The layer by layer method [1] become important for preparation of bioelectrocatalytic electrode. However, only recently it was applied to obtain biocathodes potentially applicable in biofuel cell [2, 3]. Our group shown that the nanoparticulate film electrode prepared by layer by layer method combined with sol-gel process exhibits mediated bioelectrocatalysis with laccase dissolved in electrolyte. The electrode modified by film assembled from osmium redox polymer and laccase layers also catalyses dioxygen reduction .
         Here we will show bioelectrocatalytic behaviour of earlier proposed electrodes modified with adsorbed bilirubin oxidase from Myrothecium sp. (EC 1.3.3.5). Clearly mediatorless bioelectrocatalysis indicates efficient electron exchange between the enzyme and carbon nanoparticles [4]. The catalytic current is significantly larger if mediator: 2,2’-azino-bis(3-ethylbenzothiazoline-6-sulfonate) is preadsorbed at the electrode. One can conlude that in the latter case two processes mediated and mediatorless occurs simultaneously. The bioelectrocatalytic current is proportional to the number of immersion and withdrawal steps in electrode preparation procedure.

[1] G. Decher, J.D. Hong, Makromol. Chem. Macromol. Symp. 46 (1991) 321.
[2] K. Szot, A. Lesniewski, J. Niedziolka, M. Jönsson, C. Rizzi, L. Gaillon, F. Marken, J. Rogalski, M. Opallo, J. Electroanal. Chem. 623 (2008) 170.
[3] R. Szamocki, V. Flexer, L. Levin, F. Forchiasin, E.J. Calvo, Electrochim. Acta 54 (2009) 1970.
[4] K. Szot, W. Nogala, J. Niedziolka-Jönssona, M. Jönsson-Niedziolka, F. Marken, J. Rogalski, C. Nunes Kirchner, G. Wittstock, M. Opallo, Electrochim. Acta 54 (2009) 4620.

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