title: A single-molecule analysis reveals morphological targets for cellulase synergy.
abstract: The mechanisms of enzyme activity on solid substrates are not well understood. Unlike enzyme catalysis in aqueous solutions, enzyme activity on surfaces is complicated by adsorption steps and structural heterogeneities that make enzyme-substrate interactions difficult to characterize. Cellulase enzymes, which catalyze the depolymerization of cellulose, show binding specificities for different cellulose surface morphologies, but the influence of these specificities on the activity of multienzyme mixtures has remained unclear. We developed a metric to quantify binding-target arrangements determined by photoactivated localization microscopy, and we used that metric to show that combinations of cellulases designed to bind within similar but nonidentical morphologies can have synergistic activity. This phenomenon cannot be explained with the binary crystalline or amorphous classifications commonly used to characterize cellulase-binding targets. Our results reveal a strategy for improving the activity of cellulolytic mixtures and demonstrate a versatile method for investigating protein organization on heterogeneous surfaces.
authors.affiliation: Energy Biosciences Institute, University of California, Berkeley, California, USA.
authors.forename: Jerome M
authors.initials: JM
authors.lastname: Fox
authors.forename: Phillip
authors.initials: P
authors.lastname: Jess
authors.forename: Rakesh B
authors.initials: RB
authors.lastname: Jambusaria
authors.forename: Genny M
authors.initials: GM
authors.lastname: Moo
authors.forename: Jan
authors.initials: J
authors.lastname: Liphardt
authors.forename: Douglas S
authors.initials: DS
authors.lastname: Clark
authors.forename: Harvey W
authors.initials: HW
authors.lastname: Blanch
mesh_terms: D000327:Adsorption; D001665:Binding Sites; D002384:Catalysis; D002480:Cellulase / Q000737:chemistry*; D002482:Cellulose / Q000737:chemistry; D032561:Cotton Fiber; D005456:Fluorescent Dyes; D006868:Hydrolysis; D008856:Microscopy, Fluorescence / Q000379:methods*; D008969:Molecular Sequence Data; D011485:Protein Binding; D013379:Substrate Specificity
chemical_list: D005456:Fluorescent Dyes; D002482:Cellulose; D002480:Cellulase
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