A new approach is described to produce nanostructured electrocatalytic membranes using a combination of three methods. Using the layer-by-layer technique, PAMAM dendrimers with cobalt hexacyanoferrates-modified gold nanoparticles were alternated with poly(vinylsulfonic acid) layers on ITO (indium tin oxide) electrodes. This film was used as substrate for immobilization of glucose oxidase in the presence of bovine serum albumin and glutaraldehyde as cross-linker. The modified electrode was successfully applied as a biosensor for the amperometric measurement of glucose, using glucose oxidase enzyme, at 0.0V vs. SCE.