Engineering Escherichia coli to biosynthesize O-polysaccharide-derived recombinant glycoconjugate vaccines against pathogenic serotypes O8 and O9a - PubMed
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- #Glycoconjugate vaccines
- #ExPEC
- #Metabolic engineering
- Engineered E. coli to produce glycoconjugate vaccines against ExPEC serotypes O8 and O9a.
- Optimized nucleotide sugar precursor supply and enhanced O-polysaccharide biosynthesis via metabolic engineering.
- CRISPR-Cas9 and λ-RED genome editing used to delete competitive and catabolic pathways.
- Overexpression of ptsA gene increased O8- and O9a-OPS yields by 2.11-fold and 2.4-fold, respectively.
- Co-expression of PglL and CTB improved conjugate yields by 2.59-fold (O8) and 4.18-fold (O9a).
- Mass spectrometry confirmed site-specific O-glycosylation at CTB Thr19 with preserved OPS structure.
- Vaccine candidate showed a favorable safety profile in mice, inducing robust Th1-biased IgG responses.
- Immunization conferred 80-90% protection against lethal challenge and reduced bacterial loads in vivo.
- Established an efficient, scalable glycoconjugate vaccine production platform.