The experiment was conducted with minitab software and applied Packett-Burman design with five factors, including temperature, pH, inoculate capacity, rotational speed and the initial amount of oil to optimize the surfactant produce conditions of oil degrading strain CT-6 (Pseudomonas aeruginosa). The results showed that when the temperature was 35℃, the original amount of oil 4.0%, pH 8.0, rotational speed 213 r/min, incubation time 152 h, the activity of surfactant which produced by strain CT-6 was best, the exhibit circle was 6.4 cm, emulsification performance reached 92%, cells hydrophobic was 41%, increased by 35.9%, 29.6%, 64.0%, respectively.