Engineering N-linked protein glycosylation with diverse O antigen lipopolysaccharide structures in Escherichia coli
Engineering N-linked protein glycosylation with diverse O antigen lipopolysaccharide structures in Escherichia coli
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DOI：
10.1073/pnas.0500044102
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发表时间：
2005-02-22
影响因子：
11.1
通讯作者：
Aebi, M
中科院分区：
文献类型：
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作者：
Feldman, MF;Wacker, M;Aebi, M
空肠弯曲菌具有通用的N-连接蛋白糖基化系统，其可以功能性地转移到大肠杆菌。在这项研究中，我们设计了E。蛋白质N-糖基化和脂多糖（LIPS）生物合成这两条不同的途径在空肠弯曲菌N-糖基化系统的关键酶PgIB将O-多糖从脂质载体（焦磷酸十一异戊烯酯）转移到受体蛋白的步骤中会聚。PgIB是唯一的蛋白质的细菌N-糖基化机器的必要和足够的转移。利用PgIB寡糖基转移酶对聚糖结构的松弛特异性来产生含有两个不同E.大肠杆菌或铜绿假单胞菌O抗原。PgIB介导的多糖转移可能是有价值的在体内生产的O多糖-蛋白质结合物用作抗菌疫苗。
Campylobacter jejuni has a general N-linked protein glycosylation system that can be functionally transferred to Escherichia coli. In this study, we engineered E. coli cells in a way that two different pathways, protein N-glycosylation and lipopolysaccharide (LIPS) biosynthesis, converge at the step in which PgIB, the key enzyme of the C jejuni N-glycosylation system, transfers Opolysaccharide from a lipid carrier (undecaprenyl pyrophosphate) to an acceptor protein. PgIB was the only protein of the bacterial N-glycosylation machinery both necessary and sufficient for the transfer. The relaxed specificity of the PgIB oligosaccharyltransferase toward the glycan structure was exploited to create novel N-glycan structures containing two distinct E. coli or Pseudomonas aeruginosa O antigens. PgIB-mediated transfer of polysaccharides might be valuable for in vivo production of O polysaccharides-protein conjugates for use as antibacterial vaccines.