Purification and Partial Characterization of Lectin from Broccolini
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Abstract:
Broccolini lectin (BL) was extracted from Broccolini with normal saline, distilled water, Tris-HCl buffer and phosphate buffered saline (PBS), followed by ammonium sulphate precipitation, and finally purified by chromatography DEAE-52 and Sephadex G-75. Partial characterization of BL was also studied. The results showed that Tris-HCl buffer had the highest extraction rate of 3.57±0.02%. Molecular mass of BL was estimated as 27 ku by SDS-PAGE. The BL contained 7.60±0.03% sugar and its clotting activity was 28. The sugar inhibition test showed that its hemagglutinating activity of erythrocyte could be inhibited by D- mannose and arabinose. Moreover, the lectin remained stable at 20~90 ℃ and its optimum pH was 7~8. Its clotting activity was significantly decreased in strong acid?and?strong alkali environment. The lectin reaction was also influenced by divalent cations Ca2+, in which its hemagglutinating activity was increased.
