TY - JOUR
T1 - Enhanced Detection of Estrogen-like Compounds by Genetically Engineered Yeast Sensor Strains
AU - Abu-Rmailah, Nidaa
AU - Moscovici, Liat
AU - Riegraf, Carolin
AU - Atias, Hadas
AU - Buchinger, Sebastian
AU - Reifferscheid, Georg
AU - Belkin, Shimshon
N1 - Publisher Copyright:
© 2024 by the authors.
PY - 2024/4
Y1 - 2024/4
N2 - The release of endocrine-disrupting compounds (EDCs) to the environment poses a health hazard to both humans and wildlife. EDCs can activate or inhibit endogenous endocrine functions by binding hormone receptors, leading to potentially adverse effects. Conventional analytical methods can detect EDCs at a high sensitivity and precision, but are blind to the biological activity of the detected compounds. To overcome this limitation, yeast-based bioassays have previously been developed as a pre-screening method, providing an effect-based overview of hormonal-disruptive activity within the sample prior to the application of analytical methods. These yeast biosensors express human endocrine-specific receptors, co-transfected with the relevant response element fused to the specific fluorescent protein reporter gene. We describe several molecular manipulations of the sensor/reporter circuit in a Saccharomyces cerevisiae bioreporter strain that have yielded an enhanced detection of estrogenic-like compounds. Improved responses were displayed both in liquid culture (96-well plate format) as well as in conjunction with sample separation using high-performance thin-layer chromatography (HPTLC). The latter approach allows for an assessment of the biological effect of individual sample components without the need for their chemical identification at the screening stage.
AB - The release of endocrine-disrupting compounds (EDCs) to the environment poses a health hazard to both humans and wildlife. EDCs can activate or inhibit endogenous endocrine functions by binding hormone receptors, leading to potentially adverse effects. Conventional analytical methods can detect EDCs at a high sensitivity and precision, but are blind to the biological activity of the detected compounds. To overcome this limitation, yeast-based bioassays have previously been developed as a pre-screening method, providing an effect-based overview of hormonal-disruptive activity within the sample prior to the application of analytical methods. These yeast biosensors express human endocrine-specific receptors, co-transfected with the relevant response element fused to the specific fluorescent protein reporter gene. We describe several molecular manipulations of the sensor/reporter circuit in a Saccharomyces cerevisiae bioreporter strain that have yielded an enhanced detection of estrogenic-like compounds. Improved responses were displayed both in liquid culture (96-well plate format) as well as in conjunction with sample separation using high-performance thin-layer chromatography (HPTLC). The latter approach allows for an assessment of the biological effect of individual sample components without the need for their chemical identification at the screening stage.
KW - endocrine-disrupting compounds (EDCs)
KW - enhanced green fluorescent protein (EGFP)
KW - high-performance thin-layer chromatography (HPTLC)
KW - wastewater treatment plants (WWTPs)
KW - yeast-based estrogen bioreporters
UR - http://www.scopus.com/inward/record.url?scp=85191343101&partnerID=8YFLogxK
U2 - 10.3390/bios14040193
DO - 10.3390/bios14040193
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C2 - 38667186
AN - SCOPUS:85191343101
SN - 2079-6374
VL - 14
JO - Biosensors
JF - Biosensors
IS - 4
M1 - 193
ER -