Novel bacterial bioassay for a high-throughput screening of 4-hydroxyphenylpyruvate dioxygenase inhibitors

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.
Document type :
Journal articles
Complete list of metadatas

https://hal-univ-perp.archives-ouvertes.fr/hal-01174775
Contributor : Olivier Savoyat <>
Submitted on : Thursday, July 9, 2015 - 5:07:36 PM
Last modification on : Thursday, January 11, 2018 - 6:23:12 AM

Identifiers

Collections

Citation

Emilie Rocaboy-Faquet, Thierry Noguer, Sana Romdhane, Cédric Bertrand, Franck Emmanuel Dayan, et al.. Novel bacterial bioassay for a high-throughput screening of 4-hydroxyphenylpyruvate dioxygenase inhibitors. Applied Microbiology and Biotechnology, Springer Verlag, 2014, 98 (16), pp.7243-7252. ⟨10.1007/s00253-014-5793-5⟩. ⟨hal-01174775⟩

Share

Metrics

Record views

251