Popis: |
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. Keywords: Biosensor, Fluorescence, NADH, Frequency, Amplitude, Sensing |