Related Experiment Video
Updated: Jun 19, 2026
High-throughput Screening of Recalcitrance Variations in Lignocellulosic Biomass: Total Lignin, Lignin Monomers, and Enzymatic Sugar Release
Published on: September 15, 2015
Direct quantitative determination of adsorbed cellulase on lignocellulosic biomass with its application to study
Zhiguang Zhu1, Noppadon Sathitsuksanoh, Y-H Percival Zhang
1Biological Systems Engineering Department, Virginia Polytechnic Institute and State University (Virginia Tech), 210-A Seitz Hall, Blacksburg, Virginia 24061, USA.
Abstract:
Effective hydrolysis of pretreated lignocellulose mediated by cellulase requires an in-depth understanding of cellulase adsorption and desorption. Here we developed a simple method for determining the adsorbed cellulase on cellulosic materials or pretreated lignocellulose, which involves (i) hydrolysis of adsorbed cellulase in the presence of 10 M of NaOH at 121 degrees C for 20 min, and (ii) the ninhydrin assay for the amino acids released from the hydrolyzed cellulase. The major lignocellulosic components (i.e., cellulose, hemicellulose, and lignin) did not interfere with the ninhydrin assay. A number of cellulase desorption methods were investigated: pH change as well as the use of detergents, high salt solution, and polyhydric alcohols. The pH adjustment to 13.0 and the elution by 72% ethylene glycol at neutral pH were among the most efficient approaches for desorbing the adsorbed cellulase. For the recycling of active cellulase, a modest pH adjustment to 10.0 may be a low-cost viable method to desorb active cellulase. It was found that more than 90% of cellulase for hydrolysis of the pretreated corn stover could be recycled by washing at pH 10.0.
