Stibnite (Sb2S3) is the predominant form of the antimony (Sb) in nature and the main component of antimony tailings. The process of biodissolution of stibnite under neutral or alkaline conditions is one of the most prevalent geochemical processes of antimony release. Recent studies have shown that microorganisms such as Acidithiobacillus (A.) ferrooxidans can promote the biodissolution process of stibnite under extreme acidic conditions, but the process is affected by other substances in the environment, such as its associated minerals pyrite (FeS2) and arsenopyrite (FeAsS). Therefore, this study is devoted to investigating the mechanism of influence pyrite (FeS2) or arsenopyrite (FeAsS) on the biodissolution of stibnite. Through the analysis of environmental physicochemical properties, mineral surface morphology, elemental morphology transformations, electrochemistry, transcriptomics, and molecular morphology simulations, we found that: (1) the presence of FeS2/FeAsS can promote the biodissolution process of stibnite, accelerate the oxidation of Sb and sulfur (S); (2) A. ferrooxidans can enhance the electrochemical activity of the surface of the associated minerals; (3) the expression of functional genes of A. ferrooxidans has increased with the presence of associated minerals, such as electron transfer and metabolism-related functional genes. These results prove the presence of the associated minerals could accelerate the process of antimony release and elucidate the mechanism of stibnite dissolution mediated by the role of pyrite/arsenopyrite-A. ferrooxidans, which are of great theoretical significance for understanding the geochemical cycle of antimony in sulfide mining areas and for the development of antimony pollution treatment and remediation technologies.