In this study, the novel sulfur autotrophic electrodialysis ion-exchange membrane bioreactor (SEDIMB) with high denitrification performance was developed without the addition of carbon source. Sulfur autotrophic denitrification occurs at the same time of nitrate enrichment by electric field to achieve harmless nitrate transformation in water. The final nitrate concentration in water chamber was 2.16 mg-N/L (removal efficiency of 94.49 %) at an initial nitrate concentration of 40 mg-N/L, with the maximum nitrate concentration in biological chamber of 4.09 mg-N/L. Nitrate removal in water chamber of the SEDIMB conformed to the first-order kinetic model (R2 = 0.98) and reaction rate constant K1 = 0.0030 (1/min). The effect of environmental variables (i.e., voltage, initial nitrate concentration and water body) on the SEDIMB denitrification performance was investigated. Increasing voltage could promote nitrate migration in water chamber, and when the voltage was 7 V, initial nitrate concentrations of 20 ∼ 60 mg-N/L was effectively removed (all > 93.54 %). Moreover, the change of water body (i.e., tap water, lake water and deionized water) did not affect denitrification performance of SEDIMB. Microbial community structure analysis indicated that Betaproteobacteria were dominant in the SEDIMB. Membrane characterizations (ATR-IR, SEM and CLSM) confirmed that the bacteria in biological chamber tended to accumulate and grow on one side of the anion exchange membrane (AEM), while could not affect the selective permeability of AEM.