| Literature DB >> 26724475 |
Mark Dörr1, Michael P C Fibinger1, Daniel Last1, Sandy Schmidt1, Javier Santos-Aberturas1, Dominique Böttcher1, Anke Hummel1, Clare Vickers1, Moritz Voss1, Uwe T Bornscheuer2.
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.Keywords: activity assay; enzymes; growth conditions; high-throughput screening; protein engineering; robotic platform
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Year: 2016 PMID: 26724475 DOI: 10.1002/bit.25925
Source DB: PubMed Journal: Biotechnol Bioeng ISSN: 0006-3592 Impact factor: 4.530