Pickering emulsion (PE) is used as a sustaining delivery system to prevent food-borne pathogenic contamination through slow release of bactericidal agents. But it is still challenging in timely bactericidal effect and antagonistic effect on the efficacy of the bio-active agents in the presence of surfactants. Smarter strategies for controlling food contamination requires a surfactant-free system which can be smart sensed and targeted release of bactericidal agents. In the present study, we prepared a thymol loaded protein-based PE, and microscopic, low-field nuclear magnetic resonance, rheological measurements, etc. Proved that the PE was stable at protein concentration of 10% and oil ratio at 0.5. And this thymol loaded PE showed comparative antibacterial effect through accumulation of reactive oxygen species, induction of membrane lipid peroxidation. Moreover, the potential of the PE for “smart sensing and targeted releasing” was assessed by targeting Escherichia coli, Staphylococcus aureus and Aspergillus flavus. Our results are of great interest for developing novel intelligent surfactant-free delivery system for the application of thymol, and also provide a basis for a safe and biocompatible antimicrobial additive in food industry.