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Updated: Mar 6, 2026

Homogeneous Glycoconjugate Produced by Combined Unnatural Amino Acid Incorporation and Click-Chemistry for Vaccine Purposes
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Biosynthesis of a Polysaccharide-based Glycoconjugate Vaccine against Neonatal Meningitis-Causing Escherichia coli

Yuhui Wang1,2,3,4, Haodi Liu1,2,3,4, Yahao Wang1,2,3,4

  • 1National Glycoengineering Research Center, Shandong University, Qingdao, Shandong 266237, China.

ACS Synthetic Biology
|March 4, 2026
PubMed

Insights

A novel glycoengineered Escherichia coli strain efficiently produces O45 O-polysaccharide for a glycoconjugate vaccine. This vaccine shows high protection against neonatal meningitis-causing E. coli (NMEC) infections.

Area of Science:

  • Microbiology
  • Glycobiology
  • Vaccine Development

Background:

  • Extraintestinal pathogenic Escherichia coli O45 (O45) is a growing multidrug-resistant threat.
  • Neonatal meningitis-causing E. coli (NMEC) poses a significant public health challenge.

Purpose of the Study:

  • To develop a glycoengineered E. coli strain for efficient biosynthesis of O45 O-polysaccharide (OPS45).
  • To create an OPS45-based glycoconjugate vaccine for protection against NMEC infections.

Main Methods:

  • Glycoengineering of E. coli for enhanced OPS45 production, including rare sugars 6-deoxy-l-talose and N-acetylfucosamine.
  • Oligosaccharyltransferase-mediated conjugation of OPS45 to cholera toxin B subunit (CTB).
  • Structural confirmation of OPS45 using NMR and LC-MS/MS analysis of the CTB-OPS45 conjugate.

Main Results:

  • Optimized OPS45 production in the glycoengineered E. coli strain.
  • Generation of a homogeneous CTB-OPS45 glycoconjugate with improved antigen loading (11.31 ± 0.59 mg/L).
  • Immunization in mice induced strong O45-specific IgG responses and provided 90% protection against lethal NMEC infection.

Conclusions:

  • The integrated biosynthesis and conjugation approach enables rapid, efficient production of a well-defined glycoconjugate vaccine.
  • The developed vaccine demonstrates significant potential for combating resistant NMEC and O45 infections.
  • This strategy offers a promising platform for developing vaccines against other challenging bacterial pathogens.
Keywords:
Escherichia coli O45O-polysaccharidecholera toxin B subunitglycoconjugate vaccinemetabolic engineeringoligosaccharyltransferase

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