Vorgias C E, Lemaire H G, Wilson K S
European Molecular Biology Laboratory, c/o DESY, Hamburg, Germany.
Protein Expr Purif. 1991 Oct-Dec;2(5-6):330-8. doi: 10.1016/1046-5928(91)90091-v.
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.
本文介绍了一种简便的新方法，用于纯化在大肠杆菌中过表达的半乳糖激酶、1-磷酸半乳糖尿苷酰转移酶和UDP-半乳糖4-差向异构酶。该方法比文献中描述的任何其他方法都更短，有助于大量纯化这三种重组酶，且纯度和比活性都很高。通过凝胶过滤柱色谱和等电聚焦对纯化的gal操纵子酶进行了生化表征，并测定了它们对底物的Km值。
