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AssigneesResearch Corporation Technologies, Inc., Universiteit Gent, Vib Vzw
InventorsRoland Contreras, Nico L. M. Callewaert, Steven C. J. Geysens, Vladimir Kaigorodov, Vervecken Wouter
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The present invention provides genetically engineered strains of methylotrophic yeast including Pichia and especially Pichia pastoris capable of producing proteins with reduced or modified glycosylation. Methods of producing glycoproteins with reduced and/or modified glycosylation using such genetically engineered strains of Pichia are also provided. Vectors, which comprise coding sequences for α-1,2-mannosidase I, glucosidase II, GlcNAc-tranferase I and mannosidase II or comprising OCH1 disrupting sequence, for transforming methylotrophic yeasts are contemplated by the present invention. Kit for providing the comtemplated vectors are also included in this invention.
A method of reducing glycosylation on proteins produced from methylotrophic yeast, comprising providing a methylotrophic yeast strain, which strain is transformed with a nucleic acid coding for a protein which comprises a GlcNAc-transferase I (GnT1) or an enzymatically active fragment thereof wherein said GnT1 is a human, rat or rabbit GnT1, and which strain is also transformed with a nucleic acid coding for a protein which comprises an α-1,2-mannosidase or an enzymatically active fragment thereof, wherein said α-1,2-mannosidase is T. reesei α-1,2-mannosidase, and wherein said α-1,2-mannosidase and said enzymatically active fragment are engineered to contain an ER localization signal and convert Man8GlcNAc2 to Man5GlcNAc2;
Dependencies are inferred from the claim text.