title: Competitive sorption kinetics of inhibited endo- and exoglucanases on a model cellulose substrate.
abstract: For the first time, the competitive adsorption of inhibited cellobiohydrolase I (Cel7A, an exoglucanase) and endoglucanase I (Cel7B) from T. longibrachiatum is studied on cellulose. Using quartz crystal microgravimetry (QCM), sorption histories are measured for individual types of cellulases and their mixtures adsorbing to and desorbing from a model cellulose surface. We find that Cel7A has a higher adsorptive affinity for cellulose than does Cel7B. The adsorption of both cellulases becomes irreversible on time scales of 30-60 min, which are much shorter than those typically used for industrial cellulose hydrolysis. A multicomponent Langmuir kinetic model including first-order irreversible binding is proposed. Although adsorption and desorption rate constants differ between the two enzymes, the rate at which each surface enzyme irreversibly binds is identical. Because of the higher affinity of Cel7A for the cellulose surface, when Cel7A and Cel7B compete for surface sites, a significantly higher bulk concentration of Cel7B is required to achieve comparable surface enzyme concentrations. Because cellulose deconstruction benefits significantly from the cooperative activity of endoglucanases and cellobiohydrolases on the cellulose surface, accounting for competitive adsorption is crucial to developing effective cellulase mixtures.
authors.affiliation: Department of Chemical and Biomolecular Engineering, University of California-Berkeley, Berkeley, California 94720-1462, USA.
authors.forename: Samuel A
authors.initials: SA
authors.lastname: Maurer
authors.forename: Claire N
authors.initials: CN
authors.lastname: Bedbrook
authors.forename: Clayton J
authors.initials: CJ
authors.lastname: Radke
mesh_terms: D000327:Adsorption; D044602:Cellulases / Q000037:antagonists & inhibitors / Q000737:chemistry* / Q000378:metabolism; D002482:Cellulose / Q000737:chemistry*; D043366:Cellulose 1,4-beta-Cellobiosidase / Q000037:antagonists & inhibitors / Q000737:chemistry* / Q000378:metabolism; D006868:Hydrolysis; D007700:Kinetics; D008956:Models, Chemical; D013379:Substrate Specificity; D013499:Surface Properties; D014242:Trichoderma / Q000201:enzymology
chemical_list: D002482:Cellulose; D044602:Cellulases; D043366:Cellulose 1,4-beta-Cellobiosidase