The effects of Al2O3 gate insulator grown by atomic layer deposition (ALD) system on the two-dimensional electron gas (2DEG) transport characteristics in AlGaN/GaN metal-insulator-semiconductor high electron mobility transistor (MISHEMT) were investigated. The shape of the electron lateral distribution in the quantum well at AlGaN/GaN interface was found to be slightly influenced by the Al2O3 thin layer. The drift mobility (μd) of the electrons in ALD-Al2O3/AlGaN/GaN MISHEMT is increased due to the surface passivation effects of the included dielectric layer. The higher dynamic channel current of the MISHEMT indicates that the electron saturation velocity (vsat) is also increased. These results show the improvement of the transport characteristics of 2DEG in Al2O3/AlGaN/GaN MISHEMT by the excellent properties of the Al2O3 grown by ALD.