Graphene, is a carbon allotrope, which is widely used as a substrate for various catalysts due to its interesting physicochemical properties. In the present study, graphene oxide sheets were prepared from graphite, then, the graphene oxide surface was modified by a low-temperature method using sulfur and copper atoms to obtain pseudo-enzyme Cu/S/Graphene prosthetic group. The current density passing through Cu/S/Graphene catalyst was four times higher than that passing through graphite. The novel copper-based catalyst had an extraordinary performance for oxygen reduction reaction (ORR) due to the unique bio-inspired and stoichiometric structure. The results of Raman and Dispersive X-ray spectroscopy confirmed the presence of ultra-low content of copper (2%) and sulfur (1%) atoms on the graphene surface. Thermogravimetric analysis indicated a strong interaction between nanoparticles and graphene layers. The number of electrons transferred for ORR varied from 3.98 to 4.16 in a wide range of over-potentials indicating an effective 4-electron pathway form O-2 to H2O. The Tafel slopes indicated insignificant amount of formed copper oxide on the catalyst surface. The catalyst showed excellent electrochemical durability and its half-wave potential (E-1/2) was exhibited a negative shift only 8.2 mV after 10000 cycles. (C) 2019 Hydrogen Energy Publications LLC. Published by Elsevier Ltd. All rights reserved.