To optimize the performance of the pulsed xenon lamp sterilization device, the influence of spectral range and specifications of lamps on the sterilization effect is studied based on a self-developed high-energy microsecond pulse power supply and xenon lamps with different specifications. The results show that in the UV-visible spectrum of a xenon lamp with an arc length of 50 mm and a pressure of 50 kPa, the UV accounts for 38.5% and the UVC accounts for 17.6%. Increasing the arc length and decreasing the pressure can both increase the spectral intensity, and the latter can also increase the ratio of UV. The xenon lamp with an arc length of 100 mm and a pressure of 50 kPa can basically inactivate all Escherichia coli in 3 s with a discharge energy of 20 J. The sterilization rate is positively correlated with arc length and discharge energy of the lamp, negatively correlated with pressure. All bands of xenon lamp radiation have sterilization effects, with UV accounting for 87.7% in log value and the wavelength band less than 280 nm accounting for 64.6%. The AFM images show that pulsed xenon lamp changed the morphology and mechanical properties of Escherichia coli, hence the bacteria shrank, their surface roughness, elasticity, and adhesion increased.