Engineering N-linked protein glycosylation with diverse O antigen lipopolysaccharide structures in Escherichia coli

Proc Natl Acad Sci U S A. 2005 Feb 22;102(8):3016-21. doi: 10.1073/pnas.0500044102. Epub 2005 Feb 9.


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 (LPS) biosynthesis, converge at the step in which PglB, the key enzyme of the C. jejuni N-glycosylation system, transfers O polysaccharide from a lipid carrier (undecaprenyl pyrophosphate) to an acceptor protein. PglB was the only protein of the bacterial N-glycosylation machinery both necessary and sufficient for the transfer. The relaxed specificity of the PglB oligosaccharyltransferase toward the glycan structure was exploited to create novel N-glycan structures containing two distinct E. coli or Pseudomonas aeruginosa O antigens. PglB-mediated transfer of polysaccharides might be valuable for in vivo production of O polysaccharides-protein conjugates for use as antibacterial vaccines.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Bacterial Vaccines / immunology
  • Escherichia coli / metabolism*
  • Escherichia coli Proteins / metabolism*
  • Glycosylation
  • Hexosyltransferases / genetics
  • Hexosyltransferases / physiology*
  • Lipopolysaccharides / biosynthesis
  • Lipoproteins / metabolism
  • Membrane Proteins / genetics
  • Membrane Proteins / physiology*
  • Membrane Transport Proteins
  • O Antigens / metabolism*
  • Protein Engineering
  • Vaccines, Conjugate / immunology


  • AcrA protein, E coli
  • Bacterial Vaccines
  • Escherichia coli Proteins
  • Lipopolysaccharides
  • Lipoproteins
  • Membrane Proteins
  • Membrane Transport Proteins
  • O Antigens
  • Vaccines, Conjugate
  • Hexosyltransferases
  • dolichyl-diphosphooligosaccharide - protein glycotransferase