Bilirubin oxidase bound to multi-walled carbon nanotube-modified gold

We report on direct electron transfer (DET) reactions of bilirubin oxidase at multi-walled carbon nanotube-(MWCNT) modified gold electrodes. MWCNTs are very suitable for protein immobilisation and provide surface groups that can be used for the stable fixation on electrodes. They can also effectively replace the natural substrate of BOD - bilirubin, the electron donor for oxygen reduction. The bioelectrocatalytic oxygen reduction was recorded using linear sweep voltammetry (LSV) with BOD covalently linked to the nanotubes. The start potential of the bioelectrocatalytic oxygen reduction at pH 7 and a scan rate of 10 mV/s was determined to be 485 +/- 10 mVvs. Ag/AgCl, 1 M KCl (720 mV vs. SHE). Current densities up to 500 mu A/cm(2) were detected in an air-saturated buffer at room temperature (25 +/- 5 degrees C). Experiments with a rotating disk electrode (RDE) indicate a diffusion controlled electrode reaction. A k(s) value in the range of 80-100s(-1) could be estimated. The DET could also be observed directly by the redox conversion of a copper centre of BOD under anaerobic conditions. A peak pair with a formal potential of 680 +/- 10 mV vs. SHE was found. The T1 site is probably addressed by the electrode as indicated by several experimental studies. (C) 2008 Elsevier Ltd. All rights reserved.