Magnetic Parkia pendula seed gum as matrix for Concanavalin A lectin immobilization and its application in affinity purification.

Moacyr J.B.M Rêgo,Eduardo I.C Beltrão,Sérgio A Bezerra,Sinara M Almeida,Luiz B Carvalho Júnior

doi:10.1590/0001-3765201420130316

Moacyr J.B.M Rêgo, Eduardo I.C Beltrão + Show 3 more

Open Access

https://doi.org/10.1590/0001-3765201420130316

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Abstract

The present work aimed to magnetize Parkia pendula seeds gum and use it as a matrix for Concanavalin A covalent immobilization. This composite was applied in affinity purification of glycoconjugates. Parkia pendula seeds were hydrated and the gum provenient from the supernatant was precipitated and washed with ethanol and dried. The gum was magnetized in co-precipitation using solutions of Fe+2 and Fe+3. Matrix activation was accomplished with NaIO4. Magnetized Parkia pendula seeds gum with covalently immobilized Concanavalin A was used as an affinity matrix for the recognition of bovine serum fetuin glycoprotein. Fetuin elution was carried out with a solution of glucose (300mM) and evaluated through SDS-PAGE. The efficiency of lectin immobilization and fetuin purification were 63% and 14%, respectively. These results indicate that the composite produced is a promising magnetic polysaccharide matrix for lectins immobilization. Thus, such system can be applied for affinity purification allowing an easy recovery by magnetic field.

Highlights

Many biomolecules have been isolated using separation techniques based on the interaction of biospecific molecules
The lectin extracted from Canavalia ensiformis, named Concanavalin A (Con A) has been extensively used in the isolation, fractioning and structural characterization of glycoproteins (Bucur et al 2004, Uygun et al 2012) and other important glycoconjugates bearing glucose and/or mannose residues (Fraguas et al 2004)
Anderson and Pinto (1985) evaluated the gum exudates from Parkia bicolor and P. biglobosa, and the gum extracted from the seed pods of P. pendula

Summary

Introduction

Many biomolecules have been isolated using separation techniques based on the interaction of biospecific molecules Among these molecules of particular interest there is a class named lectins that are sugar-specific and cell-agglutinating proteins of non-immune origin (Sharon 2007). Among the matrices used in protein immo bilization, those which contain carbohydrates have become the focus of intense interest in biotechnology (Fraguas et al 2003, 2004, Mislovicová et al 2004, Angeli et al 2009) To increase their performance, matrices have been magnetized to decrease the processing time of samples, the utilization of chemicals as well as to facilitate separation and process automation (Pan et al 2005, Angeli et al 2009)

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