title: Nanocaged enzymes with enhanced catalytic activity and increased stability against protease digestion.
abstract: Cells routinely compartmentalize enzymes for enhanced efficiency of their metabolic pathways. Here we report a general approach to construct DNA nanocaged enzymes for enhancing catalytic activity and stability. Nanocaged enzymes are realized by self-assembly into DNA nanocages with well-controlled stoichiometry and architecture that enabled a systematic study of the impact of both encapsulation and proximal polyanionic surfaces on a set of common metabolic enzymes. Activity assays at both bulk and single-molecule levels demonstrate increased substrate turnover numbers for DNA nanocage-encapsulated enzymes. Unexpectedly, we observe a significant inverse correlation between the size of a protein and its activity enhancement. This effect is consistent with a model wherein distal polyanionic surfaces of the nanocage enhance the stability of active enzyme conformations through the action of a strongly bound hydration layer. We further show that DNA nanocages protect encapsulated enzymes against proteases, demonstrating their practical utility in functional biomaterials and biotechnology.
authors: Center for Molecular Design and Biomimetics, the Biodesign Institute at Arizona State University, Tempe, Arizona 85287, USA.|School of Molecular Sciences, Arizona State University, Tempe, Arizona 85287, USA.
authors: Zhao
authors: Z
authors: Zhao
authors: Department of Chemistry, Center for Computational and Integrative Biology, Rutgers University-Camden, Camden, New Jersey 08102, USA.
authors: Jinglin
authors: J
authors: Fu
authors: Department of Chemistry, Single Molecule Analysis Group, University of Michigan, Ann Arbor, Michigan 48109, USA.
authors: Soma
authors: S
authors: Dhakal
authors: Department of Chemistry, Single Molecule Analysis Group, University of Michigan, Ann Arbor, Michigan 48109, USA.
authors: Alexander
authors: A
authors: Johnson-Buck
authors: Center for Molecular Design and Biomimetics, the Biodesign Institute at Arizona State University, Tempe, Arizona 85287, USA.
authors: Minghui
authors: M
authors: Liu
authors: Department of Chemistry, Center for Computational and Integrative Biology, Rutgers University-Camden, Camden, New Jersey 08102, USA.
authors: Ting
authors: T
authors: Zhang
authors: School of Molecular Sciences, Arizona State University, Tempe, Arizona 85287, USA.|Center for Innovations in Medicine, the Biodesign Institute at Arizona State University, Tempe, Arizona 85287, USA.
authors: Neal W
authors: NW
authors: Woodbury
authors: Center for Molecular Design and Biomimetics, the Biodesign Institute at Arizona State University, Tempe, Arizona 85287, USA.|School of Molecular Sciences, Arizona State University, Tempe, Arizona 85287, USA.
authors: Yan
authors: Y
authors: Liu
authors: Department of Chemistry, Single Molecule Analysis Group, University of Michigan, Ann Arbor, Michigan 48109, USA.
authors: Nils G
authors: NG
authors: Walter
authors: Center for Molecular Design and Biomimetics, the Biodesign Institute at Arizona State University, Tempe, Arizona 85287, USA.|School of Molecular Sciences, Arizona State University, Tempe, Arizona 85287, USA.
authors: Hao
authors: H
authors: Yan
mesh_terms: D002384:Catalysis; D004247:DNA / Q000648:ultrastructure*; D004795:Enzyme Stability; D004798:Enzymes / Q000378:metabolism*; D005949:Glucose Oxidase / Q000378:metabolism; D005954:Glucosephosphate Dehydrogenase / Q000378:metabolism; D006735:Horseradish Peroxidase / Q000378:metabolism; D045022:Lactate Dehydrogenases / Q000378:metabolism; D008291:Malate Dehydrogenase / Q000378:metabolism; D046529:Microscopy, Electron, Transmission; D008856:Microscopy, Fluorescence; D049329:Nanostructures / Q000648:ultrastructure*; D001616:beta-Galactosidase / Q000378:metabolism
chemical_list: D004798:Enzymes; D004247:DNA; D045022:Lactate Dehydrogenases; D008291:Malate Dehydrogenase; D005954:Glucosephosphate Dehydrogenase; D005949:Glucose Oxidase; D006735:Horseradish Peroxidase; D001616:beta-Galactosidase