Steroid estrogens (SEs), especially 17β-estradiol (E2), can be a serious threat to the health of organisms. The removal of E2 from the natural environment is mainly carried out by microbial degradation partly mediated by biochar, which contains quinone structures. In this study, reed straw biochar samples made at four different heat treatment temperatures (HTTs) were used to mediate E2 microbial degradation by Shewanella oneidensis MR-1. The removal efficiency of E2 (95%) was highest in the presence of HTT – 500 °C biochar under anaerobic conditions after 120 h of microbial degradation. The effect of biochar on promoting microbial degradation was far more superior under anaerobic conditions than under aerobic conditions. The redox-activity and types of surface functional groups of biochar reveal that biochar can accept electrons generated by microorganisms in a timely manner. This mechanism promotes the metabolic process of cells and microbial degradation of E2 (exponential increase). Biochar particles rather than biochar-derived water-soluble organic compounds are responsible for this stimulating effect. These results highlight the impact that biochar has on microbial degradation of trace pollutants in a natural environment.