ImobilizaÃÃo covalente de concanavalina a em goma de sementes de Parkia pendula magnetizada e sua aplicaÃÃo biotecnolÃgica

AUTOR(ES)
DATA DE PUBLICAÇÃO

2006

RESUMO

Protein immobilization in water insoluble supports is an important tool in biotechnology. Meanwhile, support magnetization increases its potential application since a magnetic field can be used to separate the support from the reaction system and its reuse afterwards. Among the proteins immobilized special attention have been given for lectins, mainly Concanavalin A (Con A) witch presents an affinity for residues of glucose, mannose and their derivatives. Natural plant gums are sources of polysaccharides. Parkia pendula seeds produce a gum (PpeG) witch has been investigated as support for lectin immobilization. The present work aimed to magnetize PpeG and immobilize Con A on it to be used as affinity chromatography matrix in the purification of glycoconjugates. P. pendula seeds were hydrated and the gum from the supernatant was precipitated and washed with ethanol being dried at 37ÂC. The gum was magnetized with the co-precipitation method using solutions of Fe+2 and Fe+3. Support activation was accomplished with NaIO4 for Con A immobilization. MagPpeG-Con A was used as affinity chromatography matrix for the recognition of bovine serum fetuin glycoprotein. Fetuin elution was carried out with a solution of glucose (300mM) and investigated through SDS-PAGE. The efficiency of Con A immobilization was 63%. After elution 14% of fetuin was recovered. These results indicate that PpeG is a promising polysaccharide support for immobilization of lectins and application as affinity chromatography matrix.

ASSUNTO(S)

parkia pendula magnetismo biotecnolÃgica bioquimica

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