| abstract |
method for the production of fucosylated oligosaccharides using a genetically modified prokaryotic host cell, the method comprising the steps of: - providing a prokaryotic host cell that has been genetically modified, so that at least (i) the activity of a fructose-6-phosphate converting enzyme, which in the unmodified host cell has a regular level, wherein said fructose-6-phosphate converting enzyme is selected from the group consisting of phosphofructokinase, glucose-6-phosphate isomerase , fructose-6-phosphate aldolase, a transketolase or a transaldolase, and / or increasing the activity of a fructose-1,6-bisphosphate phosphatase; (ii) at least one gene encoding an enzyme necessary for the de novo synthesis of GDP-fucose is overexpressed in the host cell, wherein the at least one gene encoding an enzyme necessary for the de novo synthesis of GDP-fucose is a gene encoding a ManA enzyme that catalyzes the isomerization of fructose-6-phosphate to mannose-6-phosphate, a gene encoding phosphomannomutase, a gene encoding mannose-1-phosphate guanosyltransferase, a gene encoding GDP-mannose-4 , 6-dehydratase or a gene encoding GDP-L-fucose synthase; (iii) an exogenous gene encoding an alpha-1,2-fucosyltransferase and / or alpha-1,3-fucosyltransferase is expressed in the host cell; - cultivating and / or developing said genetically modified host cell in a culture medium from a carbon and / or energy source, which is selected from at least one of the following: glycerol, succinate, malate, pyruvate, lactate, ethanol, citrate; and - providing lactose to the culture medium; thus producing the fucosylated oligosaccharide obtainable from the medium in which the host cell is cultured. |