Glycomimicry: display of fucosylation on the lipo-oligosaccharide of recombinant Escherichia coli K12

Glycoconj J. 2011 Jan;28(1):39-47. doi: 10.1007/s10719-010-9322-1. Epub 2011 Feb 1.

Abstract

We recently described the design of Escherichia coli K12 and Salmonella enterica sv Typhimurium to display the gangliomannoside 3 (GM3) antigen on the cell surface. We report here the fucosylation of modified lipooligosaccharide in a recombinant E.coli strain with a truncated lipid A core due to deletion of the core glycosyltransferases genes waaO and waaB. This truncated structure was used as a scaffold to assemble the Lewis Y motif by consequent action of the heterologously expressed β-1,4 galactosyltransferase LgtE (Neisseria gonorrheae), the β-1,3 N-acetylglucosaminyltransferase LgtA and the β-1,3 galactosyltransferase LgtB from Neisseria meningitidis, as well as the α-1,2 and α-1,3 fucosyltransferases FutC and FutA from Helicobacter pylori. We show the display of the Lewis Y structure by immunological and chemical analysis.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Base Sequence
  • Carbohydrate Sequence
  • DNA Primers
  • Enzyme-Linked Immunosorbent Assay
  • Escherichia coli K12 / genetics
  • Escherichia coli K12 / metabolism*
  • Fucose / metabolism*
  • Lipid Metabolism*
  • Methylation
  • Molecular Mimicry*
  • Molecular Sequence Data
  • Oligosaccharides / metabolism*
  • Recombination, Genetic*
  • Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization

Substances

  • DNA Primers
  • Oligosaccharides
  • Fucose