Department of Chemistry, Graduate School of Science, The University of Tokyo, Tokyo, Japan; email:
Annu Rev Biochem. 2022 Jun 21;91:221-243. doi: 10.1146/annurev-biochem-040320-103817.
Genetic code reprogramming has enabled us to ribosomally incorporate various nonproteinogenic amino acids (npAAs) into peptides in vitro. The repertoire of usable npAAs has been expanded to include not only l-α-amino acids with noncanonical sidechains but also those with noncanonical backbones. Despite successful single incorporation of npAAs, multiple and consecutive incorporations often suffer from low efficiency or are even unsuccessful. To overcome this stumbling block, engineering approaches have been used to modify ribosomes, EF-Tu, and tRNAs. Here, we provide an overview of these in vitro methods that are aimed at optimal expansion of the npAA repertoire and their applications for the development of de novo bioactive peptides containing various npAAs.
遗传密码重编程使我们能够在核糖体上将各种非天然氨基酸（npAAs）掺入肽中。可用的 npAA 种类已扩展到不仅包括具有非典型侧链的 l-α-氨基酸，还包括具有非典型骨架的氨基酸。尽管 npAA 的单次掺入已获得成功，但多次和连续掺入往往效率低下，甚至不成功。为了克服这一障碍，人们已经使用工程方法来修饰核糖体、EF-Tu 和 tRNA。在这里，我们提供了这些体外方法的概述，这些方法旨在最佳扩展 npAA 库，并将其应用于开发含有各种 npAA 的新型生物活性肽。
