Thiobacillus denitrificans, a kind of autotrophic facultative bacteria, can oxidize sulfide into elemental sulfur or sulfate when nitrate was adopted as its electron accepter and carbon dioxide as its carbon resource under anoxic or anaerobic environment. In this way, nitrate is converted into nitrogen. In addition, Thiobacillus denitrificans can accumulate sulfur extracellularly. In this study, in a process of simultaneous desulfurization and denitrification, a strain of Thiobacillus denitrificans is employed as sulfur-producer in the treatment of wastewater containing sulfide and nitrate. The key factors affecting this process are investigated through batch tests. The experimental results indicate that the sulfide concentration and the ratio of sulfide to nitrate (S2-/NO3-) in the influent are the key factors, and their suitable values are suggested to be 5/3 and no more than 300 mg center dot L-1, respectively, in order to achieve high conversion of sulfur.