1,4‐Benzoquinone Derivatives for Enhanced Bioelectrocatalysis by Fructose Dehydrogenase from Gluconobacter Japonicus: Towards Promising D‐Fructose Biosensor Development

Abstract

D‐fructose amount in food and beverages should be carefully controlled in order to avoid its overconsumption by human, which can lead to various metabolic diseases. Thus, the development of a low‐cost and portable (bio)sensors, which realize the on‐site monitoring of D‐fructose, is still highly required. In this work, we proposed several reagentless bioelectrochemical systems (BioESs) based on fructose dehydrogenase EC1.1.99.11 from Gluconobacter japonicus (FDH) and on 2‐arylamine‐1,4‐benzoquinone (ABQ) derivatives as a platform for the development of D‐fructose electrochemical biosensor. To design a reliable and easily reproducible BioES, we used the simple physical sorption as the electrode modification strategy for ABQs and FDH immobilization that is suitable for low‐cost mass production of the biosensors by standard microfabrication techniques. To choose the most suitable ABQ compounds for BioESs design, we proposed a novel theoretical approach of potential ET mediator evaluation through its electrochemical properties. A set of newly synthesized and of previously applied ABQs was characterized both electrochemically and using the density functional theory (DFT). It was shown that results obtained by quantum chemical analysis were in a good agreement with the ABQ characteristics obtained in electrochemical measurements. We suggest that the calculated local ionization energy values and theoretical redox potential obtained by DFT are a powerful tool for prediction of ABQs electrochemical properties. The performance of the BioESs with FDH under study was determined by electrochemical and electronic properties of ABQ derivatives and FDH orientation and stabilization on the electrode surface. The most promising BioES was based on carbon paste electrodes and 2‐(3‐nitro(phenyl)amino)‐ cyclohexa‐2,5‐dien‐1,4‐dione as ET mediator, which accelerated FDH catalysis and improved its stability better, then other studied ABQs. The bioelectrocatalytic process was characterized by initial apparent maximum current density of 7.1 μA cm−2 at the optimal conditions (+400 mV vs. Ag/AgCl, McIlvaine's buffer, pH 5.0 and 20 °C). The findings of this research open new possibilities for development of cost‐effective biosensors with FDH and, potentially, with other redox enzymes.

Bioamperometric Systems with Fructose Dehydrogenase From Gluconobacter japonicus for D‐Tagatose Monitoring

“…The elimination of compound 1 provided means of the correlating exponential functions with better statistic parameters. Our previous work [14] revealed that the BioAS with compound 1 exhibited extremely high reactivity in the case of oxidation of D‐fructose whilst in the case of D‐tagatose the same BioAS exhibited a moderate response. Such a difference shows that the compound 1 interacts with the active center of the enzyme and/or substrate intermolecularly in the way that high FDH reactivity in D‐tagatose oxidation is not achieved.…”

“…D‐Fructose is structurally similar to D‐tagatose, and both sugars differ only in the disposition of one hydroxyl group at the 4 th carbon atom [3a]. To check if FDH is able to catalyze the oxidation of D‐tagatose, we chose recently introduced by our research group reagentless bioamperometric systems (BioASs) with FDH [14].…”

“…ABQs were synthesized following protocols described here [15]. The BioASs were made following consequent modification of the carbon paste electrodes (CPE, Ø 2.4 mm, made in the Institute of Biochemistry, Life Sciences Center, Vilnius University) by the ABQs and FDH (from Gluconobacter industrius (japonicus) , E.C.1.1.99.11, which was purchased from “Sigma‐Aldrich” (Germany)) as described elsewhere [14]. The obtained BioESs were stored in McIlvaine's buffer (pH 4.6) at 4 °C between the experiments.…”

“…The BioASs operate on the basis of mediated electron transfer, using 2-arylamine-1,4-benzoquinone derivatives (ABQs) (Figure S1) [14][15] as electron transfer mediators. In our previous study [14], we demonstrated that the FDH catalysis in these BioASs depended not only on electrochemical properties of ABQ derivatives, but also on the differences in FDH orientation on the electrode surface, modified with ABQ compounds.…”

Multisensor Systems and Arrays for Medical Applications Employing Naturally-Occurring Compounds and Materials

“…Both organic and inorganic compounds can serve as mediators. Although redox mediation is well known, in recent years, several new types such mediators were designed and studied [ 62 , 63 ]. As stated above, mediators are also used in MSAs and MSSs [ 43 , 59 , 64 ].…”