RESUMEN
Methods of monitoring of estrogenicity in water were gathered, compared, and tested within the context of their practical use as measurement and design tools, in the development of a process of degradation of estrogenic endocrine disruptors. In this work, the focus was put on in vitro assays, with the use of analytical techniques as additional analysis when possible. Practically, from a literature review, four methods that seemed most suitable to practical use required in a process development were tested: the Yeast Estrogen Screen assay, the Lyticase-assisted Yeast Estrogen Screen assay (LYES), the MMV-LUC assay and the HPLC-UV analytical method. Dose-response curves in response to estrogenic standard 17ß-estradiol were compared. Bisphenol A estrogenicity was measured by the methods as well. The model for the calculation of estradiol equivalents as measurements units was adapted. The methods were assessed in terms of ranges of detection, time of experiment, cost, ease of the experiment, reproducibility, etc. Based on that assessment, the LYES assay was selected and successfully applied to the monitoring of estrogenicity removal from 17ß-estradiol and bisphenol A. More precisely, the bioassay allowed the acquisition of kinetic curves for a laboratory-scaled process of estrogenicity removal by immobilized enzymes in a continuous packed-bed reactor. The LYES assay was found to have a real methodological potential for scale-up and design of a treatment process. The HPLC-UV method showed good complementarity with the LYES assay for the monitoring of bisphenol A concentrations in parallel with estrogenicity, reporting no significant estrogenicity from degradation byproducts, among others.
Asunto(s)
Disruptores Endocrinos/análisis , Monitoreo del Ambiente/métodos , Estrógenos/análisis , Contaminantes Químicos del Agua/análisis , Compuestos de Bencidrilo/análisis , Bioensayo , Cromatografía Líquida de Alta Presión , Estradiol/análisis , Genes Reporteros , Glucano Endo-1,3-beta-D-Glucosidasa/farmacología , Humanos , Luciferasas/genética , Luciferasas/metabolismo , Luminiscencia , Células MCF-7 , Complejos Multienzimáticos/farmacología , Péptido Hidrolasas/farmacología , Fenoles/análisis , Receptores de Estrógenos/genética , Receptores de Estrógenos/metabolismo , Elementos de Respuesta , Saccharomyces cerevisiae/genética , Saccharomyces cerevisiae/metabolismo , Vitelogeninas/genética , Purificación del Agua , beta-Galactosidasa/metabolismoRESUMEN
Phellinus robustus produced both laccase (700-4,000 U l(-1)) and manganese peroxidase (MnP) (1,000-11,300 U l(-1)) in fermentation of nine food wastes, whereas Ganoderma adspersum produced only laccase (600-34,000 U l(-1)). Glucose provided high laccase and MnP activity of P. robustus but repressed enzyme production by G. adspersum. Ammonium sulphate and ammonium tartrate increased the P. robustus laccase yield (3-fold), whereas the accumulation of MnP was not enhanced by additional nitrogen.
