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  • Harnessing prefoldin proteins for enzyme assemblies and fabrication of metalloprotein nanowires
    (2023) Lam, Nga
    Thesis
    The self-assembly of proteins into intricate high-order structures can be harnessed for the precise positioning of functional molecules in nanotechnology. The organisation of enzymes on protein scaffolds has been previously shown to enhance enzyme catalytic activity. Additionally, the alignment of metal-binding proteins, known as metalloproteins, on filamentous proteins has been exploited to produce electrically conductive nanowires. The focus of this thesis is on the development of improved biosynthetic strategies for the creation of multifunctional nanomaterials by harnessing the self-assembly of filamentous proteins. Central to the engineering in this thesis is prefoldin, which is a molecular chaperone from archaea with the ability to self-assemble into hetero-hexameric complexes and filamentous structures. Prefoldin proteins exhibits high thermal stability and have engineerable interfaces for bioconjugation of functional proteins. Therefore, the research goal of this thesis was to engineer robust and modular protein scaffolds for the precise position of enzymes and alignment of electrically conductive subunits to create biocatalysis and bioelectronic systems. The first aim of the research was to construct a protein scaffold from a hexameric self-assembling protein and immobilise enzymes on the protein scaffold to examine for enhanced sequential catalytic reactions. The second aim explored the capability of prefoldin filaments to align various metalloproteins in proximity over large distances for electron transfer. Distinct metalloproteins were exploited to create nanowires with various electronic properties for applications in bioelectronic devices. The third aim developed a strategy to localise redox enzymes at either end of the metalloprotein nanowires and potentially demonstrate energy transfer along the nanowire between enzymes undergoing redox reactions. The successful achievement of these aims establishes a biostrategy to use a controllable and modular prefoldin protein scaffold for the fabrication of biocatalysis and bioelectronic devices.