The hydrophobicity of, coal limits the adsorption of microorganisms and the efficiency of biological desulfurization was influenced, while the surfactant can enhance the interaction of the four phases, air, water, bacteria, and coal grain. In this experiment, three types of surfactant, anionic (SDS), cationic (DTAB), and nonionic (Tween 20), were investigated at 30 degrees C, and the cell concentration, pH value, leaching time, and coal biodesulfurization rate of the coal desulfurization system were detected. It is shown that the total desulfurization rate approached 29.7% in 16 days when 1100 mg/L Tween 20 was added in the shaking test, which represented the optimum efficiency. Further column leaching desulfurizating experiments showed that the desulfurization rate of the experimental group with Tween 20 (12.75%) was notably higher than that of the control group (8.32%). Along with the processing of desulfurization, the cell concentration decreased at first, and then rapidly increased, and finally stayed constant. The pH value of coal water slurry increased. The leaching time increased with the process of the desulfurization system. We concluded that the nonionic surfactant has a significant effect on coal biodesulfurization.