Electrodes for non-enzymatic glucose sensing based on gold nanostructures onto graphene paper (GP-AuNPs) have been obtained inducing dewetting, by laser annealing, of 8 nm-thick Au layer deposited by sputtering. Molten-phase dewetting of gold layer, which produces the formation of spherical nanoparticles (AuNPs), was achieved by nanosecond laser annealing using a pulsed (12 ns) Nd: yttrium aluminum garnet YAG laser operating at 532 nm. The Surface of the electrode presents gold rich regions consisting of graphene nanoplatelets covered by spherical AuNPs. The sizes of AuNPs are in the range of 10–150 nm. Glucose was detected at a potential of 0.17 V vs SCE, which corresponds to the intense peak of two electrons oxidation. Highest sensitivity of 600 µA mM−1 cm−2 of glucose detection was obtained. The resulting sensitivity, detection limit and linear range of glucose detection are very promising since comparable to the actual state of art results for nanostructured gold electrodes which are, however, produced by complex multi-steps processes.