Method for the production of fucosyllactose in bacterial cells
The present invention relates to a cell to be stably cultured in a medium, which cell is adjusted for the production of oligosaccharides, the cell being transformed to comprise at least one nucleic acid sequence coding for an enzyme involved in oligosaccharide synthesis. In addition the cell is transformed to comprise at least one nucleic acid sequence coding for a protein of the sugar efflux transporter family, a functional homolog or derivative thereof. Further, the invention concerns a method for the production of oligosaccharides involving above cell.
1. A bacterial cell transformed to comprise at least one nucleic acid sequence coding for fucosyltransferase, and transformed to comprise at least one heterologous nucleic acid sequence coding for a protein of the sugar efflux transporter (SET) family, wherein the protein of the SET family is overexpressed in said bacterial cell.
2. The bacterial cell according to claim 1 , wherein the cell is an Escherichia coli cell.
3. The bacterial cell according to claim 1 , wherein the protein of the sugar efflux transporter family is SetA.
4. The bacterial cell according to claim 1 , wherein it is further transformed to comprise at least one nucleic acid sequence coding for a protein facilitating or promoting the import of educts required for oligosaccharide synthesis, wherein the protein is selected from the group consisting of lactose transporter, fucose transporter, sialic acid transporter, galactose transporter, mannose transporter, N-acetylglucosamine transporter, N-acetylgalactosamine transporter, ABC-transporter, transporter for a nucleotide-activated sugar, and transporter for a nucleobase, nucleoside or nucleotide.
5. The bacterial cell according to claim 1 , wherein it is further transformed to comprise at least one nucleic acid sequence coding for a protein selected from the group consisting of nucleotidyltransferase, guanylyltransferase, uridylyltransferase, Fkp, L-fucose kinase, fucose-1-phosphate guanylyltransferase, CMP-sialic acid synthetase, galactose kinase, galactose-1-phosphate uridylyltransferase, glucose kinase, glucose-1-phosphate uridylyltransferase, mannose kinase, mannose-1-phosphate guanylyltransferase, GDP-4-keto-6-deoxy-D-mannose reductase, glucosamine kinase, glucosamine-phosphate acetyltransferase, N-acetyl-glucosamin-phosphate uridylyltransferase, UDP-N-acetylglucosamine 4-epimerase, UDP-N-acetyl-glucosamine 2-epimerase.
6. The bacterial cell according to claim 1 , wherein it comprises a catabolic pathway for selected mono-, di- or oligosaccharides, which is at least partially inactivated, the mono-, di- or oligosaccharides being involved in and/or required for the synthesis of oligosaccharides.