Kidney Bean Lectin Research Articles

Entomotoxic Properties of White Kidney Bean and Soybean Lectins and their Effects against Two Digestive Enzymes of the Spiny Bollworm, Earias insulana (Boisd.)

Entomotoxic Properties of White Kidney Bean and Soybean Lectins and their Effects against Two Digestive Enzymes of the Spiny Bollworm, Earias insulana (Boisd.)

Insight of pH-shifting as an effective pretreatment to reduce the antigenicity of lectin from red kidney bean (Phaseolus vulgaris L.) combining with autoclaving treatments: The structure investigation

Combined effects of pH-shifting and an autoclaving cycle (121 °C, 15 min) on red kidney bean lectin (RKBL) were investigated using intrinsic and extrinsic fluorescence, UV, FTIR, DSC, SEC, dot-blot analysis and in vitro digestibility. Spectroscopic studies suggested that the protein refolding was stable after 3 h incubation with the hydrophobic exposure after pH-shifting, and hydrophobicity was significantly increased with the formation of more looser structure, which would influence the structural stability of known epitopes. In details, the increase of β-turn and reduction of random coil was related with the lower denaturation enthalpy, while the protein aggregation was also observed in acidic treated samples after autoclaving. Lower antigenicity and good digestibility suggested the exposure of enzyme cutting sites, and confirmed the effectivity of pH-shifting prior to the autoclaving. Then the results would be beneficial to the development of hypoallergenic kidney bean foods.

Detection of Lectin Protein Allergen of Kidney Beans (Phaseolus vulgaris L.) and Desensitization Food Processing Technology.

With the increase of food allergy events related to not properly cooked kidney beans (Phaseolus vulgaris L.), more and more researchers are paying attention to the sensitization potential of lectin, one of the major storage and defensive proteins with the specific carbohydrate-binding activity. The immunoglobulin E (IgE), non-IgE, and mixed allergic reactions induced by the lectins were inducted in the current paper, and the detection methods of kidney bean lectin, including the purification strategies, hemagglutination activity, specific polysaccharide or glycoprotein interactions, antibody combinations, mass spectrometry methods, and allergomics strategies, were summarized, while various food processing aspects, such as the physical thermal processing, physical non-thermal processing, chemical modifications, and biological treatments, were reviewed in the potential of sensitization reduction. It might be the first comprehensive review on lectin allergen detection from kidney bean and the desensitization strategy in food processing and will provide a basis for food safety control.

Combined effects of pH and thermal treatments on IgE-binding capacity and conformational structures of lectin from black kidney bean (Phaseolus vulgaris L.).

Combined effects of pH and thermal treatments on black kidney bean lectin (BKBL) were investigated by response surface methodology (RSM). Low-pH (1.0, 2.0, 3.0) incubation decreased hemagglutination activity (HA) and IgE-binding capacity, but the activities would be restored when the lectin was treated by pH shifting to 7.2. Conformational structure analyses indicated that low-pH induced protein unfolding and pH-shifting treatment resulted in a limited structural rearrangement. Mild heating, such as 60°C for 3min, slightly increased the HA and IgE-binding activities of pH shifted BKBL, but no obvious effects in the pH 1.0 incubated BKBL. High-temperature and long-time treatment might induce the protein aggregation, further decreased HA and IgE-binding capacities. RSM results showed both IgE-binding capacity and HA were the lowest under the combination of pH 1.0 incubation with 80°C heating for 15min or pH shifting from 1.0 to 7.2 with 100°C heating for 10min.

A novel oriented antibody immobilization based voltammetric immunosensor for allergenic activity detection of lectin in kidney bean by using AuNPs-PEI-MWCNTs modified electrode

As a well-known allergenic indicator in kidney beans, lectins have always been the serious threats for human health. Herein, we introduced a new label-free voltammetric immunosensor for the direct determination of kidney bean lectin (KBL) with potential allergenic activity. Gold nanoparticles-polyethyleneimine-multiwalled carbon nanotubes nanocomposite was one-pot synthesized and modified onto the glass carbon electrode to enhance catalytic currents of oxygen reduction reaction. The KBL polyclonal antibody, acquired from rabbit immunization, was orientedly immobilized on the electrode modified with recombinant staphylococcal protein A via fragment crystallizable (Fc) region of antibody. Under the optimized condition, the immunosensor displayed a good linear response (R2 = 0.978) to KBL with a range from 0.05 to 100 μg/mL and a detection limit of 0.023 μg/mL. Simultaneously, the immunosensor exhibited well selectivity, interference-resistant ability, stability (4 °C) and reproducibility. Compared with the conventional enzyme-linked immunosorbent assay (ELISA) method, the immunosensor was successfully applied to quantify allergenic activity of lectin in raw and cooked (boiled for 30 min) kidney bean milk samples. This new approach provides new perspectives both for rapid quantification of lectin in kidney beans-derived foodstuffs and as a real-time monitoring tool for the allergenic potential during the whole production and consumption process.

White kidney bean lectin exerts anti-proliferative and apoptotic effects on cancer cells

A 60-kDa glucosamine binding lectin, white kidney bean lectin (WKBL), was purified from Phaseolus vulgaris cv. white kidney beans, by application of anion exchange chromatography on Q-Sepharose, affinity chromatography on Affi-gel blue gel, and FPLC-size exclusion on Superdex 75. The anti-proliferative activity of WKBL on HONE1 cells and HepG2 cells was stronger than the activity on MCF7 cells and WRL68 cells (IC50 values for a 48-h treatment with WKBL on HONE1 cells: 18.8μM; HepG2 cells: 19.7μM; MCF7 cells: 26.9μM; and WRL68 cells: >80μM). The activity could be reduced by addition of glucosamine, which occupies the binding sites of WKBL, indicating that carbohydrate binding is crucial for the activity. Annexin V-FITC and PI staining, JC-1 staining and Hoechst 33342 staining revealed that apoptosis was induced on WKBL-treated HONE1 cells and HepG2 cells, but not as obviously on MCF7 cells. Cell cycle analysis also showed a slight cell cycle arrest on HONE1 cells after WKBL treatment. Western blotting suggested that WKBL induced apoptosis of HONE1 cells occurred through the extrinsic apoptosis pathway, with detection of increased level of active caspase 3, 8 and 9.

Identification and Characterization of Phytohemagglutinins from White Kidney Beans (Phaseolus vulgaris L., var. Beldia) in the Rat Small Intestine.

Although kidney bean (Phaseolus vulgaris L.) lectin toxicity is widely known, its effects in the gastrointestinal tract require further study. This investigation aimed to identify and characterize phytohemagglutinins (PHAs) in the small intestine and sera of rats following oral challenge with ground white beans. Twenty young, adult male rats were divided randomly into two groups of 10 animals each. The control group underwent gavage with a suspension of 300 mg of rodent pellet flour. The experimental group was administered a 300 mg Beldia bean flour suspension (BBFS). After 10 days of daily treatment, jejunal rinse liquid (JRL) and ileum rinse liquid and secretions, as well as sera, were collected. All biological fluids were screened for lectin reactivity using competitive inhibition ELISA, Ouchterlony double immunodiffusion, and immunoelectrophoresis techniques. The results revealed the presence of immunogenic intraluminal PHAs 3-4 h after the oral intake of the BBFS in the JRLs as well as in the jejunal and ileal secretions; however, no PHA was detectable in the rat sera. Ingestion of raw Beldia beans may lead to interaction between PHAs and the mucosa of the small intestine, potentially resulting in an inflammatory response.

The effect of early-life kidney bean lectin administration on pig performance in the peri-weaning period – a safety study

The present study aimed to assess the safety issues of supplying kidney bean lectin extract early in life to piglets. Four litters totalling or 30 piglets were used. On postnatal day (PD) 10-11, thirty piglets were divided into 3 experimental groups (T1, T2, and T3), each consisting of 10 animals. On PD 10 or 11, group T1 (control) received water. Groups T2 and T3 received, respectively, a dose of 640 (recommended) and 6400 haemagglutination/piglet (10 x recommended) of kidney bean extract orally in a water suspension. On PD 42, all of the pigs were euthanized, and 6 piglets from each group were randomly taken for necropsy, sampling tissue for histology and bacteriological analyses of the lower gut. Blood was withdrawn before euthanasia for haematology and biochemistry. There were no significant differences in mortality, feed intake or body weight gains among the groups. All examined blood parameters were within the physiological range. No significant effects of the two doses of kidney bean extract on tissue morphology were observed in the post mortem inspection and histology. No significant differences among the groups in the number of aerobic and anaerobic bacteria were found. Concluding, a one-month tolerance study with the recommended and 10-fold recommended dose of kidney bean extract revealed no negative effects on piglet health or performance.

Effect of Nigella sativa alcoholic extract and oil, as well as Phaseolus vulgaris (kidney bean) lectin on the ultrastructure of Trichomonas vaginalis trophozoites

Trichomonas vaginalis is a parasitic protozoan that is the aetiological agent of trichomoniasis, the most common non-viral sexually transmitted disease worldwide. Currently, the compound of choice for the treatment of T. vaginalis infections is metronidazole, however, it has many side effects and an increase in metronidazole-resistant trichomoniasis has been observed. Medicinal plants could be a source of new antiprotozoal drugs with high activity, low toxicity and lower price. The present work was carried out to investigate the therapeutic potential of Nigella sativa alcoholic extract and oil, as well as Phaseolus vulgaris (kidney bean) lectin and their in vitro activity on the ultrastructure of T. vaginalis trophozoites in comparison to metronidazole, as detected by transmission electron microscope. Both N. sativa oil and P. vulgaris lectin showed high toxic effect as evidenced by severe cell damage with cytoplasmic and nuclear destruction, while the effect of N. sativa alcoholic extract was moderate. Therefore, these two extracts could offer an effective, cheaper and more safe alternative for metronidazole in treatment of trichomoniasis.

Red kidney bean (Phaseolus vulgaris) lectin stimulation increases the number of enterochromaffin cells in the small intestine of suckling piglets

Abstract The quantities and distribution patterns of serotonin-immunoreactive (serotonin-IR) enterochromaffin cells (EC) were studied immunohistochemically in the small intestine of suckling piglets stimulated with red kidney bean lectin, and in nonstimulated, control animals. The co-expression patterns of serotonin with somatostatin (SOM) or corticotropin releasing-factor (CRF) were also studied. After the lectin treatment, the increased numbers of EC were noted in the duodenum of experimental animals. Lectin stimulation did not change the proportions of EC in the jejunum and ileum. In the duodenal epithelium of the lectin-stimulated piglets, the vast majority of serotonin-IR EC were distributed at the basis of crypts. After the lectin administration, the proportions of serotonin-IR/SOM-IR EC were statistically similar in all sections of the small intestine. No upregulation of CRF was found in duodenal, jejunal, and ileal EC of lectin-treated animals. The findings demonstrated that red kidney bean lectin increased the serotonin reservoir in the duodenum, and thus may be an effective stimulant of the gut maturation in suckling mammals.

Effect of ionizing radiation on structural and functional attributes of red kidney bean (Phaseolus vulgaris L.) lectin

Gamma radiation inactivation of phytohemagglutinin (PHA) of red kidney bean in purified form as well as in seeds in different moisture conditions was demonstrated at different doses viz. 1, 10, 30, 50, 100 and 200 kGy. Irradiation of PHA in dry state (99.5% dry) or in 50% moisture condition showed structural intactness as studied by SDS–PAGE, size exclusion chromatography (SEC), fluorescence measurement, CD spectroscopy below 30 kGy, while a 50% reduction in hemagglutinin and mitogenic activity was observed in the dose range of 30–50 kGy. Radiation inactivation of purified PHA was more pronounced when irradiated in aqueous solution form resulting in complete destruction of secondary and tertiary structure as well as function at a dose of 10 kGy. Radiation processing of dry and soaked seeds of kidney bean exhibited 50% loss in functional activity of PHA at the doses of 50 and 30 kGy, respectively. Practicality of implication of the radiation processing for inactivation of this antinutrient in legume seeds is discussed.

Immunolocalization of NOS, VIP, galanin and SP in the small intestine of suckling pigs treated with red kidney bean (Phaseolus vulgaris) lectin

Lectins belong to a family of glycoproteins that can act both beneficially and detrimentally on the morphology of the small intestine. The aim of the study was to determine whether experimental treatment with red kidney bean (Phaseolus vulgaris) lectin influences the chemical code of the small intestine nervous system of suckling pigs. The immunolocalization sites of vasoactive intestinal polypeptide (VIP), nitric oxide synthase (NOS), substance P (SP) and galanin were determined in control and lectin-treated animals. In all segments of the small intestine (duodenum, jejunum, ileum), the subpopulations of VIP-, NOS-, SP- and galanin-immunoreactive (IR) myenteric neurons were unchanged. After lectin stimulation, increased proportions of NOS-IR and decreased numbers of VIP-IR submucous neurons/mucosa innervating nerve fibers were observed in the duodenum, jejunum and ileum. In lectin-treated animals down-regulation of submucous neurons expressing SP and up-regulation of galanin-IR submucous neurons were seen in the duodenum and jejunum (but not in the ileum). The distribution patterns of NOS-IR, galanin-IR and SP-IR nerve fibers supplying the duodenum, jejunum and ileum of the lectin-treated animals showed no substantial differences in relation to control piglets. We conclude that exposure to red kidney bean (P. vulgaris) lectin substantially changes the chemical content of VIP, NOS, SP and galanin in submucous neurons of the small intestine. These results are in line with previous findings outlining the key role(s) of these substances in enteric neuroplasticity processes and may constitute the basis for further functional studies on maturation of the gut.

Isolation of a Glucosamine Binding Leguminous Lectin with Mitogenic Activity towards Splenocytes and Anti-Proliferative Activity towards Tumor Cells

A dimeric 64-kDa glucosamine-specific lectin was purified from seeds of Phaseolus vulgaris cv. “brown kidney bean.” The simple 2-step purification protocol involved affinity chromatography on Affi-gel blue gel and gel filtration by FPLC on Superdex 75. The lectin was absorbed on Affi-gel blue gel and desorbed using 1M NaCl in the starting buffer. Gel filtration on Superdex 75 yielded a major absorbance peak that gave a single 32-kDa band in SDS-PAGE. Hemagglutinating activity was completely preserved when the ambient temperature was in the range of 20°C–60°C. However, drastic reduction of the activity occurred at temperatures above 65°C. Full hemagglutinating activity of the lectin was observed at an ambient pH of 3 to 12. About 50% activity remained at pH 0–2, and only residual activity was observed at pH 13–14. Hemagglutinating activity of the lectin was inhibited by glucosamine. The brown kidney bean lectin elicited maximum mitogenic activity toward murine splenocytes at 2.5 µM. The mitogenic activity was nearly completely eliminated in the presence of 250 mM glucosamine. The lectin also increased mRNA expression of the cytokines IL-2, TNF-α and IFN-γ. The lectin exhibited antiproliferative activity toward human breast cancer (MCF7) cells, hepatoma (HepG2) cells and nasopharyngeal carcinoma (CNE1 and CNE2) cells with IC50 of 5.12 µM, 32.85 µM, 3.12 µM and 40.12 µM respectively after treatment for 24 hours. Flow cytometry with Annexin V and propidum iodide staining indicated apoptosis of MCF7 cells. Hoechst 33342 staining also indicated formation of apoptotic bodies in MCF7 cells after exposure to brown kidney bean lectin. Western blotting revealed that the lectin-induced apoptosis involved ER stress and unfolded protein response.

580 Isolation and characterization of a new human lung cancer cell line derived from a metastatic limph node

A 60-kDa glucosamine binding lectin, white kidney bean lectin (WKBL), was purified from Phaseolus vulgaris cv. white kidney beans, by application of anion exchange chromatography on Q-Sepharose, affinity chromatography on Affi-gel blue gel, and FPLC-size exclusion on Superdex 75. The anti-proliferative activity of WKBL on HONE1 cells and HepG2 cells was stronger than the activity on MCF7 cells and WRL68 cells (IC50 values for a 48-h treatment with WKBL on HONE1 cells: 18.8 μM; HepG2 cells: 19.7 μM; MCF7 cells: 26.9 μM; and WRL68 cells: >80 μM). The activity could be reduced by addition of glucosamine, which occupies the binding sites of WKBL, indicating that carbohydrate binding is crucial for the activity. Annexin V-FITC and PI staining, JC-1 staining and Hoechst 33342 staining revealed that apoptosis was induced on WKBL-treated HONE1 cells and HepG2 cells, but not as obviously on MCF7 cells. Cell cycle analysis also showed a slight cell cycle arrest on HONE1 cells after WKBL treatment. Western blotting suggested that WKBL induced apoptosis of HONE1 cells occurred through the extrinsic apoptosis pathway, with detection of increased level of active caspase 3, 8 and 9.

581 Characterization of anaplastic astrocytoma xenografts derived from cancer stem cells in nude mice

A 60-kDa glucosamine binding lectin, white kidney bean lectin (WKBL), was purified from Phaseolus vulgaris cv. white kidney beans, by application of anion exchange chromatography on Q-Sepharose, affinity chromatography on Affi-gel blue gel, and FPLC-size exclusion on Superdex 75. The anti-proliferative activity of WKBL on HONE1 cells and HepG2 cells was stronger than the activity on MCF7 cells and WRL68 cells (IC50 values for a 48-h treatment with WKBL on HONE1 cells: 18.8 μM; HepG2 cells: 19.7 μM; MCF7 cells: 26.9 μM; and WRL68 cells: >80 μM). The activity could be reduced by addition of glucosamine, which occupies the binding sites of WKBL, indicating that carbohydrate binding is crucial for the activity. Annexin V-FITC and PI staining, JC-1 staining and Hoechst 33342 staining revealed that apoptosis was induced on WKBL-treated HONE1 cells and HepG2 cells, but not as obviously on MCF7 cells. Cell cycle analysis also showed a slight cell cycle arrest on HONE1 cells after WKBL treatment. Western blotting suggested that WKBL induced apoptosis of HONE1 cells occurred through the extrinsic apoptosis pathway, with detection of increased level of active caspase 3, 8 and 9.

Generation of IFN-γ-producing cells that recognize the major piroplasm surface protein in Theileria orientalis-infected bovines

Theileriosis is a tick-borne protozoan disease caused by Theileria species. The Theileria species are classified into two groups depending on the cell type in which they proliferate and the clinical symptoms. The first group consists of lymphoproliferative Theileria species (T. parva and T. annulata), which mainly proliferate in lymphocytes, causing uncontrolled lymphocyte proliferation. The other group consists of a nonlymphoproliferative Theileria species (T. orientalis, also known as T. sergenti) that proliferates in erythrocytes and causes hemolytic anemia. Based on reports of generation of antigen-specific CD4(+) and CD8(+) T cells in lymphoproliferative theileriosis, we investigated whether T cells specific to the T. orientalis antigen are present in the nonlymphoproliferative form of the disease. In this study, we developed a new assay based on an enzyme-linked immunospot (ELISpot) to detect interferon-gamma (IFN-gamma)- and interleukin-10 (IL-10)-secreting cells in a series of cryogenically preserved bovine peripheral blood mononuclear cells (PBMCs). We first determined that IFN-gamma- and IL-10-secreting T cells were present in PBMCs by stimulating them with phytohemagglutinin L (PHA-L=red kidney bean lectin L, known as T cell stimulator), and then determined whether T. orientalis-specific T cells are present in T. orientalis-infected bovines. Peptides derived from T. orientalis major piroplasm surface protein (MPSP) were used as a T. orientalis-specific stimulator in the ELISpot assay, and peptides from glycoprotein B (gB) of the bovine herpes virus-1 (BHV-1) were used as a BHV-1-specific stimulator as a control for monitoring the immune response. Compared with results obtained using the BHV-1 (gB peptides)-specific IFN-gamma ELISpot assay to assess BHV-1-immunized Holsteins, prominent T. orientalis MPSP peptide-specific IFN-gamma and IL-10 positive spots were detected in T. orientalis-infected Holsteins but weak positive responses were exhibited by T. orientalis-infected Angus and Japanese Black cattle. As far as we are aware, this is the first report to show direct evidence of the presence of T. orientalis-specific T cells in T. orientalis-infected bovines using an antigen-specific ELISpot assay system and that T. orientalis-specific, IFN-gamma- and IL-10-producing T cells are produced in T. orientalis-infected Holsteins.

Extraction and Purification of a Lectin from Red Kidney Bean and Preliminary Immune Function Studies of the Lectin and Four Chinese Herbal Polysaccharides

Reversed micelles were used to extract lectin from red kidney beans and factors affecting reverse micellar systems (pH value, ionic strength and extraction time) were studied. The optimal conditions were extraction at pH 4–6, back extraction at pH 9–11, ion strength at 0.15 M NaCl, extraction for 4–6 minutes and back extraction for 8 minutes. The reverse micellar system was compared with traditional extraction methods and demonstrated to be a time-saving method for the extraction of red kidney bean lectin. Mitogenic activity of the lectin was reasonably good compared with commercial phytohemagglutinin (extracted from Phaseolus vulgaris) Mitogenic properties of the lectin were enhanced when four Chinese herbal polysaccharides were applied concurrently, among which 50 μg/mL Astragalus mongholicus polysaccharides (APS) with 12.5 μg/mL red kidney bean lectin yielded the highest mitogenic activity and 100 mg/kg/bw APS with 12.5 mg/kg/bw red kidney bean lectin elevated mouse nonspecific immunity.

ESR determination of the thermal stability of Mn2+ ion binding sites in different legume lectins

The legume lectins are a large family of homologous carbohydrate-binding proteins. Their carbohydrate specificity and quaternary structure vary widely. The carbohydrate binding activity of legume lectins depends on the simultaneous presence of calcium and transition-metal ions, especially the Mn2+ ion. In the present work, thermal stabilities of Mn2+ binding sites in pea, lentil, soybean and kidney-bean lectins have been studied by electron spin resonance spectroscopy in the temperature range of 120–400 K in different atmospheres. The evolution of parameters, such as the line width, peak-to-peak intensity associated with the hyperfine transition [m I (+5/2) → m′ I (−5/2)] at the lowest magnetic field side of the central electron transition [m S (+1/2) → m′ S (−1/2)] of the Mn+2 ion, g-factor and hyperfine splitting was evaluated. Annealing was also performed at high temperatures (353, 373, 383, 403 and 443 K) and the oxidation activation energy of Mn2+ ions in the indicated legumes was determined.

Biological Properties and Characterization of Lectin from Red Kidney Bean (Phaseolus Vulgaris)

Red kidney beans (Phaseolus vulgaris) contain significant amounts of lectins which have both beneficial and detrimental biological properties. Lectins are carbohydrate-binding glycoproteins that can react specifically with human blood cells, preferentially agglutinate malignant cells, and undergo mitogenic stimulation of lymphocytes. Some lectins are resistant to heat and proteolytic enzymes and can enter the circulatory system intact. Phytohaemagglutinin (PHA)—a lectin isolated from the red kidney bean—consists of four subunits with a molecular weight of 125 kDa. This bioactive compound has been partially purified by affinity chromatography using Affi-gel Blue. PHA has been shown to inhibit the viral enzymes, immunodeficiency virus type 1 reverse transcriptase (HIV-1 RT), and α- and β-glucosidases. This paper will review the chemical properties, biological activity, distribution, isolation, and heath benefits of red kidney bean lectin.

Biochemical and Genomic Analysis of Neoculin Compared to Monocot Mannose-Binding Lectins

Neoculin occurring in an edible tropical fruit is a heterodimeric protein which has both sweetness and a taste-modifying activity that converts sourness to sweetness. Both the primary and the overall tertiary structures of neoculin resemble those of monocot mannose-binding lectins. This study investigated differences in biochemical properties between neoculin and the lectins. Structural comparison between the mannose-binding sites of lectins and the corresponding regions of neoculin showed that there is at least one amino acid substitution at each site in neoculin, suggesting a reason for the lack of its mannose-binding ability. This was consistent with hemagglutination assay data demonstrating that neoculin had no detectable agglutinin activity. DNA microarray analysis indicated that neoculin had no significant influence on gene expression in Caco-2 cell, whereas kidney bean lectin (Phaseolus vulgaris agglutinin) greatly influenced various gene expressions. These data strongly suggest that neoculin has no lectin-like properties, encouraging its practical use in the food industry.