Protein glycosylation modification in methylotrophic yeast
AssigneesVib Vzw, Universiteit Gent, Research Corporation Technologies, Inc.
InventorsRoland Contreras, Nico L.M. Callewaert, Steven C.J. Geysens
Life of this patent
Estimating the term…
Abstract
The present invention provides genetically engineered strains of Pichia capable of producing proteins with reduced glycosylation. In particular, the genetically engineered strains of the present invention are capable of expressing either or both of an α-1,2-mannosidase and glucosidase II. The genetically engineered strains of the present invention can be further modified such that the OCH1 gene is disrupted. Methods of producing glycoproteins with reduced glycosylation using such genetically engineered stains of Pichia are also provided.
Claims12
A genetically engineered methylotrophic yeast strain, which lacks a functional enzyme involved in production of high mannose structures, and expresses at least one exogenous enzyme for production of Man5GlcNAc2, wherein said enzyme involved in production of high mannose structures is α-1,6-mannosyltransferase encoded by the OCH1 gene, said at least one enzyme for production of Man5GlcNAc2 is α-1,2-mannosidase or an enzymatically active fragment thereof.
A method for producing a glycoprotein with reduced hyperglycosylation in methylotrophic yeast, comprising providing a methylotrophic yeast strain, which lacks a functional enzyme involved in production of high mannose structures, and expresses at least one exogenous enzyme for production of Man5GlcNAc2, wherein said enzyme involved in production of high mannose structures is α-1,6-mannosyltransferase encoded by the OCH1 gene, said at least one enzyme for production of Man5GlcNAc2 is α-1,2-mannosidase or an enzymatically active fragment thereof.
Dependencies are inferred from the claim text.