Houssen Wael E
Marine Biodiscovery Centre, Chemistry Department, University of Aberdeen, Aberdeen, UK.
Institute of Medical Sciences, University of Aberdeen, Aberdeen, UK.
Methods Mol Biol. 2019;2012:193-210. doi: 10.1007/978-1-4939-9546-2_11.
Cyclic peptides are an emerging class of therapeutics that can modulate targets not amenable to traditional small molecule intervention (e.g., protein-protein interactions). However, N-to-C macrocyclization of peptides is a challenging and often a low yielding chemical transformation. Several macrocyclases from cyanobactin biosynthetic clusters have been used to catalyze this reaction.This chapter provides practical guidance to the processes of heterologous expression and purification of these enzymes as well as performing in vitro biochemical reactions. Finally, approaches to recover the final product from an enzymatic reaction mixture are also discussed.
环肽是一类新兴的治疗药物，可调节传统小分子干预难以作用的靶点（如蛋白质-蛋白质相互作用）。然而，肽的N端到C端大环化是一项具有挑战性且通常产率较低的化学转化过程。来自蓝细菌素生物合成簇的几种大环化酶已被用于催化此反应。本章为这些酶的异源表达、纯化过程以及进行体外生化反应提供了实用指南。最后，还讨论了从酶促反应混合物中回收最终产物的方法。
