Production of Lewis x tetrasaccharides by metabolically engineered Escherichia coli.

Claire Dumon, Claude Bosso, Jean Pierre Utille, Alain Heyraud, Eric Samain

Centre de Recherches sur les Macromolécules Végétales, ICMG, FR CNRS-UJF 2607, B P 53, 38041 Grenoble Cedex 9, France.

Chembiochem : a European journal of chemical biology 2006 Feb

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Two tetrasaccharides carrying the trisaccharidic Lewis x motif on a GlcNAc or a Gal residue were produced on the gram-scale by high-cell-density cultures of metabolically engineered Escherichia coli strains that overexpressed the Helicobacter pylori futA gene for alpha-3 fucosyltransferase and the Neisseria meningitidis lgtB gene for beta-4 galactosyltransferase. The first compound Galbeta-4(Fucalpha-3)GlcNAcbeta-4GlcNAc was produced by glycosylation of chitinbiose, which was endogenously generated in the bacterial cytoplasm by the successive action of the rhizobial chitin-synthase NodC and the Bacillus circulans chitinase A1, whose genes were additionally expressed in the E. coli strain. The second compound, Galbeta-4(Fucalpha-3)GlcNAcbeta-3Gal, was produced from exogenously added Gal by a strain that was deficient in galactokinase activity and overexpressed the additional N. meningitidis lgtA gene for beta-3 N-acetylglucosaminyltransferase.

Citation

Claire Dumon, Claude Bosso, Jean Pierre Utille, Alain Heyraud, Eric Samain. Production of Lewis x tetrasaccharides by metabolically engineered Escherichia coli. Chembiochem : a European journal of chemical biology. 2006 Feb;7(2):359-65