Protein conformational rearrangement triggered by adsorption to the hydrophobic interface of oil droplets has long been considered as a key factor in emulsification. In this study, an alkaline pH-shifting-driven conformational adaptation enhanced interfacial proteins was used to improve their stability against heat-induced flocculation of myosin emulsions. We used the unfolded myosin at pH 12 to emulsify soy oil and then readjusted the pH of the emulsion to neutral. The corresponding myosin emulsion (0.5% w/v protein, 10% v/v soy oil, and 0.6 M NaCl) almost not flocculated when heated at 75 °C for 30 min. Moreover, after thermal treatment, the particle size of the emulsion was not significantly increased ( P > 0.05) and the emulsion did not exhibit a creaming phenomenon after a week. Based on the circular dichroism and Fourier transform infrared analysis, we speculated the superiority of the emulsion is closely related to the alkaline pH-shifting-driven conformational adaptation enhanced interfacial protein. Additionally, the resulting steric stabilization in overcoming the attractive hydrophobic forces between denatured protein molecules coated droplets might be the main factor for the inhibition of heat-induced flocculation of the emulsion. Our research may have important implications for the formulation of protein-stabilized oil-in-water emulsions.