羊毛硫肽的高通量工程改造方法新进展

羊毛硫肽(lanthipeptide)是由核糖体合成并经翻译后修饰产生的肽类天然产物,具有丰富的分子结构和多样的生物活性。新型羊毛硫肽是活性药物的重要来源,可以通过基因组挖掘和工程改造获得。羊毛硫肽前体肽由基因编码,同时其合成酶具有较高的底物杂泛性。基于这些特征,可以对羊毛硫肽的生物合成过程开展高通量工程改造,从而快速获得新的羊毛硫肽衍生物。综述了近些年高通量构建和筛选羊毛硫肽衍生物的新方法:介绍了非天然氨基酸引入、组合式生物合成、嵌合前导肽等文库构建技术;讨论了细胞表面展示、反向双杂交、细胞自裂解、无细胞(cell-free)体系等方法在结构与活性筛选中的应用;对基于自动化合成生物技术开展羊毛硫肽的规模化工程改造进行了展望。

Recent Advances in the High-throughput Engineering of Lanthipeptides

Lanthipeptides are a major class of ribosomally synthesized and posttranslationally modified peptides(Ri PPs)with diverse molecular structures and biological activities.New lanthipeptides obtained by genome mining and engineering are an important source of drug leads.Lanthipeptides are particularly amenable to bioengineering because their precursor peptides are encoded by genes and biosynthetic enzymes often exhibit high promiscuity,which is helpful for the efficient construction of lanthipeptides derivatives.This paper reviews the recent advances in high-throughput creation and screening of lanthipeptide derivatives.For mutant library creation,we discuss the introduction of noncanonical amino acids(ncAAs),combinatorial biosynthesis,and chimeric-leader approach for creating hybrid Ri PPs.Then,we introduce large-scale structural and activity screening of lanthipeptide mutants assisted by cell surface display,reverse two-hybrid system,cellular autolysis,cell-free system,and microfluidics.Finally,we present future perspectives on the use of synthetic biology automation to streamline lanthipeptide bioengineering.