The carbohydrate chains decorating cell membranes and secreted proteins participate in a range of important biological processes. However, their ultimate significance and possible therapeutic potential have not been fully explored due to the lack of economical methods for their production. This study is an example of the use of a genetically engineered bacterial strain in the preparation of diverse oligosaccharides. Based on an ex vivo biosynthetic pathway, an artificial gene cluster was constructed by linking the genes of five associated enzymes on a plasmid vector. This plasmid was inserted into the E. coli NM522 strain to form globotriose-producing cells (‘superbug’ pLDR20-CKTUF). The specific strain was conveniently applied to the synthesis of globotriose trisaccharide and its derivatives, as potential neutralizers for Shiga toxin. This work demonstrates a novel and economical method for generating ligand diversity for carbohydrate drug development.
装饰细胞膜和分泌蛋白的
碳水化合物链参与一系列重要的
生物过程。然而,由于缺乏经济的生产方法,它们的最终意义和可能的治疗潜力尚未得到充分探索。这项研究是使用
基因工程菌株制备多种
寡糖的一个例子。基于离体
生物合成途径,通过将五种相关酶的
基因连接在
质粒载体上构建了人工
基因簇。将该质粒插入大肠杆菌 NM522 菌株中,形成球三糖生产细胞(“超级细菌”pLDR20-CKTUF)。该特定菌株可方便地应用于球三糖三糖及其衍
生物的合成,作为志贺毒素的潜在中和剂。这项工作展示了一种为
碳水化合物药物开发产生
配体多样性的新颖且经济的方法。