Primary Research On ConA’s Separation, Purification From Canavalia Gladiata (Jacq.) DC And Its Glycan Structure

Lectins are proteins or glycoproteins that can specifically and reversibly bind with carbohydrates, but without any enzymatic or antibiotic activity. Canavalia gladiata (Jacq.) DC is a very common food in China and other countries. Concanavalin A is widely used in biochemistry and analytic chemistry of glycan. Recently researchers found that oligosaccharides were the very important components for ConA’s hemagglutination activity. In this study, we focused on the isolation, purification, physicochemical properties, structural characterization and biological activities of a glycoprotein ConA from seeds of Canavalia gladiata (Jacq.) DC. The main results were summarized as follows:1. Isolation, purification ConA from Canavalia gladiata (Jacq.) DCCrude ConA was obtained from the seed of Canavalia gladiata (Jacq.) DC by water extraction and salt precipitation. Then the ConA was purified by DEAE-Cellulose column chromatography followed by Sephadex G-100column chromatography from the crude glycoprotein.2. Physicochemical properties of glycoprotein ConAThe physicochemical properties of ConA were determined by HPLC, GC, ESI-MS and spectrophotometer (Table1). 2. Structural characterization of the glycan of ConAThe glycans were obtained by Pronase E digestion. Through methylation analysis, partial hydrolysis and GC-MS, the results indicated that ConA-OL was a oligosaccharide of Ara with different degree. The backbone of the oligosaccharide was proposed as Ara-Ara-Ara repeating unit by1-3glycobond.