Xylose chemostat isolates of Saccharomyces cerevisiae show altered metabolite and enzyme levels compared with xylose, glucose, and ethanol metabolism of the original strain

Juha Pekka Pitkänen, Eija Rintala, Aristos Aristidou, Laura Ruohonen, Merja Penttilä

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48 Citations (Scopus)

Abstract

The efficient conversion of xylose-containing biomass hydrolysate by the ethanologenic yeast Saccharomyces cerevisiae to useful chemicals such as ethanol still remains elusive, despite significant efforts in both strain and process development. This study focused on the recovery and characterization of xylose chemostat isolates of a S. cerevisiae strain that overexpresses xylose reductase- and xylitol dehydrogenase-encoding genes from Pichia stipitis and the gene encoding the endogenous xylulokinase. The isolates were recovered from aerobic chemostat cultivations on xylose as the sole or main carbon source. Under aerobic conditions, on minimal medium with 30 g l-1 xylose, the growth rate of the chemostat isolates was 3-fold higher than that of the original strain (0.15 h-1 vs 0.05 h-1). In a detailed characterization comparing the metabolism of the isolates with the metabolism of xylose, glucose, and ethanol in the original strain, the isolates showed improved properties in the assumed bottlenecks of xylose metabolism. The xylose uptake rate was increased almost 2-fold. Activities of the key enzymes in the pentose phosphate pathway (transketolase, transaldolase) increased 2-fold while the concentrations of their substrates (pentose 5-phosphates, sedoheptulose 7-phosphate) decreased correspondingly. Under anaerobic conditions, on minimal medium with 45 g l-1 xylose, the ethanol productivity (in terms of cell dry weight; CDW) of one of the isolates increased from 0.012 g g -1 CDW h-1 to 0.017 g g-1 CDW h-1 and the yield from 0.09 g g-1 xylose to 0.14 g g-1 xylose, respectively.

Original languageEnglish
Pages (from-to)827-837
Number of pages11
JournalApplied Microbiology and Biotechnology
Volume67
Issue number6
DOIs
Publication statusPublished - 1 Jun 2005
MoE publication typeA1 Journal article-refereed

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Xylose
Saccharomyces cerevisiae
Ethanol
Enzymes
D-Xylulose Reductase
Transaldolase
Transketolase
Pentoses
xylose-glucose
Pentose Phosphate Pathway
Aldehyde Reductase
Pichia
Biomass
Genes
Carbon
Yeasts
Phosphates
Weights and Measures
Growth

Cite this

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title = "Xylose chemostat isolates of Saccharomyces cerevisiae show altered metabolite and enzyme levels compared with xylose, glucose, and ethanol metabolism of the original strain",
abstract = "The efficient conversion of xylose-containing biomass hydrolysate by the ethanologenic yeast Saccharomyces cerevisiae to useful chemicals such as ethanol still remains elusive, despite significant efforts in both strain and process development. This study focused on the recovery and characterization of xylose chemostat isolates of a S. cerevisiae strain that overexpresses xylose reductase- and xylitol dehydrogenase-encoding genes from Pichia stipitis and the gene encoding the endogenous xylulokinase. The isolates were recovered from aerobic chemostat cultivations on xylose as the sole or main carbon source. Under aerobic conditions, on minimal medium with 30 g l-1 xylose, the growth rate of the chemostat isolates was 3-fold higher than that of the original strain (0.15 h-1 vs 0.05 h-1). In a detailed characterization comparing the metabolism of the isolates with the metabolism of xylose, glucose, and ethanol in the original strain, the isolates showed improved properties in the assumed bottlenecks of xylose metabolism. The xylose uptake rate was increased almost 2-fold. Activities of the key enzymes in the pentose phosphate pathway (transketolase, transaldolase) increased 2-fold while the concentrations of their substrates (pentose 5-phosphates, sedoheptulose 7-phosphate) decreased correspondingly. Under anaerobic conditions, on minimal medium with 45 g l-1 xylose, the ethanol productivity (in terms of cell dry weight; CDW) of one of the isolates increased from 0.012 g g -1 CDW h-1 to 0.017 g g-1 CDW h-1 and the yield from 0.09 g g-1 xylose to 0.14 g g-1 xylose, respectively.",
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Xylose chemostat isolates of Saccharomyces cerevisiae show altered metabolite and enzyme levels compared with xylose, glucose, and ethanol metabolism of the original strain. / Pitkänen, Juha Pekka; Rintala, Eija; Aristidou, Aristos; Ruohonen, Laura; Penttilä, Merja.

In: Applied Microbiology and Biotechnology, Vol. 67, No. 6, 01.06.2005, p. 827-837.

Research output: Contribution to journalArticleScientificpeer-review

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AU - Pitkänen, Juha Pekka

AU - Rintala, Eija

AU - Aristidou, Aristos

AU - Ruohonen, Laura

AU - Penttilä, Merja

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N2 - The efficient conversion of xylose-containing biomass hydrolysate by the ethanologenic yeast Saccharomyces cerevisiae to useful chemicals such as ethanol still remains elusive, despite significant efforts in both strain and process development. This study focused on the recovery and characterization of xylose chemostat isolates of a S. cerevisiae strain that overexpresses xylose reductase- and xylitol dehydrogenase-encoding genes from Pichia stipitis and the gene encoding the endogenous xylulokinase. The isolates were recovered from aerobic chemostat cultivations on xylose as the sole or main carbon source. Under aerobic conditions, on minimal medium with 30 g l-1 xylose, the growth rate of the chemostat isolates was 3-fold higher than that of the original strain (0.15 h-1 vs 0.05 h-1). In a detailed characterization comparing the metabolism of the isolates with the metabolism of xylose, glucose, and ethanol in the original strain, the isolates showed improved properties in the assumed bottlenecks of xylose metabolism. The xylose uptake rate was increased almost 2-fold. Activities of the key enzymes in the pentose phosphate pathway (transketolase, transaldolase) increased 2-fold while the concentrations of their substrates (pentose 5-phosphates, sedoheptulose 7-phosphate) decreased correspondingly. Under anaerobic conditions, on minimal medium with 45 g l-1 xylose, the ethanol productivity (in terms of cell dry weight; CDW) of one of the isolates increased from 0.012 g g -1 CDW h-1 to 0.017 g g-1 CDW h-1 and the yield from 0.09 g g-1 xylose to 0.14 g g-1 xylose, respectively.

AB - The efficient conversion of xylose-containing biomass hydrolysate by the ethanologenic yeast Saccharomyces cerevisiae to useful chemicals such as ethanol still remains elusive, despite significant efforts in both strain and process development. This study focused on the recovery and characterization of xylose chemostat isolates of a S. cerevisiae strain that overexpresses xylose reductase- and xylitol dehydrogenase-encoding genes from Pichia stipitis and the gene encoding the endogenous xylulokinase. The isolates were recovered from aerobic chemostat cultivations on xylose as the sole or main carbon source. Under aerobic conditions, on minimal medium with 30 g l-1 xylose, the growth rate of the chemostat isolates was 3-fold higher than that of the original strain (0.15 h-1 vs 0.05 h-1). In a detailed characterization comparing the metabolism of the isolates with the metabolism of xylose, glucose, and ethanol in the original strain, the isolates showed improved properties in the assumed bottlenecks of xylose metabolism. The xylose uptake rate was increased almost 2-fold. Activities of the key enzymes in the pentose phosphate pathway (transketolase, transaldolase) increased 2-fold while the concentrations of their substrates (pentose 5-phosphates, sedoheptulose 7-phosphate) decreased correspondingly. Under anaerobic conditions, on minimal medium with 45 g l-1 xylose, the ethanol productivity (in terms of cell dry weight; CDW) of one of the isolates increased from 0.012 g g -1 CDW h-1 to 0.017 g g-1 CDW h-1 and the yield from 0.09 g g-1 xylose to 0.14 g g-1 xylose, respectively.

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