A series of heteropoly acid (HPA) based Al2O3 catalysts with three-dimensional ordered (3DOM) structure were synthesized by colloidal crystal template method. Interconnected macropores (250 nm) could be clearly observed by scanning electron microscope (SEM) and transmission electron microscope (TEM). Mesopores could be detected by N2 adsorption-desorption isotherms which further confirmed the 3DOM structural characteristics of catalyst. Moreover, Keggin-type HPW was highly dispersed in the Al2O3 framework, which suggested by powder X-ray diffraction (XRD) and Fourier transform infrared spectra (FT-IR) results. The oxidation desulfurization (ODS) performance of 3DOM H3PW12O40/Al2O3 of refractory sulphur compounds was evaluated in the presence of hydrogen peroxide. It oxidized 98.5％ of dibenzothiophene (DBT) into corresponding sulfone within 3 h, which exhibited superior ODS performance than corresponding mesoporous and microporous H3PW12O40/Al2O3 catalyst. The enhancement of ODS efficiency is related to the improvement of mass transfer of DBT in the pore channel resulting from the interconnected 3DOM structure. Furthermore, the as-prepared catalyst still demonstrates outstanding cycle performance after 6 runs, which could be easily recovered from the model fuel.