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Lectin extracted from Amaranthus hypochondriacus was purified using an affinity column with an agarose-fetuin matrix specific to the lectin of interest. Purification was confirmed by SDS-PAGE, revealing a single protein band with a molecular mass of 34.4 kDa. A hemagglutination assay showed that the lectin had a higher affinity for human type A erythrocytes, and its hemagglutinating activity was inhibited only by fetuin, not by mono-, di-, or trisaccharides. This demonstrated the lectin's selectivity for the N-acetylgalactosamine present on the surface of type A erythrocytes and fetuin. Amaranth lectin exhibited antioxidant activity, which was attributed to the phenolic compounds, amino acids, and specific peptides within the protein structure that are known for their antioxidant properties. Infrared (IR) spectroscopy provided a structural analysis and confirmed lectin glycosylation, a crucial factor in its stability and its ability to bind specific glycans on cell surfaces. Cu2+, Mn2+, and Zn2+ ions were found in the lectin, and these ions were strongly bound to the protein, as dialysis against ethylenediaminetetraacetic acid (EDTA) did not remove them. pH and temperature influenced lectin stability, with higher hemagglutinating activity observed at pH 7, and it remained thermostable at 25 °C. [ABSTRACT FROM AUTHOR]

As they manifest specifically and reversibly, lectins are proteins or glycoproteins with the characteristic of agglutinating erythrocytes. Given that grain legume lectins can represent 10% of protein content and can have various biological functions, they are extensively studied. The objective of this work was to purify and partially characterize the lectins of Phaseolus vulgaris black, var surco and vara (LBBS and LBBV). Both lectin types were purified by affinity chromatography on stroma matrix, which agglutinated human erythrocytes type A, B, and O, as well as rabbit, hamster, pig, and chicken erythrocytes. Native-PAGE was employed for molecular mass determination, yielding 109.36 and 112.68 kDa for BBS and BBV, respectively. Further analyses revealed that these lectins are tetrameric glycoproteins that require Ca+2, Mn+2 and Mg+2 ions for exhibiting their hemagglutinating function, which can be inhibited by fetuin. Moreover, optimal pH was established for both lectins (10.5 for LBBS and 7−9 for LBBV), while their activity was temperature-dependent and ceased above 70 °C. Finally, the observed differences in the biochemical characteristics and bioactive functions were ascribed to the different physiological characteristics of each seed, as well as the protein itself. [ABSTRACT FROM AUTHOR]