title: Scaled-up expression of human alpha 2,6(N)sialyltransferase in Saccharomyces cerevisiae.
abstract: Expression of recombinant full length human alpha 2,6(N)sialyltransferase has been scaled-up in S. cerevisiae in a 150-l bioreactor yielding 47 U at a concentration of 0.31 U/l. The protein specific activity as measured in reconstituted yeast lyophilisate was 0.8 mU/mg protein. The recombinant enzyme exhibited similar Michaelis constants as previously determined for the native rat enzyme. By immunoblotting the enzyme was shown to be heterogeneous by size (44-48 kD) and N-glycosylated. We conclude that recombinant alpha 2,6(N)sialyltransferase expressed in S. cerevisiae is retained in the endoplasmic reticulum as a fully active enzyme.
authors.affiliation: Institute of Physiology, University of Zurich, Switzerland.
authors.forename: L
authors.initials: L
authors.lastname: Borsig
authors.forename: S X
authors.initials: SX
authors.lastname: Ivanov
authors.forename: G F
authors.initials: GF
authors.lastname: Herrmann
authors.forename: U
authors.initials: U
authors.lastname: Kragl
authors.forename: C
authors.initials: C
authors.lastname: Wandrey
authors.forename: E G
authors.initials: EG
authors.lastname: Berger
mesh_terms: D015153:Blotting, Western; D006801:Humans; D007700:Kinetics; D008970:Molecular Weight; D011994:Recombinant Proteins / Q000302:isolation & purification / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000378:metabolism; D012799:Sialyltransferases / Q000096:biosynthesis* / Q000378:metabolism
chemical_list: D011994:Recombinant Proteins; D012799:Sialyltransferases