Yang Junzhu, Wang Chen, Lu Yuan
Key Laboratory of Industrial Biocatalysis, Ministry of Education, Department of Chemical Engineering, Tsinghua University, Beijing 100084, China.
ACS Synth Biol. 2022 Apr 15;11(4):1408-1416. doi: 10.1021/acssynbio.1c00641. Epub 2022 Mar 23.
The cell-free protein synthesis (CFPS) system is a typical protein production platform in the field of synthetic biology. However, there are limitations in controlling protein synthesis in the CFPS system. Compared with the traditional method of adding chemicals, temperature is an ideal control switch to achieve precise spatiotemporal control with few side effects. Hence, the design of a temperature-controlled cell-free protein synthesis (tcCFPS) system based on was carried out with the repressor cI protein in this study. The corresponding tcCFPS achieved a 143-fold dynamic protein expression level at 37 °C than that at 30 °C. Besides, the artificial cell controlled by temperature was constructed to expand the applications of tcCFPS. This study provides a new effective method for active protein synthesis in a cell-free system and a potential means of drug synthesis and delivery.
无细胞蛋白质合成（CFPS）系统是合成生物学领域中一种典型的蛋白质生产平台。然而，CFPS系统在控制蛋白质合成方面存在局限性。与添加化学物质的传统方法相比，温度是一种理想的控制开关，能够在副作用较少的情况下实现精确的时空控制。因此，本研究利用阻遏蛋白cI设计了一种基于温度控制的无细胞蛋白质合成（tcCFPS）系统。相应的tcCFPS在37°C时的蛋白质动态表达水平比在30°C时提高了143倍。此外，构建了受温度控制的人工细胞以拓展tcCFPS的应用。本研究为无细胞系统中的活性蛋白质合成提供了一种新的有效方法，以及一种潜在的药物合成与递送手段。
