期刊
CHEMBIOCHEM
卷 7, 期 2, 页码 359-365出版社
WILEY-V C H VERLAG GMBH
DOI: 10.1002/cbic.200500293
关键词
biosynthesis; carbohydrates; glycoconjugates; glycosyltransferases; metabolic engineering
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 IgtB gene for beta-4 galactosyltransferase. The first compound Gal beta-4(Fuc alpha-3)GlcNAc beta-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, Gal beta-4(Fuc alpha-3)GlcNAc beta-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-acetylglucosominyltransferase.
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