Fast Knoevenagel Condensations Catalyzed by an Artificial Schiff-Base-Forming Enzyme
| Autor: | Xavier Garrabou, Basile I. M. Wicky, Donald Hilvert |
|---|---|
| Rok vydání: | 2016 |
| Předmět: |
Models
Molecular 010402 general chemistry 01 natural sciences Biochemistry Catalysis chemistry.chemical_compound Colloid and Surface Chemistry Fructose-Bisphosphate Aldolase Organic chemistry Methylene Schiff Bases chemistry.chemical_classification Aldehydes Schiff base Molecular Structure 010405 organic chemistry Lysine Computational Biology General Chemistry Directed evolution 0104 chemical sciences Enzyme chemistry Biocatalysis Knoevenagel condensation Directed Molecular Evolution |
| Zdroj: | Journal of the American Chemical Society. 138:6972-6974 |
| ISSN: | 1520-5126 0002-7863 |
| DOI: | 10.1021/jacs.6b00816 |
| Popis: | The simple catalytic motifs utilized by enzymes created by computational design and directed evolution constitute a potentially valuable source of chemical promiscuity. Here we show that the artificial retro-aldolase RA95.5-8 is able to use a reactive lysine in a hydrophobic pocket to accelerate promiscuous Knoevenagel condensations of electron-rich aldehydes and activated methylene donors. Optimization of this activity by directed evolution afforded an efficient enzyme variant with a catalytic proficiency of 5 × 10(11) M(-1) and a10(8)-fold catalytic advantage over simple primary and secondary amines. Divergent evolution of de novo enzymes in this way could be a promising strategy for creating tailored biocatalysts for many synthetically useful reactions. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|