An anaerobic sequencing batch reactor containing immobilized biomass (AnSBBR) was used to produce biomethane by treating the effluent from another AnSBBR used to produce biohydrogen from glucose- (AR-EPHG) and sucrose-based (AR-EPHS) wastewater. In addition, biomethane was also produced from sucrose-based synthetic wastewater (AR-S) in a single AnSBBR to compare the performance of biomethane production in two steps (acidogenic and methanogenic) in relation to a one-step operation. The system was operated at 30 °C and at a fixed stirring rate of 300 rpm. For AR-EPHS treatment, concentrations were 1,000, 2,000, 3,000, and 4,000 mg chemical oxygen demand (COD) L(-1) and cycle lengths were 6 and 8 h. The applied volumetric organic loads were 2.15, 4.74, 5.44, and 8.22 g COD L(-1) day(-1). For AR-EPHG treatment, concentration of 4,000 mg COD L(-1) and 4-h cycle length (7.21 g COD L(-1) day(-1)) were used. For AR-S treatment, concentration was 4,000 mg COD L(-1) day(-1) and cycle lengths were 8 (7.04 g COD L(-1) day(-1)) and 12 h (4.76 g COD L(-1) day(-1)). The condition of 8.22 g COD L(-1) day(-1) (AR-EPHS) showed the best performance with respect to the following parameters: applied volumetric organic load of 7.56 g COD L(-1) day(-1), yield between produced methane and removed organic material of 0.016 mol CH4 g COD(-1), CH4 content in the produced biogas of 85 %, and molar methane productivity of 127.9 mol CH4 m(-3) day(-1). In addition, a kinetic study of the process confirmed the trend that, depending on the biodegradability characteristics of the wastewaters used, the two-step treatment (acidogenic for biohydrogen production and methanogenic for biomethane production) has potential advantages over the single-step process.