Bio-surfactant is a new type of surfactant that is produced in microbial metabolism. Adding bio-surfactant during composting process, especially to those contain some toxic substances, has been proved to be a promising way. In this study, Strains Ⅲ (2), a bacterial with high activity to produce bio-surfactant, were isolated firstly. Following comparison experiments with and without adding Strains Ⅲ (2), namely Run 1 and Run R, were conducted, respectively. The experimental results showed that, by adding Strains Ⅲ (2),the surface tension could reduce from 46.5 mN/m to 39.8 mN/m and the corresponding time to maintain the surface tension under 50 mN/m could prolong from 60 h to 90 h. The oxygen uptake rate and total accumulated oxygen consumption with Stains Ⅲ (2) were both higher than those without Strains Ⅲ (2), while the accumulation of H2S in outlet gas was reduced to around 50% of Run R. Moreover, two additional experiments were also carried out to examine the effects of strains coming from different systems. One is adding Strains Ⅲ (2)with a dose of 0.4% (Run 2), and the other is seedling commercial Strains at the same conditions, the composting experiments showed that: Run 2 was more effective than Run 3, because the commercial Strains can be suppressed significantly in a complex composting system with different pH, high temperature and some of metals. The bio-surfactant was also added into the solid waste, which contained some toxic substances, the corresponding results showed that the remove rate of Hg and sodium pentachlorophenolate(PCP-Na) could be improved highly. Thus, the microenvironment, reactionrate and composting quality could be enhanced effectively by adding bio-surfactant to the composting process.