Metabolic flux screening of Saccharomyces cerevisiae single knockout strains on glucose and galactose supports elucidation of gene function

Vidya Velagapudi, Christoph Wittmann, Konstantin Schneider, Elmar Heinzle (Corresponding Author)

Research output: Contribution to journalArticleScientificpeer-review

24 Citations (Scopus)

Abstract

New methods for an extended physiological characterization of yeast at a microtiter plate scale were applied to 27 deletion mutants of Saccharomyces cerevisiae cultivated on glucose and galactose as sole carbon sources. In this way, specific growth rates, specific rates of glucose consumption and ethanol production were determined. Flux distribution, particularly concerning branching into the pentose phosphate pathway was determined using a new 13C-labelling method using MALDI-ToF-mass spectrometry. On glucose, the growth was predominantly fermentative whereas on galactose respiration was more active with correspondingly lower ethanol production. Some deletion strains showed unexpected behavior providing very informative data about the function of the corresponding gene. Deletion of malic enzyme gene, MAE1, did not show any significant phenotype when grown on glucose but a drastically increased branching from glucose 6-phosphate into the pentose phosphate pathway when grown on galactose. This allows the conclusion that MAE1 is important for the supply of NADPH during aerobic growth on galactose.
Original languageEnglish
Pages (from-to)395-404
JournalJournal of Biotechnology
Volume132
Issue number4
DOIs
Publication statusPublished - 2007
MoE publication typeA1 Journal article-refereed

Fingerprint

Galactose
Yeast
Glucose
Saccharomyces cerevisiae
Screening
Genes
Fluxes
Pentoses
Pentose Phosphate Pathway
Phosphates
Ethanol
Growth
Glucose-6-Phosphate
Matrix-Assisted Laser Desorption-Ionization Mass Spectrometry
NADP
Labeling
Mass spectrometry
Mass Spectrometry
Respiration
Carbon

Keywords

  • Saccharomyces cerevisiae
  • Malic enzyme
  • Microtiter plate cultivation
  • MALDI-ToF-mass spectrometry
  • Phenotypic analysis

Cite this

Velagapudi, Vidya ; Wittmann, Christoph ; Schneider, Konstantin ; Heinzle, Elmar. / Metabolic flux screening of Saccharomyces cerevisiae single knockout strains on glucose and galactose supports elucidation of gene function. In: Journal of Biotechnology. 2007 ; Vol. 132, No. 4. pp. 395-404.
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abstract = "New methods for an extended physiological characterization of yeast at a microtiter plate scale were applied to 27 deletion mutants of Saccharomyces cerevisiae cultivated on glucose and galactose as sole carbon sources. In this way, specific growth rates, specific rates of glucose consumption and ethanol production were determined. Flux distribution, particularly concerning branching into the pentose phosphate pathway was determined using a new 13C-labelling method using MALDI-ToF-mass spectrometry. On glucose, the growth was predominantly fermentative whereas on galactose respiration was more active with correspondingly lower ethanol production. Some deletion strains showed unexpected behavior providing very informative data about the function of the corresponding gene. Deletion of malic enzyme gene, MAE1, did not show any significant phenotype when grown on glucose but a drastically increased branching from glucose 6-phosphate into the pentose phosphate pathway when grown on galactose. This allows the conclusion that MAE1 is important for the supply of NADPH during aerobic growth on galactose.",
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Metabolic flux screening of Saccharomyces cerevisiae single knockout strains on glucose and galactose supports elucidation of gene function. / Velagapudi, Vidya; Wittmann, Christoph; Schneider, Konstantin; Heinzle, Elmar (Corresponding Author).

In: Journal of Biotechnology, Vol. 132, No. 4, 2007, p. 395-404.

Research output: Contribution to journalArticleScientificpeer-review

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T1 - Metabolic flux screening of Saccharomyces cerevisiae single knockout strains on glucose and galactose supports elucidation of gene function

AU - Velagapudi, Vidya

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AB - New methods for an extended physiological characterization of yeast at a microtiter plate scale were applied to 27 deletion mutants of Saccharomyces cerevisiae cultivated on glucose and galactose as sole carbon sources. In this way, specific growth rates, specific rates of glucose consumption and ethanol production were determined. Flux distribution, particularly concerning branching into the pentose phosphate pathway was determined using a new 13C-labelling method using MALDI-ToF-mass spectrometry. On glucose, the growth was predominantly fermentative whereas on galactose respiration was more active with correspondingly lower ethanol production. Some deletion strains showed unexpected behavior providing very informative data about the function of the corresponding gene. Deletion of malic enzyme gene, MAE1, did not show any significant phenotype when grown on glucose but a drastically increased branching from glucose 6-phosphate into the pentose phosphate pathway when grown on galactose. This allows the conclusion that MAE1 is important for the supply of NADPH during aerobic growth on galactose.

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KW - Malic enzyme

KW - Microtiter plate cultivation

KW - MALDI-ToF-mass spectrometry

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