Improved cellulose conversion to bio-hydrogen with thermophilic bacteria and characterization of microbial community in continuous bioreactor

• Continuous cellulosic-hydrogen fermentation was conducted at 55 °C.
• Hydrogen yield was improved to 15.4 mol kg−1 of consumed-cellulose.
• The cellulosic hydrogen bacteria were close to Clostridia and Enterobacter genus.
• The mixed microflora produced H2 within a wide range of temperatures (35 °C–65 °C).
• Activation energy of cellulose and glucose were 103 and 98.8 kJ mol−1, respectively.

Thermophilic hydrogen fermentation of cellulose was evaluated by a long term continuous experiment and batch experiments. The continuous experiment was conducted under 55 °C using a continuously stirred tank reactor (CSTR) at a hydraulic retention time (HRT) of 10 day. A stable hydrogen yield of 15.4 ± 0.23 mol kg−1 of cellulose consumed was maintained for 190 days with acetate and butyrate as the main soluble byproducts. An analysis of the 16S rRNA sequences showed that the hydrogen-producing thermophilic cellulolytic microorganisms (HPTCM) were close to Thermoanaerobacterium thermosaccharolyticum, Clostridium sp. and Enterobacter cloacae. Batch experiment demonstrated that the highest H2 producing activity was obtained at 55 °C and the ultimate hydrogen yield and the metabolic by-products were influenced greatly by temperatures. The effect of temperature variation showed that the activation energy for cellulose and glucose were estimated at 103 and 98.8 kJ mol−1, respectively.