Critical evaluation of sampling techniques for residual glucose determination in carbon‐limited chemostat culture of Saccharomyces cerevisiae
- 4 June 2003
- journal article
- research article
- Published by Wiley in Biotechnology & Bioengineering
- Vol. 83 (4) , 395-399
- https://doi.org/10.1002/bit.10683
Abstract
In this paper, three sampling techniques for rapid quenching of cellular metabolism and subsequent separation of cells from fermentation broth are compared: (i) quick freezing of fermentation broth directly in liquid nitrogen; (ii) quenching metabolism by exposing the fermentation broth to stainless steel beads (4‐mm diameter) in a filter syringe precooled to −18°C; and (iii) withdrawal of the filtrate through a 0.45‐μm filter attached to a syringe and a needle inserted directly into the fermentor. It was concluded that use of liquid nitrogen as a quenching method to rapidly arrest cellular metabolism, for quantitative analysis of extracellular glucose, is not a very reliable method and that the filter syringe steel beads work very well. © 2003 Wiley Periodicals, Inc. Biotechnol Bioeng 83: 395–399, 2003.Keywords
This publication has 9 references indexed in Scilit:
- Improved rapid sampling for in vivo kinetics of intracellular metabolites in Saccharomyces cerevisiaeBiotechnology & Bioengineering, 2001
- Statistical reconciliation of the elemental and molecular biomass composition of Saccharomyces cerevisiaeBiotechnology & Bioengineering, 2001
- Improved protocols for quantitative determination of metabolites from biological samples using high performance ionic-exchange chromatography with conductimetric and pulsed amperometric detectionEnzyme and Microbial Technology, 2000
- Glucose Uptake Kinetics and Transcription of HXTGenes in Chemostat Cultures of Saccharomyces cerevisiaeJournal of Biological Chemistry, 1999
- Sampling techniques and comparative extraction procedures for quantitative determination of intra- and extracellular metabolites in filamentous fungiFEMS Microbiology Letters, 1998
- A rapid and reliable method for metabolite extraction in yeast using boiling buffered ethanolYeast, 1997
- In vivo analysis of metabolic dynamics inSaccharomyces cerevisiae : I. Experimental observationsBiotechnology & Bioengineering, 1997
- In Vivo Analysis of Glucose-Induced Fast Changes in Yeast Adenine Nucleotide Pool Applying a Rapid Sampling TechniqueAnalytical Biochemistry, 1993
- Kinetics of growth and glucose transport in glucose‐limited chemostat cultures of Saccharomyces cerevisiae CBS 8066Yeast, 1989