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Background  Integrating hydrogen-producing bacteria with complementary capabilities, dark-fermentative bacteria (DFB) and photo-fermentative bacteria (PFB), is a promising way to completely recover bioenergy from waste biomass. However, the current coupled models always suffer from complicated pretreatment of the effluent from dark-fermentation or imbalance between dark and photo-fermentation, respectively. In this work, an integrated dark and photo-fermentative reactor (IDPFR) was developed to completely convert an organic substrate into bioenergy.    Results  In the IDPFR,  Ethanoligenens harbinese  B49 and  Rhodopseudomonas faecalis  RLD-53 were separated by a membrane into dark and photo chambers, while the acetate produced by  E. harbinese  B49 in the dark chamber could freely pass through the membrane into the photo chamber and serve as a carbon source for  R. faecalis  RLD-53. The hydrogen yield increased with increasing working volume of the photo chamber, and reached 3.38 mol H 2 /mol glucose at the dark-to-photo chamber ratio of 1:4. Hydrogen production by the IDPFR was also significantly affected by phosphate buffer concentration, glucose concentration, and ratio of dark-photo bacteria. The maximum hydrogen yield (4.96 mol H 2 /mol glucose) was obtained at a phosphate buffer concentration of 20 mmol/L, a glucose concentration of 8 g/L, and a ratio of dark to photo bacteria of 1:20. As the glucose and acetate were used up by  E. harbinese  B49 and  R. faecalis  RLD-53, ethanol produced by  E. harbinese  B49 was the sole end-product in the effluent from the IDPFR, and the ethanol concentration was 36.53 mmol/L with an ethanol yield of 0.82 mol ethanol/mol glucose.    Conclusions  The results indicated that the IDPFR not only circumvented complex pretreatments on the effluent in the two-stage process, but also overcame the imbalance of growth and metabolic rate between DFB and PFB in the co-culture process, and effectively enhanced cooperation between  E. harbinense  B49 and  R. faecalis  RLD-53. Moreover, simultaneous hydrogen and ethanol production were achieved by coupling  E. harbinese  B49 and  R. faecalis  RLD-53 in the IDPFR. According to stoichiometry, the hydrogen and ethanol production efficiencies were 82.67% and 82.19%, respectively. Therefore, IDPFR was an effective strategy for coupling DFB and PFB to fulfill efficient energy recovery from waste biomass.

Simultaneous hydrogen and ethanol production from cascade utilization of mono-substrate in integrated dark and photo-fermentative reactor