Construction and in vivo assembly of a catalytically proficient and hyperthermostable de novo enzyme

Watkins, Daniel W., Jenkins, Jonathan M. X., Grayson, Katie J., Wood, Nicola, Steventon, Jack W., Le Vay, Kristian K., Goodwin, Matthew I., Mullen, Anna S., Bailey, Henry J., Crump, Matthew P., MacMillan, Fraser, Mulholland, Adrian J., Cameron, Gus, Sessions, Richard B., Mann, Stephen and Anderson, J. L. Ross (2017) Construction and in vivo assembly of a catalytically proficient and hyperthermostable de novo enzyme. Nature Communications, 8 (1). ISSN 2041-1723

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Abstract

Although catalytic mechanisms in natural enzymes are well understood, achieving the diverse palette of reaction chemistries in re-engineered native proteins has proved challenging. Wholesale modification of natural enzymes is potentially compromised by their intrinsic complexity, which often obscures the underlying principles governing biocatalytic efficiency. The maquette approach can circumvent this complexity by combining a robust de novo designed chassis with a design process that avoids atomistic mimicry of natural proteins. Here, we apply this method to the construction of a highly efficient, promiscuous, and thermostable artificial enzyme that catalyzes a diverse array of substrate oxidations coupled to the reduction of H2O2. The maquette exhibits kinetics that match and even surpass those of certain natural peroxidases, retains its activity at elevated temperature and in the presence of organic solvents, and provides a simple platform for interrogating catalytic intermediates common to natural heme-containing enzymes.

Item Type: Article
Faculty \ School: Faculty of Medicine and Health Sciences > Norwich Medical School
Faculty of Science
Faculty of Science > School of Chemistry
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Depositing User: Pure Connector
Date Deposited: 05 Sep 2017 05:07
Last Modified: 14 Nov 2020 00:57
URI: https://ueaeprints.uea.ac.uk/id/eprint/64743
DOI: 10.1038/s41467-017-00541-4

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