title: Revealing Additional Stereocomplementary Pairs of Old Yellow Enzymes by Rational Transfer of Engineered Residues.
abstract: Every year numerous protein engineering and directed evolution studies are published, increasing the knowledge that could be used by protein engineers. Here we test a protein engineering strategy that allows quick access to improved biocatalysts with very little screening effort. Conceptually it is assumed that engineered residues previously identified by rational and random methods induce similar improvements when transferred to family members. In an application to ene-reductases from the Old Yellow Enzyme (OYE) family, the newly created variants were tested with three compounds, revealing more stereocomplementary OYE pairs with potent turnover frequencies (up to 660 h-1) and excellent stereoselectivities (up to >99 %). Although systematic prediction of absolute enantioselectivity of OYE variants remains a challenge, "scaffold sampling" was confirmed as a promising addition to protein engineers' collection of strategies.
authors.affiliation: Department of Chemistry, Philipps-Universität Marburg, Hans-Meerwein-Strasse 4, 35032, Marburg, Germany.
authors.forename: Nathalie
authors.initials: N
authors.lastname: Nett
authors.affiliation: Department of Chemistry, Philipps-Universität Marburg, Hans-Meerwein-Strasse 4, 35032, Marburg, Germany.
authors.forename: Sabine
authors.initials: S
authors.lastname: Duewel
authors.affiliation: Department of Chemistry, Philipps-Universität Marburg, Hans-Meerwein-Strasse 4, 35032, Marburg, Germany.
authors.forename: Alexandra Annelis
authors.initials: AA
authors.lastname: Richter
authors.affiliation: Department of Chemistry, Philipps-Universität Marburg, Hans-Meerwein-Strasse 4, 35032, Marburg, Germany.
authors.forename: Sabrina
authors.identifier: 0000-0002-4715-7123
authors.initials: S
authors.lastname: Hoebenreich
mesh_terms: D000179:Acrylates / Q000737:chemistry; D001224:Aspartic Acid / Q000737:chemistry; D000081005:Cyclohexane Monoterpenes; D003510:Cyclohexanes / Q000737:chemistry; D004795:Enzyme Stability; D005998:Glycine / Q000737:chemistry; D007700:Kinetics; D039821:Monoterpenes / Q000737:chemistry; D016296:Mutagenesis; D009252:NADPH Dehydrogenase / Q000737:chemistry* / Q000235:genetics*; D015202:Protein Engineering; D013237:Stereoisomerism; D013912:Threonine / Q000737:chemistry
chemical_list: D000179:Acrylates; D000081005:Cyclohexane Monoterpenes; D003510:Cyclohexanes; D039821:Monoterpenes; D013912:Threonine; D001224:Aspartic Acid; C006923:carvone; C473568:3-methyl-2-cyclohexen-1-one; D009252:NADPH Dehydrogenase; D005998:Glycine
keywords: asymmetric synthesis; ene reductases; enzyme-based stereocontrol; protein engineering; residue transfer