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An Enzymatic Active Site Embedded in a DNA Nanostructure

Tadija Kekic, Yasaman Ahmadi, Ivan Barisic
doi: https://doi.org/10.1101/804518
Tadija Kekic
1Molecular Diagnostics, Center for Health and Bioresources, AIT Austrian Institute of Technology GmbH, Giefinggasse 4, 1210 Vienna, Austria
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Yasaman Ahmadi
1Molecular Diagnostics, Center for Health and Bioresources, AIT Austrian Institute of Technology GmbH, Giefinggasse 4, 1210 Vienna, Austria
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Ivan Barisic
1Molecular Diagnostics, Center for Health and Bioresources, AIT Austrian Institute of Technology GmbH, Giefinggasse 4, 1210 Vienna, Austria
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  • For correspondence: Ivan.Barisic@ait.ac.at
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ABSTRACT

Artificial enzymes hold a great potential in the field of biotechnology. The currently available design strategies are limited to recruit and organise secondary and tertiary structural elements, and functional motifs of native enzymes. Here, we demonstrate an approach towards the bottom-up development of an artificial enzyme based on DNA nanotechnology. Structural analyses of the FlaI ATPase were used to identify a set of amino acids essential for the catalytic reaction. The selected amino acids were used to design four peptide-oligonucleotide conjugates (POCs). The POCs were integrated into a DNA nanostructure and reconstituted the catalytic site of FlaI. DNA origami technology was employed to maintain the relative orientations of the amino acids and their positions analogous to those in the protein catalytic site. The substrate turnover rates were found to be comparable in our artificial nanozyme (kcat = 0.3622 s-1) and the native enzyme (kcat ≈ 0.016 s-1). The nanozymes could be recycled using an ultrafiltration protocol and reused in multiple experiments allowing a reliable reproduction of the measurements. The emulation of enzymatic activity, by this novel and computable technological framework, can be of high relevance in many different areas.

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Posted January 13, 2020.
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An Enzymatic Active Site Embedded in a DNA Nanostructure
Tadija Kekic, Yasaman Ahmadi, Ivan Barisic
bioRxiv 804518; doi: https://doi.org/10.1101/804518
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An Enzymatic Active Site Embedded in a DNA Nanostructure
Tadija Kekic, Yasaman Ahmadi, Ivan Barisic
bioRxiv 804518; doi: https://doi.org/10.1101/804518

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