title: Overexpression and purification of the galactose operon enzymes from Escherichia coli.
abstract: A convenient new procedure for the purification of galactokinase, galactose-1-phosphate uridyltransferase, and UDP-galactose 4-epimerase overexpressed in Escherichia coli is presented. The procedure is shorter than any other described in the literature and facilitates the purification of the three recombinant enzymes in considerable amounts and at high purity and specific activity. The purified gal operon enzymes were biochemically characterized by gel-filtration column chromatography and isoelectric focusing, and the Km values for their substrates were determined.
authors.affiliation: European Molecular Biology Laboratory, c/o DESY, Hamburg, Germany.
authors.forename: C E
authors.initials: CE
authors.lastname: Vorgias
authors.forename: H G
authors.initials: HG
authors.lastname: Lemaire
authors.forename: K S
authors.initials: KS
authors.lastname: Wilson
mesh_terms: D002850:Chromatography, Gel; D002852:Chromatography, Ion Exchange; D004926:Escherichia coli / Q000201:enzymology* / Q000235:genetics*; D005686:Galactokinase / Q000235:genetics / Q000302:isolation & purification; D005690:Galactose / Q000235:genetics*; D015870:Gene Expression; D007525:Isoelectric Focusing; D007700:Kinetics; D009876:Operon; D011994:Recombinant Proteins / Q000235:genetics / Q000302:isolation & purification; D014534:UDPglucose 4-Epimerase / Q000235:genetics / Q000302:isolation & purification; D005694:UTP-Hexose-1-Phosphate Uridylyltransferase / Q000235:genetics / Q000302:isolation & purification
chemical_list: D011994:Recombinant Proteins; D005686:Galactokinase; D005694:UTP-Hexose-1-Phosphate Uridylyltransferase; D014534:UDPglucose 4-Epimerase; D005690:Galactose