Isolation and partial characterization of Bromelia hemisphaerica protease by affinity chromatography.
Hemisphaericin, the protease from Bromelia hemisphaerica fruit juice was isolated by affinity chromatography in one step, using a mercurial sepharose derivative. The enzyme behaves as a single component in immunodifussion, immunoelectrophoresis and polyacrylamide electrophoresis in the presence of SDS and 2-mercaptoethanol. Association and dissociation of active components were evidenced in electrophoresis at pH 3.6 and at pH 8.6. Immunoelectrophoresis analyses also disclosed a certain degree of internal immunological heterogeneity. The results are explained by the presence of an enzyme subunit, of about 8000 daltons, endowed with polymeric properties induced by the pH and oxidative environment.
Unidad de Medicina Experimental, Facultad de Medicina, Universidad Nacional Autónoma de México.
N
N
Ochoa
C
C
Agundis
F
F
Córdoba
D001963:Bromelains / Q000276:immunology; D002241:Carbohydrates / Q000032:analysis; D002846:Chromatography, Affinity; D003429:Cross Reactions; D003546:Cysteine Endopeptidases / Q000276:immunology / Q000302:isolation & purification* / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel; D046911:Macromolecular Substances; D010944:Plants / Q000201:enzymology*
D002241:Carbohydrates; D046911:Macromolecular Substances; D001963:Bromelains; D003546:Cysteine Endopeptidases; C021345:hemisphaericin