Cui Yi, Chen Xinjie, Wang Ze, Lu Yuan
Key Laboratory of Industrial Biocatalysis, Ministry of Education, Department of Chemical Engineering, Tsinghua University, Beijing 100084, China.
College of Life Sciences, Shenyang Normal University, Shenyang 110034, Liaoning, China.
Biodes Res. 2022 Aug 30;2022:9847014. doi: 10.34133/2022/9847014. eCollection 2022.
The cell-free protein synthesis (CFPS) system, as a technical core of synthetic biology, can simulate the transcription and translation process in an open environment without a complete living cell. It has been widely used in basic and applied research fields because of its advanced engineering features in flexibility and controllability. Compared to a typical crude extract-based CFPS system, due to defined and customizable components and lacking protein-degrading enzymes, the protein synthesis using recombinant elements (PURE) system draws great attention. This review first discusses the elemental composition of the PURE system. Then, the design and preparation of functional proteins for the PURE system, especially the critical ribosome, were examined. Furthermore, we trace the evolving development of the PURE system in versatile areas, including prototyping, synthesis of unnatural proteins, peptides and complex proteins, and biosensors. Finally, as a state-of-the-art engineering strategy, this review analyzes the opportunities and challenges faced by the PURE system in future scientific research and diverse applications.
无细胞蛋白质合成（CFPS）系统作为合成生物学的一项技术核心，能够在无完整活细胞的开放环境中模拟转录和翻译过程。因其在灵活性和可控性方面具有先进的工程特性，该系统已在基础研究和应用研究领域得到广泛应用。与典型的基于粗提物的CFPS系统相比，由于其成分明确且可定制，并且缺乏蛋白质降解酶，使用重组元件的蛋白质合成（PURE）系统备受关注。本文综述首先讨论了PURE系统的元素组成。然后，研究了用于PURE系统的功能蛋白的设计与制备，尤其是关键的核糖体。此外，我们追溯了PURE系统在多个领域的不断发展，包括原型设计、非天然蛋白质、肽和复杂蛋白质的合成以及生物传感器。最后，作为一种前沿的工程策略，本文综述分析了PURE系统在未来科学研究和各种应用中面临的机遇与挑战。
