Use of differential dye-ligand chromatography with affinity elution for enzyme purification: 6-phosphogluconate dehydratase from Zymomonas mobilis.
Using differential dye-ligand chromatography and affinity elution with a substrate analog, 6-phosphogluconate dehydratase (EC 4.2.1.12) has been isolated from extracts of Zymomonas mobilis in a one-step procedure with 50% recovery. The specific activity of freshly isolated enzyme was 245 units mg-1. The enzyme contains iron, and it is rapidly inactivated in oxidizing conditions. It is inhibited by glycerophosphates, most strongly by the D-alpha-isomer which structurally corresponds to half of the substrate molecule.
D000476:Alkanesulfonates; D017738:Alkanesulfonic Acids; D001421:Bacteria, Anaerobic / Q000201:enzymology*; D002021:Buffers; D002845:Chromatography / Q000379:methods*; D002846:Chromatography, Affinity / Q000379:methods; D004396:Coloring Agents; D006836:Hydro-Lyases / Q000302:isolation & purification*; D007700:Kinetics; D009025:Morpholines
D000476:Alkanesulfonates; D017738:Alkanesulfonic Acids; D002021:Buffers; D004396:Coloring Agents; D009025:Morpholines; C004550:2-(N-morpholino)ethanesulfonic acid; D006836:Hydro-Lyases; C022362:phosphogluconate dehydratase