Production of Lewis x Tetrasaccharides by Metabolically Engineered Escherichia coli

• Published in: Chembiochem : a European journal of chemical biology Vol. 7; no. 2; pp. 359 - 365
• Main Authors: Dumon, Claire, Bosso, Claude, Utille, Jean Pierre, Heyraud, Alain, Samain, Eric
• Format: Journal Article
• Language: English
• Published: Weinheim Wiley-VCH Verlag 06-02-2006; WILEY-VCH Verlag; WILEY‐VCH Verlag
• Subjects: Bacillus circulans; biosynthesis; Carbohydrate Conformation; Carbohydrate Sequence; carbohydrates; Escherichia coli; Escherichia coli - chemistry; Escherichia coli - metabolism; glycoconjugates; glycosyltransferases; Helicobacter pylori; metabolic engineering; Molecular Sequence Data; Neisseria meningitidis; Oligosaccharides - biosynthesis; Oligosaccharides - chemistry; Oligosaccharides - isolation & purification; Trisaccharides - chemistry
• ISSN: 1439-4227; 1439-7633; 1439-4227
• DOI: 10.1002/cbic.200500293

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 α-3 fucosyltransferase and the Neisseria meningitidis lgtB gene for β-4 galactosyltransferase. The first compound Galβ-4(Fucα-3)GlcNAcβ-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, Galβ-4(Fucα-3)GlcNAcβ-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 β-3 N-acetylglucosaminyltransferase.