植物天门冬酰胺连接酶在大环肽类药物应用中的研究进展  

Advances in peptidyl Asx-specific ligases for the application of cyclic peptides

作　　者：申欣 刘忞之 杨燕 王伟 SHEN Xin;LIU Min-zhi;YANG Yan;WANG Wei(State Key Laboratory of Bioactive Substance and Function of Natural Medicines,NHC Key Laboratory of Natural Drug Biosynthesis,Institute of Materia Medica,Peking Union Medical College and Chinese Academy of Medical Sciences,Beijing 100050,China)

机构地区：[1]中国医学科学院、北京协和医学院药物研究所,天然药物活性物质与功能国家重点实验室/国家卫生健康委员会天然药物生物合成重点实验室,北京100050

出　　处：《药学学报》2023年第9期2656-2668,共13页Acta Pharmaceutica Sinica

基　　金：国家自然科学基因项目(82073757,81903487);中国医学科学院医学与健康科技创新工程项目(2021-I2M-1-029);北京协和医学院合成生物学学科建设专项资助(201920100801)。

摘　　要：植物中的天门冬酰胺内肽酶(asparaginyl endopeptidases,AEPs)属于半胱氨酸蛋白酶家族,以酶原的形式在酸性液泡中发生自激活,在种子储存蛋白成熟、蛋白质降解、细胞程序性死亡和宿主防御中发挥着重要的生理作用。最近从几种富含环肽的植物中分离鉴定了促进环肽成熟的生物加工酶-天门冬酰胺连接酶(peptidyl Asxspecific ligases,PALs),由AEPs进化而来,可位点特异性催化天冬酰胺或天冬氨酸肽键的生成,由于相对无痕和宽泛的底物谱以及催化效率高等优势,已经在多肽和蛋白质的环化和修饰方面起到重要作用,是提高多肽类药物稳定性的有力工具。本文介绍了AEPs在植物中的生理功能,综述了PALs的发现、结构特点、催化机制和蛋白质工程,以及限制其应用的原因和未来的发展趋势。此外,还重点阐述了PALs在环肽生物合成以及在大环肽类药物开发中的应用,旨在为生物催化合成环多肽类药物提供一种新思路。Asparaginyl endopeptidases(AEPs)in plants belong to the family of cysteine protease that undergo self-activation in the form of zymogen in acidic vacuole and play important physiological roles in maturation of seed storage proteins,protein degradation,programmed cell death and host defense.Bioprocessing enzymes(peptidyl Asx-specific ligases,PALs)that promote the maturation of cyclotides have recently been isolated and identified from several cyclotide-rich plants.PALs derived from AEPs can site-specifically catalyze the formation of asparagine or aspartate peptide bonds.Due to the advantages of relatively traceless peptide bonds and broad substrate spectrum and high catalytic efficiency,they have been playing important roles in the cyclization and modification of peptides and proteins,and are powerful tools for improving the stability of peptide drugs.This review describes the physiological functions of AEPs in plants and summarizes the discoveries,structural characteristics,catalytic mechanism and protein engineering of PALs,as well as the limitation of their applications and future trends.In addition,the applications of PALs in cyclotides biosynthesis and the development of macrocyclic peptides are highlighted,with the aim of providing a new idea for the biocatalytic synthesis of cyclic peptides.

关 键 词：天门冬酰胺内肽酶 天门冬酰胺连接酶 大环肽 酶促环化 位点特异性连接 

分 类 号：R931[医药卫生—生药学]