title: New Concepts for Increasing the Efficiency in Directed Evolution of Stereoselective Enzymes.
abstract: Directed evolution of stereo- and regioselective enzymes constitutes a prolific source of catalysts for asymmetric transformations in organic chemistry. In this endeavor (iterative) saturation mutagenesis at sites lining the binding pocket of enzymes has emerged as the method of choice, but uncertainties regarding the question of how to group many residues into randomization sites and how to choose optimal upward pathways persist. Two new approaches promise to beat the numbers problem effectively. One utilizes a single amino acid as building block for the randomization of a 10-residue site, the other also employs only one but possibly different amino acid at each position of a 9-residue site. The small but smart libraries provide highly enantioselective epoxide hydrolase or lipase mutants, respectively.
authors.affiliation: Max-Planck-Institut für Kohlenforschung, Kaiser-Wilhelm-Platz 1, 45470, Mülheim an der Ruhr, Germany.|Fachbereich Chemie, Philipps-Universität Marburg, Hans-Meerwein-Strasse 4, 35032, Marburg, Germany.
authors.forename: Zhoutong
authors.initials: Z
authors.lastname: Sun
authors.affiliation: Department of Organic Chemistry, Arrhenius Laboratory, Stockholm University, 106 91, Stockholm, Sweden.
authors.forename: Ylva
authors.initials: Y
authors.lastname: Wikmark
authors.affiliation: Department of Organic Chemistry, Arrhenius Laboratory, Stockholm University, 106 91, Stockholm, Sweden. jeb@organ.su.se.
authors.forename: Jan-E
authors.initials: JE
authors.lastname: Bäckvall
authors.affiliation: Max-Planck-Institut für Kohlenforschung, Kaiser-Wilhelm-Platz 1, 45470, Mülheim an der Ruhr, Germany. reetz@mpi-muelheim.mpg.de.|Fachbereich Chemie, Philipps-Universität Marburg, Hans-Meerwein-Strasse 4, 35032, Marburg, Germany. reetz@mpi-muelheim.mpg.de.
authors.forename: Manfred T
authors.initials: MT
authors.lastname: Reetz
mesh_terms: D000596:Amino Acids / Q000737:chemistry* / Q000378:metabolism; D002384:Catalysis; D019020:Directed Molecular Evolution / Q000379:methods*; D004851:Epoxide Hydrolases / Q000737:chemistry* / Q000378:metabolism; D008049:Lipase / Q000737:chemistry* / Q000378:metabolism; D008958:Models, Molecular; D016296:Mutagenesis; D013237:Stereoisomerism
chemical_list: D000596:Amino Acids; D008049:Lipase; D004851:Epoxide Hydrolases
keywords: directed evolution; enzymes; high-throughput screening; saturation mutagenesis; stereoselectivity