Purification of malic enzyme by affinity chromatography on immobilized (N6-(6-aminohexyl)-adenosine 2′,5′-bisphosphate

Abstract

A simple and rapid method for the purification of NADP-dependent malic enzyme from chicken liver is described. A crude tissue extract was chromatographed on an affinity column containing immobilized N6-(6-aminohexyl)-adenosine 2′,5′-bisphosphate. Malic enzyme bound to the ligand and was eluted specifically by a gradient of NADP. Then the enzyme was purified further by gel filtration. The entire procedure requires 50 hr and provides the enzyme in about 40% yield. The purified enzyme migrated as one band during disc electrophoresis in polyacrylamide gel.