Sialyltransferases for the production of sialylated oligosaccharides
AssigneeInbiose N.V.
InventorsJoeri Beauprez, Thomas Decoene, Annelies Vercauteren, Tom Verhaeghe
Life of this patent
Estimating the term…
Abstract
The present invention is in the technical field of synthetic biology, metabolic engineering and cell cultivation. The present invention relates to newly identified sialyltransferases having alpha-2,3-sialyltransferase activity on the galactose residue of lactose. The invention also describes methods for the production of a 3′sialylated oligosaccharide using any one of the newly identified sialyltransferases as well as the purification of the 3′sialylated oligosaccharide. The present invention also provides a cell for production of the 3′sialylated oligosaccharide and the use of the cell in a cultivation or incubation.
Claims17
A method of producing a 3′sialylated oligosaccharide, the method comprising:
- contacting a sialyltransferase with a mixture comprising a donor comprising a sialic acid residue and an acceptor selected from an oligosaccharide or a disaccharide under conditions wherein the sialyltransferase catalyzes transfer of the sialic acid residue from the donor to the acceptor, thereby producing the 3′sialylated oligosaccharide,
- wherein the sialyltransferase has α-2,3-sialyltransferase activity on the galactose (Gal) residue of lactose and comprises an amino acid sequence that has at least 80.0% amino acid sequence identity to the full-length amino acid sequence of SEQ ID NO:
- 5,
- wherein the contacting occurs in a cell-free system or a cell that is heterologous to the sialyltransferase, and,
- optionally, separating the produced 3′sialylated oligosaccharide.
A method of producing a 3′sialylated oligosaccharide, the method comprising:
- i) providing a cell, heterologously expressing a sialyltransferase, wherein the sialyltransferase has α-2,3-sialyltransferase activity on the Gal residue of lactose and comprises an amino acid sequence that has at least 80.0% amino acid sequence identity to the full-length amino acid sequence of SEQ ID NO:
- 5,
- ii) providing CMP-sialic acid, optionally wherein the CMP-sialic acid is produced by the cell,
- iii) providing an oligosaccharide or disaccharide, optionally wherein the oligosaccharide or disaccharide is produced by the cell,
- iv) cultivating and/or incubating the cell under conditions
- a) permissive to express the sialyltransferase, optionally permissive to produce the CMP-sialic acid and/or the oligosaccharide or disaccharide, and
- b) wherein the sialyltransferase catalyzes the transfer of a sialic acid residue from the CMP-sialic acid to an acceptor resulting in producing the 3′sialylated oligosaccharide, and
- v) optionally, separating the 3′sialylated oligosaccharide from the cultivation or incubation.
Dependencies are inferred from the claim text.