Enzymatic Hydrolysis Strategies for Cellulosic Sugars Production to Obtain Bioethanol from Eucalyptus globulus Bark

Cellulosic sugars production for the valorization of lignocellulosic biomass residues in an industrial site has economic benefits and is promising if integrated into a biorefinery. Enzymatic hydrolysis (EH) of pretreated Eucalyptus globulus bark, an industrial residue of low-economic value widely available in Portuguese pulp and paper mills, could be an excellent approach to attain resource circularity and pulp mill profitability. This work evaluated the potential for improving cellulosic sugars concentrations by operating with high solids loading and introducing the additives Triton X-100, PEG 4000 and Tween 80 using a commercial enzymatic consortium with a dosage of 25 FPU g(carbohydrates)(-1). Additives did not improve enzymatic hydrolysis performance, but the effect of increasing solids loading to 14% (w/v) in batch operation was accomplished. The fed-batch operation strategy was investigated and, when starting with 11% (w/v) solids loading, allowed the feeding of 3% (w/v) fresh feedstock sequentially at 2, 4 and 6 h, attaining 20% (w/v) total solids loading. After 24 h of operation, the concentration of cellulosic sugars reached 161 g L-1, corresponding to an EH conversion efficiency of 76%. Finally, the fermentability of the fed-batch hydrolysate using the Ethanol Red((R)) strain was evaluated in a 5 L bioreactor scale. The present results demonstrate that Eucalyptus globulus bark, previously pretreated by kraft pulping, is a promising feedstock for cellulosic sugars production, allowing it to become the raw material for feeding a wide range of bioprocesses.