Inbiose N.v.

Gert Peters

Joeri Beauprez

Annelies Vercauteren

Pieter Coussement

Dries Van Herpe

Thomas Decoene

Nausicaä Lannoo

Sofie Aesaert

Kristof Vandewalle

This disclosure is in the technical field of synthetic biology and metabolic engineering. More particularly, this disclosure is in the technical field of fermentation of metabolically engineered microorganisms. This disclosure describes engineered micro-organisms able to synthesize sialylated compounds via an intracellular biosynthesis route. These micro-organisms can dephosphorylate N-acetylglucosamine-6-phosphate to N-acetyl glucosamine and convert the N-acetylglucosamine to N-acetylmannosamine. These micro-organisms also have the ability to convert N-acetylmannosamine to N-acetyl-neuraminate. Furthermore, this disclosure provides a method for the large scale in vivo synthesis of sialylated compounds, by culturing a microorganism in a culture medium, optionally comprising an exogenous precursor such as, but not limited to lactose, lactoNbiose, N-acetyllactosamine and/or an aglycon, wherein the microorganism intracellularly dephosphorylates N-acetylglucosamine-6-phosphate to N-acetylglucosamine, converts N-acetyl-glucosamine to N-acetylmannosamine and convert the latter further to N-acetyl-neuraminate.

In vivo synthesis of sialylated compounds

The disclosure is in the technical field of synthetic biology and metabolic engineering. More particularly, the disclosure is in the technical field of cultivation or fermentation of metabolically engineered cells. The disclosure describes a cell metabolically engineered for production of a mixture of at least four different neutral non-fucosylated oligosaccharides. Furthermore, the disclosure provides a method for the production of a mixture of at least four different neutral non-fucosylated oligosaccharides by a cell as well as the purification of at least one of the oligosaccharides from the cultivation.

Production of a mixture of neutral non-fucosylated oligosaccharides by a cell

The present invention is in the technical field of synthetic biology and metabolic engineering. More particularly, the present invention is in the technical field of fermentation of metabolically engineered microorganisms. The present invention describes engineered microorganisms able to synthesize sialylated compounds via an intracellular biosynthesis route. These microorganisms can dephosphorylate N-acetylglucosamine-6-phosphate to N-acetylglucosamine and convert the N-acetylglucosamine to N-acetylmannosamine. These microorganisms also have the ability to convert N-acetylmannosamine to N-acetyl-neuraminate. Furthermore, the present invention provides a method for the large scale in vivo synthesis of sialylated compounds, by culturing a microorganism in a culture medium, optionally comprising an exogenous precursor such as, but not limited to lactose, lactoNbiose, N-acetyllactosamine and/or an aglycon, wherein said microorganism intracellularly dephosphorylates N-acetylglucosamine-6-phosphate to N-acetylglucosamine, converts N-acetylglucosamine to N-acetylmannosamine and convert the latter further to N-acetyl-neuraminate.

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Ghent, Belgium

Gert Peters