The use of Carbohydrate Binding Modules (CBMs) to monitor changes in fragmentation and cellulose fibre surface morphology during Cellulase and Swollenin induced deconstruction of lignocellulosic substrates [Microbiology]
December 19th, 2014 by Gourlay, K., Hu, J., Arantes, V., Penttila, M., Saddler, J. N.
Although the actions of many of the hydrolytic enzymes involved in cellulose hydrolysis are relatively well understood, the contributions that amorphogenesis-inducing proteins might contribute to cellulose deconstruction are still relatively undefined. Earlier work has shown that disruptive proteins, such as the non-hydrolytic non-oxidative protein Swollenin, can open up and disaggregate the less-ordered regions of lignocellulosic substrates. Within the cellulosic fraction, relatively disordered, amorphous regions known as dislocations are known to occur along the length of the fibres. It was postulated that Swollenin might act synergistically with hydrolytic enzymes to initiate biomass deconstruction within these dislocation regions. Carbohydrate Binding Modules (CBMs) that preferentially bind to cellulosic substructures were fluorescently labelled. They were imaged, using Confocal Microscopy, to assess the distribution of crystalline and amorphous cellulose at the fibre surface, as well as to track changes in surface morphology over the course of enzymatic hydrolysis and fibre fragmentation. Swollenin was shown to promote targeted disruption of the cellulosic structure at fibre dislocations.