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References (25)
-
H. Sahm, H. J. Rehm (1993)
Biotechnology, 1
-
G. Ruijter, J. Visser (1996)
Determination of intermediary metabolites in Aspergillus niger.Journal of Microbiological Methods, 25
-
F. Gamo, F. Portillo, C. Gancedo (1993)
Characterization of mutations that overcome the toxic effect of glucose on phosphoglucose isomerase less strains of Saccharomyces cerevisiae.FEMS microbiology letters, 106 3
-
H. Bergmeyer (2019)
Methods of Enzymatic Analysis -
H. U. Bergmeyer (1985)
Methods in Enzymatic Analysis, vol. VI and VII -
E. Boles, F. Zimmermann, J. Heinisch (1993)
Different signals control the activation of glycolysis in the yeast Saccharomyces cerevisiaeYeast, 9
-
D. Weuster‐Botz, A. Graaf (1996)
Reaction engineering methods to study intracellular metabolite concentrations.Advances in biochemical engineering/biotechnology, 54
-
(1979)
Saccharo - myces cerevisiae -
C. Franco, J. Smith, D. Berry (1984)
Effect of Nitrogen and Phosphate on the Levels of Intermediates in Bakers' Yeast Grown in Continuous CultureMicrobiology, 130
-
A. Graaf, R. Wittig, U. Probst, J. Strohhaecker, S. Schoberth, H. Sahm (1992)
Continuous-flow NMR bioreactor for in vivo studies of microbial cell suspensions with low biomass concentrationsJournal of Magnetic Resonance, 98
-
A. Hartbrich, G. Schmitz, D. Weuster‐Botz, A. Graaf, C. Wandrey (2000)
Development and application of a membrane cyclone reactor for in vivo NMR spectroscopy with high microbial cell densities.Biotechnology and bioengineering, 51 6
-
K. D. Entian, F. K. Zimmermann, I. Scheel (1977)
A partial defect in carbon catabolite repression mutants of Saccharomyces cerevisiae with reduced hexose phosphorylation, 156
-
(1974)
In Bergmeyer, H -
W. Koning, K. Dam (1992)
A method for the determination of changes of glycolytic metabolites in yeast on a subsecond time scale using extraction at neutral pH.Analytical biochemistry, 204 1
-
J. Bailey (1991)
Toward a science of metabolic engineeringScience, 252
-
U. Theobald, W. Mailinger, M. Reuss, M. Rizzi (1993)
In vivo analysis of glucose‐induced fast glycolytic changes in yeast adenine nucleotide pool applying a rapid sampling technique, 214
-
J. Gancedo, C. Gancedo (1973)
Concentrations of intermediary metabolites in yeast.Biochimie, 55 2
-
C. Verduyn, E. Postma, W. Scheffers, J. Dijken (1992)
Effect of benzoic acid on metabolic fluxes in yeasts: A continuous‐culture study on the regulation of respiration and alcoholic fermentationYeast, 8
-
J. François, E. Schaftingen, H. Hers (1984)
The mechanism by which glucose increases fructose 2,6-bisphosphate concentration in Saccharomyces cerevisiae. A cyclic-AMP-dependent activation of phosphofructokinase 2.European journal of biochemistry, 145 1
-
Karl Weibel, Juan-R. Mor, Armin Fiechter (1974)
Rapid sampling of yeast cells and automated assays of adenylate, citrate, pyruvate and glucose-6-phosphate pools.Analytical biochemistry, 58 1
-
G. Stephanopoulos (1994)
Metabolic engineering.Current opinion in biotechnology, 5 2
-
M. Ciriacy, I. Breitenbach (1979)
Physiological Effects of Seven Different Blocks in Glycolysis in Saccharomyces cerevisiaeJournal of Bacteriology, 139
-
U. Theobald, W. Mailinger, M. Reuss, M. Rizzi (1993)
In vivo analysis of glucose-induced fast changes in yeast adenine nucleotide pool applying a rapid sampling technique.Analytical biochemistry, 214 1
-
J. François, E. Van Schaftingen, H. G. Hers (1984)
The mechanism by which glucose increases fructose‐2,6‐bisphosphate concentration in Saccharomyces cerevisiae, 145
-
H. Westerhoff, D. Kell (1996)
What bio technologists knew all along...?Journal of theoretical biology, 182 3
- Publisher
- Wiley
- Copyright
- Copyright © 1997 Wiley Subscription Services
- ISSN
- 0749-503X
- eISSN
- 1097-0061
- DOI
- 10.1002/(SICI)1097-0061(199711)13:14<1347::AID-YEA176>3.0.CO;2-O
- pmid
- 9392079
- Publisher site
- See Article on Publisher Site
Abstract
A simple and reliable method for the efficient inactivation of metabolism and for quantitative metabolite extraction from yeast cells is presented. It is based on the use of a boiling solution made of 75% ethanol (volume/final volume) buffered with 70 mm‐Hepes (final concentration), pH 7·5, to guarantee the stability throughout the whole procedure of a large variety of metabolites, including all glycolytic intermediates, nucleotides, pyridine nucleotides and organic acids compounds. The extraction is fast, requiring only 3 min incubation of yeast cells in the ethanol‐buffered mixture maintained at 80°C. It can be carried out either directly by spraying the cells into the boiling mixture, or after quenching the whole culture in 60% methanol kept at −40°C. Extracts are subsequently concentrated by evaporation under partial vacuum and the residue is resuspended in a small volume of water. This concentration step and the use of a highly sensitive analytical method allow us to quantify metabolites in less than 10 mg dry weight cells. This method, which can be applied to other fungi, could be very helpful for the determination of true metabolites in mutants generated through the EUROFAN programme and for metabolic flux analysis. © 1997 John Wiley & Sons, Ltd.
Journal
Yeast – Wiley
Published: Jan 1, 1997
Keywords: ; ;