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

Daniel W Watkins, Jonathan M X Jenkins, Katie J Grayson, Nicola Wood, Jack W Steventon, Kristian Le Vay, Matthew I Goodwin, Anna S Mullen, Henry J Bailey, Matthew P Crump, Fraser MacMillan, Adrian J Mulholland, Gus Cameron, Richard B Sessions, Stephen Mann, J L Ross Anderson*

*Corresponding author for this work

Research output: Contribution to journalArticle (Academic Journal)

33 Citations (Scopus)
463 Downloads (Pure)

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.
Original languageEnglish
Article number358
Number of pages9
JournalNature Communications
Volume8
DOIs
Publication statusPublished - 25 Aug 2017

Structured keywords

  • Bristol BioDesign Institute
  • BrisSynBio

Keywords

  • SYNTHETIC BIOLOGY

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    Student Theses

    Computational design, construction, and characterisation of artificial peroxidases

    Author: Jenkins, J., 19 Mar 2019

    Supervisor: Anderson, R. (Supervisor) & Mulholland, A. (Supervisor)

    Student thesis: Doctoral ThesisDoctor of Philosophy (PhD)

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    Cite this

    Watkins, D. W., Jenkins, J. M. X., Grayson, K. J., Wood, N., Steventon, J. W., Le Vay, K., Goodwin, M. I., Mullen, A. S., Bailey, H. J., Crump, M. P., MacMillan, F., Mulholland, A. J., Cameron, G., Sessions, R. B., Mann, S., & Anderson, J. L. R. (2017). Construction and in vivo assembly of a catalytically proficient and hyperthermostable de novo enzyme. Nature Communications, 8, [358]. https://doi.org/10.1038/s41467-017-00541-4