Abstract The sulfate-rich wastewater from rubber smoked sheet industry could generate hydrogen sulfide (H2S) under anaerobic condition, which created bad smell to the community and might cause toxicity and damage to the environment. The H2S can be removed from the biogas by sulfur-oxidizing bacteria (SOB) with the ability to converted H2S to sulfate. Sulfate-reducing bacteria (SRB) could remove sulfate in wastewater before anaerobic treatment for biogas production. The microbial sludge from wastewater and anaerobic digestion system was collected and test for sulfate and H2S removal efficiency. Anaerobic microbial sludge has a high ability to produced methane from gelatin with a specific methane production rate of 92.4 ml CH4 gVSS-1 day-1. Anaerobic microbial sludge has lower methane production when gelatin and sulfate used as a substrate with a specific methane production rate of 81.4 ml CH4 gVSS-1 day-1. The biomethane potential, hydrogen sulfide removal and sulfate removal in anaerobic digestion system by addition of enriched cultures of SOB and SRB were investigated. The methane yield of SRB consortium was 60.1 ml CH4/gCOD with 20% sulfate reduction from wastewater and no sulfide reduction. The methane yield of SOB consortium was 41.9 ml CH4/gCOD with no sulfate and sulfide reduction from wastewater. The addition of SRB consortium could increase methane production by reducing sulfate concentration in wastewater consequently to a reduced concentration of H2S in biogas.