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하수처리장의 내분비계장애물질에 대한 Yeast Two-hybrid Assay와 Enzyme-Linked Immunosorbent Assay에 의한 에스트로겐활성도 평가
이병천(Byoung Cheun Lee),나진성(Jin Sung Ra),김상돈(Sang Don Kim),Kawai Hukiko,이철희(Chul Hee Lee) 大韓環境工學會 2007 대한환경공학회지 Vol.29 No.7
가정계열과 공단계열로 분리하여 처리되는 하수처리장에서 에스트로겐 활성을 평가하기 위하여 yeast two-hybrid assay와 enzyme-linked immunosorbent assay(ELISA)를 이용하여 내분비계장애물질의 농도와 활성도를 측정하였다. 그 결과 가정계열 유입수 중에서 estrone(E1), 17β-estradiol(e₂), 17α-ethinylestradiol(Ee₂) 그리고 APE의 농도는 각각 최대 167.1, 39.7, 7.3, 145.4 ng/L까지 검출되었다. 활성슬러지법에 의한 처리로, 17β-estradiol의 평균제거율은 77.5%, 고도처리 공정인 모래여과와 오존산화를 거친 후에는 80.8%까지 제거되는 것으로 나타났다. 동시에 Yeast two-hybrid assay로 각 내분비계장애물질의 농도-반응곡선으로부터 반응식을 구하여, 에스트로겐 활성에 미치는 각 물질의 기여도를 분석한 결과, 가정계의 활성슬러지법에 의한 처리수에서 estrone, 17β-estradiol, 17α-ethinylestradiol, APE가 각각 70.7, 23.3, 3.7, 2.32%로 나타났다. 즉, 생물학적 처리공정을 통해 배출된 처리수의 에스트로겐 활성에 영향을 미치는 주된 기여물질은 estrone과 17β-estradiol인 것으로 나타났다. Several endocrine disrupting chemicals(EDCs) were monitored to evaluate the estrogenic activities and the concentrations by yeast two-hybrid assay and enzyme-linked immunosorbent assay(ELISA) in sewage treatment plant(STP) which consist of industrial and domestic line. In the influent of domestic line, estrone, 17β-estradiol, 17α-ethinylestradiol and alkylphenolethoxylate(APE) were detected up to 167.1, 39.7, 7.3 and 145.4 ng/L, respectively. The average removal efficiency of 17β-estradiol after the activated sludge process was 77.5% and further removed to 80.8% after the sand filtration-ozonation step. These results suggests that the activated sludge process has limited potential to remove the estrogenic activity effectively. The contributions of the estrogenic chemicals to the estrogenic activities were 70.7, 23.3, 3.7 and 2.3% for estrone, 17β-estradiol, 17α-ethinylestradiol and APE, respectively, in the domestic line effluents. Therefore, 17β-estradiol and estrone contributed most of the estrogenic activity in the domestic line effluents.
에스트로겐과 다이옥신 수용체 효모를 이용한 내분비계장애 영향 평가
이병천 ( Byoung Cheun Lee ),김수진 ( Sue Jin Kim ),윤준헌 ( Jun Heon Yoon ),김은주 ( Eun Ju Kim ),( Duong NC ),엄익춘 ( Ig Chun Eom ),( Shiraishi F. ),최경희 ( Kyung Hee Choi ) 한국환경분석학회 2010 환경분석과 독성보건 Vol.13 No.2
Estrogenic and dioxin activities in river water and effluents of sewage treatment plants were evaluated by different in vitro assays. The expression of β-galatosidase in yeast cells (Yeast two-hybrid assay) was used as an indicator of pollutants effects. Estrogenic activities were estimated using medaka estrogen receptor (mER) and human estrogen receptor (hER) yeast while the dioxin activities were measured by Aryl hydrocarbon receptor (AhR) yeast. Five samples were collected including two river water (RW) samples and three effluents (MW) samples. Target chemicals were extracted using a series of solvents which are hexane/dichloromethane, acetone/dichloromethane and methanol. Standard dose-response curves were established with 3 estrogenic chemicals (estradiol, nonylphenol and bisphenol A) and 3 dioxin-like chemicals (naphthoflavone, comestrol and equol). The results showed that both estrogen receptors (i.e., mER and hER) were suitable to be applied into the assessment of micropollutants` estrogenicity, in which, the sensitivity of mER to estrogenic activity was higher than that of hER. AhR also showed a suitable sensitivity to target dioxins. Estrogenic activity of MW-1 was highest among the samples in both mER and hER yeast assays. The estrogenic activities of two other effluents samples, however, were lower than that of MW-1 and other RW samples. This indicates that river water may receive estrogenic-polluted water other than the effluents of STPs. Similarly, dioxin activities in RW samples were higher than that of effluents. It is suggested that the untreated agriculture wastewater could be the reason. This study contributes to the establishment and application of risk assessment of micropollutant in the environment.