title: Enhanced cellulose degradation using cellulase-nanosphere complexes.
abstract: Enzyme catalyzed conversion of plant biomass to sugars is an inherently inefficient process, and one of the major factors limiting economical biofuel production. This is due to the physical barrier presented by polymers in plant cell walls, including semi-crystalline cellulose, to soluble enzyme accessibility. In contrast to the enzymes currently used in industry, bacterial cellulosomes organize cellulases and other proteins in a scaffold structure, and are highly efficient in degrading cellulose. To mimic this clustered assembly of enzymes, we conjugated cellulase obtained from Trichoderma viride to polystyrene nanospheres (cellulase:NS) and tested the hydrolytic activity of this complex on cellulose substrates from purified and natural sources. Cellulase:NS and free cellulase were equally active on soluble carboxymethyl cellulose (CMC); however, the complexed enzyme displayed a higher affinity in its action on microcrystalline cellulose. Similarly, we found that the cellulase:NS complex was more efficient in degrading natural cellulose structures in the thickened walls of cultured wood cells. These results suggest that nanoparticle-bound enzymes can improve catalytic efficiency on physically intractable substrates. We discuss the potential for further enhancement of cellulose degradation by physically clustering combinations of different glycosyl hydrolase enzymes, and applications for using cellulase:NS complexes in biofuel production.
authors.affiliation: Physical and Life Sciences, Lawrence Livermore National Laboratory, Livermore, California. blanchette2@llnl.gov
authors.forename: Craig
authors.initials: C
authors.lastname: Blanchette
authors.forename: Catherine I
authors.initials: CI
authors.lastname: Lacayo
authors.forename: Nicholas O
authors.initials: NO
authors.lastname: Fischer
authors.forename: Mona
authors.initials: M
authors.lastname: Hwang
authors.forename: Michael P
authors.initials: MP
authors.lastname: Thelen
mesh_terms: D056804:Biofuels; D002266:Carboxymethylcellulose Sodium / Q000737:chemistry*; D002480:Cellulase / Q000737:chemistry*; D004800:Enzymes, Immobilized / Q000737:chemistry*; D005656:Fungal Proteins / Q000737:chemistry*; D054118:Nanospheres / Q000737:chemistry*; D014242:Trichoderma / Q000201:enzymology*
chemical_list: D056804:Biofuels; D004800:Enzymes, Immobilized; D005656:Fungal Proteins; D002480:Cellulase; D002266:Carboxymethylcellulose Sodium