Hydrogenases are the key enzymes for hydrogen metabolism in many microorganisms. Due to the high efficiency and specificity they developed for H 2 oxidation, they have been recently used as biocatalysts for an efficient H 2 /O 2 biofuel cell. In this work we explore new properties of an O 2 -, CO-and T8-resistant hydrogenase. Enzyme immobilization on carbon nanotube modified electrodes is studied and optimized for long-term stabilization of the direct hydrogen catalytic oxidation process. The role of co-immobilized redox mediator is finally discussed in view of H 2 /O 2 fuel cell performance enhancement.