ausblenden:
Schlagwörter:
Binding Sites
Biotechnology/*methods
Catalysis
Models, Molecular
Oxidoreductases Acting on CH-CH Group Donors/*chemistry/*genetics
Protein Conformation
Protein Engineering/*methods
Protons
Stereoisomerism
Substrate Specificity
Tyrosine/chemistry/genetics
Zusammenfassung:
An improved understanding of enzymes' catalytic proficiency and stereoselectivity would further enable applications in chemistry, biocatalysis and industrial biotechnology. We use a chemical probe to dissect individual catalytic steps of enoyl-thioester reductases (Etrs), validating an active site tyrosine as the cryptic proton donor and explaining how it had eluded definitive identification. This information enabled the rational redesign of Etr, yielding mutants that create products with inverted stereochemistry at wild type-like turnover frequency.
