Yeast-Based Fluorescent Sensors for the Simultaneous Detection of Estrogenic and Androgenic Compounds, Coupled with High-Performance Thin Layer Chromatography.

Liat Moscovici,Carolin Riegraf,Hadas Atias,Nidaa Abu-Rmailah,Shimshon Belkin,Sebastian Buchinger,Georg Reifferscheid,Dror Shakibai

doi:10.3390/bios10110169

Abstract

The persistence of endocrine disrupting compounds (EDCs) throughout wastewater treatment processes poses a significant health threat to humans and to the environment. The analysis of EDCs in wastewater remains a challenge for several reasons, including (a) the multitude of bioactive but partially unknown compounds, (b) the complexity of the wastewater matrix, and (c) the required analytical sensitivity. By coupling biological assays with high-performance thin-layer chromatography (HPTLC), different samples can be screened simultaneously, highlighting their active components; these may then be identified by chemical analysis. To allow the multiparallel detection of diverse endocrine disruption activities, we have constructed Saccharomyces cerevisiae-based bioreporter strains, responding to compounds with either estrogenic or androgenic activity, by the expression of green (EGFP), red (mRuby), or blue (mTagBFP2) fluorescent proteins. We demonstrate the analytical potential inherent in combining chromatographic compound separation with a direct fluorescent signal detection of EDC activities. The applicability of the system is further demonstrated by separating influent samples of wastewater treatment plants, and simultaneously quantifying estrogenic and androgenic activities of their components. The combination of a chemical separation technique with an optical yeast-based bioassay presents a potentially valuable addition to our arsenal of environmental pollution monitoring tools.

Highlights

Endocrine disrupting chemicals (EDCs) are exogenous agents with structural similarity to endogenous hormones, that may interfere with natural hormonal activity by blocking, competing or mimicking natural hormones [1]
We have described a set of Saccharomyces cerevisiae fluorescent protein-based bioreporters for the detection of two classes of endocrine disrupting chemicals in complex samples, in combination with sample separation by high-performance thin-layer chromatography (HPTLC), with limit of detection (LOD) values similar or lower to those reported for other cellular assays (Table 2)
We have presented a panel of sensitive, specific, and robust fluorescent bioreporter S. cerevisiae strains for the detection of compounds exerting estrogenic and androgenic effects in complex samples, and have demonstrated their efficacy in the analysis of both model compounds and multi-component wastewater samples

Summary

Introduction

Endocrine disrupting chemicals (EDCs) are exogenous agents with structural similarity to endogenous hormones, that may interfere with natural hormonal activity by blocking, competing or mimicking natural hormones [1]. Some EDCs were suggested to act as carcinogenic agents [3] These adverse health effects, in some cases triggered by exposure to Biosensors 2020, 10, 169; doi:10.3390/bios10110169 www.mdpi.com/journal/biosensors. Biosensors 2020, 10, 169 parts-per-billion level concentrations [4,5,6], have raised concern among public health authorities This concern is exacerbated when considering the possible manifestation of certain health effects associated with exposure to EDCs across generations [2]. Besides being a human health risk, the release of EDCs to aquatic systems poses a significant ecological threat. Continuous exposure to such chemicals may affect local species and ecosystems. For example, that chronic exposure to EDCs causes abnormalities in the reproductive system of certain fish species [1,7,8,9]

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