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Analytical and Bioanalytical Chemistry

, Volume 407, Issue 26, pp 8109–8120 | Cite as

Development of a recombinant Arxula adeninivorans cell bioassay for the detection of molecules with progesterone activity in wastewater

  • Alexandre Chamas
  • Annabel Nieter
  • Ha Thi Minh Pham
  • Martin Giersberg
  • Karina Hettwer
  • Steffen Uhlig
  • Kirsten Simon
  • Keith Baronian
  • Gotthard KunzeEmail author
Research Paper

Abstract

This study describes the development of a bioassay to detect the presence of progesterone and progesterone-like molecules in wastewater samples. The basis of the bioassay is the integration of the human progesterone receptor gene into the yeast Arxula adeninivorans for the constitutive synthesis of the receptor. After incubation, binding of the analyte to the receptor induces the production of a reporter protein. Two reporter proteins were compared for detection parameters such as half-maximal activity (EC50), limit of detection (LoD) and limit of quantification (LoQ). When the extracellular phytase K was used, an EC50 value of 155 ng L−1 and a LoD of 27 ng L−1 progesterone were obtained after 4 h incubation, while use of the fluorescent dsRED as the reporter protein, resulted in an EC50 of 320 ng L−1 and a LoD of 65 ng L−1 after 20 h incubation. Use of phytase K as the reporter protein offers decreased incubation time and increased sensitivity; however the dsRED reporter system is less labor-intensive. Additionally, the affinity of known agonists and antagonists of the human progesterone receptor was determined. The utility of this bioassay was confirmed by measuring total progesterone equivalent concentration of samples from a wastewater treatment plant. The A. adeninivorans-based transactivation assay was able to measure concentrations of about 311 ng L−1 in the influent stream but could not detect progesterone activity in effluent. One key feature of the assay is the robustness of A. adeninivorans, which allows sample measurement without any sample preparation.

Keywords

Progesterone Arxula adeninivorans Transactivation assay Reed-bed wastewater treatment plant 

Notes

Acknowledgments

We would like to thank Ruth Franz for her technical support. The research work was supported by grant from the BMWi (Grant No. KF2131610MD0 and KF2131626SB4).

Conflict of interest

The authors declare no conflict of interest.

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Copyright information

© Springer-Verlag Berlin Heidelberg 2015

Authors and Affiliations

  • Alexandre Chamas
    • 1
  • Annabel Nieter
    • 1
  • Ha Thi Minh Pham
    • 1
    • 2
  • Martin Giersberg
    • 1
  • Karina Hettwer
    • 3
  • Steffen Uhlig
    • 3
  • Kirsten Simon
    • 4
  • Keith Baronian
    • 5
  • Gotthard Kunze
    • 1
    Email author
  1. 1.Leibniz Institute of Plant Genetics and Crop Plant Research (IPK)GaterslebenGermany
  2. 2.Institute of BiotechnologyVietnam Academy of Science and TechnologyCau GiayVietnam
  3. 3.QuoData GmbHDresdenGermany
  4. 4.New diagnostics GmbHFreisingGermany
  5. 5.School of Biological SciencesUniversity of CanterburyChristchurchNew Zealand

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