Purification of Pinguinain by Gel Filtration

Abstract

THE separation and characterization of macromolecules have been recently simplified by the introduction of a useful technique which has been variously termed gel filtration¹, molecular sieve chromatography², restricted diffusion chromatography³, or exclusion chromatography⁴. The formulation of the mechanism of operation of these columns has been discussed by Ackers⁵. Since the presence of more than one protein component in proteolytic enzymes isolated from plant tissue is not an unusual finding, this method should be useful in their separation. Recently, based on filtration experiments, alcohol fractionation techniques and free electrophoresis, Messing et al.^6,7 have identified two proteolytic components in the juice obtained from the Cuban variety of the fruit of Bromelia pinguin L.