title: Fully automatized high-throughput enzyme library screening using a robotic platform.
abstract: A fully automatized robotic platform has been established to facilitate high-throughput screening for protein engineering purposes. This platform enables proper monitoring and control of growth conditions in the microtiter plate format to ensure precise enzyme production for the interrogation of enzyme mutant libraries, protein stability tests and multiple assay screenings. The performance of this system has been exemplified for four enzyme classes important for biocatalysis such as Baeyer-Villiger monooxygenase, transaminase, dehalogenase and acylase in the high-throughput screening of various mutant libraries. This allowed the identification of novel enzyme variants in a sophisticated and highly reliable manner. Furthermore, the detailed optimization protocols should enable other researchers to adapt and improve their methods. Biotechnol. Bioeng. 2016;113: 1421-1432. © 2016 Wiley Periodicals, Inc.
authors.affiliation: Department of Biotechnology and Enzyme Catalysis, Institute of Biochemistry, Greifswald University, Felix-Hausdorff-Str. 4, Greifswald, 17489, Germany.
authors.forename: Mark
authors.initials: M
authors.lastname: Dörr
authors.affiliation: Department of Biotechnology and Enzyme Catalysis, Institute of Biochemistry, Greifswald University, Felix-Hausdorff-Str. 4, Greifswald, 17489, Germany.
authors.forename: Michael P C
authors.initials: MP
authors.lastname: Fibinger
authors.affiliation: Department of Biotechnology and Enzyme Catalysis, Institute of Biochemistry, Greifswald University, Felix-Hausdorff-Str. 4, Greifswald, 17489, Germany.
authors.forename: Daniel
authors.initials: D
authors.lastname: Last
authors.affiliation: Department of Biotechnology and Enzyme Catalysis, Institute of Biochemistry, Greifswald University, Felix-Hausdorff-Str. 4, Greifswald, 17489, Germany.
authors.forename: Sandy
authors.initials: S
authors.lastname: Schmidt
authors.affiliation: Department of Biotechnology and Enzyme Catalysis, Institute of Biochemistry, Greifswald University, Felix-Hausdorff-Str. 4, Greifswald, 17489, Germany.
authors.forename: Javier
authors.initials: J
authors.lastname: Santos-Aberturas
authors.affiliation: Department of Biotechnology and Enzyme Catalysis, Institute of Biochemistry, Greifswald University, Felix-Hausdorff-Str. 4, Greifswald, 17489, Germany.
authors.forename: Dominique
authors.initials: D
authors.lastname: Böttcher
authors.affiliation: Department of Biotechnology and Enzyme Catalysis, Institute of Biochemistry, Greifswald University, Felix-Hausdorff-Str. 4, Greifswald, 17489, Germany.
authors.forename: Anke
authors.initials: A
authors.lastname: Hummel
authors.affiliation: Department of Biotechnology and Enzyme Catalysis, Institute of Biochemistry, Greifswald University, Felix-Hausdorff-Str. 4, Greifswald, 17489, Germany.
authors.forename: Clare
authors.initials: C
authors.lastname: Vickers
authors.affiliation: Department of Biotechnology and Enzyme Catalysis, Institute of Biochemistry, Greifswald University, Felix-Hausdorff-Str. 4, Greifswald, 17489, Germany.
authors.forename: Moritz
authors.initials: M
authors.lastname: Voss
authors.affiliation: Department of Biotechnology and Enzyme Catalysis, Institute of Biochemistry, Greifswald University, Felix-Hausdorff-Str. 4, Greifswald, 17489, Germany. uwe.bornscheuer@uni-greifswald.de.
authors.forename: Uwe T
authors.initials: UT
authors.lastname: Bornscheuer
mesh_terms: D057205:Automation, Laboratory / Q000295:instrumentation / Q000379:methods; D057075:Enzyme Assays / Q000295:instrumentation / Q000379:methods; D004867:Equipment Design; D057166:High-Throughput Screening Assays / Q000295:instrumentation / Q000379:methods; D015202:Protein Engineering / Q000295:instrumentation / Q000379:methods; D012371:Robotics / Q000295:instrumentation*; D054852:Small Molecule Libraries; D000637:Transaminases
chemical_list: D054852:Small Molecule Libraries; D000637:Transaminases
keywords: activity assay; enzymes; growth conditions; high-throughput screening; protein engineering; robotic platform