METHOD FOR INCREASING MANNOSE PHOSPHORYLATION BY USING YLMPO1 OVEREXPRESSION

摘要

The present invention relates to a method for increasing mannose phosphorylation by using YlMpo1 overexpression and, more specifically, to a method for preparing a bi-mannosylphosphorylated glycan-added protein by overexpressing YlMpo1 proteins in a recombinant yeast in which a yeast-specific glycan biosynthetic pathway is removed. The bi-mannosylphosphorylated glycan, prepared by the method of the present invention, is easily converted into a bi-phosphorylated glycan to which two mannose-6-phosphates are added by an enzymatic or chemical method, and then a trimming step is performed by a mannosidase treatment, so as to be converted into a glycan (P₂-Man₆GlcNAc₂) having the structure, which has high binding affinity for a mannose-6-phosphate receptor and has a superior lysosomal targeting ability. Therefore, the present invention can overexpress the YlMpo1 in a yeast, in which a yeast-specific glycan biosynthetic pathway is removed, so as to produce a glycoprotein to which a high content of the bi-mannosylphosphorylated glycan is added, thereby providing a glycoprotein having a superior lysosomal targeting ability.