Acidic wastewater containing high sulfate is produced by various industries such as battery manufacturing, chemical manufacturing, mining and metallurgy, and acid mine drainage, which pose detrimental effects on the environment and living organisms. Biological sulfate reduction employing sulfate-reducing bacteria is a promising technique to treat such wastewater. Mixed bacterial culture collected from a wastewater treatment plant of IIT Guwahati was grown in an acclimatization reactor for 24 days at neutral pH, with sulfate concentration of 500 mg/l, and COD/SO42− ratio of 1.0. Following this a laboratory-scale semi-batch reactor was fabricated to treat sulfate-rich acidic wastewater by stepwise decrement of pH from 7.0 to 3.45, increment of sulfate concentration of 500 to 2500 mg/L, and varying COD/SO42− ratio of 1.0 and 0.7. The reactor was operated for 128 days with an HRT of 4 days using glucose as the sole carbon source. The best performance was achieved in phase VIII with an influent pH rise from 3.45 to 7.15 due to the production of bicarbonate alkalinity, and sulfate and COD removal of 70.2, and 89.3%, respectively. In phase IX, the influent sulfate concentration increased to 2500 mg/L after which the system performance drastically declined due to the synergic effect of both higher sulfate concentration and low pH and continued to fall further which is considered as a system failure. This study demonstrates that biological sulfate reduction using glucose offers a sustainable solution for mitigating environmental hazards posed by industrial effluents containing low pH and high sulfate concentration.