Proteome analysis of recombinant xylose-fermenting Saccharomyces cerevisiae

Laura Salusjärvi, Marjo Poutanen, Juha Pekka Pitkänen, Heini Koivistoinen, Aristos Aristidou, Nisse Kalkkinen, Laura Ruohonen, Merja Penttilä

Research output: Contribution to journalArticleScientificpeer-review

46 Citations (Scopus)

Abstract

Introduction of an active xylose utilization pathway into Saccharomyces cerevisiae, which does not naturally ferment pentose sugars, is likely to have a major impact on the overall cellular metabolism as the carbon introduced to the cells will now flow through the pentose phosphate pathway. The metabolic responses in the recombinant xylose-fermenting S. cerevisiae were studied at the proteome level by comparative two-dimensional gel electrophoresis of cellular proteins within a pH range of 3-10. Glucose-limited chemostat cultivations and corresponding chemostat cultivations performed in media containing xylose as the major carbon source were compared. The cultivations were studied in aerobic and anaerobic metabolic steady states and in addition at time points 5, 30 and 60 min after the switch-off of oxygen supply. We identified 22 proteins having a significant abundance difference on xylose compared to glucose, and 12 proteins that responded to change from aerobic to anaerobic conditions on both carbon sources. On xylose in all conditions studied, major changes were seen in the abundance of alcohol dehydrogenase 2 (Adh2p), acetaldehyde dehydrogenases 4 and 6 (Ald4p and Ald6p), and DL-glycerol 3-phosphatase (Gpp1p). Our results give indications of altered metabolic fluxes especially in the acetate and glycerol pathways in cells growing on xylose compared to glucose.

Original languageEnglish
Pages (from-to)295-314
Number of pages20
JournalYeast
Volume20
Issue number4
DOIs
Publication statusPublished - 1 Mar 2003
MoE publication typeA1 Journal article-refereed

Fingerprint

Xylose
Proteome
proteome
xylose
Yeast
Saccharomyces cerevisiae
Proteins
Chemostats
Glucose
Pentoses
Carbon
pentoses
glycerol-1-phosphatase
Glycerol
glucose
glycerol
carbon
acetaldehyde dehydrogenase
Oxygen supply
Pentose Phosphate Pathway

Keywords

  • Ethanol
  • Metabolic engineering
  • Pentose phosphate pathway
  • Proteome
  • Proteomics
  • Saccharomyces cerevisiae
  • Two-dimensional gel electrophoresis
  • Xylose

Cite this

Salusjärvi, L., Poutanen, M., Pitkänen, J. P., Koivistoinen, H., Aristidou, A., Kalkkinen, N., ... Penttilä, M. (2003). Proteome analysis of recombinant xylose-fermenting Saccharomyces cerevisiae. Yeast, 20(4), 295-314. https://doi.org/10.1002/yea.960
Salusjärvi, Laura ; Poutanen, Marjo ; Pitkänen, Juha Pekka ; Koivistoinen, Heini ; Aristidou, Aristos ; Kalkkinen, Nisse ; Ruohonen, Laura ; Penttilä, Merja. / Proteome analysis of recombinant xylose-fermenting Saccharomyces cerevisiae. In: Yeast. 2003 ; Vol. 20, No. 4. pp. 295-314.
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Salusjärvi, L, Poutanen, M, Pitkänen, JP, Koivistoinen, H, Aristidou, A, Kalkkinen, N, Ruohonen, L & Penttilä, M 2003, 'Proteome analysis of recombinant xylose-fermenting Saccharomyces cerevisiae', Yeast, vol. 20, no. 4, pp. 295-314. https://doi.org/10.1002/yea.960

Proteome analysis of recombinant xylose-fermenting Saccharomyces cerevisiae. / Salusjärvi, Laura; Poutanen, Marjo; Pitkänen, Juha Pekka; Koivistoinen, Heini; Aristidou, Aristos; Kalkkinen, Nisse; Ruohonen, Laura; Penttilä, Merja.

In: Yeast, Vol. 20, No. 4, 01.03.2003, p. 295-314.

Research output: Contribution to journalArticleScientificpeer-review

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T1 - Proteome analysis of recombinant xylose-fermenting Saccharomyces cerevisiae

AU - Salusjärvi, Laura

AU - Poutanen, Marjo

AU - Pitkänen, Juha Pekka

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AU - Aristidou, Aristos

AU - Kalkkinen, Nisse

AU - Ruohonen, Laura

AU - Penttilä, Merja

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AB - Introduction of an active xylose utilization pathway into Saccharomyces cerevisiae, which does not naturally ferment pentose sugars, is likely to have a major impact on the overall cellular metabolism as the carbon introduced to the cells will now flow through the pentose phosphate pathway. The metabolic responses in the recombinant xylose-fermenting S. cerevisiae were studied at the proteome level by comparative two-dimensional gel electrophoresis of cellular proteins within a pH range of 3-10. Glucose-limited chemostat cultivations and corresponding chemostat cultivations performed in media containing xylose as the major carbon source were compared. The cultivations were studied in aerobic and anaerobic metabolic steady states and in addition at time points 5, 30 and 60 min after the switch-off of oxygen supply. We identified 22 proteins having a significant abundance difference on xylose compared to glucose, and 12 proteins that responded to change from aerobic to anaerobic conditions on both carbon sources. On xylose in all conditions studied, major changes were seen in the abundance of alcohol dehydrogenase 2 (Adh2p), acetaldehyde dehydrogenases 4 and 6 (Ald4p and Ald6p), and DL-glycerol 3-phosphatase (Gpp1p). Our results give indications of altered metabolic fluxes especially in the acetate and glycerol pathways in cells growing on xylose compared to glucose.

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KW - Proteomics

KW - Saccharomyces cerevisiae

KW - Two-dimensional gel electrophoresis

KW - Xylose

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Salusjärvi L, Poutanen M, Pitkänen JP, Koivistoinen H, Aristidou A, Kalkkinen N et al. Proteome analysis of recombinant xylose-fermenting Saccharomyces cerevisiae. Yeast. 2003 Mar 1;20(4):295-314. https://doi.org/10.1002/yea.960