Interfacial molecular interactions of cellobiohydrolase Cel7A and its variants on cellulose.

Background: Molecular-scale mechanisms of the enzymatic breakdown of cellulosic biomass into fermentable sugars are still poorly understood, with a need for independent measurements of enzyme kinetic parameters. We measured binding times of cellobiohydrolase Cel7A (Cel7A) on celluloses using wild-type Cel7A (WT), the catalytically deficient mutant Cel7A E212Q (E212Q) and their proteolytically isolated catalytic domains (CD) (WT and E212Q, respectively). The binding time distributions were obtained from time-resolved, super-resolution images of fluorescently labeled enzymes on cellulose obtained with total internal reflection fluorescence microscopy.

Mechanistic kinetic models of enzymatic cellulose hydrolysis-A review.

Bioconversion of lignocellulose forms the basis for renewable, advanced biofuels, and bioproducts. Mechanisms of hydrolysis of cellulose by cellulases have been actively studied for nearly 70 years with significant gains in understanding of the cellulolytic enzymes. Yet, a full mechanistic understanding of the hydrolysis reaction has been elusive.