New sialyltransferases for in vivo synthesis of 3'sl and 6'sl
OwnerManos PAPADAKISinventor
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Abstract
The present disclosure relates to the production of mixtures of sialylated Human Milk Oligosaccharides (HMOs), in particular 3′SL and 6′SL and genetically modified cells and their use in said production, where the cells express a heterologous sialyltransferase with dual α-2,3-sialyltransferase/α-2,6-sialyltransferase activity.
Claims22
A genetically modified cell comprising a recombinant nucleic acid sequence encoding an enzyme with dual α-2,3-sialyltransferase/α-2,6-sialyltransferase activity wherein the cell is capable of producing at least 20% 3′SL and at least 20% 6′SL of the total molar HMO content produced by the cell.
A nucleic acid construct comprising recombinant nucleic acid sequence encoding a sialyltransferase with dual α-2,3-sialyltransferase/α-2,6-sialyltransferase activity, wherein said recombinant nucleic acid sequence is selected from the group consisting of:
- a. Chepa comprising the nucleic acid sequence of SEQ ID NO:
- 7 or a nucleic acid sequence with at least 80% identity to SEQ ID NO:
- 7,
- b. Cinf1 comprising of the nucleic acid sequence of SEQ ID NO:
- 8 or a nucleic acid sequence with at least 80%, identity to SEQ ID NO:
- 8,
- c. Ccol2 comprising the nucleic acid sequence of SEQ ID NO:
- 9 or a nucleic acid sequence with at least 80% identity to SEQ ID NO:
- 9,
- d. Cjej1 comprising the nucleic acid sequence of SEQ ID NO:
- 10 or a nucleic acid sequence with at least 80%, identity to SEQ ID NO:
- 10,
- e. Poral_2 comprising the nucleic acid sequence of SEQ ID NO:
- 11 or a nucleic acid sequence with at least 80% identity to SEQ ID NO:
- 11, and
- f. CstII comprising the nucleic acid sequence of SEQ ID NO:
- 12 or a nucleic acid sequence with at least 80% identity to SEQ ID NO:
- 12,
- wherein the sialyltransferase encoding sequence is under the control of a promoter sequence selected from the group consisting of PglpF, Plac, PmglB_70UTR or a PlgpA_70UTR or a PlgpT_70UTR (SEQ ID NOs:
- 17, 18, 28, 29, 30) and variants thereof.
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Dependencies are inferred from the claim text.