In this study, homogeneously dispersed gold particles growth onto carbon felt were fabricated by electrodeposition method followed by a thermal treatment at 1000 °C under nitrogen. The thermal treatment induced the dewetting of gold and the formation of well-crystallized gold particles that exhibited large surface area. The structural properties of the resulted Au@CF material were evaluated by SEM, XRD and TGA. We studied the electrocatalytic properties of this new gold material through the abiotic glucose oxidation in alkaline medium and the enzymatic dioxygen electroreduction by the enzyme bilirubin oxidase. Finally, we showed the potentiality of the resulting Au@CF material to build a 3-dimensional glucose hybrid biofuel cell by assemblying an abiotic anode with an enzymatic cathode. The system exhibited high electrochemical performance with an open circuit voltage of 0.71 V and a maximum power density of 310 μW cm−2 at 352 mV (by taking into account the projected surface area), in spite of a low gold loading (0.2 wt%). The advance presented in this work is the efficiency of the synthesis technique to get a new free-standing material for electrocatalysis based on gold particles with high reactive surface area for electron transfer and macropores for diffusion transport