We report on the fabrication and characterisation of mediator-less sugar and oxygen sensitive biodevices based on three-dimensonal gold nanoparticle-modified electrodes with immobilised sugar oxidising and oxygen reducing enzymes. To create sugar and oxygen sensitive biodevices, Corynascus thermophilus cellobiose dehydrogenase (CtCDH) and Myrothecium verrucaria bilirubin oxidase (MvBOx) were used, respectively. For CtCDH modified bioelectrodes maximal current densities of 28 µA cm^-2 and 40 µA cm^-2 could be obtained in the presence of 120 mM glucose and 5 mM lactose, respectively. It was shown that bioelectrocatalytic oxidation of sugars on CtCDH-based electrodes was limited by the  activity of the enzyme. Contrary, in the case of MvBOx modified biodevices, a maximal current density equal to 110 µA cm^-2 was obtained in air saturated solution due to mass transfer limitation.

By connecting the electrodes together a membrane- and mediator-less sugar/oxygen biofuel cell (BFC) was created operating in buffers and human physiological liquids. The following characteristics of the mediator-, separator- and membrane-less, miniature BFC, were obtained: open-circuit voltages of 0.68 and 0.65 V, maximum power densities of 15 mW cm^-2 and 3 mW cm^-2 at 0.52 V and 0.45 V of cell voltage, in phosphate buffer and human blood, respectively. The estimated half-lifes of biodevices were measured to be 24 h and 8 h in sugar-containing buffers and human physiological liquids, respectively. Thus, a mediatorless sugar/oxygen BFC with significantly improved basic characteristics compared to previously designed biodevices^1,2 could be constructed because of the usage of three-dimensional gold nanoparticle-modified electrodes.

1. V. Coman, R. Ludwig, W. Harreither, D. Haltrich, L. Gorton, T. Ruzgas, and S. Shleev. (2010) A direct electron transfer-based glucose/oxygen biofuel cell operating in human serum. Fuel Cells. 10(1), 9-16.2. V. Coman, C. Vaz-Domínguez, R. Ludwig, W. Harreither, D. Haltrich, A. L. De Lacey, T. Ruzgas, L. Gorton and S. Shleev. (2008) A membrane-, mediator-, cofactor-less glucose/oxygen biofuel cell. Phys. Chem. Chem. Phys. 10(40), 6093–6096.

Acknowledgements: The authors thank Amano Enzyme Inc. for Amano 3 preparation of M. verrucaria bilirubin oxidase. The authors would like to also thank Petri Gudmundsson and Karin Nilsson (Malmö University, Sweden) for their kind help with blood collection. Ms. Wang would like to thank the Finnish Graduate School of Chemical Engineering (GSCE) for the financial support to her PhD study and the scholarship for her research visit at Lund University in 2010. The work has been supported financially by the European Commission (FP7 project NMP4-SL-2009-229255) and the Swedish Research Council (projects 2007-4124, 2009-3266 and 2010-5031).

1. PRESENTATION: Gold nanoparticle-modified enzyme-based sugar and oxygen sensitive electrodes for biosensing and biofuel cell applications, Microsoft Office Document, 0MB

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