Chemical and Biological Engineering, Northwestern University Evanston, IL, USA ; Chemistry of Life Processes Institute, Northwestern University Evanston, IL, USA.
Chemical and Biological Engineering, Northwestern University Evanston, IL, USA ; Chemistry of Life Processes Institute, Northwestern University Evanston, IL, USA ; Robert H. Lurie Comprehensive Cancer Center, Northwestern University Chicago, IL, USA ; Institute of Bionanotechnology in Medicine, Northwestern University Chicago, IL, USA.
Front Chem. 2014 Jun 10;2:34. doi: 10.3389/fchem.2014.00034. eCollection 2014.
Incorporating non-standard amino acids (NSAAs) into proteins enables new chemical properties, new structures, and new functions. In recent years, improvements in cell-free protein synthesis (CFPS) systems have opened the way to accurate and efficient incorporation of NSAAs into proteins. The driving force behind this development has been three-fold. First, a technical renaissance has enabled high-yielding (>1 g/L) and long-lasting (>10 h in batch operation) CFPS in systems derived from Escherichia coli. Second, the efficiency of orthogonal translation systems (OTSs) has improved. Third, the open nature of the CFPS platform has brought about an unprecedented level of control and freedom of design. Here, we review recent developments in CFPS platforms designed to precisely incorporate NSAAs. In the coming years, we anticipate that CFPS systems will impact efforts to elucidate structure/function relationships of proteins and to make biomaterials and sequence-defined biopolymers for medical and industrial applications.
将非标准氨基酸（NSAAs）掺入蛋白质中可以赋予其新的化学性质、结构和功能。近年来，无细胞蛋白质合成（CFPS）系统的改进为 NSAAs 准确有效地掺入蛋白质开辟了道路。这一发展的驱动力有三方面。首先，技术复兴使得源自大肠杆菌的 CFPS 系统能够实现高产（>1 g/L）和长时（批处理操作>10 小时）运行。其次，正交翻译系统（OTS）的效率得到了提高。第三，CFPS 平台的开放性带来了前所未有的控制和设计自由度。在这里，我们回顾了最近为精确掺入 NSAAs 而设计的 CFPS 平台的发展。在未来几年，我们预计 CFPS 系统将影响阐明蛋白质结构/功能关系的努力，并为医学和工业应用制造生物材料和序列定义的生物聚合物。
