Taxonomic group: bacteria / Proteobacteria
(Phylum: Proteobacteria)
Associated disease: infection due to Escherichia coli [ICD11:
XN6P4 
]
The structure was elucidated in this paperNCBI PubMed ID: 37739281Publication DOI: 10.1016/j.ijbiomac.2023.126993Journal NLM ID: 7909578Publisher: Butterworth-Heinemann
Correspondence: D. Huang <huangdi
nankai.edu.cn>; B. Liu <liubin1981
nankai.edu.cn>
Institutions: Key Laboratory of Molecular Microbiology and Technology, Ministry of Education, Nankai University, Tianjin, China, TEDA Institute of Biological Sciences and Biotechnology, Tianjin Key Laboratory of Microbial Functional Genomics, Nankai University, Tianjin, China, National Glycoengineering Research Center, Shandong University, Qingdao, Shandong 266237, China, N. D. Zelinsky Institute of Organic Chemistry, Russian Academy of Sciences, 119991 Moscow, Russian Federation
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. Uropathogenic Escherichia 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-pathogenic E. coli MG1655 to achieve robust, cost-effective de novo biosynthesis of O21 O-antigen polysaccharide-based glycoprotein against UPEC O21. Specifically, this glycoengineered E. coli MG1655 was manipulated for high-efficient glucose-glycerol co-utilization and for the gene cluster installation and O-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.
polysaccharides, glycoprotein, protein glycosylation, O21 O-antigen gene cluster, UDP-sugar precursors, Urinary tract infections (UTI)
Structure type: polymer chemical repeating unit
Location inside paper: Fig. 1A
Compound class: O-polysaccharide, O-antigen
Contained glycoepitopes: IEDB_130648,IEDB_135813,IEDB_136044,IEDB_137340,IEDB_137472,IEDB_137473,IEDB_141794,IEDB_141807,IEDB_142487,IEDB_142488,IEDB_146664,IEDB_151531,IEDB_190606,IEDB_221845,IEDB_983931,SB_165,SB_166,SB_187,SB_192,SB_195,SB_21,SB_6,SB_7,SB_88
Methods: 13C NMR, 1H NMR, NMR-2D, PCR, SDS-PAGE, sugar analysis, ELISA, Western blotting, genetic methods, HPLC, GPC, immunization, delipidation, glycoengineering, RP-LC-MS/MS, isolation, cultivation
Biosynthesis and genetic data: wclN, wclP, wclO, wclQ (encode the glycosyltransferases)
NCBI Taxonomy refs (TaxIDs): 562,
2162911Reference(s) to other database(s): GTC:G92807QP, GlycomeDB:
3591
Show glycosyltransferases
There is only one chemically distinct structure:
Found 
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