The pathogenicity of bacteria is closely related to their tolerance to environmental stress. When the pathogenic Vibrio bacteria is exposed to external stressosome, a protein-signaling hub within Vibrio bacteria senses and transmits stress signals through reversible phosphorylation. In order to explore the influence of ultra-high pressure (UHP) on the conformation of RsbT protein kinase in Vibrio and on its downstream signaling transduction, three pressure points of 100 MPa, 300 MPa and 500 MPa are used in this experiment. Each pressure point is kept for 5 minutes. After western blotting, in the RsbT protein kinase, phosphorylation ability decreases as the pressure increases. The RsbT conformation alters and its phosphorylation ability almost disappears when the pressure is above 300 MPa. This study offers new theoretical support for the influence of pressure on RsbT protein kinase conformation and biological activity in stressosome, which provide insights into treatment of food-bone bacteria using UHP.