Method of producing sialylated oligosaccharides

C - Chemistry – Metallurgy – 12 – P

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C12P 19/18 (2006.01) C12N 1/21 (2006.01) C12N 9/04 (2006.01) C12N 9/10 (2006.01) C12N 9/12 (2006.01) C12P 19/26 (2006.01)

Patent

CA 2645141

The present invention relates to a method for the large scale in vivo synthesis of sialylated oligosaccharides, culturing a microorganism in a culture medium, optionally comprising an exogenous precursor such as lactose, wherein said microorganism comprises heterologous genes encoding a CMP-Neu5Ac synthetase, a sialic acid synthase, a GlcNAc-6-phosphate 2 epimerase and a sialyltransferase, and wherein the endogenous genes coding for sialic acid aldolase (NanA) and for ManNac kinase (NanK) have been deleted or inactivated. The invention also relates to these micoorganisms which are capable of producing internally activated sialic acid.

L'invention concerne un procédé pour la synthèse in vivo à grande échelle d'oligosaccharides sialylés. Ce procédé consiste à cultiver un micro-organisme dans un milieu de culture comprenant éventuellement un précurseur exogène de type lactose; ledit micro-organisme comprenant des gènes hétérologues codant pour une CMP-Neu5Ac synthétase, une synthase d'acide sialique, une GlcNAc-6-phosphate 2 épimérase et une sialyltransférase; et les gènes endogènes codant pour une aldolase d'acide sialique (NanA) et pour une ManNac kinase (NanK) ayant été supprimés ou inactivés. L'invention concerne également lesdits micro-organismes qui sont capables de produire de l'acide sialique activé de manière interne.

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