Effect of Deglycosylation of Cellobiose Dehydrogenases on the Enhancement of Direct Electron Transfer with Electrodes

Abstract

Cellobiose dehydrogenase (CDH) is a monomeric extracellular flavocytochrome composed of a catalytic dehydrogenase domain (DH_CDH) containing flavin adenine dinucleotide (FAD), a cytochrome domain (CYT_CDH) containing heme <i>b</i>, and a linker region connecting the two domains. In this work, the effect of deglycosylation on the electrochemical properties of CDH from Phanerochaete chrysosporium (<i>Pc</i>CDH) and Ceriporiopsis subvermispora (<i>Cs</i>CDH) is presented. All the glycosylated and deglycosylated enzymes show direct electron transfer (DET) between the CYT_CDH and the electrode. Graphite electrodes modified with deglycosylated <i>Pc</i>CDH (d<i>Pc</i>CDH) and <i>Cs</i>CDH (d<i>Cs</i>CDH) have a 40–65% higher <i>I</i>_max value in the presence of substrate than electrodes modified with their glycosylated counterparts. <i>Cs</i>CDH trapped under a permselective membrane showed similar changes on gold electrodes protected by a thiol-based self-assembled monolayer (SAM), in contrast to <i>Pc</i>CDH for which deglycosylation did not exhibit any different electrocatalytical response on SAM-modified gold electrodes. Glycosylated <i>Pc</i>CDH was found to have a 30% bigger hydrodynamic radius than d<i>Pc</i>CDH using dynamic light scattering. The basic bioelectrochemistry as well as the bioelectrocatalytic properties are presented