title: Glycan engineering of proteins with whole living yeast cells expressing rat liver alpha2,3-sialytransferase in the porous cell wall.
abstract: The N-glycans of recombinant proteins produced via the secretory pathway of cultured mammalian cells are often undersialylated, and insect cells lack sialytransferases. Undersialylated glycoproteins are rapidly cleared from the circulation, compromising the effect of pharmaceuticals. We show that incubation with living Saccharomyces cerevisiae cells expressing the catalytic ectodomain of rat liver alpha2,3-sialyltransferase (ST3Ne) in the porous cell wall resulted in sialylation of glycoproteins. The Km values of the yeast enzyme for several substrates were similar to those of recombinant ST3Ne from insect cells and of authentic ST3N. The yeast strain provides an inexpensive self-perpetuating source of ST3N activity for glycan engineering of recombinant proteins.
authors.affiliation: Institute of Biotechnology, University of Helsinki, Finland.
authors.forename: E
authors.initials: E
authors.lastname: Sievi
authors.forename: J
authors.initials: J
authors.lastname: Helin
authors.forename: R
authors.initials: R
authors.lastname: Heikinheimo
authors.forename: M
authors.initials: M
authors.lastname: Makarow
mesh_terms: D000818:Animals; D001212:Asialoglycoproteins / Q000737:chemistry; D020134:Catalytic Domain; D002473:Cell Wall / Q000201:enzymology*; D007700:Kinetics; D008099:Liver / Q000201:enzymology*; D019158:N-Acetylneuraminic Acid / Q000737:chemistry; D011134:Polysaccharides / Q000737:chemistry*; D015202:Protein Engineering; D051381:Rats; D011994:Recombinant Proteins / Q000737:chemistry / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000737:chemistry*; D012799:Sialyltransferases / Q000737:chemistry* / Q000378:metabolism; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D000094663:beta-Galactoside alpha-2,3-Sialyltransferase
chemical_list: D001212:Asialoglycoproteins; D011134:Polysaccharides; D011994:Recombinant Proteins; D012799:Sialyltransferases; D019158:N-Acetylneuraminic Acid; D000094663:beta-Galactoside alpha-2,3-Sialyltransferase