Chromatographic purification and characterization of B-phycoerythrin from Porphyridium cruentum -: Semipreparative high-performance liquid chromatographic separation and characterization of its subunits

A fast preparative two-step chromatographic method for purification of B-phycoerythrin from Porphyridium cruentum is described. This biliprotein was homogeneous as determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis yielding three closely migrating bands corresponding to its three subunits. Baseline separation of its alpha-, beta- and gamma -subunits was achieved by a reversed-phase HPLC gradient semipreparative method with a C, large-pore column and a solvent system consisting of 0.05% trifluoroacetic acid (TFA) in water and 0.05% TFA in acetonitrile. B-Phycoerythrin in different aggregation states and its subunits have been spectroscopically characterized. Hexameric B-phycoerythrin has similar secondary and tertiary structure than dissociated B-phycoerythrin determined by circular dichroism. (C) 2001 Elsevier Science B.V. All rights reserved.