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Applied and Environmental Microbiology, March 2005, p. 1515-1521, Vol. 71, No. 3
0099-2240/05/$08.00+0     doi:10.1128/AEM.71.3.1515-1521.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

Intracellular pH Distribution in Saccharomyces cerevisiae Cell Populations, Analyzed by Flow Cytometry

Minoska Valli,1 Michael Sauer,1 Paola Branduardi,2 Nicole Borth,1 Danilo Porro,2 and Diethard Mattanovich1*

Institute of Applied Microbiology, University of Natural Resources and Applied Life Sciences, Vienna, Austria,1 Department of Biotechnology and Biosciences, University of Milan-Bicocca, Milan, Italy2

Received 6 September 2004/ Accepted 18 October 2004

Intracellular pH has an important role in the maintenance of the normal functions of yeast cells. The ability of the cell to maintain this pH homeostasis also in response to environmental changes has gained more and more interest in both basic and applied research. In this study we describe a protocol which allows the rapid determination of the intracellular pH of Saccharomyces cerevisiae cells. The method is based on flow cytometry and employs the pH-dependent fluorescent probe carboxy SNARF-4F. The protocol attempts to minimize the perturbation of the system under study, thus leading to accurate information about the physiological state of the single cell. Moreover, statistical analysis performed on major factors that may influence the final determination supported the validity of the optimized protocol. The protocol was used to investigate the effect of external pH on S. cerevisiae cells incubated in buffer. The results obtained showed that stationary cells are better able than exponentially grown cells to maintain their intracellular pH homeostasis independently of external pH changes. Furthermore, analysis of the intracellular pH distribution within the cell populations highlighted the presence of subpopulations characterized by different intracellular pH values. Notably, a different behavior was observed for exponentially grown and stationary cells in terms of the appearance and development of these subpopulations as a response to a changing external pH.

* Corresponding author. Mailing address: Institute of Applied Microbiology, University of Natural Resources and Applied Life Sciences, Muthgasse 18, A-1190 Vienna, Austria. Phone: 43 1 36006 6569. Fax: 43 1 3697615. E-mail: diethard.mattanovich{at}boku.ac.at.

Applied and Environmental Microbiology, March 2005, p. 1515-1521, Vol. 71, No. 3
0099-2240/05/$08.00+0     doi:10.1128/AEM.71.3.1515-1521.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

  • Weigert, C., Steffler, F., Kurz, T., Shellhammer, T. H., Methner, F.-J. (2009). Application of a Short Intracellular pH Method to Flow Cytometry for Determining Saccharomyces cerevisiae Vitality. Appl. Environ. Microbiol. 75: 5615-5620 [Abstract] [Full Text]  
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  • Oh-oka, K., Nakatogawa, H., Ohsumi, Y. (2008). Physiological pH and Acidic Phospholipids Contribute to Substrate Specificity in Lipidation of Atg8. J. Biol. Chem. 283: 21847-21852 [Abstract] [Full Text]  
  • Valli, M., Sauer, M., Branduardi, P., Borth, N., Porro, D., Mattanovich, D. (2006). Improvement of Lactic Acid Production in Saccharomyces cerevisiae by Cell Sorting for High Intracellular pH. Appl. Environ. Microbiol. 72: 5492-5499 [Abstract] [Full Text]  


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