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 fucosyllactose the method comprises cultivating a genetically modified Escherichia coli having the ability to produce the fucosyllactose under suitable cultivation conditions in a medium comprising L-fucose and an acceptor substrate, wherein said acceptor substrate is lactose, and wherein the Escherichia coli has been transformed to express
i) a gene encoding a bacterial bifunctional fucose kinase/fucose-1-phosphate guanylyltransferase, wherein the bacterial bifunctional fucose kinase/fucose-1-phosphate guanylyltransferase gene is an fkp gene of Bacteroides fragilis , and
ii) a gene encoding a fucosyltransferase from Helicobacter,
wherein at least one gene encoding a protein that functions in the catabolic pathway for fucose, selected from the group consisting of a fucose-1-phosphate aldolase gene (fucA gene), a fucose isomerase gene and a fuculose kinase gene, is inactivated in the Escherichia coli.
2. The method of claim 1 , wherein the fucosyllactose is 2′-fucosyllactose, 3-fucosyllactose or lactodifucotetraose.
3. The method of claim 1 , wherein the fucA gene encoding fucose-1-phosphate aldolase is deleted.
4. The method of claim 1 , wherein the fucosyllactose is 2′-fucosyllactose and the fucosyltransferase is an alpha-1,2-fucosyltransferase encoded by the Helicobacter pylori NCTC364 fucT2 gene.
5. The method of claim 1 , the fucosyllactose is 3-fucosyllactose and the fucosyltransferase is an alpha-1,3-fucosyltransferase encoded by the Helicobacter pylori 26695 futA gene.