BIOLUMINOMETRIC ASSAY OF CELL ATP DETECTION UNDER STRESS CONDITIONS AND IN THE ASSESSMENT OF CLEANNESS OF THE FOOD PRODUCING COMPANY

Abstract

Detection and tracking of microorganism’s behavior in natural environments and under stressed conditions have been nowadays accelerated by the requirement for the risk assessment associated with environmental contamination. Luminescence-based techniques, for they cost efficiency, fastness, reliability and reproducibility are particularly appropriate for such studies. We used bioluminescence sensing assay to determine ATP levels in the yeast Schizosaccharomyce pombe. Determination of adenosine concentration per cell was assessed in the yeast cells counted prior to the experiment. To test the methodology, model organism was exposed to life-threatening concentration of heavy metals cadmium and nickel for three hours. Afterwards, changes in the ATP levels were detected and compared to the control. Strikingly, our results revealed that such conditions lead to considerable increase in the production of ATP by mitochondria compared to untreated cells. Normal level of ATP in the control increased from the basal 9.92x10^-16 M to 2.90x10^-15 M after Cd exposure and to 7.58x10^-15 M upon Ni treatment. The methodology was subsequently used for assessment of cleanness of margarine producing company devices after regular sanitation procedure. Detection of only negligible amount of the ATP signal confirmed good cleaning strategy in the tested food processing company.