Novel bacterial bioassay for a high-throughput screening of 4-hydroxyphenylpyruvate dioxygenase inhibitors |
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Authors: | Emilie Rocaboy-Faquet Thierry Noguer Sana Romdhane Cédric Bertrand Franck Emmanuel Dayan Lise Barthelmebs |
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Affiliation: | 1. Institut de Modélisation et d’Analyse en Géo-Environnement et Santé, Université Perpignan Via Domitia, EA 4218, 66860, Perpignan, France 2. Laboratoire de Chimie des Biomolécules et de l’Environnement, Université Perpignan Via Domitia, EA 4215, 66860, Perpignan, France 3. Natural Products Utilization Research Unit, ARS, United States Department of Agriculture, University of Mississippi, Oxford, MS, 38677, USA
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Abstract: | Plant 4-hydroxyphenylpyruvate dioxygenase (HPPD) is the molecular target of a range of synthetic β-triketone herbicides that are currently used commercially. Their mode of action is based on an irreversible inhibition of HPPD. Therefore, this inhibitory capacity was used to develop a whole-cell colorimetric bioassay with a recombinant Escherichia coli expressing a plant HPPD for the herbicide analysis of β-triketones. The principle of the bioassay is based on the ability of the recombinant E. coli clone to produce a soluble melanin-like pigment, from tyrosine catabolism through p-hydroxyphenylpyruvate and homogentisate. The addition of sulcotrione, a HPPD inhibitor, decreased the pigment production. With the aim to optimize the assay, the E. coli recombinant clone was immobilized in sol–gel or agarose matrix in a 96-well microplate format. The limit of detection for mesotrione, tembotrione, sulcotrione, and leptospermone was 0.069, 0.051, 0.038, and 20 μM, respectively, allowing to validate the whole-cell colorimetric bioassay as a simple and cost-effective alternative tool for laboratory use. The bioassay results from sulcotrione-spiked soil samples were confirmed with high-performance liquid chromatography. |
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