Ruohonen L, Salusjärvi L, Oresic M, Maaheimo H, Rintala E, Simolin H et al. Integrative approaches for studying pentose metabolism in Saccharomyces cerevisiae. 2004. Paper presented at Metabolic Engineering V, Lake Tahoe, United States.