Fully automatized high-throughput enzyme library screening using a robotic platform
Fully automatized high-throughput enzyme library screening using a robotic platform
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DOI：
10.1002/bit.25925
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发表时间：
2016-07-01
影响因子：
3.8
通讯作者：
Bornscheuer, Uwe T.
中科院分区：
文献类型：
--
作者：
Doerr, Mark;Fibinger, Michael P. C.;Bornscheuer, Uwe T.
建立了一个全自动机器人平台，以促进蛋白质工程目的的高通量筛选。该平台能够在微滴板格式中对生长条件进行适当的监测和控制，以确保精确的酶生产，用于酶突变文库的询问，蛋白质稳定性测试和多种分析筛选。该系统在Baeyer-Villiger单加氧酶、转氨酶、脱卤酶和酰化酶等四种对生物催化重要的酶类的高通量筛选中得到了验证。这使得以复杂和高度可靠的方式鉴定新的酶变体成为可能。此外，详细的优化协议应该使其他研究人员能够适应和改进他们的方法。Biotechnol。Bioeng。2016;113: 1421 - 1432。(c) 2016 Wiley Periodicals, Inc.；
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. (c) 2016 Wiley Periodicals, Inc.