Production of human milk oligosaccharides in microbial hosts with engineered import / export
AssigneeChr. Hansen A/S
InventorsStefan Jennewein, Dirk Wartenberg
Life of this patent
Estimating the term…
Abstract
The present invention relates to methods for the production of oligosaccharides in genetically modified bacterial host cells, as well as to the genetically modified host cells used in the methods. The genetically modified host cell comprises at least one recombinant glycosyltransferase, and at least one nucleic acid sequence coding for a protein enabling the export of the oligosaccharide.
Claims19
A method for production of an oligosaccharide by a genetically modified microbial host cell, wherein the oligosaccharide is a lacto-N-tetraose comprising a lacto-N-triose II (LNT-II;
- GlcNAc(b1-3)Gal(b1-4)Gluc) as a core trisaccharide, the method comprising:
- providing a genetically modified microbial host cell that comprises:
- at least one recombinant glycosyltransferase selected from the group consisting of a β-1,3-N-acetylglucosaminyltransferase and a β-1,3-galactosyltransferase;
- increased expression or activity of at least one homologous or heterologous sugar export protein capable of exporting the oligosaccharide;
- and
- decreased expression or inactivation of at least one export protein that exports LNT-II from the host cell;
- cultivating the host cell in a medium under conditions permissive for the production of the oligosaccharide, whereby the oligosaccharide is exported into the medium at an increased level compared to an unmodified host cell, and
- obtaining the oligosaccharide from the medium.
A genetically modified microbial host cell for the production of lacto-N-tetraose comprising a lacto-N-triose II (LNT-II;
- GlcNAc(β1-3)Gal(β1-4)Gluc) as a core trisaccharide, wherein the host cell comprises
- at least one recombinant glycosyltransferase, selected from the group consisting of a β-1,3-N-acetylglucosaminyltransferase and a β-1,3-galactosyltransferase;
- at least one homologous or heterologous nucleic acid sequence coding for a sugar export protein capable exporting of the acto-N-tetraose into a culture medium the host cell is cultivated in wherein said sugar export protein capable of exporting the lacto-N-tetraose belongs to the class of secondary active transporters, and wherein said homologous or heterologous nucleic acid sequence coding for the sugar export protein capable of exporting the lacto-N-tetraose is overexpressed or under control of a promoter sequence enabling the overexpression of said nucleic acid sequence;
- and
- inactivated expression or activity of at least one endogenous sugar export protein that exports LNT-II from the host cell.
Dependencies are inferred from the claim text.