Purification and Characterization of a Lectin from the Indian Cultivar of French Bean Seeds

Abstract

A lectin specific for glucuronic acid and galacturonic acid has been isolated from seeds of the French bean Phaseolus vulgaris using a procedure that involved affinity chromatography on Affi-gel blue gel, fast protein liquid chromatography (FPLC)-ion exchange chromatography on Mono S, and FPLC-gel filtration on Superdex 200. The lectin was comprised of two 32-kDa subunits with substantial N-terminal sequence similarity to other Phaseolus lectins. The hemagglutinating activity of the lectin was stable within the pH range of 1-13 and the temperature range of 10-60 degrees C. The lectin neither exhibited any antiproliferative activity against tumor cells nor stimulated nitric oxide production by murine peritoneal macrophages at doses as high as 1mM, The lectin failed to evoke any mitogenic response from murine splenocytes as measured by [(3)H-methyl]-thymidine incorporation and did not inhibit the activity of HIV-1 reverse transcriptase. The lectin had no antifungal activity.

Isolation of a Hemagglutinin with Potent Antiproliferative Activity and a Large Antifungal Defensin from Phaseolus vulgaris cv. Hokkaido Large Pinto Beans

“…They were stable at 80 and 90°C, respectively, and retained their hemagglutinating activity after treatment with solutions at pH 1−14 and 1−13, respectively. 19,27 The high thermal stability may be due to the absence of disulfide linkages, which become unstable at elevated temperatures or attributed to the absence of heat-sensitive tryptophan residues. 34 Some lectins had poor thermostability and pH stability, such as French bean cultivar no.…”

A Hemagglutinin from Northeast Red Beans with Immunomodulatory Activity and Anti-proliferative and Apoptosis-inducing Activities Toward Tumor Cells

“…Similar protocols were widely used in purification of other P. vulgaris lectins and hemagglutinins, such as lectins from French bean Indian cultivar and cultivar no. 12 and Anasazi beans [35–37], as well as those from other plant species such as the lectins from graviola seeds, white Bauhinia seeds and Luetzelburgia auriculata cotyledons [38–40]. The protocol used in present study allowed fast and simple purification of the target protein.…”

“…For example, lectins from Indian cultivar of French bean and Anasazi bean had pronounced thermostability (up to 90 °C and 80 °C, respectively) and pH stability (1 – 13 and 1 – 14, respectively) [35, 37], while lectin from French bean cultivar no. 35 was less heat stable (up to 50 °C) and pH stable (6 – 8) [43].…”

“…NRBH, the PHAs from cultivar 12, 35 and Indian cultivar of French beans [35, 36, 43], and Hokkaido red beans are other examples [21]. On the other hand, the sugar binding PHAs from different P. vulgaris cultivars exhibited a variety of carbohydrate-binding specificities.…”

Affinity Chromatography of Lectins

“…In addition to Sepharose and Sephadex, commercially available Affi-Gel blue gel (Bio-Rad), composed of beaded, cross-linked agarose, has been used as an adsorbent matrix (Lin & Ng, 2008;Shao et al, 2011;Sharma et al, 2010;Wong et al, 2010). Further, naturally occurring cross-linked polysaccharides, which are inexpensive and simple to prepare with epichlorhydrin or divinyl sulphone, have also been extensively used as adsorbent matrices for isolating galactose-specific lectins (Lis & Sharon, 1981).…”