Microorganisms genetically engineered to have modified N-glycosylation activity
AssigneesOxyrane UK Limited, Vib Vzw, Universiteit Gent
InventorsNico Luc Marc Callewaert, Wouter Vervecken, Karen Jacqueline Marcel De Pourcq, Steven Christian Jozef Geysens, Mouna Guerfal
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Abstract
Described herein are methods and genetically engineered cells useful for producing an altered N-glycosylation form of a target molecule. Also described are methods and molecules with altered N-glycosylation useful for treating a variety of disorders such as metabolic disorders.
Claims54
A method of producing an altered N-glycosylation form of a target protein, the method comprising:
- providing a Yarrowia lipolytica or an Arxula adeninivorans cell genetically engineered to express a protein capable of promoting mannosyl phosphorylation of N-glycans;
- and
- introducing into the cell a nucleic acid encoding a target protein, wherein the cell produces the target protein in an altered N-glycosylation form, and
- wherein the protein capable of promoting mannosyl phosphorylation of N-glycans is selected from the group consisting of:
- a wild-type MNN4 polypeptide or a biologically active variant of a wild-type MNN4 polypeptide;
- a wild-type PNO1 polypeptide or a biologically active variant of a wild-type PNO1 polypeptide;
- and
- a wild-type MNN6 polypeptide.
An isolated Yarrowia lipolytica or Arxula adeninivorans cell genetically engineered to express a protein capable of promoting mannosyl phosphorylation of N-glycans, wherein the cell is capable of producing a target polypeptide in an altered glycosylation form when a nucleic acid encoding the target polypeptide is introduced into the cell, and
- wherein the protein capable of promoting mannosyl phosphorylation of N-glycans is selected from the group consisting of:
- a wild-type MNN4 polypeptide or a biologically active variant of a wild-type MNN4 polypeptide;
- a wild-type PNO1 polypeptide or a biologically active variant of a wild-type PNO1 polypeptide;
- and
- a wild-type MNN6 polypeptide.
A substantially pure culture of Yarrowia lipolytica or Arxula adeninivorans cells, wherein the culture comprises a cell genetically engineered to express a protein capable of promoting mannosyl phosphorylation of N-glycans, wherein the cell is capable of producing a target polypeptide in an altered glycosylation form when a nucleic acid encoding the target polypeptide is introduced into the cell, and
- wherein the protein capable of promoting mannosyl phosphorylation of N-glycans is selected from the group consisting of:
- a wild-type MNN4 polypeptide or a biologically active variant of a wild-type MNN4 polypeptide;
- a wild-type PNO1polypeptide or a biologically active variant of a wild-type PNO1polypeptide;
- and a wild-type MNN6 polypeptide.
Dependencies are inferred from the claim text.