Hydrogen-producing acetogens (HPA) have a transitional role in anaerobic wastewater treatment. Thus, bioaugmentation with HPA cultures can enhance the chemical oxygen demand (COD) removal efficiency and CH4 yield of anaerobic wastewater treatment. Cultures with high degradation capacities for propionic acid and butyric acid were obtained through continuous subculture in enrichment medium and were designated as Z08 and Z12. Bioaugmentation with Z08 and Z12 increased CH4 production by glucose removal to 1.58. Bioaugmentation with Z08 and Z12 increased the COD removal rate in molasses wastewater from 71.60% to 85.84%. The specific H2 and CH4 yields from COD removal increased by factors of 1.54 and 1.63, respectively. Results show that bioaugmentation with HPA-dominated cultures can improve CH4 production from COD removal. Furthermore, hydrogen-producing acetogenesis was identified as the rate-limiting step in anaerobic wastewater treatment.