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Fast Knoevenagel Condensations Catalyzed by an Artificial Schiff-Base-Forming Enzyme.
Journal article

Fast Knoevenagel Condensations Catalyzed by an Artificial Schiff-Base-Forming Enzyme.

  • Garrabou X Laboratory of Organic Chemistry, ETH Zurich , 8093 Zurich, Switzerland.
  • Wicky BI Laboratory of Organic Chemistry, ETH Zurich , 8093 Zurich, Switzerland.
  • Hilvert D Laboratory of Organic Chemistry, ETH Zurich , 8093 Zurich, Switzerland.
  • 2016-05-20
Published in:
  • Journal of the American Chemical Society. - 2016
Aldehydes
Biocatalysis
Computational Biology
Directed Molecular Evolution
Fructose-Bisphosphate Aldolase
Lysine
Models, Molecular
Molecular Structure
Schiff Bases
English 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 a >10(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.
Language
  • English
Open access status
closed
Identifiers
Persistent URL
https://folia.unifr.ch/global/documents/241475
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