Optimization of enzymatic saccharification of industrial wastes using a thermostable and halotolerant endoglucanase through Box-Behnken experimental design.

This study describes the production, characterization and application of an endoglucanase from Penicillium roqueforti using lignocellulosic agro-industrial wastes as the substrate during solid-state fermentation. The endoglucanase was generated after culturing with different agro-industrial wastes for 96 h without any pretreatment. The highest activity was obtained at 50 °C and pH 4.0. Additionally, the enzyme showed stability in the temperature and pH ranges of 40-80 °C and 4.0-5.0, respectively. The addition of Ca2+, Zn2+, Mg2+, and Cu2+ increased enzymatic activity. Halotolerance as a characteristic of the enzyme was confirmed when its activity increased by 35% on addition of 2 M NaCl. The endoglucanase saccharified sugarcane bagasse, coconut shell, wheat bran, cocoa fruit shell, and cocoa seed husk. The Box-Behnken design was employed to optimize fermentable sugar production by evaluating the following parameters: time, substrate, and enzyme concentration. Under ideal conditions, 253.19 mg/g of fermentable sugars were obtained following the saccharification of wheat bran, which is 41.5 times higher than that obtained without optimizing. This study presents a thermostable, halotolerant endoglucanase that is resistant to metal ions and organic solvents with the potential to be applied in producing fermentable sugars for manufacturing biofuels from agro-industrial wastes.

Dilute acid pretreatment for enhancing the enzymatic saccharification of agroresidues using a Botrytis ricini endoglucanase.

The enormous amount of agroindustrial residues generated in Brazil can be used as biomass to produce fermentable sugars. This study compared the pretreatments with different proportions of dilute acid. The method involved pretreatment with 0.5%, 1%, and 1.5% (v/v) sulfuric acid, followed by hydrolysis using the halotolerant and thermostable endoglucanase from Botrytis ricini URM 5627. The physicochemical characterization of plant biomass was performed using XRD, FTIR, and SEM. The pretreatment significantly increased the production of fermentable sugars following enzymatic saccharification from wheat bran, sugarcane bagasse, and rice husk: 153.67%, 91.98%, and 253.21% increment in sugar production; 36.39 mg⋅g-1 ± 1.23, 39.55 mg⋅g-1 ± 1.70, and 42.53 mg⋅g-1 ± 7.61 mg⋅L-1 of glucose; and 3.26 ± 0.35 mg⋅g-1 , 3.61mg⋅g-1 ± 0.74 and 3.59 mg⋅g-1 ± 0.80 of fructose were produced, respectively. In conclusion, biomass should preferably be pretreated before the enzymatic saccharification using B. ricini URM 5627 endoglucanase.