Artificial redox mediators (ARM) have been proven to accelerate the azo dye anaerobic bio-reduction (ADAB) but the mechanisms involved are still unclear. Previous studies do seldom focus on the production of methane during the ADAB, particularly if supplemented with ARM. Our studies revealed that the supplement of ARM could significantly accelerate the decolorization rate, recover the inhibited methanogenesis and decrease extracellular polymeric substance secretion in an ADAB system. Supplement of an ARM only enhanced the preexisting metabolic pathway of the ADAB system. Significant differences in metabolic pathway and microbial community were found between traditional methanogenic system and ADAB system by high-throughput sequencing technique. The ADAB system performed an "over-requirement for electron donor" phenomenon and the requirement amount can be altered by regulating ARM dosage. Response surface methodology was then successfully employed to optimize the dosage of ARM and possible additional electron donor required for ADAB under different conditions.