Dörr Mark, Fibinger Michael P C, Last Daniel, Schmidt Sandy, Santos-Aberturas Javier, Böttcher Dominique, Hummel Anke, Vickers Clare, Voss Moritz, Bornscheuer Uwe T
Department of Biotechnology and Enzyme Catalysis, Institute of Biochemistry, Greifswald University, Felix-Hausdorff-Str. 4, Greifswald, 17489, Germany.
Biotechnol Bioeng. 2016 Jul;113(7):1421-32. doi: 10.1002/bit.25925. Epub 2016 Feb 3.
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.
已建立了一个全自动机器人平台,以促进用于蛋白质工程目的的高通量筛选。该平台能够以微孔板形式对生长条件进行适当的监测和控制,以确保为酶突变文库的研究、蛋白质稳定性测试和多种分析筛选精确生产酶。该系统的性能已在生物催化中重要的四类酶(如拜耳-维利格单加氧酶、转氨酶、脱卤酶和酰基转移酶)对各种突变文库的高通量筛选中得到例证。这使得能够以复杂且高度可靠的方式鉴定新型酶变体。此外,详细的优化方案应能使其他研究人员调整和改进他们的方法。《生物技术与生物工程》2016年;113卷:1421 - 1432页。© 2016威利期刊公司