Enzyme Catalytic Activity Emulated Within DNA-based Nanodevice

Artificial enzymes hold great potential in the field of biotechnology. We present an approach towards the bottom-up development of an artificial enzyme using the DNA origami technology. A set of peptide-oligonucleotide conjugates, designed to recreate the structure of an active site of a native protein, was placed in the central area of a large, self-folding DNA origami shell structure. The peptide-oligonucleotide conjugates were placed in a predefined positions by the integration on the angle-adjustable, linear constructs that protruded from the nanostructure. We demonstrate a workflow to obtain the essential elements of the protein9s active site; to design and assemble the nanodevice; to measure, quantify and inactivate the catalytic activity; and to reuse our structures in multiple experiments. By use of the high-resolution spectroscopy, we demonstrated a significant increase in the product accumulation that originates from the correctly assembled emulated active sites.