2.9. Mise au point d'outils de criblage de biocatalyseurs de réactions de Diels-Alder bimoléculaire
W. Ghattas (en collaboration avec laboratoire COBRA-Rouen, LabEx CHARM3AT et LabEx Synorg)
There are no easy-to-use rapid identification techniques of systems capable of catalyzing Diels-Alder reactions. To this end, we develop sequential reactions to detect the formation of cycloadducts through a chemoselective post-cycloaddition labelling strategy with a fluorogenic probe. Selected reactions must meet specific requirements in order to detect Diels-Alderases in situ: 1) Diene (S1) and dienophile (S2) substrates should react in presence of the enzyme (Reaction 1) to provide substrate S3, which should react subsequently with the substrate S4 to generate fluorescent product P (Reaction 2). For the success of this analytical strategy, it is important to avoid any false positives hence, neither S1 nor S2 should be able to react neither with S3 nor with S4. Such a post-cycloaddition labelling method interferes as little as possible with the catalytic process.