Monitoring the functionality and stress response of yeast cells using flow cytometry

Abstract

Throughout fermentation, yeast faces continuously changing medium conditions and reacts by adapting its metabolism. The adaptation is a critical process and is dependent on the accurate functioning of the cell. A stable membrane potential, which is, among other roles, responsible for protecting the yeast from low pH, is an important attribute for evaluating functionality. Other factors are storage products such as glycogen, trehalose, and neutral lipids, as well as mitochondrial activity and the integrity of the DNA. These parameters can be complemented by the analysis of viability, cell cycle, intracellular pH, and reactive oxygen species in the cell. The correlation of all these factors provides valuable information for evaluating the performance of a yeast population during fermentation. In order to demonstrate the analytical capabilities of flow cytometry, a Saccharomyces cerevisiae yeast strain was observed in a modified growth medium for 384 h (16 days). The results confirm observations made with other methods and reports from the literature. However, with flow cytometry, it is possible to gain deeper insight into stress response and adaptation behavior of yeast at a cellular level. The causality from the formation of oxygen-radicals to cell death, for example, can be shown, as well as the dependency of the intracellular pH on the stability of the membrane. The proposed bio-monitoring system has the potential to provide applicable information as a process control tool for wineries.