Enhanced Saccharification of Purple Alfalfa via Sequential Pretreatment with Acidified Ethylene Glycol and Urea/NaOH

Purple Alfalfa is an inexpensive, abundant, readily available lignocellulosic material. This work was attempted to develop an efficient combination pretreatment by sequential HClO4–ethyl glycol–H2O (1.2:88.8:10, w/w/w) extraction at 130 °C in 0.5 h and urea/NaOH (urea 12 wt%, NaOH 7 wt%) soaking at −20 °C for 0.5 h for the pretreatment of purple alfalfa. The porosity, morphology, and crystallinity of pretreated purple alfalfa were characterized with SEM, FM, XRD, and FTIR. This combination pretreatment had a significant influence on hemicellulose removal and delignification. The above changes could enhance cellulose accessibility to enzymes and improve the enzymatic digestibility of cellulose. High yields of reducing sugars from pretreated purple alfalfa were obtained at 93.4%. In summary, this combination pretreatment has high potential application in the future.

Understanding the effects of different residual lignin fractions in acid-pretreated bamboo residues on its enzymatic digestibility

Background During the dilute acid pretreatment process, the resulting pseudo-lignin and lignin droplets deposited on the surface of lignocellulose and inhibit the enzymatic digestibility of cellulose in lignocellulose. However, how these lignins interact with cellulase enzymes and then affect enzymatic hydrolysis is still unknown. In this work, different fractions of surface lignin (SL) obtained from dilute acid-pretreated bamboo residues (DAP-BR) were extracted by various organic reagents and the residual lignin in extracted DAP-BR was obtained by the milled wood lignin (MWL) method. All of the lignin fractions obtained from DAP-BR were used to investigate the mechanism for interaction between lignin and cellulase using surface plasmon resonance (SPR) technology to understand how they affect enzymatic hydrolysis Results The results showed that removing surface lignin significantly decreased the yield for enzymatic hydrolysis DAP-BR from 36.5% to 18.6%. The addition of MWL samples to Avicel inhibited its enzymatic hydrolysis, while different SL samples showed slight increases in enzymatic digestibility. Due to the higher molecular weight and hydrophobicity of MWL samples versus SL samples, a stronger affinity for MWL (KD = 6.8–24.7 nM) was found versus that of SL (KD = 39.4–52.6 nM) by SPR analysis. The affinity constants of all tested lignins exhibited good correlations (r > 0.6) with the effects on enzymatic digestibility of extracted DAP-BR and Avicel. Conclusions This work revealed that the surface lignin on DAP-BR is necessary for maintaining enzyme digestibility levels, and its removal has a negative impact on substrate digestibility.