Effect of deglycosylation of cellobiose dehydrogenase applied to 3rd generation biosensors and biofuel cells

Roberto Ortiz 1Hirotoshi Matsumura 1,2Kiyohiko Igarashi 2Federico Tasca 1Roland Ludwig 3Lo Gorton 1

1. Department of Analytical Chemistry and Biochemistry, Lund University,, P.O Box 124,, Lund SE-22100, Sweden
2. The University of Tokyo, Dept. Biomat. Sci., Grad. Sch. of Agri. and Life Sci., Bunkyo-ku, Tokyo 113-8657, Japan
3. BOKU-University of Natural Resources and Life Sciences, Institute of Food Technology, Muthgasse 18, Vienna 1190, Austria

Abstract
Cellobiose dehydrogenase (CDH) is an extracellular highly glycosylated two domain redox enzyme, which is one of the very few redox enzymes that shows direct electron transfer (DET) properties with electrodes. The catalytically active domain contains flavin adenine dinucleotide (FAD) and the other contains heme b. For the native glycosylated enzyme only the heme domain shows DET properties reflected by that the redox conversion of the heme group is shown in cyclic voltammetry (CV). For the native enzyme DET of the FAD domain is not show. Recently we initiated studies of deglycolysated CDH aiming at obtaining more efficient DET properties in line with previous reports for horseradish peroxidase [1,2] and glucose oxidase [3]. Ceriporiopsis subvermispora (Cs) and Phanerochaete chrysosporium (Pc) CDH were used in this study. When deglycosylated CDH was used instead CV revealed signals from both the heme and FAD. However, in the presence of substrate catalytic currents in CV only emanate at potentials close that of the heme for both CDHs.

When investigated adsorbed on graphite electrodes CsCDH (16% glycosylation) its deglycosylated equivalent shows at least 3 times as high catalytic currents as its glycosylated counterpart. PcCDH (9% glycosylation) shows twice higher currents for its deglycosylated form. A similar behavior is observed on thiol modified gold electrodes for CsCDH but not for PcCDH. In this last case both the glycosylated and deglycosylated variants show equally high catalytic currents. The improvement in current response on graphite is due to a higher amount of deglycosylated enzyme immobilized on the graphite electrode. The basic bioelectrochemistry as well as the bioelectrocatalytic properties will be shown as well as applications of CDH modified electrodes as 3rd generation biosensors and as bioanodes in biofuel cells.

 

[1] G. Presnova, V. Grigorenko, A. Egorov, T. Ruzgas, A. Lindgren, L. Gorton, T. Börchers, Faraday Discus., 116 (2000) 281-289.

[2] E. E. Ferapontova, V. G. Grigorenko, A. M. Egorov, T. Börchers, T. Ruzgas, L. Gorton, Biosens. Bioelectron., 16 (2001) 147 - 157.

[3] O. Courjean, F. Gao, N. Mano, Angew. Chem. Int. Ed., 2009, 48, 5897 –5899.

 

This work was financially supported by the European Commission, “3D-Bionanodevice” NMP4-SL-2009-229255.
 

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Presentation: Poster at SMCBS'2011 International Workshop, by Lo Gorton
See On-line Journal of SMCBS'2011 International Workshop

Submitted: 2011-08-31 17:42
Revised:   2011-08-31 17:43