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Analysis of N-acylhomoserine lactone dynamics in continuous cultures of Pseudomonas putida IsoF by use of ELISA and UHPLC/qTOF-MS-derived measurements and mathematical models

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Abstract

In this interdisciplinary approach, the dynamics of production and degradation of the quorum sensing signal 3-oxo-decanoylhomoserine lactone were studied for continuous cultures of Pseudomonas putida IsoF. The signal concentrations were quantified over time by use of monoclonal antibodies and ELISA. The results were verified by use of ultra-high-performance liquid chromatography. By use of a mathematical model we derived quantitative values for non-induced and induced signal production rate per cell. It is worthy of note that we found rather constant values for different rates of dilution in the chemostat, and the values seemed close to those reported for batch cultures. Thus, the quorum-sensing system in P. putida IsoF is remarkably stable under different environmental conditions. In all chemostat experiments, the signal concentration decreased strongly after a peak, because emerging lactonase activity led to a lower concentration under steady-state conditions. This lactonase activity probably is quorum sensing-regulated. The potential ecological implication of such unique regulation is discussed.

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Acknowledgments

We enormously appreciated the great dedication to immunochemical analysis of AHL compounds of Dr Petra Krämer, who died unexpectedly in the early phase of these experiments. Furthermore we want to thank Dr Elisabeth Kremmer, Institute of Molecular Immunology, Helmholtz Zentrum München, for providing the monoclonal antibodies. Maria Vittoria Barbarossa was supported by ERC starting grant no. 259559.

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Authors and Affiliations

  1. Research Unit Microbe-Plant Interactions, Helmholtz Zentrum München, German Research Centre for Environmental Health (GmbH), Ingolstädter Landstrasse 1, 85764, Neuherberg, Germany

    Katharina Buddrus-Schiemann, Martin Rieger, Marlene Mühlbauer, Michael Rothballer, Michael Schmid & Anton Hartmann

  2. Bolyai Institute, University of Szeged, Aradi vértanúk tere 1, 6720, Szeged, Hungary

    Maria Vittoria Barbarossa

  3. Centre for Mathematical Sciences, Technische Universität München, Boltzmannstrasse 3, 85748, Garching, Germany

    Christina Kuttler

  4. Institute of Computational Biology, Helmholtz Zentrum München, German Research Centre for Environmental Health (GmbH), Ingolstädter Landstrasse 1, 85764, Neuherberg, Germany

    Burkhard A. Hense

  5. Research Unit Analytical BioGeoChemistry, Helmholtz Zentrum München, German Research Centre for Environmental Health (GmbH), Ingolstädter Landstrasse 1, 85764, Neuherberg, Germany

    Jenny Uhl, Juliano R. Fonseca & Philippe Schmitt-Kopplin

  6. Analytical Food Chemistry, Technische Universität München, Alte Akademie 10, 85354, Freising-Weihenstephan, Germany

    Philippe Schmitt-Kopplin

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  1. Katharina Buddrus-Schiemann
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  2. Martin Rieger
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  3. Marlene Mühlbauer
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Correspondence to Anton Hartmann.

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Buddrus-Schiemann, K., Rieger, M., Mühlbauer, M. et al. Analysis of N-acylhomoserine lactone dynamics in continuous cultures of Pseudomonas putida IsoF by use of ELISA and UHPLC/qTOF-MS-derived measurements and mathematical models. Anal Bioanal Chem 406, 6373–6383 (2014). https://doi.org/10.1007/s00216-014-8063-6

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