Evaluation and validation of a novel Arxula adeninivorans estrogen screen (nAES) assay and its application in analysis of wastewater, seawater, brackish water and urine

Kaiser, Christian; Uhlig, Steffen; Gerlach, Torsten; Körner, Martina; Simon, Kirsten; Kunath, Kirstin; Florschütz, Kristina; Baronian, Keith; Kunze, Gotthard

doi:10.1016/j.scitotenv.2010.08.050

Cited by 27 publications

(28 citation statements)

References 52 publications

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“…The four parameters A, B, C, and D indicate the unknown model parameters, which have to be estimated. A represents the mean value of the corrected extinction effects y ij without progesterone effects (at spike level 0, bottom curve point), D is the mean value of y ij with maximum progesterone effect (curve plateau), C describes the mean effect concentration (curve point of inflection-EC 50 ) at which the progesterone effect attains half of its maximum value (A+D)/2, and the parameter B is proportional to the slope at C [25]. Calculation of these parameters and the statistical analysis was carried out using BioVal® software (QuoData GmbH, Dresden, Germany).…”

Section: Calculation Of the Progesterone Activity And Statistical Anamentioning

confidence: 99%

“…Some in vitro whole cell biosensors have focused on estrogen-or androgen-related compounds, for example the YES assay (using S. cerevisiae as the detection element) [24] or the A-YES assay (using A. adeninivorans) [25] and their counterparts such as YAS [26,27] and A-YAS assays for androgen binding substances [28]. For progesterone detection, similar assays have been developed using mammalian or yeast cells as host for the production of the progesterone receptor and an enzyme or a fluorescent protein as reporter [29][30][31][32][33][34].…”

Section: Introductionmentioning

confidence: 99%

“…The effect of a wide range of substances as well as the antagonistic properties of some EDCs was then investigated. Because a potential application of the assay is in water analysis, different samples from a wastewater treatment plant (WWTP) were assayed without extraction of the compounds to demonstrate the utility of the system and the robustness of A. adeninivorans [37,25]. …”

Section: Introductionmentioning

confidence: 99%

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

et al. 2015

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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.

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Section: Calculation Of the Progesterone Activity And Statistical Anamentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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

et al. 2015

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“…A. adeninivorans G1212/YRC102-hER␣-phyK was engineered to co-express the human estrogen receptor (hER˛) gene and the inducible phytase (phyK, derived from Klebsiella sp. ASR1) reporter gene under control of a promoter containing estrogen response elements (EREs) [6]. In the presence of estrogenic substances, such as 17␤-estradiol (E2), the phyK gene is expressed and recombinant phytase is secreted into the media.…”

Section: Introductionmentioning

confidence: 99%

“…The yeast can grow in minimal and rich media supplemented up to 20% NaCl without any changes in physiology [7,8]. Kaiser et al also reported that the robustness of A. adeninivorans cells enable measurements in samples with up to 5% NaCl [6].…”

Section: Introductionmentioning

confidence: 99%

Application of modified Arxula adeninivorans yeast cells in an online biosensor for the detection of estrogenic compounds in wastewater samples

Pham

Kunath²,

Gehrmann

et al. 2013

Sensors and Actuators B: Chemical

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Simultaneous detection of three sex steroid hormone classes using a novel yeast‐based biosensor

Chamas

Pham

Jähne³

et al. 2017

Biotech & Bioengineering

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A biosensor detecting estrogens, progestogens, and androgens in complex samples and in a single step is described. Three Arxula adeninivorans yeast strains were created, each strain producing a different recombinant human hormone receptor and a different fluorescent reporter protein. These strains were then mixed to create G1212/YRC102-hHR-fluo, the biological component of the biosensor. During incubation with G1212/YRC102-hHR-fluo, hormones present in a sample bind to their target receptor, which leads to the production of a specific fluorescent protein. Three fluorescence scans of the yeast suspension determine which fluorescence protein has been produced, thus revealing which hormone receptor (estrogen, progesterone, and androgen) has been activated by the hormones or hormone mimics present in the sample. The biosensor has similar sensitivities to the existing A. adeninivorans cell-based assays. The detection of the three hormone classes in one single experiment reduces the labor and time required to assay for the three hormone classes. The biosensor was also trialed with animal serum samples for the detection of progestogens, androgens, and estrogens and gave results that correlated well with ELISA analysis in case of progestogens. These results highlight the potential usefulness of the biosensor for comprehensive determination of hormone status in samples from veterinary origin. Biotechnol. Bioeng. 2017;114: 1539-1549. © 2017 Wiley Periodicals, Inc.

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Evaluation and validation of a novel Arxula adeninivorans estrogen screen (nAES) assay and its application in analysis of wastewater, seawater, brackish water and urine

Cited by 27 publications

References 52 publications

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

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

Application of modified Arxula adeninivorans yeast cells in an online biosensor for the detection of estrogenic compounds in wastewater samples

Simultaneous detection of three sex steroid hormone classes using a novel yeast‐based biosensor

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