To explore the interaction between lipids and proteins during emulsion film formation, the linoleic acid concentration effects on the physicochemical properties of soybean β-conglycinin (7S) films were studied. The viscosity and size of oil droplets in the film-forming solution gradually increased with the increasing linoleic acid concentration. As the linoleic acid concentration increased, the number of oil droplets on the film surfaces and elongation at break of films gradually increased, whereas the tensile strength decreased. The films containing 20% linoleic acid had the highest water vapor permeability value, which was decreased by increasing or decreasing the linoleic acid concentration. According to the molecular dynamics simulation and chemical interactions, 7S could be adsorbed at the linoleic acid interface and bind stably, resulting in the decreased ionic and hydrogen bonds but the increased hydrophobic interactions and covalent bonds among proteins in the films.