[Objective] To study the mechanisms of mutual promotion between chemolithoauto-trophic sulfur-oxidizing bacteria and chemoheterotrophic bacteria under co-culture based on carbon metabolism. [Methods] Ion chromatography was employed to determine the concentrations of S₂O₃^2‒ (thiosulfate) and SO₄^2‒ (sulfate). Bacterial growth dynamics were monitored by the dilution plate method. Extracellular carbon characteristics were analyzed via total organic carbon analyzer measurement and LC-MS. Cellular morphology was observed by scanning electron microscopy. The relative mRNA levels of related genes were quantified by RT-qPCR. [Results] During the growth process, sulfur-oxidizing bacteria continuously fixed inorganic carbon and secreted organics, providing a stable carbon source for the growth of heterotrophic bacterium. In return, heterotrophic bacteria significantly enhanced the sulfur-oxidizing and carbon-fixing capabilities of sulfur-oxidizing bacteria. This was evidenced by the significantly up-regulated expression of the enzyme gene soxB involved in sulfur oxidation and the RubisCO gene cbbL involved in carbon fixation. Additionally, the production of extracellular polymeric substances was induced, which enhanced the biofilm formation. [Conclusion] This study elucidated the interaction mechanisms between sulfur-oxidizing bacteria and heterotrophic bacteria, particularly the significant enhancement of the carbon-fixing capability of sulfur-oxidizing bacteria. The findings provide a new perspective for the enrichment culture of chemolithoautotrophic bacteria and for understanding the carbon fixation mechanisms of autotrophic sulfur-oxidizing bacteria in microbial communities. Additionally, this study offers theoretical support for the low-carbon and efficient treatment of wastewater.