Isolation and partial characterization of bromelia hemisphaerica protease by affinity chromatography
Por:
Ochoa N., Agundis C., Córdoba F.
Publicada:
1 ene 1987
Resumen:
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-mercap_toethanol. 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. Tropical America presents a variety of plants comprising bromeliaceae, one of which is Bromelia hemisphaerica, commonly known in Mexico as “Timbirichi”. The elongated fruit measures about 8 cm in length, and 3–4 cm in width, and contains a juice displaying proteolytic activity. The protease was named — Hemisphaericin (12). Hemisphaericin has thiol groups present in the active site as is the case with bromelain (1) and pinguinain (2), two other proteases from bromeliaceae. Since sulphydryl groups in proteins combine readily with heavy metal radicals such as mercury attached to insoluble matrix (3), the property was utilized in this work for the isolation, purification and characterization of Hemisphaericin. © 1987, Taylor & Francis Group, LLC. All rights reserved.
Filiaciones:
Ochoa N.:
Unidad de Medicina Experimental, Facultad de Medicina, Universidad Nacional Autónoma de Mexico, Centro de Investigaciones Biológicas UABJO, Apdo. Postal 872, Oaxaca 68000, Mexico
Agundis C.:
Unidad de Medicina Experimental, Facultad de Medicina, Universidad Nacional Autónoma de Mexico, Centro de Investigaciones Biológicas UABJO, Apdo. Postal 872, Oaxaca 68000, Mexico
Córdoba F.:
Unidad de Medicina Experimental, Facultad de Medicina, Universidad Nacional Autónoma de Mexico, Centro de Investigaciones Biológicas UABJO, Apdo. Postal 872, Oaxaca 68000, Mexico
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