Effect-Targeted Mapping of Potential Estrogenic Agonists and Antagonists in Wastewater via a Conformation-Specific Reporter-Mediated Biosensor
Jisui Tan, Fangxu Li, Lanhua Liu, Jing Zhang, Ping Gui, Miao He, Xiaohong Zhou
Abstract
Wastewater treatment plants (WWTPs) are regarded as the main sources of estrogens that reach the aquatic environment. Hence, continuous monitoring of potential estrogenic-active compounds by a biosensor is an appealing approach. However, existing biosensors cannot simultaneously distinguish and quantify estrogenic agonists and antagonists. To overcome the challenge, we developed an estrogen receptor-based biosensor that selectively screened estrogenic agonists and antagonists by introducing rationally designed agonist/antagonist conformation-specific reporters. The double functional conformation-specific reporters consist of a Cy5.5-labeled streptavidin moiety and a peptide moiety, serving as signal recognition and signal transduction elements. In addition, the conformation recognition mechanism was further validated at the molecular level through molecular docking. Based on the two-step “turn-off” strategy, the biosensor exhibited remarkable sensitivity, detecting 17β-estradiol-binding activity equivalent (E 2 -BAE) at 7 ng/L and 4-hydroxytamoxifen-binding activity equivalent (4-OHT-BAE) at 91 ng/L. To validate its practicality, the biosensor was employed in a case study involving wastewater samples from two full-scale WWTPs across different treatment stages to map their estrogenic agonist and antagonist binding activities. Comparison with the yeast two-hybrid bioassay showed a strong liner relationship ( r 2 = 0.991, p < 0.0001), indicating the excellent accuracy and reliability of this technology in real applications.