Novel method for detecting and quantifying phenol with transient response of glycolytic oscillations of synchronised yeast cells

Grzegorz Pasternak,Martin M Hanczyc

doi:10.1016/j.sbsr.2019.100259

Abstract

Yeast displays cyclic changes of autofluorescence of NADH in living, metabolising cells, known as glycolytic oscillations. In this study, we have demonstrated the proof of concept for exploiting this mechanism for a novel oscillation-based biosensor. The metabolism of Sacharomyces cerevisae BY4742 cells were synchronised and treated with various concentrations of phenol and with glucose and nitrobenzene additions for comparison. The established method allowed detection of phenol within 0.6–26.6 mM concentration range resulting in signal frequency of 0.29–2.03*10−2 Hz, correspondingly. The analysed frequency was negatively and linearly correlated with phenol concentration (R = 0.98). Positive (glucose) and negative (nitrobenzene) control did not caused significant oscillations indicating selectivity of the proposed mechanism. The proposed method is fast (27 min), easy to conduct and can be performed also with other types of yeast strains which makes it accessible for application with local strains and in developing countries. This is the first time, when the natural oscillatory mechanism of yeasts was demonstrated for the biosensing purposes.

Highlights

The role of oscillatory behaviour in biology ranges from periodic cell growth and division to pattern formation in embryogenesis
Simple models may capture the qualitative oscillations of cellular metabolism [6,7], this mechanism has never been exploited for the biosensing purposes
We show that the frequency and amplitude of glycolytic oscillations in yeast cells can be sensitive to potential pollutants and toxins in the environment

Summary

Introduction

The role of oscillatory behaviour in biology ranges from periodic cell growth and division to pattern formation in embryogenesis. In particular it was shown that fluorescence spectrophotometry can be used to monitor the fluctuations of NADH in intact yeast cells [11]. This is typically done by adding glucose to starved yeast cells. A downstream membrane-permeable metabolite, acetaldehyde, is thought to the coordinating agent [15,16,17,18] along with a role for glucose as well [19] Using such cell density threshold dynamics, yeast cells exchange information of their metabolic state [20]

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Results

Conclusion