Abstract: Thyme essential oil antibacterial nanoemulsions (tPTNs) was prepared by a two-step method of homogenization and ultrasound and constructed by TGase-modified potato protein (37 ℃, 1 h) and its antibacterial activity and application in pork preservative was investigated in this study. Results indicated that the diameters of inhibition zone of tPTNs against Staphylococcus aureus (S. aureus) and Escherichia coli (E. coli) were 17.98 and 14.28 mm, respectively. The bacteriostatic curve showed tPTNs exhibited excellent antibacterial activity. The tPTNs significantly inhibited the increase of pH of pork inoculated with S. aureus (5.75~6.14) and E. coli (5.69~6.20) during 7-day storage. The growth of two kinds of bacteria (6.00~7.07 lg CFU/g, 6.12~6.95 lg CFU/g) in pork treated with tPTNs were significantly slow. The oxidation of lipid (0.006~0.27 mg/g, 0.037~0.23 mg/g) and protein (4.46~2.80 μmol/mL, 4.01~2.84 μmol/mL) were effectively inhibited in pork treated with tPTNs. Results of water holding capacity and texture test indicated that the tenderness, hardness and other properties of pork treated with tPTNs led to little changes during storage, possessing the improved storage stability and the higher sensory evaluation score. In conclusion, this study provides certain theoretical guidance for the preparation of essential oil antibacterial nanoemulsion and its application in the preservation of chilled pork, and possesses broad development and application prospects in the field of meat preservation.