蛋白质的自动连续进化
Automated Continuous Evolution of Proteins .
机构信息
Department of Biomedical Engineering, University of California, Irvine, California 92697, United States.
Biological Design Center, Boston University, Boston, Massachusetts 02215, United States.
出版信息
ACS Synth Biol. 2020 Jun 19;9(6):1270-1276. doi: 10.1021/acssynbio.0c00135. Epub 2020 May 12.
Abstract
We present automated continuous evolution (ACE), a platform for the hands-free directed evolution of biomolecules. ACE pairs OrthoRep, a genetic system for continuous targeted mutagenesis of user-selected genes , with eVOLVER, a scalable and automated continuous culture device for precise, multiparameter regulation of growth conditions. By implementing real-time feedback-controlled tuning of selection stringency with eVOLVER, genes of interest encoded on OrthoRep autonomously traversed multimutation adaptive pathways to reach desired functions, including drug resistance and improved enzyme activity. The durability, scalability, and speed of biomolecular evolution with ACE should be broadly applicable to protein engineering as well as prospective studies on how selection parameters and schedules shape adaptation.
摘要
我们提出了自动化连续进化（ACE）平台，用于生物分子的无手动定向进化。ACE 将 OrthoRep（一种用于用户选择基因的连续靶向诱变的遗传系统）与 eVOLVER（一种可扩展和自动化的连续培养设备，用于精确、多参数调节生长条件）配对。通过使用 eVOLVER 实时反馈控制选择严格性的调整，OrthoRep 上编码的目标基因自动遍历多突变适应性途径，达到所需的功能，包括耐药性和提高酶活性。ACE 进行生物分子进化的耐久性、可扩展性和速度应该广泛适用于蛋白质工程以及关于选择参数和方案如何塑造适应性的未来研究。
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