Microbial fuel cells (MFCs) are devices that use microorganisms to produce electricity. Pseudomonas aeruginosa is a rod-shaped bacterium and the only bacterial species that produces pyocyanin (PYO). In MFCs, the pyocyanin can play an important role as an electron shuttle. PYO production can be improved in glycerol enriched culture media. Glycerol has become a huge market problem because it is de main biodiesel byproduct. Its use as fuel in MFC is being promising. In the present study we evaluated in a MFC the current output achieved by pure cultures of P. aeruginosa cultivated in King broth medium using Nafion® 117 as proton exchange membrane. Results have shown that a maximum current output of 15.3 μAcm-2 can be achieved when using the chosen culture medium. In parallel, experiments involving the oxygen reduction reaction showed that Nafion® membranes can lose the activity due to the medium composition.