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Large-scale in vivo synthesis of the carbohydrate moieties of gangliosides GM1 and GM2 by metabolically engineered Escherichia coli.
- Source :
-
Chembiochem : a European journal of chemical biology [Chembiochem] 2003 May 09; Vol. 4 (5), pp. 406-12. - Publication Year :
- 2003
-
Abstract
- Two metabolically engineered Escherichia coli strains have been constructed to produce the carbohydrate moieties of gangliosides GM2 (GalNAcbeta-4(NeuAcalpha-3)Galbeta-4Glc; Gal = galactose, Glc = glucose, Ac = acetyl) and GM1 (Galbeta-3GalNAcbeta-4(NeuAcalpha-3)Galbeta-4Glc. The GM2 oligosaccharide-producing strain TA02 was devoid of both beta-galactosidase and sialic acid aldolase activities and overexpressed the genes for CMP-NeuAc synthase (CMP = cytidine monophosphate), alpha-2,3-sialyltransferase, UDP-GlcNAc (UDP = uridine diphosphate) C4 epimerase, and beta-1,4-GalNAc transferase. When this strain was cultivated on glycerol, exogenously added lactose and sialic acid were shown to be actively internalized into the cytoplasm and converted into GM2 oligosaccharide. The in vivo synthesis of GM1 oligosaccharide was achieved by taking a similar approach but using strain TA05, which additionally overexpressed the gene for beta-1,3-galactosyltransferase. In high-cell-density cultures, the production yields for the GM2 and GM1 oligosaccharides were 1.25 g L(-1) and 0.89 g L(-1), respectively.
- Subjects :
- Carbohydrate Conformation
Carbohydrate Epimerases genetics
Carbohydrate Epimerases metabolism
Carbohydrate Sequence
Genetic Engineering
Glycosyltransferases genetics
Glycosyltransferases metabolism
Lactose chemistry
Lactose metabolism
Molecular Sequence Data
N-Acetylneuraminic Acid analogs & derivatives
N-Acetylneuraminic Acid metabolism
N-Acylneuraminate Cytidylyltransferase genetics
N-Acylneuraminate Cytidylyltransferase metabolism
Oligosaccharides chemistry
Escherichia coli genetics
Escherichia coli metabolism
G(M1) Ganglioside biosynthesis
G(M1) Ganglioside chemistry
G(M2) Ganglioside biosynthesis
G(M2) Ganglioside chemistry
Oligosaccharides biosynthesis
Subjects
Details
- Language :
- English
- ISSN :
- 1439-4227
- Volume :
- 4
- Issue :
- 5
- Database :
- MEDLINE
- Journal :
- Chembiochem : a European journal of chemical biology
- Publication Type :
- Academic Journal
- Accession number :
- 12740812
- Full Text :
- https://doi.org/10.1002/cbic.200200540