Titanium alloy (Ti-6Al-4V) having a bimodal “harmonic structure”, which consists of the coarse-grained structure surrounded by the network structure with fine grains, was fabricated by mechanical milling (MM) and spark plasma sintering (SPS) to achieve high strength and good plasticity. The microstructure of the MM-processed powder and the sintered compacts were characterized using a micro-Vickers hardness tester, scanning electron microscope (SEM) and an electron backscatter diffraction technique (EBSD). Harmonic structure was created in the sintered Ti-6Al-4V compacts prepared from the MM-processed powders having fine grains at its surface. The Ti-6Al-4V alloy with harmonic structure exhibited high tensile strength and good plasticity. The effects of the harmonic structure on the 4-points bending fatigue properties of Ti-6Al-4V alloy was investigated under the stress ratio R = 0.1 in ambient air without any controls of the temperature. The compacts with harmonic structure exhibited higher fatigue strength compared to the conventional coarse-grained material prepared from as-received initial powders. This was because the Ti-6Al-4V alloy with harmonic structure had higher tensile strength and hardness. Moreover, fatigue fracture mechanism of the Ti-6Al-4V alloy with harmonic structure was discussed from viewpoints of fractography and crystallography. As results of observing and analyzing the fracture surfaces, the Ti-6Al-4V alloy with harmonic structure failed from the coarse grain in the harmonic structure in the surface fracture mode.