Purification and Properties of d-Glycerate 3-Kinase from Escherichia coli

Abstract

Abstract The adaptive d-glycerate 3-kinase from glycerate-adapted Escherichia coli has been purified 4200-fold in 37% yield and has been crystallized. The procedure, starting with the crude, cell-free extract, consisted of treatment with protamine sulfate, ammonium sulfate, passage through a diethylaminoethyl Sephadex column, and precipitation with ethanol. The reaction product, 3-phosphoglycerate, has been identified by paper chromatography and specific enzymatic assay. Some characteristics of the enzyme which show clearly its differences from other glycerate kinases are described.