The effects of anthraquinone-2,6-disulfonate (AQDS) on microbial populations and the biocatalysing characteristics of AQDS on denitrifying strain GW1 are discussed. The results showed that microbial population abundances were 30, 6, 12 and 18% during the different periods (1st, 5th, 10th and 20th days). Strain GW1 utilized AQDS as an electron acceptor in the respiration chain and reduced AQDS to hydroquinone (QH2). The nitrate removal efficiency of strain GW1 was increased 1.14-1.63 fold with 0.04-0.32 mmol/L AQDS. A linear correlation was found between the nitrate removal reaction constant k[gNO3(-)- N/(gVSS x d)] (where VSS is volatile suspended solids) and AQDS concentration (CAQDS), which was k = 19.332 C(AQDS) + 11.115 (R2 = 0.9749). The stabilized oxidation-reduction potential (ORP) values with AQDS were lower 22-92 mv than the control during the denitrification process. The concentration of nitrite sequentially accumulated up to around 250 mg/L when nitrate have removed completely. The results suggested that AQDS as redox mediator was capable of biocatalysing the nitrate and nitrite removal rate.