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US 11,926,858 B2Granted
Oligosaccharide production
AssigneeGlycom A/S
InventorsMargit Pedersen, Manos Papadakis, Peter Becker, Eric Samain, Pauline Peltier-Pain, Katrine Bych, Ted Johanson, Elise Champion, Gyula Dekany
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Abstract
The invention relates to a genetically modified microorganism for making a oligosaccharide, preferably of 3-8 monosaccharide units, more preferably of 3-5 monosaccharide units, particularly a HMO, which comprises one or more genes encoding a sucrose utilization system, so the microorganism can use sucrose as a carbon and energy source.
Claims20
A genetically modified microorganism for making an oligosaccharide or a glycoside of said oligosaccharide, comprising:
- two or more recombinant genes encoding two or more glycosyl transferases which can transfer a glycosyl residue of an activated sugar nucleotide to a carbohydrate acceptor or a glycoside of said acceptor,
- a biosynthetic pathway for making said activated sugar nucleotide from sucrose, and
- one or more genes encoding a heterologous PTS-dependent sucrose utilization system, so that said cell is capable to use sucrose as a carbon source for making said activated sugar nucleotide and as energy source for making said oligosaccharide,
- wherein the carbohydrate acceptor is not sucrose,
- wherein said genetically modified microorganism is an E. coli cell of LacZ− genotype,
- said oligosaccharide is lacto-N-tetraose (LNT),
- said activated sugar nucleotide is UDP-Gal and UDP-GlcNAc,
- said two or more glycosyl transferase comprises β-1,3-N-acetyl glucosaminyl transferase and β-1,3-galactosyl transferase,
- and said heterologous PTS-dependent sucrose utilization system comprises proteins encoded by scr genes scrY for a sucrose porin, scrA for a PTS permease, scrB for a sucrose-6-phosphate hydrolase and scrR for a repressor protein.
6 more elements
Dependencies are inferred from the claim text.