Overexpression and purification of the galactose operon enzymes from Escherichia coli |
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| Authors: | C E Vorgias H G Lemaire K S Wilson |
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| Institution: | European Molecular Biology Laboratory, c/o DESY, Hamburg, Germany. |
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| 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. |
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