PRODUCTION OF FUCOSYLATED OLIGOSACCHARIDES IN BACILLUS
Disclosed are non-sporulating Bacillus cells for the production of a fucosylated oligosaccharide as well as a method for producing a fucosylated oligosaccharide, wherein said Bacillus cell has been genetically engineered to possess a lactose permease, a GDP-fucose biosynthesis pathway and a fucosyltransferase.
1 . A non-sporulating Bacillus cell for the production of a fucosylated oligosaccharide, wherein said cell has been genetically engineered to possess a lactose permease, a GDP-fucose biosynthesis pathway and a fucosyltransferase.
2 . The Bacillus cell according to claim 1 , wherein the sporulation has been impaired by deletion or functional inactivation of one or more of the genes encoding Spo0A, sigma E and sigma F.
3 . The Bacillus cell according to claim 1 , wherein the lactose permease is E. coli LacY or a functional variant thereof.
4 . The Bacillus cell according to claim 1 , wherein the GDP-fucose biosynthesis pathway is a de novo GDP-fucose biosynthesis pathway and/or a salvage pathway.
5 . The Bacillus cell according to claim 1 , wherein the fucosyltransferase is a lactose-accepting fucosyltransferase, optionally a fucosyltransferase selected from the group of α-1,2-fucosyltransferases and α-1,3-fucosoyltransferases.
6 . The Bacillus cell according to claim 1 , wherein said Bacillus cell lacks any β-galactosidase activity or possesses reduced β-galactosidase activity as compared to a wild-type progenitor Bacillus cell of the same species.
7 . The Bacillus cell according to claim 6 , wherein the Bacillus cell has been genetically engineered by deletion or functional inactivation of at least one of the genes selected from the group consisting of yesZ and ganA.
8 . The Bacillus cell according to claim 1 , wherein said Bacillus cell is a Bacillus subtilis cell.
9 . A product comprising a Bacillus cell according to claim 1 for production of a fucosylated oligosaccharide.
10 . A method for production of a fucosylated oligosaccharide, comprising
providing a non-sporulating Bacillus cell as defined in claim 1 ;
cultivating said Bacillus cell in a medium and under conditions that are permissive for production of a fucosylated oligosaccharide; and optionally
retrieving the fucosylated oligosaccharide from the medium and/or the Bacillus cell.
11 . The method according to claim 10 , wherein the medium comprises lactose.
12 . The method according to claim 10 , wherein the medium comprises L-fucose.
13 . A fucosylated oligosaccharide produced by a method according to claim 10 .
14 . A product comprising a fucosylated oligosaccharide according to claim 13 for manufacture of a nutritional composition, optionally of an infant formula.
15 . A nutritional composition comprising a fucosylated oligosaccharide according to claim 13 .