Commenced in January 2007
Paper Count: 30077
In Vitro Antibacterial and Antifungal Effects of a 30 kDa D-Galactoside-Specific Lectin from the Demosponge, Halichondria okadai
Abstract:The present study has been taken to explore the screening of in vitro antimicrobial activities of D-galactose-binding sponge lectin (HOL-30). HOL-30 was purified from the marine demosponge Halichondria okadai by affinity chromatography. The molecular mass of the lectin was determined to be 30 kDa with a single polypeptide by SDS-PAGE under non-reducing and reducing conditions. HOL-30 agglutinated trypsinized and glutaraldehydefixed rabbit and human erythrocytes with preference for type O erythrocytes. The lectin was subjected to evaluation for inhibition of microbial growth by the disc diffusion method against eleven human pathogenic gram-positive and gram-negative bacteria. The lectin exhibited strong antibacterial activity against gram-positive bacteria, such as Bacillus megaterium and Bacillus subtilis. However, it did not affect against gram-negative bacteria such as Salmonella typhi and Escherichia coli. The largest zone of inhibition was recorded of Bacillus megaterium (12 in diameter) and Bacillus subtilis (10 mm in diameter) at a concentration of the lectin (250 μg/disc). On the other hand, the antifungal activity of the lectin was investigated against six phytopathogenic fungi based on food poisoning technique. The lectin has shown maximum inhibition (22.83%) of mycelial growth of Botrydiplodia theobromae at a concentration of 100 μg/mL media. These findings indicate that the lectin may be of importance to clinical microbiology and have therapeutic applications.
Digital Object Identifier (DOI): doi.org/10.5281/zenodo.1055569Procedia APA BibTeX Chicago EndNote Harvard JSON MLA RIS XML ISO 690 PDF Downloads 1925
 Goldstein, I. J. (2002). Lectin structure-activity: the story is never over. J Agric Food Chem, 50, 6583-6585.
 Vazquez, L., Alpuche, J., Maldonado, G., Agundis, C., Morales, A. P., and Zenteno, E. (2009). Immunity mechanisms in crustaceans. Innate Immun, 15, 179-188.
 Gabius, H. J., Unverzagt, C., and Kayser, K. (1998). Beyond plant lectin histochemistry: preparation and application of markers to visualize the cellular capacity for protein-carbohydrate recognition. Biotech Histochem 73, 263-277.
 Stabili, I., Pagliara, P., and Roch, P. (1996). Antibacterial activity in the coelomocytes of the sea urchin Paracentrotus lividus. Comp Biochem Physiol, 113B, 639-644.
 Wang, C., Wang, Y., Huffman, N. T., Cui, C., Yao, X., and Midura, S. (2009). Confocal laser raman microspectroscopy of biomineralization foci in UMR 106 osteoblastic cultures reveals temporally synchronized protein changes preceding and accompanying mineral crystal deposition. J Biol Chem, 284, 7100-7113.
 Mali, B., Ried, J. S., Frohme, M., and Frank, U. (2006). Structural but not functional conservation of an immune molecule: a tachylectin-like gene in Hydractinia. Dev Comp Immunol, 30, 275-281.
 Kvennefors, E. C. E., Leggat, W., Guldberg, O. H., Degan, B. M., and Barnes, A. C. (2008). An ancient and variable mannose-binding lectin from the coral Acropora millepora binds both pathogens and symbionts. Dev Comp Immunol, 32, 1582-1592.
 Canesi, L., Gallo, G., Gavioli, M., and Pruzzo, C. (2002). Bacteriahemocyte interactions and phagocytosis in marine bivalves. Microbe Res Tech, 57, 469-476.
 Austin, B. (1988). Marine Microbiology. University Press, Cambridge, U. K., 166-171.
 Muller, W. E. G. (2001). How was metazoan threshold crossed: the hypothetical Urmetazoan. Comp Biochem Physiol, 129A, 433-460.
 Vogel, S. (1977). Current-induced flow through living sponges in nature. Proc Natl Acad Sci, 74, 2069-2071.
 Gonzales, J. M., and Moran, M. A. (1997). Numerical dominance of a group of marine bacteria in the alpha-subclass of the class Proteobacteria in coastal seawater. Appl Environ Microbiol, 63, 4237-4242.
 Proksch, P. (1994). Defensive roles for secondary metabolites from marine sponges and sponge-feeding nudibranchs. Toxicon, 32, 639-655.
 Muller, W. E. G., Blumbach, B., and Muller, I. M. (1999). Evolution of the innate and adaptive immune systems: Relationships between Potential Immune Molecules in the Lowest Metazoan Phylum and Those in Vertebrates1. Transplantation, 68, 1215-1227.
 Bretting, H., Jacobs, G., Donadey, C., and Vacelet, J. (1983). Immunohistochemical studies on the distribution and the function of the D-galactose-specific lectins in the sponge Axinella polypoides (Schmidt). Cell Tiss Res, 229, 551-571.
 Schroder, H. C., Ushijima, H., Krasko, A., Gamulin, V., Thakur, N. L., Diehl-Seifert, B., Muller, I. M., and Muller, W. E. G. (2003). Emergence and disappearance of an immune molecule, an antibacterial lectin, in basal metazoan; A tachylectin-related protein in the sponge Suberites domuncula. J Biol Chem, 278, 32810 -32817.
 Tachibana, K., Scheuer, P. J., Tsukitani, Y., Kikuchi, H., Engen, D. V., et al. (1981). Okadaic acid, a cytotoxic polyether from two marine sponges of the genus Halichondria. J Am Chem Soc, 103, 2469-2471.
 Kawagishi, H,. Yamawaki, M., Isobe, S., Usui, T., Kimura, A., and Chiba, S. (1994). Two lectins from the marine sponge Halichondria okadai: an N-acetyl-sugar-specific lectin (HOL-I) and an Nacetyllactosamine- specific lectin (HOL-II). J Biol Chem, 269, 1375- 1379.
 Kawsar, S. M. A., Fujii, Y., Matsumoto, R., Ichikawa, T., Tateno, H., Hirabayashi, J., et al. (2008). Isolation, purification, characterization and glycan-binding profile of a D-galactoside specific lectin from the marine sponge, Halichondria okadai. Comp Biochem Physiol, 150B, 349-357.
 Smith, P. K., Krohn, R. I., Hermanson, G. T., Mallia, A. K., Gartner, F. H., Provenzano, M. D., Fujimoto, E. K., Goeke, N. M., Olson, B. J., and Klenk, D. C. (1985). Measurement of protein using bicinchoninic acid. Anal Biochem , 150, 76-85.
 Wiechelman, K. J., Braun, R. D., and Fitzpatrick, J. D. (1988). Investigation of the bicinchoninic acid protein assay: identification of the groups responsible for color formation. Anal Biochem, 175, 231-237.
 Laemmli, U. K. (1970). Cleavage of structural proteins during assembly of the head of bacteriophage T4. Nature, 227, 680-685.
 Matsui, T. (1984). D-galactoside specific lectins from coelomocytes of the echiuran, Urechis unicinctus. Biol Bull, 166, 178-188.
 Bauer, A. W., Kirby, M. M., Sherris, J. C., and Turck, M. (196). Antibiotic susceptibility testing by a standardized single disc method. Am J Clin Path, 45, 493-496.
 Grover, R. K., and Moore, J. D. (1962). Toximetric studies of fungicides against the brown rot organisms, Sclerotinia fructicola and S. laxa. Phytopathology, 52, 876-880.
 Miah, M. A.T., Ahmed, H. U., Sharma, N. R., Ali, A., and Miah, S. A. (1990). Antifungal activity of some plant extracts. Bang J Bot, 19, 5-10.
 Pfeifer, K., Haasemann, M., Gamulin, V., Bretting, H., Fahrenholz, F., and Muller, W. E. G. (1993). S-type lectins occur also in invertebrates: high conservation of the carbohydrate recognition domain in the lectin genes from the marine sponge Geodia cydonium. Glycobiology, 3, 179- 184.
 Schroder, H. C., Boreiko, A., Korzhev, M., Tahir, M. N., Tremel, W., Eckert, C., Ushijima, H., Muller, I. M., and Muller, W. E. G. (2006). Coexpression and functional interaction of silicatein with galectin: matrixguided formation of siliceous spicules in the marine demosponge, Suberites domuncula. J Biol Chem, 281, 12001-12009.
 Miarons, P. B., and Fresno, M. (2000). Lectins from tropical sponges; purification and characterization of lectins from genus Aplysina. J Biol Chem, 275, 29283-29289.
 Kurata, O., and Hatai, K. (2002). Activation of carp leukocytes by a galactose-binding protein from Aphanomyces piscicida. Dev Comp Immunol, 26, 461-469.
 Oliveira, M. D. L., Andrade, C. A. S., Magalhaes, N. S. S., Coelho, L. C. B. B., etal. (2008). Purification of a lectin from Eugenia uniflora L. seeds and its potential antibacterial activity. Lett Appl Microbiol, 46, 371-376.
 Broekaert, W. F., Van, P. J., Leyn, F., Joos, H., and Peumans, W. (1998). A chitin-binding lectin from stinging rettle rhizomes with antifungal properties. Science, 245, 1100-1102.
 Dhainaut, A., and Scaps, P. (2001). Immune defense and biological responses induced by toxics in Annelida. Can J Zool, 79, 233-253.
 Paul, V. J., and Puglisi, M. P. (2004). Chemical mediation of interactions among marine organisms. Nat Prod Rep, 21, 189-209.
 Kelly, S. R., Garo, E., Jensen, P. R., Fenical, W., and Pawlik, J. R. (2005). Effects of caribbean sponge secondary metabolites on bacterial surface colonization. Aquat Microb Ecol, 40, 191-203.