Engineering N-linked protein glycosylation with diverse O antigen lipopolysaccharide structures in Escherichia coli.
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.
Feldman
Mario F
MF
Institute of Microbiology, Department of Biology, Swiss Federal Institute of Technology, ETH-Hönggeberg, CH-8093 Zurich, Switzerland.
Wacker
Michael
M
Hernandez
Marcela
M
Hitchen
Paul G
PG
Marolda
Cristina L
CL
Kowarik
Michael
M
Morris
Howard R
HR
Dell
Anne
A
Valvano
Miguel A
MA
Aebi
Markus
M
D001428:Bacterial Vaccines / Q000276:immunology; D004926:Escherichia coli / Q000378:metabolism*; D029968:Escherichia coli Proteins / Q000378:metabolism*; D006031:Glycosylation; D006602:Hexosyltransferases / Q000235:genetics / Q000502:physiology*; D008070:Lipopolysaccharides / Q000096:biosynthesis; D008074:Lipoproteins / Q000378:metabolism; D008565:Membrane Proteins / Q000235:genetics / Q000502:physiology*; D026901:Membrane Transport Proteins; D019081:O Antigens / Q000378:metabolism*; D015202:Protein Engineering; D018074:Vaccines, Conjugate / Q000276:immunology
C083497:AcrA protein, E coli; D001428:Bacterial Vaccines; D029968:Escherichia coli Proteins; D008070:Lipopolysaccharides; D008074:Lipoproteins; D008565:Membrane Proteins; D026901:Membrane Transport Proteins; D019081:O Antigens; D018074:Vaccines, Conjugate; D006602:Hexosyltransferases; C025620:dolichyl-diphosphooligosaccharide - protein glycotransferase