The purification of orotidine-5'-phosphate decarboxylase from yeast by affinity chromatography.
We have prepared an affinity column for the purification of orotidine-5'-phosphate decarboxylase from yeast. The column effects a 3200-fold purification from yeast homogenate in one pass; simple additional steps produce enzyme that has been purified 6700-fold and is not contaminated by any other protein that can be detected by sodium dodecyl sulfate-acrylamide gel electrophoresis. Overall, 35% of the activity present in the yeast is recovered as pure enzyme. The resin for the column is synthesized by attaching the ethylenediamine amide of 5-(2-carboxyethyl)-6-azauridine 5'-phosphate to carboxymethyl-agarose.
D002262:Carboxy-Lyases / Q000302:isolation & purification*; D002846:Chromatography, Affinity / Q000379:methods; D009964:Orotidine-5'-Phosphate Decarboxylase / Q000302:isolation & purification*; D011485:Protein Binding; D012441:Saccharomyces cerevisiae / Q000201:enzymology*; D012685:Sepharose
D012685:Sepharose; D002262:Carboxy-Lyases; D009964:Orotidine-5'-Phosphate Decarboxylase