Production of Lewis x tetrasaccharides by metabolically engineered Escherichia coli.
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.
Dumon
Claire
C
Centre de Recherches sur les Macromolécules Végétales, ICMG, FR CNRS-UJF 2607, B P 53, 38041 Grenoble Cedex 9, France.
Bosso
Claude
C
Utille
Jean Pierre
JP
Heyraud
Alain
A
Samain
Eric
E
D002236:Carbohydrate Conformation; D002240:Carbohydrate Sequence; D004926:Escherichia coli / Q000737:chemistry / Q000378:metabolism*; D016256:Lewis X Antigen / Q000031:analogs & derivatives; D008969:Molecular Sequence Data; D009844:Oligosaccharides / Q000096:biosynthesis* / Q000737:chemistry / Q000302:isolation & purification; D014312:Trisaccharides / Q000737:chemistry*
D016256:Lewis X Antigen; D009844:Oligosaccharides; D014312:Trisaccharides; C110530:galactosyl-(1,4)-fucopyranosyl-(1,3)-N-acetylglucosamine