Resumo

MARTINEZ, Patricia; PABON, Marta Lucía; GOMEZ, Diego Armando  e  ALBARRACIN, Sonia Luz. Purification and carbohydrate determination of vicilin from Canavalia ensiformis. Rev. colomb. cienc. quim. farm. [online]. 2011, vol.40, n.1, pp.5-26. ISSN 0034-7418.

This study reports the purification and carbohydrate determination of vicilin from Canavalia ensiformis ( Jack bean). The vicilin was purified by isoelectric precipitation at pH 6.4 and 4.8, ion exchange chromatography (DEAE-Sephadex A-50) and affinity chromatography (Con A -Sepharosa 4B). Protein identity was confirmed by mass spectrometry (MALDI-TOF-PMF) (Matrix assisted laser desorption ionizationtime of flight-Peptide mass fingerprinting) and purification was confirmed by sdspage. Oxidation of glycosylated moiety in pure vicilin demonstrates the presence of carbohydrates in the protein. The presence of carbohydrates in the pure vicilin fraction was confirmed by enzymatic deglycosylation by Peptide N-glycosidase F (PNGasa F) and by Dubois' method, finding a 4.03% carbohydrate content in such protein. These results demonstrate for the first time the presence of glycosides in C. ensiformis vicilin. The purification process of vicilin seed of Canavalia Ensiformis used in this study allowed us to obtain a protein with a high degree of purity, superior to the ones described in the literature.

Palavras-chave : vicilin; Canavalia ensiformis; glycosylated form of vicilin; leguminosae.

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