Reducing non-productive adsorption of cellulase and enhancing enzymatic hydrolysis of lignocelluloses by noncovalent modification of lignin with lignosulfonate.

Four fractions of one commercial sodium lignosulfonate (SXP) with different molecular weight (MW) and anionic polymers were studied to reduce non-productive adsorption of cellulase on bound lignin in a lignocellulosic substrate. SXP with higher MW had stronger blocking effect on non-productive adsorption of a commercial Trichoderma reesi cellulase cocktail (CTec2) on lignin measured by quartz crystal microgravimetry with dissipation monitoring. Linear anionic aromatic polymers have strong blocking effect, but they would also reduce CTec2 adsorption on cellulose to decrease the enzymatic activity. The copolymer of lignin and polyethylene glycol (AL-PEG1000) has strong enhancement in enzymatic hydrolysis of lignocelluloses, because it not only improves the cellulase activity to cellulose, but also blocks the non-productive cellulase adsorption on lignin. Apart from improving the cellulase activity to cellulose, the enhancements of enzymatic hydrolysis of lignocellulose by adding AL-PEG1000 and SXPs are the result of the decreased cellulase non-productive adsorption on lignin.

pH-Induced lignin surface modification to reduce nonspecific cellulase binding and enhance enzymatic saccharification of lignocelluloses.

We studied the mechanism of the significant enhancement in the enzymatic saccharification of lignocelluloses at an elevated pH of 5.5-6.0. Four lignin residues with different sulfonic acid contents were isolated from enzymatic hydrolysis of lodgepole pine pretreated by either dilute acid (DA) or sulfite pretreatment to overcome recalcitrance of lignocelluloses (SPORL). The adsorption isotherms of a commercial Trichoderma reesi cellulase cocktail (CTec2) produced by these lignin residues at 50 °C were measured in the pH range of 4.5-6.0. The zeta potentials of these lignin samples were also measured. We discovered that an elevated pH significantly increased the lignin surface charge (negative), which causes lignin to become more hydrophilic and reduces its coordination affinity to cellulase and, consequently, the nonspecific binding of cellulase. The decreased nonspecific cellulase binding to lignin is also attributed to enhanced electrostatic interactions at elevated pH through the increased negative charges of cellulase enzymes with low pI. The results validate the hypothesis that the increases in enzymatic saccharification efficiencies at elevated pH for different pretreated lignocelluloses are solely the result of decreased nonspecific cellulase binding to lignin. This study contradicts the well-established concept that the optimal pH is 4.8-5.0 for enzymatic hydrolysis using Trichoderma reesi cellulose, which is widely accepted and exclusively practiced in numerous laboratories throughout the world. Because an elevated pH can be easily implemented commercially without capital cost and with minimal operating cost, this study has both scientific importance and practical significance.