Engineering Thermostability in Artificial Metalloenzymes to Increase Catalytic Activity
| Autor: | Megan V. Doble, Henriëtte J. Rozeboom, Misun Lee, Paul C. J. Kamer, Lorenz Obrecht, Emma Branigan, Henk-Jan Joosten, James H. Naismith, Amanda G. Jarvis, Dick B. Janssen |
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| Přispěvatelé: | Biotechnology |
| Jazyk: | angličtina |
| Rok vydání: | 2021 |
| Předmět: |
chemistry.chemical_classification
bioengineering biocatalysis 010405 organic chemistry Rational design General Chemistry Protein engineering bioinformatics 010402 general chemistry 01 natural sciences Combinatorial chemistry Catalysis 0104 chemical sciences Turnover number Enzyme chemistry Biocatalysis protein thermostability artificial metalloenzyme hydroformylation Hydroformylation Thermostability |
| Zdroj: | Doble, M V, Obrecht, L, Joosten, H, Lee, M, Rozeboom, H J, Branigan, E, Naismith, J H, Janssen, D B, Jarvis, A G & Kamer, P C J 2021, ' Engineering Thermostability in Artificial Metalloenzymes to Increase Catalytic Activity ', ACS Catalysis, vol. 11, no. 6, pp. 3620-3627 . https://doi.org/10.1021/acscatal.0c05413 ACS Catalysis, 11(6), 3620-3627. AMER CHEMICAL SOC |
| ISSN: | 2155-5435 |
| DOI: | 10.1021/acscatal.0c05413 |
| Popis: | Protein engineering has shown widespread use in improving the industrial application of enzymes and broadening the conditions they are able to operate under by increasing their thermostability and solvent tolerance. Here, we show that protein engineering can be used to increase the thermostability of an artificial metalloenzyme. Thermostable variants of the human steroid carrier protein 2L, modified to bind a metal catalyst, were created by rational design using structural data and a 3DM database. These variants were tested to identify mutations that enhanced the stability of the protein scaffold, and a significant increase in melting temperature was observed with a number of modified metalloenzymes. The ability to withstand higher reaction temperatures resulted in an increased activity in the hydroformylation of 1-octene, with more than fivefold improvement in turnover number, whereas the selectivity for linear aldehyde remained high up to 80%. |
| Databáze: | OpenAIRE |
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