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Application of a tyrosinase microreactor – detector in a flow injection configuration for the determination of affinity and dynamics of inhibitor binding

par Vandeput, Marie ;Patris, Stéphanie ;Silva, Hugo;Parsajoo, Cobra ;Dejaegher, Bieke ;Arcos-Martínez, Julia Maria;Kauffmann, Jean-Michel
Référence Sensors and actuators B Chemical, 248, page (385-394)
Publication Publié, 2017-04-04

Article révisé par les pairs

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Résumé :

An original self-contained enzymatic microreactor − detector in a thin layer flow-through configuration has been developed for the screening of tyrosinase inhibitors. The microreactor- detector consisted of a gold disk for tyrosinase immobilization adjacent to a glassy carbon disk as working electrode (GCE). Using L-tyrosine as substrate, the enzymatic product formed (dopaquinone) was detected amperometrically at the GCE. Experimental parameters most affecting the L-tyrosine response were optimized with a central composite design. Measurements were performed in phosphate buffer pH 6.0 with an applied potential of +50 mV vs Ag/AgCl and a flow rate of 100 μL/min. The determined Kmapp was 1.9 ± 0.1 mM. The setup permitted the determination of the efficiency and the duration of inhibition of twelve compounds known as tyrosinase inhibitors namely kojic acid, gallic acid, azelaic acid, benzoic acid, L-ascorbic acid, hydroquinone, glabridine, resveratrol, quercetin dihydrate, captopril, L-cysteine and L-glutathione reduced. L-Ascorbic acid and kojic acid were found to be the most potent inhibitors for the two studied criteria with IC50 of 32 μM and 156 μM, respectively.