Skip to main content

Characterization of chickpea (Cicer arietinum L.) lectin for biological activity

Abstract

Lectins are proteins that are subject of intense investigations. Information on lectin from chickpea (Cicer arietinum L.) with respect to its biological activities are very limited. In this study, we purified lectin from the seeds of chickpea employing DEAE-cellulose and SP-Sephadex ion exchange chromatography and identified its molecular subunit mass as 35 kDa. The free radical scavenging activity of lectin measured by the DPPH assay has IC50 of 0.88 µg/mL. Lectin exerted antifungal activity against Candida krusei, Fusarium oxysporium oxysporium, Saccharomyces cerevisiae and Candida albicans, while antibacterial activity against E. coli, B. subtilis, S. marcescens and P. aeruginosa. The minimum inhibitory concentrations were 200, 240, 160 and 140 µg for C. krusei, F. oxysporium, S. cerevisiae and C. albicans respectively. Lectin was further examined for its antiproliferative potential against cancerous cell line. The cell viability assay indicated a high inhibition activity on Ishikawa, HepG2, MCF-7 and MDA-MB-231 with IC50 value of 46.67, 44.20, 53.58 and 37.46 µg/mL respectively. These results can provide a background for future research into the benefits of chickpea lectin to pharmacological perspective.

This is a preview of subscription content, access via your institution.

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6
Fig. 7
Fig. 8

References

  1. Adams DJ (2004) Fungal cell wall chitinases and glucanases. Microbiology 150:2029–2035. https://doi.org/10.1099/mic.0.26980-0

    CAS  Article  PubMed  Google Scholar 

  2. Agrawal P, Kumar S, Jaiswal YK, Das HR, Das RH (2011) A mesorhizobium lipopolysaccharide (LPS) specific lectin (CRL) from the roots of nodulating host plant, Cicer arietinum. Biochimie 93:440–449. https://doi.org/10.1016/j.biochi.2010.10.017

    CAS  Article  PubMed  Google Scholar 

  3. Ang ASW, Cheung RCF, Dan X, Chan YS, Pan W, Ng TB (2014) Purification and characterization of a glucosamine binding antifungal lectin from Phaseolus vulgaris cv. Chinese Pinto Beans with antiproliferative activity towards nasopharyngeal carcinoma cells. Appl Biochem Biotech 172(2):672–686. https://doi.org/10.1007/s12010-013-0542-2

    CAS  Article  Google Scholar 

  4. Baker RL, Brown RL, Chen ZY, Cleveland TE, Fakhoury AM (2009) A maize lectin-like protein with antifungal activity against Aspergillus flavus. J Food Prot 72:120–127. https://doi.org/10.4315/0362-028X-72.1.120

    CAS  Article  PubMed  Google Scholar 

  5. Bersuder P, Hole M, Smith G (1998) Antioxidants from a heated histidine glucose model system I: investigation of the antioxidant role of histamine and isolation of antioxidants by high performance liquid chromatography. J Am Oil Chem Food Agr 75:181–187. https://doi.org/10.1007/s11746-998-0030-y

    CAS  Article  Google Scholar 

  6. Boleti APA, Freire MGM, Coelho MB, Silva W, Baldasso PA, Gomes VM, Marangoni S, Novello JC, Macedo ML (2007) Insecticidal and antifungal activity of a protein from Pouteria torta seeds with lectin-like properties. J Agric Food Chem 55:2653–2658. https://doi.org/10.1021/jf0636317

    Article  PubMed  Google Scholar 

  7. Bourne Y, Ayouba A, Rouge P, Cambillau C (1994) Interaction of a legume lectin with two components of the bacterial cell wall. J Biol Chem 269:9429–9435

    CAS  PubMed  Google Scholar 

  8. Cardador-Martínez A, Loarca-Piña G, Oomah BD (2002) Antioxidant activity in common beans (Phaseolus vulgaris L.). J Agric Food Chem 50:6975–6980. https://doi.org/10.1021/jf020296n

    Article  PubMed  Google Scholar 

  9. Carrasco-Castilla J, Hernández-Álvarez AJ, Jiménez-Martínez C, Jacinto-Hernández C, Alaiz M, Girón-Calle J, Vioque J, Dávila-Ortiz G (2012) Antioxidant and metal chelating activities of Phaseolus vulgaris L. var Jamapa protein isolates, phaseolin and lectin hydrolysates. Food Chem 131:1157–1164. https://doi.org/10.1016/j.foodchem.2011.09.084

    CAS  Article  Google Scholar 

  10. Chakraborti D, Sarkar A, Mondal HA, Das S (2009) Tissue specific expression of potent insecticidal, Allium sativum leaf agglutinin (ASAL) in important pulse crop, chickpea (Cicer arietinum L.) to resist the phloem feeding Aphis craccivora. Transgenic Res 18:529–544. https://doi.org/10.1007/s11248-009-9242-7

    CAS  Article  PubMed  Google Scholar 

  11. Chan YS, Wong JH, Ng TB (2011) A glucuronic acid binding leguminous lectin with mitogenic activity toward mouse splenocytes. Protein Pept Lett 18:194–202. https://doi.org/10.2174/092986611794475110

    CAS  Article  PubMed  Google Scholar 

  12. Charungchitrak S, Petsom A, Sangvanich P, Karnchanatat A (2011) Antifungal and antibacterial activities of lectin from the seeds of Archidendron jiringa Nielsen. Food Chem 126(3):1025–1032. https://doi.org/10.1016/j.foodchem.2010.11.114

    CAS  Article  Google Scholar 

  13. Chu KT, Liu KH, Ng TB (2003) Cicerarin, a novel antifungal peptide from the green chickpea. Peptides 24:659–663. https://doi.org/10.1016/S0196-9781(03)00134-7

    CAS  Article  PubMed  Google Scholar 

  14. Damme EJMV, Peumans WJ, Barre A, Rougé P (1998) Plant lectins: a composite of several distinct families of structurally and evolutionary related proteins with diverse biological roles. Crit Rev Plant Sci 17(6):575–692. https://doi.org/10.1080/07352689891304276

    Article  Google Scholar 

  15. Dziarski R, Rasenick MM, Gupta D (2000) Bacterial peptidoglycan binds to tubulin. Biochim Biophys Acta 1524:17–26. https://doi.org/10.1016/S0304-4165(00)00137-9

    CAS  Article  Google Scholar 

  16. E Lacerda RR, do Nascimento ES, de Lacerda JT, Pinto LD, Rizzi C, Bezerra MM, Pinto IR, Filho SM, Pinto VP, Filho GC, Gadelha CA, Gadelha TS (2017) Lectin from seeds of a Brazilian lima bean variety (Phaseolus lunatus L. var. cascavel) presents antioxidant, antitumour and gastroprotective activities. Int J Biol Macromol 95:1072–1081. https://doi.org/10.1016/j.ijbiomac.2016.10.097

    CAS  Article  PubMed  Google Scholar 

  17. Esteban R, Dopico B, Muñoz FJ, Romo S, Labrador E (2002) A seedling specific vegetative lectin gene is related to development in Cicer arietinum. Physiol Plant 114:619–626. https://doi.org/10.1034/j.1399-3054.2002.1140416.x

    CAS  Article  PubMed  Google Scholar 

  18. Faheina-Martins GV, da Silveira AL, Cavalcanti BC, Ramos MV, Moraes MO, Pessoa C, Araújo DAM (2012) Antiproliferative effects of lectins from Canavalia ensiformis and Canavalia brasiliensis in human leukemia cell lines. Toxicol In Vitro. https://doi.org/10.1016/j.tiv.2012.06.017

    PubMed  Google Scholar 

  19. Ghosh M (2009) Purification of a lectin-like antifungal protein from the medicinal herb, Withania somnifera. Fitoterapia 80:91–95. https://doi.org/10.1016/j.fitote.2008.10.004

    CAS  Article  PubMed  Google Scholar 

  20. Giron-Calle J, Vioque J, DelMarYust M, Pedroche J, Alaiz M, Millán F (2004) Effect of chickpea aqueous extracts, organic extracts, and protein concentrates on cell proliferation. J Med Food 7(2):122–129. https://doi.org/10.1089/1096620041224175

    Article  PubMed  Google Scholar 

  21. Gomes FS, Procópio TF, Napoleão TH, Coelho LCBB, Paiva PMG (2012) Antimicrobial lectin from Schinus terebinthifolius leaf. J Appl Microbiol 114:672–679. https://doi.org/10.1111/jam.12086

    Article  PubMed  Google Scholar 

  22. Irlanda LD, Ana Maria GP, Luz VM (2017) Legume lectins: proteins with diverse applications. Int J Mol Sci 18:1–18. https://doi.org/10.3390/ijms18061242

    Google Scholar 

  23. Jebali J, Fakhfekh E, Morgen M, Srairi-Abid N, Majdoub H, Gargouri A, El Ayeb M, Luis J, Marrakchi N, Sarray S (2014) Lebecin, a new C-type lectin like protein from Macrovipera lebetinavenom with antitumor activity against the breast cancer cell line MDA-MB231. Toxicon 86:16–27. https://doi.org/10.1016/j.toxicon.2014.04.010

    CAS  Article  PubMed  Google Scholar 

  24. Katre UV, Gaikwad SM, Bhagyawant SS, Deshpande UD, Khan MI, Suresh CG (2005) Crystallization and preliminary X-ray characterization of a lectin from Cicer arietinum (chickpea). Acta Crystallogr Sect F Struct Biol Cryst Commun 61:141–143. https://doi.org/10.1107/S1744309104032166

    CAS  Article  PubMed  Google Scholar 

  25. Kaur N, Dhuna V, Kamboj SS, Agrewala JN, Singh J (2006) A novel antiproliferative and antifungal lectin from Amaranthus viridis Linn seeds. Protein Pept Lett 13:897–905. https://doi.org/10.2174/092986606778256153

    CAS  Article  PubMed  Google Scholar 

  26. Kiss R, Camby I, Duckworth C, De Decker R, Salmon I, Pasteel JL, Danguy A, Yeaton P (1997) In vitro influence of Phaseolus vulgaris, Griffonia simpliciflia, concanavalin A, wheat germ, and peanut agglutinin on HCT-15, Lovo, d SW 837 human colorectal cancer cell growth. Gut 40:253–261. https://doi.org/10.1136/gut.40.2.253

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  27. Kolberg J, Michaelsen TE, Sletten K (1983) Properties of a lectin purified from the seeds of Cicer arietinum. Hoppe Seylers Z Physiol Chem 364(6):655–664

    CAS  Article  PubMed  Google Scholar 

  28. Kumar S, Kapoor V, Gill K, Singh K, Xess I, Das SN, Dey S (2014) Antifungal and antiproliferative protein from Cicer arietinum: a bioactive compound against emerging pathogens. Biomed Res Int 2014:387203. https://doi.org/10.1155/2014/387203

    PubMed  PubMed Central  Google Scholar 

  29. Lam SK, Ng TB (2010) First report of a haemagglutinin induced apoptotic pathway in breast cancer cells. Biosci Rep 30(5):307–317. https://doi.org/10.1042/BSR20090059

    CAS  Article  PubMed  Google Scholar 

  30. Li Y, Zhang G, Ng TB, Wang H (2010) A novel lectin with antiproliferative and HIV-1 reverse transcriptase inhibitory activities from dried fruiting bodies of the monkey head mushroom Hericium erinaceum. J Biomed Biotech 2010:716515. https://doi.org/10.1155/2010/716515

    Google Scholar 

  31. Liener IE (1962) Toxic factors in edible legumes and their elimination. Am J Clin Nutr 11:281–298

    CAS  Article  Google Scholar 

  32. Lin P, Ye X, Ng TB (2008) Purification of melibiose-binding lectins from two cultivars of chinese black soybeans. Acta Biochim Biophys Sin 40(12):1029–1038. https://doi.org/10.1111/j.1745-7270.2008.00488.x

    CAS  Article  PubMed  Google Scholar 

  33. Lis H, Sharon N (1981) Lectins in higher plants. In: Abraham M (ed) Proteins and nucleic acids: the biochemistry of plants. Academic Press, New York, pp 371–447

    Chapter  Google Scholar 

  34. Liu B, Bian HJ, Bao JK (2010) Plant lectins: potential antineoplastic drugs from bench to clinic. Cancer Lett 287(1):1–12. https://doi.org/10.1016/j.canlet.2009.05.013

    CAS  Article  PubMed  Google Scholar 

  35. Llorach R, Espín JC, Tomás-Barberán FA, Ferreres F (2003) Valorization of cauliflower (Brassica oleracea L. var. botrytis) by-products as a source of antioxidant phenolics. J Agric Food Chem 51:2181–2187. https://doi.org/10.1021/jf021056a

    CAS  Article  PubMed  Google Scholar 

  36. Lowry OH, Resebrough JJ, Farel L, Randal RJ (1951) Protein measurement with Folin phenol reagent. J Biol Chem 193:265–275

    CAS  PubMed  Google Scholar 

  37. Luo Y, Xu X, Liu J, Li J, Sun Y, Liu Z, Liu J, Van Damme E, Balzarini J, Bao J (2007) A novel mannose-binding tuber lectin from Typhonium divaricatum (L.) decne (family Araceae) with antiviral activity against HSV-II and anti-proliferative effect on human cancer cell lines. J Biochem Mol Biol 40(3):358–367. https://doi.org/10.5483/BMBRep.2007.40.3.358

    CAS  PubMed  Google Scholar 

  38. Manach C, Scalbert A, Morand C, Remesy S, Jimenez L (2004) Polyphenols: food source and bioavailability. Am J Clin Nutr 79:727–747

    CAS  Article  PubMed  Google Scholar 

  39. Ngai PH, Ng TB (2007) A lectin with antifungal and mitogenic activities from red cluster (Capsicum frutescens) seeds. Appl Microbiol Biotechnol 74:366–371. https://doi.org/10.1007/s00253-006-0685-y

    CAS  Article  PubMed  Google Scholar 

  40. Nowell PC (1960) Phytohemagglutinin: an initiator of mitosis in cultures of normal human leukocytes. Cancer Res 20(4):462–466

    CAS  PubMed  Google Scholar 

  41. Petnual P, Sangvanich P, Karnchanatat A (2010) A lectin from the rhizomes of turmeric (Curcuma longa L.) and its antifungal, antibacterial, and alpha-glucosidase inhibitory activities. Food Sci Biotechnol 19:907–916. https://doi.org/10.1007/s10068-010-0128-5

    CAS  Article  Google Scholar 

  42. Pueppke SG (1979) Distribution of lectins in the Jumbo Virginia and Spanish varieties of the peanut, Arachis hypogaea L. Plant Physiol 64:575–580. https://doi.org/10.1104/pp.64.4.575

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  43. Pusztai A (1993) Antinutritive effects of wheat-germ agglutinin and other N- acetylglucosamine-specific lectins. Br J Nutri 70:313–321

    CAS  Article  Google Scholar 

  44. Qureshi IA, Dash P, Srivastava PS, Koundal KR (2006) Purification and characterization of an N-acetyl-D-galactosamine-specific lectin from seeds of chickpea (Cicer arietinum L.). Phytochem Anal 17:350–356. https://doi.org/10.1002/pca.925

    CAS  Article  PubMed  Google Scholar 

  45. Ramnath V, Rekha PS, Kuttan G, Kuttan R (2009) Regulation of caspase-3 and Bcl-2 expression in Dalton’s lymphoma ascites cells by Abrin. Evid Based Complement Altern Med 6(2):233–238. https://doi.org/10.1093/ecam/nem099

    CAS  Article  Google Scholar 

  46. Rizzi C, Galeoto L, Zoccatelli G, Vincenzi S, Chignola R, Peruffo ADB (2003) Active soybean lectin in foods: quantitative determination by ELISA using immobilised asialofetuin. Food Res Int 36:815–821. https://doi.org/10.1016/S0963-9969(03)00076-0

    CAS  Article  Google Scholar 

  47. Roy F, Boye JI, Simpson BK (2010) Bioactive proteins and peptides in pulse crops: pea, chickpea and lentil. Food Res Int 43:432–442. https://doi.org/10.1016/j.foodres.2009.09.002

    CAS  Article  Google Scholar 

  48. Schwarz RE, Wojciechowicz DC, Picon AI, Schwarz MZ, Paty PB (1999) Wheat germ agglutinin-mediated toxicity in pancreatic cancer cells. J Cancer 80:1754–1762. https://doi.org/10.1038/sj.bjc.6690593

    CAS  Article  Google Scholar 

  49. Selitrennikoff CP (2001) Antifungal proteins. Appl Environ Microbiol 67(7):2883–2894. https://doi.org/10.1128/AEM.67.7.2883-2894.2001

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  50. Sharma U, Katre UV, Suresh CG (2015) Crystal structure of a plant albumin from Cicer arietinum (chickpea) possessing hemopexin fold and hemagglutination activity. Planta 241(5):1061–1073. https://doi.org/10.1007/s00425-014-2236-6

    CAS  Article  PubMed  Google Scholar 

  51. Sitohy M, Doheim M, Badr H (2007) Isolation and characterization of a lectin with antifungal activity from Egyptian Pisum sativum seeds. Food Chem 104(3):971–979. https://doi.org/10.1016/j.foodchem.2007.01.026

    CAS  Article  Google Scholar 

  52. Skehan P, Storeng R, Scudiero D, Monks A, McMahon J, Vistica D, Warren JT, Bokesch H, Kenney S, Boyd MR (1990) New colorimetric cytotoxicity assay for anticancer-drug screening. J Natl Cancer Inst 82(13):1107–1112. https://doi.org/10.1093/jnci/82.13.1107

    CAS  Article  PubMed  Google Scholar 

  53. Summer JB, Howell SF (1936) The identification of the hemagglutinin of the Jack bean with Concanavalin A. J Bacteriol 32:227–237

    Google Scholar 

  54. Trindade MB, Lopes JL, Soares-Costa A, Monteiro-Moreira AC, Moreira RA, Oliva ML, Beltramini LM (2006) Structural characterization of novel chitin-binding lectins from the genus Artocarpus and their antifungal activity. Biochim Biophys Acta 1764:146–152. https://doi.org/10.1016/j.bbapap.2005.09.011

    CAS  Article  PubMed  Google Scholar 

  55. Vogelsang R, Barz W (1993) Purification, characterization and differential hormonal regulation of a β-1, 3-glucanase and two chitinases from chickpea (Cicer arietinum L.). Planta 189:60–69. https://doi.org/10.1007/BF00201344

    CAS  Article  PubMed  Google Scholar 

  56. Wakankar MS, Krishnasastry MV, Jaokar TM, Patel KA, Gaikwad SM (2013) Solution and in silico studies on the recombinant lectin from Cicer arietinum seeds. Int J Biol Macromol 56:149–155. https://doi.org/10.1016/j.ijbiomac.2013.02.015

    CAS  Article  PubMed  Google Scholar 

  57. Wang Z, Chen M, Zhu Y, Qian P, Zhou Y, Wei J, Shen Y, Mijiti A, Gu A, Wang Z, Zhang H, Ma H (2017) Isolation, identification and characterization of a new type of lectin with α-amylase inhibitory activity in chickpea (Cicer arietinum L.). Protein Pept Lett. https://doi.org/10.2174/0929866524666170711120501

    PubMed Central  Google Scholar 

  58. Xia L, Ng TB (2006) A hemagglutinin with mitogenic activity from dark red kidney beans. J Chromatogr B Anal Technol Biomed Life Sci 844:213–216. https://doi.org/10.1016/j.jchromb.2006.07.042

    CAS  Article  Google Scholar 

  59. Ye XY, Ng TB (2002) Isolation of a new cyclophilin-like protein from chickpeas with mitogenic, antifungal and anti-HIV-1 reverse transcriptase activities. Life Sci 70:1129–1138. https://doi.org/10.1016/S0024-3205(01)01473-4

    CAS  Article  PubMed  Google Scholar 

  60. Ye XJ, Ng TB (2011) Antitumor and HIV-1 reverse transcriptase inhibitory activities of a hemagglutinin and a protease inhibitor from mini-black soybean. Evid Based Complement Altern Med 2011:851396. https://doi.org/10.1155/2011/851396

    Article  Google Scholar 

  61. Ye XY, Wang HX, Ng TB (2000) Dolichin, a new chitinase-like antifungal protein isolated from field beans (Dolichos lablab). Biochem Biophys Res Commun 269:155–159. https://doi.org/10.1006/bbrc.2000.2115

    CAS  Article  PubMed  Google Scholar 

  62. Ye XY, Ng TB, Tsang PWK, Wang J (2001) Isolation of a homodimeric lectin with antifungal and antiviral activities from red kidney bean (Phaseolus vulgaris) seeds. J Protein Chem 20(5):367–375. https://doi.org/10.1023/A:1012276619686

    CAS  Article  PubMed  Google Scholar 

Download references

Acknowledgement

The authors are thankful to University Grants Commission (UGC), New Delhi for funding and specially Mr. Ajay Gautam, for providing research fellowship. The authors are indebted to Dr. M. Yasin, Senior Breeder, RAK Agriculture College, Sehore for providing chickpea seed material.

Author information

Affiliations

Authors

Corresponding author

Correspondence to Sameer S. Bhagyawant.

Ethics declarations

Conflict of interest

The authors declare that there are no conflicts of interest.

Rights and permissions

Reprints and Permissions

About this article

Verify currency and authenticity via CrossMark

Cite this article

Gautam, A.K., Gupta, N., Narvekar, D.T. et al. Characterization of chickpea (Cicer arietinum L.) lectin for biological activity. Physiol Mol Biol Plants 24, 389–397 (2018). https://doi.org/10.1007/s12298-018-0508-5

Download citation

Keywords

  • Chickpea lectin
  • Sp-Sephadex
  • SRB assay
  • Human cancer cell lines