Glycoengineering directs de novo biomanufacturing of UPEC O21 O-antigen polysaccharide based glycoprotein.
Glycoengineering directs de novo biomanufacturing of UPEC O21 O-antigen polysaccharide based glycoprotein.
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DOI：
10.1016/j.ijbiomac.2023.126993
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发表时间：
2023-09
影响因子：
8.2
通讯作者：
Yuhui Wang;A. Perepelov;S. Senchenkova;Ge-Jin Lu;Xiaohan Wang;Guozhen Ma;Qian Yang;Jian Yuan-Jian-Yua
中科院分区：
文献类型：
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作者：
Yuhui Wang;A. Perepelov;S. Senchenkova;Ge-Jin Lu;Xiaohan Wang;Guozhen Ma;Qian Yang;Jian Yuan-Jian-Yua
糖蛋白，其中多糖通常附着在蛋白质上，是一类重要的生物分子，几十年来在临床治疗中广泛用作治疗剂。尿路致病性大肠杆菌 (UPEC) O21 已被确定为诱发尿路感染的血清群，全球范围内女性和幼儿中感染人数不断增加。因此，迫切需要建立针对 UPEC 感染的保护性疫苗。在这里，我们设计了非致病性E。 coliMG1655 实现针对 UPEC O21 的 O21 O 抗原多糖糖蛋白的稳健、经济有效的从头生物合成。具体来说，这种糖工程化的E。操纵大肠杆菌MG1655以实现高效葡萄糖-甘油共同利用以及基因簇安装和O-糖基化机器组装。 UDP-糖前体的关键途径也得到了加强，以强制更多的碳流向糖基供体，从而使糖蛋白滴度提高了 5.6 倍。培养条件的进一步优化产生了高达 35.34 mg/L 的糖蛋白。糖肽MS证实了糖蛋白的精确生物合成。这种糖蛋白引发抗原特异性 IgG 免疫反应，并显着减少肾脏和膀胱定植。这种基于细菌细胞的糖平台和优化策略可以为其他增值糖蛋白的生物合成提供指导。
Glycoproteins, in which polysaccharides are usually attached to proteins, are an important class of biomolecules that are widely used as therapeutic agents in clinical treatments for decades. UropathogenicEscherichia coli(UPEC) O21 has been identified as a serogroup that induces urinary tract infections, with a global increasing number among women and young children. Therefore, there is an urgent need to establish protective vaccines against UPEC infection. Herein, we engineered non-pathogenicE. coliMG1655 to achieve robust, cost-effectivede novobiosynthesis of O21 O-antigen polysaccharide-based glycoprotein against UPEC O21. Specifically, this glycoengineeredE. coliMG1655 was manipulated for high-efficient glucose-glycerol co-utilization and for the gene cluster installation andO-glycosylation machinery assembly. The key pathways of UDP-sugar precursors were also strengthened to enforce more carbon flux towards the glycosyl donors, which enhanced the glycoprotein titer by 5.6-fold. Further optimization of culture conditions yielded glycoproteins of up to 35.34 mg/L. Glycopeptide MS confirmed the preciset biosynthesis of glycoprotein. This glycoprotein elicited antigen-specific IgG immune responses and significantly reduced kidney and bladder colonization. This bacterial cell-based glyco-platform and optimized strategies can provide a guideline for the biosynthesis of other value-added glycoproteins.