细菌细胞壁肽聚糖的生物合成途径是抗生素研发的重要药物靶标，采用传统的筛选方法已经开发出四十多种抗菌药物，但是在临床应用后均出现了多重耐药性菌株。本课题在前期单一功能性表达细菌肽聚糖单体合成途径的8个组成酶的基础上，通过自主构建和优化大肠杆菌无细胞表达体系，首次实现肽聚糖单体合成途径的6个可溶性蛋白和2个膜蛋白在一个无细胞体系中同时功能性表达，深入研究膜蛋白MurG转移酶与MraY转位酶的细胞定位及相互作用。整合自动化检测平台，从组合酶学角度构建一个抗耐药性的抗菌分子筛选新模型。这一工作既能建立高通量筛选抗耐药性抗生素的新方法，又能为深入研究肽聚糖合成机制奠定坚实的理论基础和技术源头。

Biosynthetic pathway of cell wall plays an important role in identifying antibacterial targets. More than 40 antimicrobial were developed based on traditional methods, however, many multi-drug resistant bacterium were found in the clinical application. Based on the expression of all functional enzymes of peptidoglycan pathway, this project aims to co-expresse 6 soluble proteins and 2 membrane proteins in cell-free system functionally and simultaneously. We establish a new model to screen anti-resistant antibiotics and focus on the interaction between two membrane proteins in this pathway. This research not only generates new breeding method on screening anti-resistant antibiotics, but also accelerates understanding the mechanism of peptidoglycan biosynthesis.