SYNTHESIS OF FUCOSYLATED COMPOUNDS
A method for making a genetically modified cell having the ability to produce fucosylated compounds comprising the steps of transforming the cell to express a fucose kinase transforming the cell to express a fucose-1-phosphate guanylyltransferase transforming the cell to express a fucosyltransferase.
1 . A method for making a genetically modified cell having the ability to produce fucosylated compounds comprising the steps of
transforming the cell to express a fucose kinase
transforming the cell to express a fucose-1-phosphate guanylyltransferase
transforming the cell to express a fucosyltransferase.
2 . The method of claim 1 , wherein the genetically modified cell is a microorganismm selected from the group consisting of the genera Escherichia, Klebsiella, Helicobacter, Bacillus, Lactobacillus, Streptococcus, Lactococcus, Pichia, Saccharomyces and Kluyveromyces.
3 . The method of claim 1 , wherein the fucose kinase and the fucose-1-phosphate guanylyltransferase are combined in a bifunctional enzyme.
4 . The method of claim 3 , wherein the bifunctional fucose kinase/fucose-1-phosphate guanylyltransferase is selected of bifunctional fucose kinase/fucose-1-phosphate guanylyltransferase dervied from the group consisting of the genera Bacteroides, Lentisphaera, Ruminococcus, Solibacter, Arabidopsis, Oryza, Physcomitrella, Vitis, Danio, Bos, Equus, Macaca, Pan, Homo, Rattus, Mus and Xenopus.
5 . The method of claim 1 , wherein the fucosyltransferase is derived from an organism selected from the group consisting of the genera Helicobacter, Escherichia, Yersinia, Enterococcus, Shigella, Klebsiella, Salmonella, Bacteroides, Dictyosetelium, Arabidopsis, Drosophila, Homo, Bos, Mus, Rattus, Gallus, Canis and Sus.
6 . The method of claim 1 , wherein a catabolic pathway of said cell for fucose is inactivated.
7 . The method of claim 6 , wherein the catabolic pathway for fucose is inactivated by inactivating one or several genes selected from the group consisting of a fucose-1-phosphate aldolase gene, a fucose isomerase gene and a fuculose kinase gene.
8 . The method of claim 1 , wherein the fucosylated compound is a fucosyllactose, preferably 2′-fucosyllactose, 3-fucosyllactose or lactodifucotetraose.
9 . A genetically modified cell obtainable by the method of claim 1 .
10 . A method for making fucosylated compound comprising the steps of cultivating the cell of claim 9 under suitable cultivation conditions in a medium comprising fucose and an acceptor substrate.
11 . The method of claim 10 , wherein the acceptor substrate is a mono-, di- or oligosaccharide or a peptide.
12 . The method of claim 10 , wherein the acceptor substrate is lactose, 2′-fucosyllactose or 3-fucosyllactose.
13 . The method of claim 10 , wherein the fucosylated compound is a fucosyllactose, preferably 2′-fucosyllactose or 3-fucosyllactose, or lactodifucotetraose.