In this study an attempt was made to enhance anaerobic hydrogen production from beverage wastewater (BWW) by augmenting anaerobic enriched mixed cultures (EMC) with facultative Enterobacteriaceae strains (E. Coli XL1-BLUE, Enterobacter cloacae) under mesophilic temperature 37°C, pH 5.5 and substrate concentration of 10 gglucose equivalent /L. Among the bioaugmented cultures, E. cloacae+EMC led to the peak hydrogen production rate (HPR) of 2250 mL/L-d. Furthermore, the variables substrate concentration and pH was statistically optimized using response surface methodology (RSM). Results show that the predicted maximum HPR of 3096 mL/L-d was achieved under optimal operation conditions: substrate concentration 25.6 g glucose equivalent /L and pH 5.7. Verification experimental results showed HPR of 3044 mL/L-d, reveals that predicted results were closure with the experimental data and proved experimental design optimization processes are more feasible techniques for biohydrogen production. PCR-DGGE analysis indicates that Clostridium sp. and Enterobacter sp. were appeared under optimal conditions.
Keywords: Beverage wastewater, bioaugmentation, biohydrogen, enriched mixed culture, hydrogen production rate, PCRDGGE.