Hydrogen Production During Direct Cellulose Fermentation by Mixed Bacterial Culture: The Relationship Between the Key Process Parameters Using Response Surface Methodology

Dark fermentation is a promising method to produce hydrogen from lignocellulosic biomass. This study assessed the influence of temperature, phosphate buffer concentration and substrate concentration on direct hydrogen production form cellulose using response surface methodology. Mixed bacterial culture was successfully enriched on cellulose and used as an inoculum for hydrogen production. The model indicated that the highest cumulative hydrogen production (CHP) of 2.14 L H₂/Lmedium could be obtained at 13.5 gcellulose/L, 79.5 mM buffer and 32.6 °C. However, hydrogen yield is then only 0.58 mol H₂/molhexose due to low substrate conversion efficiency (SCE). Simultaneous optimization of CHP and SCE with desirability function approach resulted in the H₂ yield of 2.71 ± 0.1 mol H2/molhexose and 93.8 ± 1.8% SCE at 3.35 gcellulose/L, 69 mM buffer and 32.9 °C. Phosphate concentration above 80 mM decreased H₂ production, but had positive effect on cellulose consumption. The bacterial community analysis showed that Ruminiclostridium papyrosolvens was responsible for cellulose hydrolysis. Lachnoclostridium sp. was positively correlated with ethanol production at high phosphate buffer concentration, while Caproiciproducens sp. with caproate production at low buffer concentration. The obtained results opens the possibility of simultaneous hydrogen and caproate production from cellulosic substrates.