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Sulfur autotrophic denitrification utilizes elemental sulfur as the electron donor for nitrate removal from aquatic environments. Organic carbon could stimulate the conversion of sulfur and facilitates the S⁰-based denitrification process in the mix-trophic. In this study, the co-cultured system of sulfur reducer (Geobacter sulfurreducens) and Thiobacillus denitrificans was used to investigate that how organic carbon could boost the S⁰-based denitrification. The results showed that the rate of S⁰-based denitrification was improved with C/N ratio of 0.13 and this improvement continued even after the acetate was exhausted. Sulfur probe test and Raman analysis suggested that reduced sulfur species (S_x²⁻) were formed with the addition of organic carbon. The S_x²⁻ could recombine with element sulfur and the bioavailability of S⁰ would be improved, as a result, the rate of S⁰-based denitrification increased as well. Nitrate …