Projects per year
Abstract
Designing enzyme-like catalysts tests our understanding of sequence-to-structure/function relationships in proteins. Here, we install hydrolytic activity predictably into a completely de novo and thermo-stable α-helical barrel, which comprises 7 helices arranged around an accessible channel. We show that the lumen of the barrel accepts 21 mutations to functional polar residues. The resulting variant, which has cysteine-histidine-glutamic acid triads on each helix, hydrolyses p-nitrophenyl acetate with catalytic efficiencies matching the most-efficient redesigned hydrolases based on natural protein scaffolds. This is the first report of a functional catalytic triad engineered into a de novo protein framework. The flexibility of our system also allows the facile incorporation of unnatural side chains to improve activity and probe the catalytic mechanism. Such predictable and robust construction of truly de novo biocatalysts holds promise for applications in chemical and biochemical synthesis.
Original language | English |
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Pages (from-to) | 837-844 |
Number of pages | 8 |
Journal | Nature Chemistry |
Volume | 8 |
Issue number | 9 |
Early online date | 4 Jul 2016 |
DOIs | |
Publication status | Published - 1 Sep 2016 |
Structured keywords
- Bristol BioDesign Institute
- BrisSynBio
- BCS and TECS CDTs
Keywords
- Biocatalysis
- Hydrolases
- Protein design
Fingerprint
Dive into the research topics of 'Installing hydrolytic activity into a completely de novo protein framework'. Together they form a unique fingerprint.Projects
- 1 Finished
Profiles
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Professor R L Brady
- Fundamental Bioscience
- School of Biochemistry - Professor of Biochemistry
Person: Academic , Member
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Professor Dek N Woolfson
- The Bristol Centre for Nanoscience and Quantum Information
- Synthesis
- School of Chemistry - Professor of Chemistry and Biochemistry
- Soft Matter, Colloids and Materials
- Catalysis
- Biological and Archaeological Chemistry
Person: Academic (former), Academic , Member