Azo dye wastewater treatment is urgent necessary nowadays. Electrochemical technologies commonly enable more efﬁcient degradation of recalcitrant organic contaminants than biological methods, but those rely greatly on the energy consumption. A novel process of bioﬁlm coupled with electrolysis, i.e., bioelectrochemical system (BES), for methyl orange (MO) dye wastewater treatment was proposed and optimization of main inﬂuence factors was performed in this study. The results showed that BES had a positive effect on enhancement of color removal of MO wastewater and 81.9% of color removal efﬁciency was achieved at the optimum process parameters: applied voltage of 2.0 V, initial MO concentration of 20 mg/L, glucose loads of 0.5 g/L and pH of 8.0 when the hydraulic retention time (HRT) was maintained at 3 d, displaying an excellent color removal performance. Importantly, a wide range of effective pH, ranging from 6 to 9, was found, thus greatly favoring the practical application of BES described here. The absence of a peak at 463 nm showed that the azo bond of MO was almost completely cleaved after degradation in BES. From these results, the proposed method of biodegradation combined with electrochemical technique can be an effective technology for dye wastewater treatment and may hopefully be also applied for treatment of other recalcitrant compounds in water and wastewater.