Production of human milk oligosaccharides in microbial hosts with engineered import / export
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.
1. A method for the production of lacto-N-triose II by a genetically modified microbial host cell, comprising
providing a genetically modified microbial host cell that comprises:
at least one recombinant β-1,3-N-acetylglucosaminyltransferase, wherein the at least one recombinant β-1,3-N-acetylglucosaminyltransferase belongs to the class of lgtA of Neisseria meningitidis , and
increased expression of at least one sugar export protein capable of exporting the lacto-N-triose II, wherein the at least one sugar export protein is SetA from Cedecea neteri;
cultivating the microbial host cell in a medium under conditions permissive for the production of the lacto-N-triose II, whereby the lacto-N-triose II is exported into the medium at an increased level compared to the unmodified microbial host cell, and
obtaining the lacto-N-triose II from the medium.
2. A genetically modified microbial host cell for the production of lacto-N-triose II, wherein the microbial host cell comprises:
at least one recombinant β-1,3-N-acetylglucosaminyltransferase, wherein the at least one recombinant β-1,3-N-acetylglucosaminyltransferase belongs to the class of lgtA of Neisseria meningitidis , and
increased expression of at least one sugar export protein capable of exporting the lacto-N-triose II, wherein the at least one sugar export protein is SetA from Cedecea neteri.