A strain ISB5 with the ability to degrade the non?ionic surfactant Triton X?100 was screened from the activated sludge in the aerobic aeration tank of a sewage treatment plant in a chemical plant. Through sequencing and comparison analysis of the 16S rDNA of the strain, the strain ISB5 was identified as Pseudomonas sp. The culture conditions of strain ISB5 to degrade Triton X?100 were further investigated, and the suitable degradation conditions of the strain were obtained: Culture temperature is 30 ℃, initial concentration of Triton X?100 is 4 g/L, culture time is 28 h, shaking rate is 150 r/min, filling volume is 100.00 mL, inoculation the amount is 1.00% (volume ratio of strain ISB5 to medium). The degradation rate of TritonX?100 by the strain under this condition can reach more than 93.0%.