Granted Patent B2

US 12,618,091 · App. 18/733,787 · Granted May 5, 2026

Inventors: Stefan Jennewein (Bad Honnef, DE); Dirk Wartenberg (Bonn, DE)

Assignee: Chr. Hansen A/S

C12P19/18C12N9/1051C12N15/70C12Y204/01146

Abstract

Claims (14)

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 or activity of at least one sugar export protein capable of exporting the lacto-N-triose II, wherein the at least one sugar export protein is YjhB from E. coli or ProP from Mannheimia succiniciproducens;

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 . The method of claim 1 , wherein, the endogenous β-galactosidase gene and the endogenous glucosamine-6-phosphate deaminase gene of the genetically modified microbial host cell are inactivated or deleted, and wherein said genetically modified microbial host cell comprises a nucleic acid sequence coding for a functional lactose permease protein.

3 . The method of claim 1 , wherein the genetically modified microbial host cell comprises an increased UDP-N-acetylglucosamine and UDP-galactose or GDP-fucose or CMP-N-acetylneuraminic acid production capability as compared to a genetically unmodified host cell, wherein optionally said increased UDP-N-acetylglucosamine and UDP-galactose production capability comprises the overexpression of one or more genes encoding for proteins comprising the following activities for a: L-glutamine:D-fructose-6-phosphate aminotransferase, N-acetyl glucosamine-1-phosphate uridyltransferase/glucosamine-1-phosphate acetyl transferase, phosphoglucosamine mutase, UDP-galactose-4-epimerase, phosphoglucomutase, glucose-1-phosphate uridylyltransferase.

4 . The method of claim 1 , wherein said genetically modified microbial host cell is cultivated in the presence of glucose, sucrose, glycerol or a combination thereof, but not by addition or in the presence of N-acetylglucosamine, galactose or combination thereof.

5 . A genetically modified microbial host cell for the production of lacto-N-triose II, wherein the microbial host cell comprises:

a. 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

b. increased expression or activity of at least one sugar export protein capable of exporting the lacto-N-triose II, wherein the at least one sugar export protein is YjhB from E. coli or ProP from Mannheimia succiniciproducens.

6 . The genetically modified microbial host cell of claim 5 , wherein the endogenous β-galactosidase gene and the endogenous glucosamine-6-phosphate deaminase gene in the genetically modified microbial host cell are inactivated or deleted, and wherein said genetically modified microbial host cell comprises a nucleic acid sequence coding for a functional lactose permease protein.

7 . The genetically modified microbial host cell of claim 5 , wherein the genetically modified microbial host cell comprises an increased UDP-N-acetylglucosamine and UDP-galactose or GDP-fucose or CMP-N-acetylneuraminic acid production capability as compared to a genetically unmodified host cell, wherein optionally said increased UDP-N-acetylglucosamine and UDP-galactose production capability comprises the overexpression of one or more genes encoding for proteins comprising the following activities for a: L-glutamine:D-fructose-6-phosphate aminotransferase, N-acetyl glucosamine-1-phosphate uridyltransferase/glucosamine-1-phosphate acetyl transferase, phosphoglucosamine mutase, UDP-galactose-4-epimerase, phosphoglucomutase, glucose-1-phosphate uridylyltransferase.

Assignments (1)

Priority Claims (1)

Continuity (3)

References Cited (39)