Identification of promiscuous ene-reductase activity by mining structural databases using active site constellations

Georg Steinkellner, Christian C. Gruber, Tea Pavkov-Keller, Alexandra Binter, Kerstin Steiner, Christoph Winkler, Andrzej Lyskowski, Orsolya Schwamberger, Monika Oberer, Helmut Schwab, Kurt Faber, Peter Macheroux, Karl Gruber*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

Abstract

The exploitation of catalytic promiscuity and the application of de novo design have recently opened the access to novel, non-natural enzymatic activities. Here we describe a structural bioinformatic method for predicting catalytic activities of enzymes based on three-dimensional constellations of functional groups in active sites (‘catalophores’). As a proof-of-concept we identify two enzymes with predicted promiscuous ene-reductase activity (reduction of activated C–C double bonds) and compare them with known ene-reductases, that is, members of the Old Yellow Enzyme family. Despite completely different amino acid sequences, overall structures and protein folds, high-resolution crystal structures reveal equivalent binding modes of typical Old Yellow Enzyme substrates and ligands. Biochemical and biocatalytic data show that the two enzymes indeed possess ene-reductase activity and reveal an inverted stereopreference compared with Old Yellow Enzymes for some substrates. This method could thus be a tool for the identification of viable starting points for the development and engineering of novel biocatalysts
Original languageEnglish
Article number4150
Pages (from-to)1-9
JournalNature Communications
Volume5
DOIs
Publication statusPublished - 2014

Fields of Expertise

  • Human- & Biotechnology

Treatment code (Nähere Zuordnung)

  • Basic - Fundamental (Grundlagenforschung)

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