合成生物学策略发现新型核糖体肽Xenopeptide  

Synthetic biology strategy to discover a novel ribosomal peptide,Xenopeptide

作　　者：莫天录 曾丹丹 马溯泽 贾一飞 韩沅均 丁伟 张琪[2] Tianlu Mo;Dandan Zeng;Suze Ma;Yifei Jia;Yuanjun Han;Wei Ding;Qi Zhang(School of Health Science and Engineering,University of Shanghai for Science and Technology,Shanghai 200093,China;Department of Chemistry,Fudan University,Shanghai 200433,China;State Key Laboratory of Microbial Metabolism,School of Life Sciences&Biotechnology,Shanghai Jiao Tong University,Shanghai 200240,China)

机构地区：[1]上海理工大学健康科学与工程学院,上海200093 [2]复旦大学化学系,上海200433 [3]上海交通大学生命科学技术学院,微生物代谢国家重点实验室,上海200240

出　　处：《科学通报》2024年第10期1361-1372,共12页Chinese Science Bulletin

基　　金：国家自然科学基金(32370092);国家重点研发计划(2023YFF1103600);生物反应器国家重点实验室开放课题;微生物代谢国家重点实验室开放课题资助。

摘　　要：S-腺苷甲硫氨酸自由基(rSAM)酶家族是目前已知的最大酶超家族之一,由22000多个成员组成.这些酶在大自然中广泛存在,被认为是地球上最早的生物催化剂之一.随着大量的微生物基因组信息被解析,分析显示,微生物中大量核糖体肽类天然产物的生物合成基因簇中含有rSAM酶;其中Xenorhabdus、Yersinia和Erwinia三个属的基因组中均含有一个高度保守的rSAM酶负责其相邻核糖体肽的前体修饰,此类前体肽和rSAM酶构成的XYE系统所合成的化合物鲜有报道.本研究合成一个来源于Xenorhabdus sp.KJ12.1的XYE系统的前体肽和rSAM修饰酶基因,在大肠杆菌中进行共表达,得到新型核糖体肽Xenopeptide,通过结构解析发现,rSAM酶XenB负责分子内2个碳碳键的形成.本研究为微生物中此类化合物的深度挖掘和合成生物学改造提供了理论支撑.Ribosomally-synthesized and post-translationally-modified peptides(RiPPs)are a major class of natural products.It originates from a corresponding biosynthetic gene cluster where the genes for a precursor peptide and several posttranslational modification(PTM)enzymes are located.The precursor peptide is translated by a ribosome,recognized on the N-terminal leader peptide and modified on the C-terminal core peptide by PTM enzymes,to finally give a structurally unique mature compound.So far,the RiPPs found by researchers only account for a small part of the family,and there are a large number of compounds remaining to be discovered.The S-adenosylmethionine radical(rSAM)enzyme family is one of the largest enzyme superfamilies,containing more than 22000 members.These enzymes are widely distributed in eukarya,bacteria and archaea and are considered to be one the earliest biocatalysts on earth which perform essential and inseparable functions in cells.As more substantial microbial genomic information becomes available,a large number of RiPPs are excavated as natural products by high-throughput analyses.Genome mining in microorganisms reveals great abundance of biosynthetic gene clusters consisting of co-occurring rSAM enzyme and RiPPs’peptide genes,mostly leading to complex bioactive compounds whether reported or not.Recently,three genera,Xenorhabdus,Yersinia,and Erwinia,are unified with the definition of XYE system,for they all employ a highly conserved rSAM enzyme responsible for modifying its adjacent peptide to create a distinctive C–C or C–O bond in the final RiPPs product.Yet complex compounds synthesized through the rSAM enzyme modifying a precursor peptide in the XYE system have rarely been reported.Through algorithm analysis,we monitored a biosynthetic gene cluster in the Xenorhabdus sp.KJ12.1 comforming a similar construction of that in an XYE system.We fully synthesized the precursor peptide and the rSAM enzyme genes from Xenorhabdus sp.KJ12.1,heterologously expressed the modifying system in Escherichia

关 键 词：S-腺苷甲硫氨酸自由基酶 核糖体肽 翻译后修饰 合成生物学 

分 类 号：Q78[生物学—分子生物学]