The effect of ultrasonic treatment on emulsifying properties of wheat germ protein (WGP) was studied in this paper. WGP was subjected to low frequency (20 kHz), high intensity ultrasonic treatment at different power (200, 400, 600, 800 W) for 10 min, or different time (2, 4, 6, 8, 10, 15, 20 min) at 400 W. The emulsifying activity index and emulsion stability index of WGP were significantly improved, and the emulsion droplet was smaller and more uniform after ultrasound treatment. Ultrasound increased the adsorbed WGP concentration at the oil-water interface and reduced the interfacial tension, which explained the improved emulsifying properties of WGP. The investigation on molecular properties and protein conformation showed that ultrasound processing increased solubility, but decreased particle size and surface charge of WGP. Ultrasound processing resulted in the unfolding of the protein molecular structure indicated by the increase of surface hydrophobicity and surface free sulfhydryl group levels, and the decrease of intrinsic fluorescence intensity. Correlation analysis showed that the changes in WGP solubility, particle size, and surface hydrophobicity were the main driven factors for the improved emulsifying properties of WGP.