title: A simple selection strategy for evolving highly efficient enzymes.
abstract: Combining tunable transcription with an enzyme-degradation tag affords an effective means to reduce intracellular enzyme concentrations from high to very low levels. Such fine-tuned control allows selection pressure to be systematically increased in directed-evolution experiments. This facilitates identification of mutants with wild-type activity, as shown here for an engineered chorismate mutase. Numerous selection formats and cell-based screening methodologies may benefit from the large dynamic range afforded by this easily implemented strategy.
authors.affiliation: Laboratory of Organic Chemistry, ETH Zurich, Hönggerberg HCI F 339, CH-8093 Zurich, Switzerland.
authors.forename: Martin
authors.initials: M
authors.lastname: Neuenschwander
authors.forename: Maren
authors.initials: M
authors.lastname: Butz
authors.forename: Caroline
authors.initials: C
authors.lastname: Heintz
authors.forename: Peter
authors.initials: P
authors.lastname: Kast
authors.forename: Donald
authors.initials: D
authors.lastname: Hilvert
mesh_terms: D002826:Chorismate Mutase / Q000235:genetics / Q000378:metabolism; D019020:Directed Molecular Evolution / Q000379:methods*; D004925:Escherichia / Q000201:enzymology; D005816:Genetic Complementation Test; D005818:Genetic Engineering; D016296:Mutagenesis; D012641:Selection, Genetic; D013752:Tetracycline; D014158:Transcription, Genetic
chemical_list: D002826:Chorismate Mutase; D013752:Tetracycline