Optimization of Operational Parameters for Biohydrogen Production from Waste Sugarcane Leaves and Semi-Pilot Scale Process Assessment

Preshanthan Moodley, E. B. Gueguim Kana


This study modeled and optimized the operational parameters for biohydrogen production from waste sugarcane leaves and assessed hydrogen production on a semi-pilot scale. A Box-Behnken design with input variables of substrate concentration (8 to 24 g/L), inoculum concentration (10% to 50% v/v), and hydraulic retention time (HRT, 24 to 96 h) was used. A coefficient of determination (R2) of 0.90 and the predicted optimum operational set-points of 14.2 g/L substrate concentration, 32.7% inoculum concentration, and 62.8 h HRT were obtained. Experimental validation produced a biohydrogen yield of 12.8 mL H2/g fermentable sugar (FS). A semi-pilot scale process in a 13-L Infors reactor under optimized conditions gave a cumulative hydrogen volume and yield of 3740 mL and 321 mL H2 g-1 FS, respectively, with a peak hydrogen fraction of 37%. Microbial analysis from the process effluent conducted by Polymerase Chain Reaction cloning indicated the presence of hydrogen-producing bacteria belonging to Clostridium sp., Klebsiella sp., and Enterobacter sp. These findings highlight the feasibility of biohydrogen production from sugarcane waste and provide preliminary knowledge on process scale up.


Biohydrogen production; Sugarcane leaves; Bioprocess optimization; Semi-pilot scale process; Dark fermentation

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