Construction and in vivo assembly of a catalytically proficient and hyperthermostable de novo enzyme
| Autor: | J. L. Ross Anderson, Adrian J. Mulholland, Daniel W. Watkins, Jack W. Steventon, Richard B. Sessions, Henry J. Bailey, Fraser MacMillan, Jonathan M. X. Jenkins, Matthew Goodwin, Anna Mullen, Matthew P. Crump, Kristian Le Vay, Gus Cameron, Stephen Mann, Nicola Wood, Katie J. Grayson |
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| Rok vydání: | 2016 |
| Předmět: |
0301 basic medicine
Models Molecular Science General Physics and Astronomy BrisSynBio 010402 general chemistry Protein Engineering 01 natural sciences General Biochemistry Genetics and Molecular Biology Article Substrate Specificity 03 medical and health sciences Synthetic biology lcsh:Science Nuclear Magnetic Resonance Biomolecular Peroxidase chemistry.chemical_classification Multidisciplinary Binding Sites biology Artificial enzyme Bristol BioDesign Institute Substrate (chemistry) General Chemistry Protein engineering respiratory system Combinatorial chemistry 0104 chemical sciences SYNTHETIC BIOLOGY Kinetics 030104 developmental biology Enzyme chemistry Biochemistry biology.protein Substrate specificity lcsh:Q human activities |
| Zdroj: | Nature Communications 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, vol. 8, 358 . https://doi.org/10.1038/s41467-017-00541-4 Nature Communications, Vol 8, Iss 1, Pp 1-9 (2017) |
| ISSN: | 2041-1723 |
| Popis: | 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. Catalytic mechanisms of enzymes are well understood, but achieving diverse reaction chemistries in re-engineered proteins can be difficult. Here the authors show a highly efficient and thermostable artificial enzyme that catalyzes a diverse array of substrate oxidations coupled to the reduction of H2O2. |
| Databáze: | OpenAIRE |
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