New fucosyltransferases for in vivo synthesis of lnfp-iii
OwnerManos PAPADAKISinventor
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Abstract
The present disclosure relates to the production of fucosylated Human Milk Oligosaccharides (HMOs). In particular, it relates to alpha-1,3-fucosyltransferases for the production of lacto-N-fucopentaose III (LNFP-III), as well as to genetically engineered cells and their use in said production. Preferably the alpha-1,3-fucosyltransferases described herein do not fucosylate the reducing end of the acceptor oligosaccharide used as substrate for the alpha-1,3-fucosyltransferases.
Claims22
A genetically engineered cell capable of producing one or more fucosylated HMOs, comprising a recombinant nucleic acid sequence encoding a fucosyltransferase with α-1,3-fucosyltransferase activity, wherein the fucosyltransferase is selected from the group consisting of:
- a. Camp1, with an amino acid sequence according to SEQ ID NO:
- 1, or a functional homologue thereof with an amino acid sequence that is at least 80% identical to SEQ ID NO:
- 1;
- b. Parm1 with an amino acid sequence according to SEQ ID NO:
- 2, or a functional homologue thereof with an amino acid sequence that is at least 80% identical to SEQ ID NO:
- 2;
- and
- c. Hpull1 with an amino acid sequence according to SEQ ID NO:
- 3, or a functional homologue thereof with an amino acid sequence that is at least 80% identical to SEQ ID NO:
- 3, and
- d. Med1 with an amino acid sequence according to SEQ ID NO:
- 4, or a functional homologue thereof with an amino acid sequence that is at least 80% identical to SEQ ID NO:
- 4.
(canceled)
A mixture of HMOs, wherein the mixture is selected from the group consisting of:
- a. 30-55 molar % LNFP-III, 40-65 molar % LNnT and 2-8% molar % pLNnH, in total adding up to 100% molar content;
- b. 30-99 molar % LNFP-III, 0-65 molar % LNnT, 0-20 molar % 3-FL, 0-10 molar % LNDFH-III and 0-8 molar % pLNnH, in total adding up to 100% molar content;
- c. 70-99 molar % LNFP-III and 1-50 molar % LNnT in total adding up to 100% molar content;
- d. 80-99 molar % LNFP-III, 0-5 molar % LNnT and 0-10 molar % LNDFH-III, in total adding up to 100% molar content;
- and
- e. 80-99 molar % LNFP-III and 0-20 molar % 3-FL in total adding up to 100% molar content.
(canceled)
Dependencies are inferred from the claim text.