EXTRACTION, PURIFICATION AND CHARACTERIZATION OF LECTIN FROM PHASEOLUS VULGARIS L. CV. WHITE SEEDS (WHITE KIDNEY BEAN (original) (raw)
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Purification and Characterization of a Lectin fromPhaseolus vulgaris cv.(Anasazi Beans)
Journal of Biomedicine and Biotechnology, 2009
A lectin has been isolated from seeds of the Phaseolus vulgaris cv. "Anasazi beans" 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 30-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 1-14 and the temperature range of 0-80 • C. The lectin potently suppressed proliferation of MCF-7 (breast cancer) cells with an IC 50 of 1.3 μM, and inhibited the activity of HIV-1 reverse transcriptase with an IC 50 of 7.6 μM. The lectin evoked a mitogenic response from murine splenocytes as evidenced by an increase in [ 3 H-methyl]-thymidine incorporation. The lectin had no antifungal activity. It did not stimulate nitric oxide production by murine peritoneal macrophages. Chemical modification results indicated that tryptophan was crucial for the hemagglutinating activity of the lectin.
Biological Properties and Characterization of Lectin from Red Kidney Bean (Phaseolus Vulgaris)
Food Reviews International, 2008
Red kidney beans (Phaseolus vulgaris) contain significant amounts of lectins which have both beneficial and detrimental biological properties. Lectins are carbohydratebinding 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 beanconsists 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 aand b-glucosidases. This paper will review the chemical properties, biological activity, distribution, isolation, and heath benefits of red kidney bean lectin.
International Journal of Research and Analytical Reviews (IJRAR) , 2024
A protein fraction exhibiting lectin activity was identified in the extract of local Roman bean (Phaseolus vulgaris) sprouts. The galactose-specific lectins demonstrated the highest activity. The galactose-specific lectin (BS-Gal) was purified using affinity chromatography on an ES-2100-100 column. Gel electrophoresis revealed that BS-Gal consists of two protein subunits with molecular weights of 60 and 54 kDa, respectively. Various physicochemical properties of the lectin were investigated, showing optimal activity within a pH range of 7-9 and temperatures from 0 to 75 °C, beyond which its hemagglutinating activity diminishes. The lectin displayed high sensitivity to divalent ions and certain amino acids. Calcium and magnesium ions significantly contribute to the structural integrity of the lectin, as indicated by changes in hemagglutinating activity in the presence of EDTA and EGTA. Additionally, BS-Gal was found to bind to human erythrocytes expressing A and B antigens.
Molecules
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 ...
Purification and Characterization of a Lectin fromPhaseolus vulgaris cv.(Anasazi Beans)
Journal of Biomedicine and Biotechnology, 2009
A lectin has been isolated from seeds of thePhaseolus vulgaris cv.“Anasazi beans” 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 30-kDa subunits with substantial N-terminal sequence similarity to otherPhaseoluslectins. The hemagglutinating activity of the lectin was stable within the pH range of 1–14 and the temperature range of 0–80∘C. The lectin potently suppressed proliferation of MCF-7 (breast cancer) cells with anIC50of 1.3 μM, and inhibited the activity of HIV-1 reverse transcriptase with anIC50of 7.6 μM. The lectin evoked a mitogenic response from murine splenocytes as evidenced by an increase in [3H-methyl]-thymidine incorporation. The lectin had no antifungal activity. It did not stimulate nitric oxide production by murine peritoneal macrophages. Chemical modification results indicated that t...
Screening and Partial Purification of Lectin from Various Bangladeshi Plant Seeds
American Journal of Sensor Technology, 2014
Lectins are group of proteins or glycoprotein of non immunological origins, which can recognize specific carbohydrate structure. In this research work hemaggulutination was used as technique for screening Bangladeshi vegetables for lectin. Five varieties of legume plant seeds Canavalia gladiata(Sword bean),Lens culinaris (Moshordal), Peasum sativam (Motorsuti), Vigna unguiculata subsp. sesquipedalis (Borboti), Cajanus cajan (Arhordal), one Amaranthaceae Amaranthus caudate (katoadata) and one fruit Citrullus lanatus (Watermelon) species from Bangladesh were examined for lectins with chicken and human erythrocytes. Crude extracts from all the species showed agglutinating activity against the erythrocytes used. The lowest protein concentration required to produce erythrocytes agglutination varied remarkable ranging from 0.7 µg/ml to 8080 µg/ml. The strongest activities were shown in the agglutination of human blood erythrocytes by partial purification of lectin from Moshordal and chick...
Biosynthesis of lectins in developing seeds of common bean
Food Chemistry, 1990
Two cultivars of Phaseolus vulgaris L., )qor de mayo FM-RMC, showing resistance to common mosaic virus and to bean rust (Uromyces phaseoli), and FM-C, without such resistance, were used to study the synthesis of lectins during seed development as measured by affinity chromatography of protein fr~ctions. Seeds at different stages of development were sorted out by size from 4-12mm length, in addition to mature samples. The spec([ic haemagglutinating activity ( SHA ) of isolated protein fractions from mature seeds', expressed as haemagglutinating units/mg protein, and the theoretical agglutination capacity (TA C). calculated from the SHA and the total protein fr~ction, were also assessed. The peak lectin level as measured by affinity chromatography was reached at a seed size of lOmm and decreased thereafter. The lec tin con ten t and TA C of all samples of the disease-resistan t cultivar were superior to that of the non-resistant variety.
Journal of protein chemistry, 2001
A homodimeric lectin adsorbed on Affi-gel blue gel and CM-Sepharose and possessing a molecular weight of 67 kDa was isolated from red kidney beans. The hemagglutinating activity of this lectin was inhibited by glycoproteins but not by simple sugars. The lectin manifested inhibitory activity on human immunodeficiency virus-1 reverse transcriptase and alpha-glucosidase. The N-terminal sequence of the lectin exhibited some differences from previously reported lectins from Phaseolus vulgaris but showed some similarity to chitinases. It exerted a suppressive effect on growth of the fungal species Fusarium oxysporum, Coprinus comatus, and Rhizoctonia solani. The lectin had low ribonuclease and negligible translation-inhibitory activities.
International Journal of Advanced Academic Research, 2016
Lectins was extracted from processed and unprocessed black-eyed peas, green beans, soybeans, peanuts, and red kidney beans by grinding using attrition mill and then soaked in distilled water. The proteins in the supernatant were precipitated by dissolving 10g of ammonium sulphate in 100ml of the supernatants. The lectin extracts were evaluated for hemagglutinin activity using human erythrocytes from blood group A + , B + , AB + , and O + , respectively. Result revealed that lectins from unprocessed soybean, black-eyed pea and green bean are blood group selective and may be classified as blood type specific lectins. While lectins from kidney beans and peanuts agglutinated all blood groups and may be classified as panhemagglutinin lectins, i.e. they agglutinated erythrocytes from all blood groups. Result shows that blood group O is less affected by hemagglutinin activity of lectin. Reduction/loss of hemagglutinin activity was observed with reduction in concentration of the crude lectin extracts in all unprocessed legumes. Boiling (at 100 0 C) for 30minutes completely inactivated hemagglutinin activity of lectin in Black eyed peas, soybeans, green beans and red kidney beans. Slight hemagglutinin activity was seen in lectin from roasted peanuts, indicating that dry heating may be very effective method to inactivate lectin completely. Also, in this study, lectins were extracted from rice (Oryza sativa), maize (Zea mays), sorghum (Sorghum bicolor), Acha (Digitaria excells), millet (Panicum maniceum); partial purification of the extract assayed from hemugglutinin activity using 4% of human erythrocyte in lectin buffer. Extract from these selected cereals showed positive and negative agglutination reaction with blood group A + , B + , O + and AB + depending on the concentration (dilution) and the blood group. Lectin extract from maize showed no agglutination reaction when diluted in blood group A + , B + , and AB + and even when processed and diluted. While crude lectin extract from rice showed no activity in blood group A + , B + , and AB + and when unprocessed and showed lectin agglutination activity when processed by boiling (cooking) showing that heat may encourage its activity. Blood group B + , and AB + showed no activity in all dilution. Also acha showed high agglutination activity in blood group A + , B + , AB + and O + but losses activity in dilution a 2 and a 3. Crude lectin extract from processed sorghum showed