Protective effects of non-catalytic proteins on endoglucanase activity at air and lignin interfaces.

The manner in which added non-catalytic proteins during enzymatic hydrolysis of lignocellulosic substrates enhances hydrolysis mechanisms is not completely understood. Prior research has indicated that a reduction in the non-specific adsorption of enzymes on lignin, and deactivation of enzymes exposed to air-liquid interface provide rationale. This work investigated root causes including effects of the air-liquid interface on non-catalytic proteins, and effects of lignin on endoglucanase. Three different experimental designs and three variables (air-liquid interfacial area, the types of lignin (acid or enzymatic lignin), and the presence of non-enzymatic protein (bovine serum albumin [BSA] or soy proteins ) were used. The results showed that acid isolated lignin adsorbed almost all endoglucanase activity initially present in supernatant, independent of air interface conditions (25 or 250 ml flasks) with the presence of BSA preventing this effect. Endoglucanase lost 30%-50% of its activity due to an air-liquid interface in the presence of lignin while addition of non-enzymatic protein helped to preserve this enzyme's activity. Langmuir and Freundlich models applied to experimental data indicated that the adsorption increases with increasing temperature for both endoglucanase and BSA. Adsorption of the enzyme and protein were endothermic with an increase in entropy. These results, combined, show that hydrophobicity plays a strong role in the adsorption of both endoglucanase and BSA on lignin.

Effect of severity factor on the hydrothermal pretreatment of sugarcane straw.

The recalcitrant structures of sugarcane straw and related lignocellulosic biomasses require a pretreatment step to enable a better enzymatic attack during the hydrolysis. Factors like the energy consumption and the formation of inhibitors require the optimization of the pretreatment step. Thus, the influence of different severity factors (SF) on hydrothermal (also called liquid hot water, LHW) pretreatment was evaluated using a factorial design 22 with central point. The obtained results showed that low values of SF (<3.39) did not promote reasonable alteration in the sugarcane straw structures, whereas high SF values (>4.70) resulted in loss of hydrolyzed sugars, generation of inhibitors such as furfural, and formation of pseudo-lignin structures, despite high hemicellulose removal (∼97%). The residence time exhibited low influence on LHW. An optimum condition was found for the process (10 min and 195 °C) with low cellulose solubilization (9.80%) and a reasonable hemicellulose removal (85.45%).