Glycobiology, 2002, Vol. 12, No. 4 235-240
© 2002 Oxford University Press
A new fermentation process allows large-scale production of human milk oligosaccharides by metabolically engineered bacteria
2Centre de Recherches sur les Macromolécules Végétales, CNRS (affiliated with the Joseph Fourier University) BP 53, 38041 Grenoble cedex 9 France, and 3Institute for Biological Sciences, NRCC, Ottawa, Ontario, K1A 0R6, Canada
When fed to a ß-galactosidase-negative (lacZ–) Escherichia coli strain that was grown on an alternative carbon source (such as glycerol), lactose accumulated intracellularly on induction of the lactose permease. We showed that intracellular lactose was efficiently glycosylated when genes of glycosyltransferase that use lactose as acceptor were expressed. High-cell-density cultivation of lacZ– strains that overexpressed the ß1,3 N acetyl glucosaminyltransferase lgtA gene of Neisseria meningitidis resulted in the synthesis of 6 g · L–1 of the expected trisaccharide (GlcNAcß1-3Galß1-4Glc). When the ß1,4 galactosyltransferase lgtB gene of N. meningitidis was coexpressed with lgtA, the trisaccharide was further converted to lacto-N-neotetraose (Galß1-4GlcNAcß1-3Galß1-4Glc) and lacto-N-neoheaxose with a yield higher than 5 g · L–1. In a similar way, the nanA– E. coli strain that was devoid of NeuAc aldolase activity accumulated NeuAc on induction of the NanT permease and the lacZ– nanA– strain that overexpressed the N. meningitidis genes of the 
2,3 sialyltransferase and of the CMP-NeuAc synthase efficiently produced sialyllactose (NeuAc2-3Galß1-4Glc) from exogenous NeuAc and lactose.
1 To whom correspondence should be addressed
CiteULike 
Connotea 
Del.icio.us What's this?
This article has been cited by other articles:
|
|
 |
|
  E. Yavuz, S. Drouillard, E. Samain, I. Roberts, and B. Priem Glucuronylation in Escherichia coli for the bacterial synthesis of the carbohydrate moiety of nonsulfated HNK-1 Glycobiology, February 1, 2008; 18(2): 152 - 157. [Abstract] [Full Text] [PDF]
|
|