Electrochemical detection of the 2-isobuty1-3-methoxypyrazine model odorant based on odorant-binding proteins: the proof of concept - Université de Bourgogne Accéder directement au contenu
Article Dans Une Revue Bioelectrochemistry Année : 2015

Electrochemical detection of the 2-isobuty1-3-methoxypyrazine model odorant based on odorant-binding proteins: the proof of concept

Résumé

We developed an electrochemical assay for the detection of odorant molecules based on a rat odorant-binding protein (rOBP3). We demonstrated that rOBP3 cavity binds 2-methyl-1,4-naphtoquinone (MNQ), an electrochemical probe, as depicted from the decrease of its electrochemical signal, and deduced the dissociation constant, K-dMNQ = 0.5(+/- 0.2) mu M. The amount of MNQ displaced from rOBP3 by 2-isobuty1-3-methoxypyrazine (IBMP), a model odorant molecule, was measured using square-wave voltammetry. The release of MNQ by competition led to an increase of the electrochemical response. In addition, this method allowed determination of the dissociation constant of rOBP3 for IBMP, Kd(IBMP) = 0.5(+/- 0.1) mu M. A negative control was performed with a non-binding species, caffeic add (CA). The determined binding affinity values were confirmed using a fluorescent competitive binding assay and isothermal titration microcalorimetry. This electrochemical assay opens the way for designing robust, reliable and inexpensive odorant biosensors.
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Dates et versions

hal-01213726 , version 1 (08-10-2015)

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Emilie Barou, Maud Sigoillot, Marcel Bouvet, Loïc Briand, Rita Meunier-Prest. Electrochemical detection of the 2-isobuty1-3-methoxypyrazine model odorant based on odorant-binding proteins: the proof of concept. Bioelectrochemistry, 2015, 101, pp.28-34. ⟨10.1016/j.bioelechem.2014.06.006⟩. ⟨hal-01213726⟩
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