A rapid purification procedure for pyruvate kinase from the hyphal fungus Aspergillus nidulans

Abstract

Pyruvate kinase was purified from the filamentous fungus Aspergillus nidulans with a 45–55% yield. The procedure involved dye-affinity chromatography and fast protein liquid chromatography, resulting in highly active and pure enzyme in milligram quantities within 2 days. The purified enzyme, a tetramer with a subunit molecular weight of 65 000 and an isoelectric point of 4.7, was used to determine the amino acid composition.