Use of differential dye-ligand chromatography with affinity elution for enzyme purification: 2-keto-3-deoxy-6-phosphogluconate aldolase from Zymomonas mobilis.
2-Keto-3-deoxy-6-phosphogluconate aldolase (EC 4.1.2.14) has been isolated from extracts of Zymomonas mobilis using differential dye-ligand chromatography and affinity elution with product/product analog. The one-step procedure gives an enzyme with specific activity 600 units mg-1. Only 1 out of 47 dyes, Procion Yellow MX-GR, bound the enzyme completely in 20 mM phosphate buffer, pH 6.5. A column of Navy HE-R adsorbent was used first to remove most of the potentially adsorbing proteins.
D000327:Adsorption; D000446:Aldehyde-Lyases / Q000302:isolation & purification*; D000476:Alkanesulfonates; D017738:Alkanesulfonic Acids; D001421:Bacteria, Anaerobic / Q000201:enzymology*; D002845:Chromatography / Q000379:methods*; D002846:Chromatography, Affinity / Q000379:methods; D004396:Coloring Agents; D008970:Molecular Weight; D009025:Morpholines
D000476:Alkanesulfonates; D017738:Alkanesulfonic Acids; D004396:Coloring Agents; D009025:Morpholines; C004550:2-(N-morpholino)ethanesulfonic acid; D000446:Aldehyde-Lyases; C100131:phospho-2-keto-3-deoxy-gluconate aldolase