Gao Wei, Bu Ning, Lu Yuan
Department of Chemical Engineering, Tsinghua University, Beijing 100084, China.
College of Life Sciences, Shenyang Normal University, Shenyang 110034, Liaoning, China.
Sheng Wu Gong Cheng Xue Bao. 2018 Sep 25;34(9):1371-1385. doi: 10.13345/j.cjb.180203.
Cell-free unnatural protein synthesis (CFUPS) as an emerging approach for protein engineering research, has been successfully applied in basic scientific studies (e.g., protein-protein interaction, protein-nucleic acid interaction) and industrial production (e.g., pharmaceutical proteins, protein materials). CFUPS can improve the engineering freedom and the process control by allowing free addition of genetic elements and chemicals as research purposes. It also can give protein novel characteristics, structures and functions, including post-translational modification of proteins, incorporation of reaction handles, synthesis of biophysical probes and polymeric protein. This article systematically reviews the unnatural amino acid incorporation methods (global suppression, stop codon suppression, frameshift suppression and unnatural base-pairs) and the application advances of unnatural amino acids in protein modifications, biophysical probes, enzyme engineering, biomaterials and biopharmaceutical protein production. The opportunities and challenges of the CFUPS system development and the wide application of industrialization are also illustrated with details.
无细胞非天然蛋白质合成（CFUPS）作为蛋白质工程研究的一种新兴方法，已成功应用于基础科学研究（如蛋白质-蛋白质相互作用、蛋白质-核酸相互作用）和工业生产（如药用蛋白质、蛋白质材料）。CFUPS通过允许根据研究目的自由添加遗传元件和化学物质，可提高工程自由度和过程控制。它还能赋予蛋白质新颖的特性、结构和功能，包括蛋白质的翻译后修饰、反应手柄的掺入、生物物理探针的合成以及聚合蛋白质的合成。本文系统综述了非天然氨基酸掺入方法（全局抑制、终止密码子抑制、移码抑制和非天然碱基对）以及非天然氨基酸在蛋白质修饰、生物物理探针、酶工程、生物材料和生物制药蛋白质生产中的应用进展。还详细阐述了CFUPS系统开发和广泛工业化应用的机遇与挑战。
