title: Polymer-immobilized ready-to-use recombinant yeast assays for the detection of endocrine disruptive compounds.
abstract: Recombinant yeast assays (RYAs) constitute a suitable tool for the environmental monitoring of compounds with endocrine disrupting activities, notably estrogenicity and androgenicity. Conventional procedures require yeast reconstitution from frozen stock, which usually takes several days and demands additional equipment. With the aim of applying such assays to field studies and making them more accessible to less well-equipped laboratories, we have optimized RYA by the immobilization of Saccharomyces cerevisiae cells in three different polymer matrices - gelatin, Bacto agar, and Yeast Extract Peptone Dextrose agar - to obtain a ready-to-use version for the fast assessment of estrogenic and androgenic potencies of compounds and environmental samples. Among the three matrices, gelatin showed the best results for both testosterone (androgen receptor yeast strain; AR-RYA) and 17β-estradiol (estrogen receptor yeast strain; ER-RYA). AR-RYA was characterized by a lowest observed effect concentration (LOEC), EC50 and induction factor (IF) of 1nM, 2.2nM and 51, respectively. The values characterizing ER-RYA were 0.4nM, 1.8nM, and 63, respectively. Gelatin immobilization retained yeast viability and sensitivity for more than 90d of storage at 4°C. The use of the immobilized yeast reduced the assay duration to only 3h without necessity of sterile conditions. Because immobilized RYA can be performed either in multiwell microplates or glass tubes, it allows multiple samples to be tested at once, and easy adaptation to existing portable devices for direct in-field applications.
authors: Masaryk University, Faculty of Science, RECETOX, Kamenice 5, CZ-62500 Brno, Czech Republic.
authors: Michal
authors: M
authors: Bittner
authors: Masaryk University, Faculty of Science, RECETOX, Kamenice 5, CZ-62500 Brno, Czech Republic.
authors: Sergio
authors: S
authors: Jarque
authors: Masaryk University, Faculty of Science, RECETOX, Kamenice 5, CZ-62500 Brno, Czech Republic. Electronic address: hilscherova@recetox.muni.cz.
authors: Klára
authors: K
authors: Hilscherová
mesh_terms: D001681:Biological Assay / Q000379:methods*; D052244:Endocrine Disruptors / Q000032:analysis*; D004784:Environmental Monitoring / Q000379:methods; D004958:Estradiol; D011108:Polymers; D011944:Receptors, Androgen; D011960:Receptors, Estrogen; D012441:Saccharomyces cerevisiae / Q000378:metabolism*
chemical_list: C410798:AR protein, human; D052244:Endocrine Disruptors; D011108:Polymers; D011944:Receptors, Androgen; D011960:Receptors, Estrogen; D004958:Estradiol
keywords: Agar; Endocrine disruption; Environmental monitoring; Gelatin; Immobilization; Recombinant yeast assay