Increasing concern about the fate of 17alpha-ethinylestradiol (EE2) in the environment stimulates the search for alternative methods for wastewater treatment plant (WWTP) effluent polishing. The aim of this study was to establish an innovative and effective biological removal technique for EE2 by means of a nitrifier enrichment culture (NEC) applied in a membrane bioreactor (MBR). In batch incubation tests, the microbial consortium was able to remove EE2 from both a synthetic minimal medium and WWTP effluent. A maximum EE2 removal rate of 9.0 microg EE2 g(-1)biomass-VSS h(-1) was achieved (>94% removal efficiency). Incubation of the heterotrophic bacteria isolated from the NEC did not result in a significant EE2 removal, indicating the importance of nitrification as driving force in the mechanism. Application of the NEC in a MBR to treat a synthetic influent with an EE2 concentration of 83 ng EE2 L(-1) resulted in a removal efficiency of 99% (loading rates up to 208 ng EE2 L(-1)d(-1); membrane flux rate: 6.9 L m(-2) h(-1)). Simultaneously, complete nitrification was achieved at an optimal ammonium influent concentration of 1.0 mg NH(4)(+)-N L(-1). This minimal NH(4)(+)-N input is very advantageous for effluent polishing since the concomitant effluent nitrate concentrations will be low as well and it offers opportunities for the nitrifying MBR as a promising add-on technology for WWTP effluent polishing.