The supported gold nanoparticle carbon catalyst with high dispersity was prepared by sodium citrate reduced-colloid gold immobilized method, and its catalytic activity was evaluated in the liquid phase catalytic oxidation reaction of D-glucose to D-gluconic acid. The influence of the pH and reaction temperature on Au/C catalytic activity were researched. The research results showed that when the pH was between 8–9 and the reaction temperature was between 50-55 °C, the Au/C catalyst has higher catalytic activity for the liquid phase catalytic oxidation reaction of D-glucose to D-gluconic acid.