BACTERIAL AGGLUTINATION BY A LECTIN FROM THE LEAVES OF THE MEDICINAL PLANT, PIMENTA DIOICA (L.) MERR
Objective: The current investigation involves the purification, characterization of the lectin from the leaves of Pimenta dioica (L.) Merr. (Myrtaceae) a medicinal plant, and its application in bacterial typing.
Methods: A lectin was purified from the leaves by cation exchange chromatography. SDS PAGE revealed the molecular weight of the purified lectin. Biochemical characterization was carried out by performing various tests. Hemagglutination inhibition was conducted to detect the sugar specificity. Additionally, bacterial agglutination was performed to predict whether the purified lectin was able to agglutinate the bacterial strains.
Results: SDS PAGE analysis revealed the lectin to be a tetramer in the range of 43-66 kDa. The purified lectin agglutinated human, avian, and mouse erythrocytes, and was inhibited by 125 mmol of mannose and xylose. The lectin was stable at 0-60 Â° C for 30 min and was unaffected by either 2-Mercaptoethanol (2-ME) or Dithiothreitol (DTT) (50-250ÂµM). A pH of 6.0â€“8.0 was found optimum for its activity and was nearly independent of metal ions. The purified lectin contained about 20% carbohydrate as estimated by Anthrone method. Purified lectin agglutinated the Gram-negative Escherichia coli and Proteus vulgaris.
Conclusion: The isolated lectin was found to possess significant hemagglutinating activity. Due to its ability to agglutinate Gram negative bacteria such as Escherichia coli and Proteus vulgaris, it could be used for bacterial typing and for the design of bacterial filters.
2. Goldstein IJ, Hughes RC, Monsigny M, Osawa T, Sharon N. What should be called a lectin? Nature 1980;2:285-66.
3. Sharon N, Lis H. History of lectins: from hemagglutinins to biological recognition molecules. Glycobiology 2004;11:53Râ€“62R.
4. Rabia Hamid, Akbar Masood, Ishfak H Wani, Shaista Rafiq. Lectins: proteins with diverse applications. J Appl Pharm Sci 2013;3(4, Suppl 1):S93-S103.
5. Agrawal BBI, Goldstein IJ. Specific binding of Concanavalin A to cross-linked dextran gels. Biochem J 1967;96:23Câ€“15C.
6. Goldstein IJ. Lectin structure-activity: the story is never over. J Agric Food Chem 2002;50:6583â€“5.
7. Kumar KK, Lalith Prakash CK, Sumanthi J, Reddy GS, Shekar PC, Reddy B. Biological role of lectins: a review. J Orofac Sci 2012;4:20-5.
8. Nicolson GL, Blaustein J, Etzler ME. Characterization of two plant lectins from Ricinus communis and their quantitative interaction with a murine lymphoma. Biochemistry 1974; 13:196-204.
9. Lis H, Sharon N. Lectins in higher plants. In: Marcus A. Ed. The Biochemistry of Plants. Vol. 6. New York; Academic Press: 1981. p. 371-447.
10. Selitrennikoff CP. Antifungal proteins. Appl Environ Microbiol 2001;67:2883-94.
11. Keyaerts, Vijgen L, Pannecouque C, Van Damme E, Peumans W, Egberink H, et al. Plant lectins are potent inhibitors of coronaviruses by interfering with two targets in the viral replication cycle. Antivir Res 2007;75:179â€“87.
12. Maria A Souza, Fernanda C Carvalho, Luciana P Ruas, Rafael Ricci-Azevedo, Maria Cristina Roque-Barreira. The immunomodulatory effect of plant lectins: a review with emphasis on Artin M properties. Glycoconj J 2013;30:641â€“57.
13. Ramadhania ZM, Insanu M, Gunarti NS, Wirasutisna KR, Sukrasno S, Hartati R. Antioxidant activity from ten species of Myrtaceae. Asian J Pharm Clin Res 2017;10:5-7.
14. Neal MC. In Gardens of Hawaiâ€™i. Bernice P. Bishop Museum special publication 40, Bishop Museum Press: Honolulu, HI; 1965.
15. Weiss EA. Spice Crops. New York; CABI Publishing: 2002.
16. Anonymous. A wealth of India (Raw Materials). Publications Information Division, CSIR Publications. New Delhi; 1969;8:58-9.
17. Ynalvez RA, Cruz CG, Ynalvez MA. Isolation, partial purification and characterization of texas live oak (Quercus fusiformis) lectin. Adv Biosci Biotechnol 2015;6:470-84.
18. Bradford MM. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 1976;72:248-54.
19. Laemmli M Favre. Gel electrophoresis of proteins. J Mol Biol 1973;80:575â€“99.
20. Rosenberg Ian M. Protein analysis and purification: benchtop techniques. USA; Birkhauser, Boston; 1996.
21. Sadasivam S, Theymoli Balasubramanian. Practical Manual, Tamil Nadu Agricultural University, Coimbatore; 1985. p. 2.
22. Hedge JE, Hofreiter BT. In: Whistler RL, Be Miller JN. Eds. In: Carbohydrate Chemistry, New York; Academic Press: 1962. p. 17.
23. Laija SN, Mahesh S, Smitha LS, Remani P. Isolation and partial characterization of two plant lectins. Curr Res J Biol Sci 2010;2:232-7.
24. Pla A, Alonso E, Batista-Viera F, Franco Fraguas L. Screening for carbohydrate binding proteins in extracts of Uruguayan plants. Braz J Med Biol Res 2003;36:851-60.
25. Makarim Elfadil M Osman, Amna KE Awadallah, Emadeldin Hassan E Konozy. Isolation, purification and partial characterization of three lectins from tamarindus Indica seeds with a novel sugar specificity. Int J Plant Res 2016;6:13-9.
26. Syed Rashel Kabir, Md Mahamodun Nabi, Md Nurujjaman, Md Abu Reza A HM Khurshid Alam, Rokon Uz Zaman, et al. Momordica charantia seed lectin: toxicity, bacterial agglutination and antitumor properties. Appl Biochem Biotechnol 2015;175:2616â€“28.
27. William I Weis. Structural basis of recognition lectin-carbohydrate. Anna Rev Biochem 1996;65:441-73.
28. Timothy Richard Helliwell. Lectin methods and protocols. Methods Mol Med 1997;9:73-94.
29. Rocca JD. Lectins as next-generation mucoadhesives for specific targeting of the gastrointestinal tract. Drug Development Delivery 2004;4:12-9.
30. Pratheeba M, Rani KU, Ramesh B. Studies on the antimicrobial and anticancer activity of Solanum trilobatium. Asian J Pharm Clin Res 2013;7:213-9.
31. Rao PS, Navinchandra S, Jayaveera KN. An important spice, Pimenta dioica (Linn.) merill a review. Int Curr Pharm J 2012;1:221-5.
32. Santi-Gadelha T, De Almeida Gadelha CA, AragÃ£o KS, De Oliveira CC, Lima Mota MR, et al. Purification and biological effects of Araucaria angustifolia (Araucariaceae) seed lectin. Biochem Biophys Res Commun 2006;350:1050-5.
33. Komath SS, Kavitha M, Swamy MJ. Beyond carbohydrate binding: new directions in plant lectin research. Org Biomol Chem 2006;4:973-88.
34. Hirmo S, Ut M, Wadstrom T. Sialyl Glycoconjugate and proteoglycan binding microbial lectins. Electronic Lectin J: Lectins Biol Biochem Clin Biochem 1997;1287-984583-0-2.
35. Caldeon AM, Buck G, Doyle RJ. Lectin-microorganism complexes. Electronic Lectin J: Lectins Biol Biochem Clin Biochem 1997;1287-984583-0-2.
36. Raja SB, Murali MR, Kumar NK, Devaraj SN. Isolation and partial characterization of a novel lectin from Aegle marmelos fruit and its effect on adherence and invasion of Shigellae to HT29 cells. PLoS One 2011;6:1-9.