Glycosyltransferases for biosynthesis of oligosaccharides,...

C - Chemistry – Metallurgy – 12 – N

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C12N 15/54 (2006.01) C12N 9/10 (2006.01) C12N 11/02 (2006.01) C12P 19/18 (2006.01) C12P 19/26 (2006.01)

Patent

CA 2200973

The present invention is directed to nucleic acids encoding glycosyltransferases, the proteins encoded thereby, and to methods for synthesizing oligosaccharides using the glycosyltransferases of the invention. In a specific embodiment, a glycosyltransferase locus of Neisseria gonorrhoeae containing five open reading frames for five different glycosyltransferases. The functionally active glycosyltransferases of the invention are characterized by catalyzing reactions such as adding Gal .beta.14 to GlcNAc or Glc; adding Ga1NAc or GlcNAc .beta.13 to Gal; and adding Gal .alpha.14 to Gal. The glycosyltransferases of the invention are particularly suited to the syntheses of the oligosaccharides Gal.beta.14G1cNAc.beta.13Gal.beta.14Glc (a mimic of lacto-N-neotetraose), GalNAc.beta.13Gal.beta.14GlcNAc.beta.13Gal.beta.14Glc.beta.14 (a mimic ganglioside), and Gal.alpha.14Gal.beta.14Glc.beta.14HepR (a mimic of the saccharide portion of globoglycolipids).

La présente invention se rapporte à des acides nucléiques codant des glycosyltransférases, aux protéines codées par ceux-ci, et aux procédés de synthèse d'oligosaccharides à l'aide des glycosyltransférases selon l'invention. Selon un mode de réalisation spécifique, un locus de glycosyltransférase de Neisseria gonorrhoeae contient cinq cadres de lectures ouverts pour cinq glycosyltransférases différentes. Les glycosyltransférases fonctionnellement actives selon l'invention se caractérisent par des réactions de catalyse telles que l'ajout de Gal beta 1}4 à GlcNAc ou Glc; l'ajout de GalNAc ou GlcNAc beta 1}3 à Gal; et l'ajout de Gal alpha 1}4 à Gal. Les glycosyltransférases selon l'invention sont notamment appropriées à la synthèse des oligosaccharides Gal beta 1}4GlcNAc beta 1}3Gal beta 1}4Glc (un imitateur de lacto-N-néotétraose), GalNAc beta 1}3Gal beta 1}4GlcNAc beta 1}3Gal beta 1}4Glc beta 1}4 (un imitateur de ganglioside), et Gal alpha 1}4Gal beta 1}4Glc beta 1}4Hep}R (un imitateur de la partie saccharide des globoglycolipides).

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