Exploring the action of endoglucanases on bleached eucalyptus kraft pulp as potential catalyst for isolation of cellulose nanocrystals

Cellulose nanocrystals (CNCs) is a high-value and emerging bionanomaterial with an increasing number of applications. The action of endoglucanases (EGs) from fungal and bacterial sources belonging to three glycosyl hydrolase (GH) families were investigated on bleached eucalyptus kraft pulp as potential catalysts to prepare CNC. Fungal GH7EG was the most efficient in hydrolysis and fiber fragmentation without altering crystallinity and crystallite size. Fiber fragmentation promoted by fungal GH45EG was similar to that observed for GH7EG, although it released a smaller amount of sugar. Bacterial GH5EG resulted in very low hydrolysis yield and practically did not fragment the fibers, resulting in a hydrolysis residue with characteristics very similar to the initial material. GH45EG was the only EG that affected the crystallinity and crystallite size and also the only enzyme capable of isolating nanoparticles. The isolated nanoparticles had very narrow width distribution range of 6–10 nm and length distribution range of 400–600 nm. Supplementation of β-glucosidase and conventional mechanical refining as a pretreatment did not improve the release of nanoparticles. Despite catalyzing the same biochemical reaction, different EGs displayed very distinct action during hydrolysis. The reported strong binding of GH45EG's CBM to the cellulose and the lack of increased accessibility of the enzyme to new substrate likely allowed continuous hydrolysis of the few fibers available, resulting in the isolation of cellulose nanoparticles.