Tailoring a cellulolytic enzyme cocktail for efficient hydrolysis of mildly pretreated lignocellulosic biomass.

Commercially available cellulase cocktails frequently demonstrate high efficiency in hydrolyzing easily digestible pretreated biomass, which often lacks hemicellulose and/or lignin fractions. However, the challenge arises with enzymatic hydrolysis of mildly pretreated lignocellulosic biomasses, which contain cellulose, hemicellulose and lignin in high proportions. This study aimed to address this question by evaluating the supplementation of a commercial cellulolytic cocktail with accessory hemicellulases and two additives (H2O2 and Tween® 80). Statistical optimization methods were employed to enhance the release of glucose and xylose from mildly pretreated sugarcane bagasse. The optimized supplement composition resulted in the production of 304 and 124 mg g-1 DM of glucose and xylose, respectively, significantly increasing glucose release by 84% and xylose release by 94% compared to using only the cellulolytic cocktail. This enhancement might be attributed to a coordinated hemicellulases action degrading hemicellulose, creating more space for cellulase activity, potentially boosted by the presence of H2O2 and Tween® 80. However, the addition of different concentrations of H2O2 in combination with hemicellulase and Tween® 80 did not result a significant difference on sugar release, which could be attributed to the limited range of concentrations studied (5 to 65 µM). The results obtained in this study using the mix of three supplements were also compared to the addition of only hemicellulase and only Tween® 80 to the cellulolytic cocktail. A significant increase in glucose release of 39% and 41%, respectively, was observed when using the optimized combination. For xylose, the increase was 38% and 41%, respectively. This study underscores the substantial potential in optimizing enzyme cocktails for the hydrolysis of mildly pretreated lignocellulosic biomass by using enzymes and additive combinations tailored to the specific biomass composition.

Enzymatic hydrolysis cocktail optimization for the intensification of sugar extraction from sugarcane bagasse.

Lignocellulosic biomasses have a very important role as a raw material to produce biofuels and biochemicals. However, a sustainable, efficient, and economically competitive process for the release of sugars from such materials has still not been achieved. In this work, the optimization of the enzymatic hydrolysis cocktail was evaluated as an approach to maximize sugar extraction from mildly pretreated sugarcane bagasse. Different additives and enzymes, including hydrogen peroxide (H2O2), laccase, hemicellulase and the surfactants Tween 80 and PEG4000 were added to a cellulolytic cocktail with the aim of improving biomass hydrolysis. An increase of 39 % and 46 % of glucose and xylose concentrations, respectively, compared to the control (when only the cellulolytic cocktail (20 or 35 FPU g-1 dry mass), was obtained when H2O2 (0.24 mM) was added at the beginning of the hydrolysis. On the other hand, the addition of hemicellulase (81-162 µL g-1 DM) increased the production of glucose up to 38 % and xylose up to 50 %. The findings of this study reveal that it is possible to increase the extraction of sugars from mildly pretreated lignocellulosic biomass by using an appropriate enzymatic cocktail supplemented with additives. This opens up new opportunities for the development of a more sustainable, efficient, and economically competitive process for biomass fractionation.