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Synthesis of globopentaose using a novel beta1,3-galactosyltransferase activity of the Haemophilus influenzae beta1,3-N-acetylgalactosaminyltransferase LgtD.
- Source :
-
FEBS letters [FEBS Lett] 2007 Jun 12; Vol. 581 (14), pp. 2652-6. Date of Electronic Publication: 2007 May 11. - Publication Year :
- 2007
-
Abstract
- We have previously described a bacterial system for the conversion of globotriaose (Gb3) into globotetraose (Gb4) by a metabolically engineered Escherichia coli strain expressing the Haemophilus influenzae lgtD gene encoding beta1,3-N-acetylgalactosaminyltransferase [Antoine, T., Bosso, C., Heyraud, A. Samain, E. (2005) Large scale in vivo synthesis of globotriose and globotetraose by high cell density culture of metabolically engineered Escherichia coli. Biochimie 87, 197-203]. Here, we found that LgtD has an additional beta1,3-galactosyltransferase activity which allows our bacterial system to be extended to the synthesis of the carbohydrate portion of globopentaosylceramide (Galbeta-3GalNAcbeta-3Galalpha-4Galbeta-4Glc) which reacts with the monoclonal antibody defining the stage-specific embryonic antigen-3. In vitro assays confirmed that LgtD had both beta1,3-GalT and beta1,3-GalNAcT activities and showed that differences in the affinity for Gb3 and Gb4 explain the specific and exclusive formation of globopentaose.
- Subjects :
- Chromatography, Thin Layer
Globosides chemistry
Globosides metabolism
Haemophilus influenzae enzymology
Kinetics
Models, Biological
Oligosaccharides chemistry
Substrate Specificity
Bacterial Proteins metabolism
Globosides biosynthesis
N-Acetylgalactosaminyltransferases metabolism
Oligosaccharides biosynthesis
Subjects
Details
- Language :
- English
- ISSN :
- 0014-5793
- Volume :
- 581
- Issue :
- 14
- Database :
- MEDLINE
- Journal :
- FEBS letters
- Publication Type :
- Academic Journal
- Accession number :
- 17517393
- Full Text :
- https://doi.org/10.1016/j.febslet.2007.05.008