Anaerobic microecosystems designed with different concentrations of 17β-estradiol (17β-E2) (0.0–10,000.0 ng/L) were simulated in this study. The influence of different concentrations of 17β-E2 on the emissions of typical greenhouse gases (CH4 and CO2) in simulated anaerobic microecosystems is analyzed to primarily explore the relationship between 17β-E2 and such emissions in aquatic anaerobic ecosystems. The results showed that 17β-E2 could promote or significantly stimulate aquatic anaerobic micro-organisms' production of CH4. The degree and the promotion time of CH4 production were both enhanced with the increase of 17β-E2 concentration. Furthermore, under higher concentration of 17β-E2 (≥500.0 ng/L), the increasing tendency of aquatic anaerobic microbial populations' activity and the function of methanogenic activity under corresponding experimental conditions had a synchronous relationship.