Cellulose utilization by hydrogen producers remains an issue due to the low hydrogen yields reported and the pretreatment of cellulose prior to fermentation requires complex and expensive steps. Clostridium termitidis is able to breakdown cellulose into glucose and produce hydrogen. On the other hand, Clostridium beijerinckii is not able to degrade cellulose but is adept at hydrogen production from glucose; therefore, it was chosen to potentially enhance hydrogen production when co-cultured with C. termitidis on cellulose. In this study, batch fermentation tests were conducted to investigate the direct hydrogen production enhancement of mesophilic cellulolytic bacteria C. termitidis co-cultured with mesophilic hydrogen producer C. beijerinckii on cellulose at 2 g l−1 compared to C. termitidis mono-culture. Microbial kinetics parameters were determined by modeling in MATLAB. The achieved highest hydrogen yield was 1.92 mol hydrogen mol−1 hexose equivalentadded in the co-culture compared to 1.45 mol hydrogen mol−1 hexose equivalentadded in the mono-culture. The maximum hydrogen production rate of 26 ml d−1 was achieved in the co-culture. Co-culture exhibited an overall 32 % enhancement of hydrogen yield based on hexose equivalent added and 15 % more substrate utilization. The main metabolites were acetate, ethanol, lactate, and formate in the mono-culture, with also butyrate in the co-culture. Additionally, the hydrogen yield of C. beijerinckii only in glucose was 2.54 mol hydrogen mol−1 hexose equivalent. This study has proved the viability of co-culture of C. termitidis with C. beijerinckii for hydrogen production directly from a complex substrate like cellulose under mesophilic conditions.