This study was aimed to investigate the effect of ultrasonic stimulation on the growth and protein expression of Escherichia coli( E. coli) at different periods. The growth was measured by spectrophotometric method and the protein expression was detected by Tricine-SDS-PAGE analysis. Fura-2 / AM fluorescence probe was used to determine the change in intracellular Ca2 +concentration. Results showed that the cells were stimulated by ultrasound at 28 ± 2 kHz for 30 min after inoculation( period 1),under which the growth was obviously promoted by ultrasonic stimulation over the control. The protein expression was not increased by the ultrasound agitation for 5,30,and 60 min at period 1. When the ultrasonic stimulation occurred at the time of optical density at 1. 00( period 2),the cell growth and protein expression were inhibited. Results showed that concentration of Ca2 +was decreased to the lowest point by ultrasonic treatment for 30 min. This research confirmed that ultrasound could affect the growth and metabolism of E. coli,which provided a basis for the use of ultrasound to enhance the fermentation of engineering bacteria.