Alpha 2.0
Login Register
» Articles » PMID: 12464

Determination of Intermediary Metabolites in Yeast. Critical Examination of the Effect of Sampling Conditions and Recommendations for Obtaining True Levels

Overview
Journal Mol Cell Biochem
Publisher Springer
Specialty Biochemistry
Date 1976 Nov 30
PMID 12464
Citations 31
Authors
M J Saez
R Lagunas
Affiliations
Soon will be listed here.
Abstract

The effect of sampling conditions on the levels of adenine nucleotides, pyridine nucleotides, glycolytic intermediates and related metabolites in yeast has been studied. A systematic examination of the conditions for harvesting has shown that it can be best accomplished by rapid filtration. Delays in the handling for removal of the medium, as is usual in the process of obtaining a number of data reported in the literature, lead to important changes in some of the metabolites examined. It is also shown that when a washing is imperative it can be carried out with a methanol-water mixture (50/50, v/v) cooled at -40 degrees without loss of intracellular concentrations of non-readily diffusible metabolites. On the basis of this experience the outline of a generally applicable procedure is presented.

Citing Articles

Nanohaloarchaea as beneficiaries of xylan degradation by haloarchaea.

La Cono V, Messina E, Reva O, Smedile F, La Spada G, Crisafi F Microb Biotechnol. 2023; 16(9):1803-1822.

PMID: 37317055 PMC: 10443357. DOI: 10.1111/1751-7915.14272.

Optimized Fast Filtration-Based Sampling and Extraction Enables Precise and Absolute Quantification of the Central Carbon Metabolome.

Thorfinnsdottir L, Garcia-Calvo L, Bo G, Bruheim P, Rost L Metabolites. 2023; 13(2).

PMID: 36837769 PMC: 9965072. DOI: 10.3390/metabo13020150.

Fast Sampling of the Cellular Metabolome.

van Gulik W, Canelas A, Taymaz-Nikerel H, Douma R, de Jonge L, Heijnen J Methods Mol Biol. 2021; 2349:11-39.

PMID: 34718989 DOI: 10.1007/978-1-0716-1585-0_2.

Transporters at the Interface between Cytosolic and Mitochondrial Amino Acid Metabolism.

Hewton K, Johal A, Parker S Metabolites. 2021; 11(2).

PMID: 33669382 PMC: 7920303. DOI: 10.3390/metabo11020112.

Analysis of Intracellular Metabolites from Microorganisms: Quenching and Extraction Protocols.

Pinu F, Villas-Boas S, Aggio R Metabolites. 2017; 7(4).

PMID: 29065530 PMC: 5746733. DOI: 10.3390/metabo7040053.

References
1.
Rothman L, CABIB E . Regulation of glycogen synthesis in the intact yeast cell. Biochemistry. 1969; 8(8):3332-41. DOI: 10.1021/bi00836a030. View
2.
Polakis E, Bartley W . Changes in the intracellular concentrations of adenosine phosphates and nicotinamide nucleotides during the aerobic growth cycle of yeast on different carbon sources. Biochem J. 1966; 99(3):521-33. PMC: 1265036. DOI: 10.1042/bj0990521. View
3.
Atkinson D . Regulation of enzyme function. Annu Rev Microbiol. 1969; 23:47-68. DOI: 10.1146/annurev.mi.23.100169.000403. View
4.
LOWRY O, Carter J, Ward J, Glaser L . The effect of carbon and nitrogen sources on the level of metabolic intermediates in Escherichia coli. J Biol Chem. 1971; 246(21):6511-21. View
5.
CONWAY E, Downey M . An outer metabolic region of the yeast cell. Biochem J. 1950; 47(3):347-55. PMC: 1275218. DOI: 10.1042/bj0470347. View