Mediatorless electron transfer in glucose dehydrogenase/laccase system adsorbed on carbon nanotubes

Abstract

A mediatorless electron transfer in the chain of glucose dehydrogenase (GDH) and laccase (LAC) catalysing the oxidation of glucose by molecular oxygen was studied. To demonstrate mediatorless processes, the GDH from Ewingella americana was adsorbed on single-walled carbon nanotubes (SWCNT). The effective mediatorless oxidation of glucose proceeded at 0.2–0.4V vs. SCE. The electrode was most active at pH 6.1, and generated 0.8mAcm−2 biocatalytic current in the presence of 50mM glucose. The electrode showed a bell-shaped pH dependence with pKa values of 4.1 and 7.5. LAC from Trichaptum abietinum adsorbed on SWCNT exhibited mediatorless oxygen reduction at electrode potential less than 0.65V. The electrode was most active at pH 3.0–4.0 and generated 1.1mAcm−2 biocatalytic current in the presence of 0.254mM oxygen, with an apparent pKa of 1.0 and 5.4. The electrodes prepared by simultaneous adsorption of GDH and LAC on SWCNT exhibited glucose oxidation at a potential higher than 0.25V.The oxygen consumption in the chain was demonstrated using a Clark-type oxygen electrode. The dependence of oxygen consumption on glucose and lactose concentrations as well as activity of the system on pH were measured. A model of the pH dependence as well as mediatorless consecutive glucose oxidation with oxygen catalysed by LAC/GDH system is presented. This work provides a novel approach towards the synthesis of artificial multi enzyme systems by wiring oxidoreductases with SWCNT, and offers a better understanding of natural electron transfer networks and chains.