Microcarrier-based fluorescent yeast estrogen screen assay for fast determination of endocrine disrupting compounds

• Published in: Talanta (Oxford) Vol. 271; p. 125665
• Main Authors: Gregório, Bruno J.R., Ramos, Inês I., Marques, Sara S., Barreiros, Luísa, Magalhães, Luís M., Schneider, Rudolf J., Segundo, Marcela A.
• Format: Journal Article
• Language: English
• Published: Netherlands Elsevier B.V 01.05.2024
• Subjects: Biological Assay; Endocrine Disruptors - analysis; Endocrine-disrupting compounds; Environmental screening; Estradiol - analysis; Estrogens - analysis; Fluorescein di-β-d-galactopyranoside; Genes, Reporter; Humans; Microcarrier; Saccharomyces cerevisiae; Saccharomyces cerevisiae - genetics; Water; Water Pollutants, Chemical - analysis; yeast estrogen screen (YES); yeast estrogen screen (YES); Saccharomyces cerevisiae; Endocrine-disrupting compounds; Microcarrier; Environmental screening; Fluorescein di-β-d-galactopyranoside
• ISSN: 0039-9140; 1873-3573
• DOI: 10.1016/j.talanta.2024.125665

The presence of endocrine-disrupting compounds (EDCs) in water poses a significant threat to human and animal health, as recognized by regulatory agencies throughout the world. The Yeast Estrogen Screen (YES) assay is an excellent method to evaluate the presence of these compounds in water due to its simplicity and capacity to assess the bioaccessible forms/fractions of these compounds. In the presence of a compound with estrogenic activity, Saccharomyces cerevisiae cells, containing a lacZ reporter gene encoding the enzyme β-galactosidase, are induced, the enzyme is synthesised, and released to the extracellular medium. In this work, a YES-based approach encompassing the use of a lacZ reporter gene modified strain of S. cerevisiae, microcarriers as solid support, and a fluorescent substrate, fluorescein di-β-d-galactopyranoside, is proposed, allowing for the assessment of EDCs’ presence after only 2 h of incubation. The proposed method provided an EC50 of 0.17 ± 0.03 nM and an LLOQ of 0.03 nM, expressed as 17β-estradiol. The assessment of different EDCs provided EC50 values between 0.16 and 1.2 × 103 nM. After application to wastewaters, similar results were obtained for EDCs screening, much faster, compared to the conventional 45 h spectrophotometric procedure using a commercial kit, showing potential for onsite high-throughput screening of environmental contamination. [Display omitted] •Endocrine-disrupting compounds (EDCs) in water are a threat to the ecosystem health.•A fluorescent S. cerevisiae-based method for the determination of EDCs is proposed.•Microcarrier beads are employed to increase growth surface of yeast cells.•Yeast cells' lysis improves the sensitivity of the assay, reducing the incubation time.•Low LLOQ value (<1 nM) for 17β-estradiol is reached after 2 h of incubation.