title: Expression of mouse Gal beta 1,4GlcNAc alpha 2,6-sialyltransferase in an insoluble form in Escherichia coli and partial renaturation.
abstract: Mouse Gal beta 1,4GlcNAc alpha 2,6-sialyltransferase was produced in an insoluble form in Escherichia coli cells harboring expression plasmids. The insoluble protein was solubilized with 8 M urea and diluted for renaturation of the enzyme. The substrate specificity and kinetic parameters, except for the specific activity, of the renatured enzyme were similar to those of the enzyme obtained from rat liver. These results suggest that a bacterial expression system is a potentially powerful tool for the large scale production of sialyltransferases and for elucidating the molecular mechanisms of sialyltransferases.
authors.affiliation: Glyco Molecular Biology, Frontier Research Program, Institute of Physical and Chemical Research (RIKEN), Saitama, Japan.
authors.forename: T
authors.initials: T
authors.lastname: Hamamoto
authors.forename: Y C
authors.initials: YC
authors.lastname: Lee
authors.forename: N
authors.initials: N
authors.lastname: Kurosawa
authors.forename: T
authors.initials: T
authors.lastname: Nakaoka
authors.forename: N
authors.initials: N
authors.lastname: Kojima
authors.forename: S
authors.initials: S
authors.lastname: Tsuji
mesh_terms: D000818:Animals; D001483:Base Sequence; D002240:Carbohydrate Sequence; D017931:DNA Primers / Q000235:genetics; D004187:Disaccharides / Q000737:chemistry / Q000378:metabolism; D004926:Escherichia coli / Q000235:genetics; D015870:Gene Expression; D008099:Liver / Q000201:enzymology; D051379:Mice; D008969:Molecular Sequence Data; D010957:Plasmids / Q000235:genetics; D011489:Protein Denaturation; D051381:Rats; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics / Q000302:isolation & purification; D012799:Sialyltransferases / Q000096:biosynthesis* / Q000235:genetics / Q000302:isolation & purification; D012995:Solubility; D013045:Species Specificity; D013379:Substrate Specificity; D000094802:beta-D-Galactoside alpha 2-6-Sialyltransferase
chemical_list: D017931:DNA Primers; D004187:Disaccharides; D011994:Recombinant Proteins; D012799:Sialyltransferases; D000094802:beta-D-Galactoside alpha 2-6-Sialyltransferase