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Glucose has proven to be a promising fuel for renewable and sustainable fuel cells due to its high energy density. The anion-exchange membrane (AEM) direct glucose fuel cell (DGFC) attracted lots of attentions because of the higher performance than the conventional microbial fuel cell. Among the factors affecting the performance of the DGFC, the anode structure plays an important role, but presently, a general understanding of the anode electrode design is far less understood. In this work, anode structure was designed for the high-performance AEM DGFC, including the configurations of the anode electrode and the compositions of both the micro-porous layer (MPL) and catalyst layer(CL). The experimental results revealed that the fabrication of the anode electrode with catalyzed diffusion medium (CDM) method yielded better performance than that with catalyst-coated membrane (CCM) method because of lower resistance of the mass transport. It was found that the addition of an ultra-thin MPL (0.3 mg cm-2 carbon loading) between backing layer (BL) and CL improved the cell performance due to it can facilitate the electron transfer. In addition, the MEA with PTFE binder in the anode CL showed a better cell performance than that with I2 binder due to the enhanced mass transport and larger active surface area.