Traceless enzymatic protein synthesis without ligation sites constraint  

作　　者：Ruifeng Li Marcel Schmidt Tong Zhu Xinyu Yang Jing Feng Yu’e Tian Yinglu Cui Timo Nuijens Bian Wu 

机构地区：[1]CAS Key Laboratory of Microbial Physiological and Metabolic Engineering,State Key Laboratory of Microbial Resources,Institute of Microbiology,Chinese Academy of Sciences,Beijing 100101,China [2]University of Chinese Academy of Sciences,Beijing 100049,China [3]Fresenius Kabi iPSUM,I&D Center EnzyPep B.V.,Geleen 6167 RD,the Netherlands

出　　处：《National Science Review》2022年第5期104-114,共11页国家科学评论（英文版）

基　　金：supported by the National Key R&D Program of China(2019YFA0905100);the National Natural Science Foundation of China(31822002 and 31870055);the Strategic Priority Research Program(XDA24020101 and XDPB1801);the Key Research Program of Frontier Sciences(ZDBS-LY-SM014);the Biological Resources Program(KFJ-BRP-017-58)from the Chinese Academy of Sciences。

摘　　要：Protein synthesis and semisynthesis offer immense promise for life sciences and have impacted pharmaceutical innovation.The absence of a generally applicable method for traceless peptide conjugation with a flexible choice of junction sites remains a bottleneck for accessing many important synthetic targets,however.Here we introduce the PALME(protein activation and ligation with multiple enzymes)platform designed for sequence-unconstrained synthesis and modification of biomacromolecules.The upstream activating modules accept and process easily accessible synthetic peptides and recombinant proteins,avoiding the challenges associated with preparation and manipulation of activated peptide substrates.Cooperatively,the downstream coupling module provides comprehensive solutions for sequential peptide condensation,cyclization and protein N/C-terminal or internal functionalization.The practical utility of this methodology is demonstrated by synthesizing a series of bioactive targets ranging from pharmaceutical ingredients to synthetically challenging proteins.The modular PALME platform exhibits unprecedentedly broad accessibility for traceless protein synthesis and functionalization,and holds enormous potential to extend the scope of protein chemistry and synthetic biology.

关 键 词：protein synthesis traceless peptide ligation BIOCATALYSIS synthetic biology chemical biology 

分 类 号：Q51[生物学—生物化学]