Yeast strains producing mammalian-like complex N-glycans
AssigneeOxyrane UK Limited
InventorsSteven Christian Jozef Geysens, Wouter Vervecken
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Abstract
Described herein are methods and genetically engineered fungal cells useful for producing target molecules containing mammalian-like complex N-glycans or containing intermediates in a mammalian glycosylation pathway.
Claims16
A substantially pure culture of Yarrowia lipolytica cells, a substantial number of which are genetically engineered to produce glycoproteins comprising GalGlcNAc2Man3GlcNAc2 or Gal2GlcNAc2Man3GlcNAc2 N-glycans, wherein said cells are genetically engineered to be deficient in Outer CHain elongation (OCH1) activity and in Asparagine Linked Glycosylation 3 (ALG3) activity and to comprise one or more nucleic acids encoding an α-1,2-mannosidase, a nucleic acid encoding Asparagine Linked Glycosylation 6 (ALG6), and one or more nucleic acids encoding each of a GlcNAc-transferase I, a GlcNAc-transferase II, and a galactosyltransferase, wherein each of said one or more nucleic acids encoding said α-1,2-mannosidase, said GlcNAc-transferase I, said GlcNAc-transferase II, and said galactosyltransferase comprises nucleotide sequences encoding targeting sequences to target each encoded protein to an intracellular compartment, wherein said cells produce proteins comprising GalGlcNAc2Man3GlcNAc2 or Gal2GlcNAc2Man3GlcNAc2 N-glycans.
An isolated Yarrowia lipolytica cell genetically engineered to produce proteins comprising GalGlcNAc2Man3GlcNAc2 or Gal2GlcNAc2Man3GlcNAc2 N-glycans, wherein said cell is genetically engineered to be deficient in Outer CHain elongation (OCH1) activity and in Asparagine Linked Glycosylation 3 (ALG3) activity and to comprise one or more nucleic acids encoding an α-1,2-mannosidase, a nucleic acid encoding Asparagine Linked Glycosylation 6 (ALG6), and one or more nucleic acids encoding each of a GlcNAc-transferase I, a GlcNAc-transferase II, and a galactosyltransferase, wherein each of said one or more nucleic acids encoding said α-1,2-mannosidase, said GlcNAc-transferase I, said GlcNAc-transferase II, and said galactosyltransferase comprises nucleotide sequences encoding targeting sequences to target each encoded protein to an intracellular compartment, wherein said cell produces proteins comprising GalGlcNAc2Man3GlcNAc2 or Gal2GlcNAc2Man3GlcNAc2 N-glycans.
Dependencies are inferred from the claim text.