Iranian Journal of Microbiology 2011. 3(2):104-108.

A simple method for primary screening of antibacterial peptides in plant seeds
A Aliahmadi, R Roghanian, G Emtiazi, A Ghassempour

Abstract


Background and Objectives: Regarding the importance of finding new antibacterial drugs, screening of plants as a promising resource are now conducted worldwide. In this study, we report the application of a simple previously described method for screening of different plant seeds in order to find the best resources of plant antimicrobial peptides.
Materials and Methods: Total water soluble protein of 10 different plant seeds were extracted and subjected to SDS-PAGE and subsequent agar-overlay bioassays. Standard strains of Staphylococcus aureus, Enterococcus faecium and Escherichia coli were included in the bioassays. This method also was used for total proteins precipitated by Ammonium sulphate which ensure the protein nature of the test substances. Molecular size and the amounts of effective peptides were estimated using Tricin-SDS-PAGE and densitometry.
Results: Two different plant seeds showed noticeable antibacterial activities against tested Gram positive bacteria and a moderate inhibitory effect on Gram negative ones. Based on the results of Tricin-SDS-PAGE analysis which were carried out in parallel to bioassays, it was concluded that effective antibacterial substances are peptides with molecular weight of slightly larger than 5 kDa.
Conclusion: On the basis of results of agar-overlay experiments and by screening of 10 different herbal seeds, we could introduce seeds of M. sativa L. and Onobrychis sativa Lam., as great sources of putative plant antibacterial peptides. The proposed screening method can be used for screening of large number of different plant seeds and even other parts of the plant body, regarding some necessary modification in total water soluble protein extraction steps.


Keywords


Plant antibacterial peptides; Staphylococcus aureus; Enterococcus faecium; screening method

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References


Wisplinghoff H, Bischoff T, Tallent SM, Seifert H, Wenzel RP, Edmond MB. Nosocomial infections in US hospitals: analysis of 24,179 cases from a prospective nationwide surveillance study. Clin Infect Dis 2004; 39:309-317.

Cowan MM. Plant products as antimicrobial agents.Clin Microbiol Rev. 1999; 12: 564-582.

Portieles R, Ayra C, Borras O. Basic insight on plant defensins. Biotecnologia Aplicada 2006; 23: 75-78.

Thevissen K, Kristensen H, Thomma BPH J, Cammue BPA, Francois IEJA. Therapeutic potential of antifungal plants and insect defensins. Drug Discov Today 2007;12(21/22): 966-972.

Peschel A, Sahl HG. The co-evolution of host cationic antimicrobial peptides and microbial resistance. Nat Rev Microbiol 2006; 4: 529-536.

Lay TF, Brugliera F, Anderson AM. Isolation and properties of floral defensins from ornamental tobacco and petunia. Plant Physiol 2003; 131: 1283-1293.

Odintsova TI, Egorov TA, Musolyamov AK, Odintsova MS, Pukhalsky VA, Grishin EV. Seed defensins from T. Kiharae and related species: Genome localization of defensin-encoding genes. Biochemie 2007; 89: 605-612.

Wang HX, Ng TB. An antifungal peptide from red lentil seeds. Peptides 2007; 28: 547-552.

Ko S-K, Ahn C. Bactriocin production by Lactococcus lactis KCA2386 isolated from white kimchi. Food Sci Biotechnol 2000; 9: 263-296.

Bradford, MM. Rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 1976; 72: 248-254.

Schagger H, Von Jagow G. Tricine-sodium dodecyl sulfate-polyacrylamide gel electrophoresis for the separation of proteins in the range from 1 to 100kDa. Anal Biochem 1987; 166: 368-379.

Tavares LS, Santos MO, Viccini, LF, Moreira JS, Miller RNG, Franco OL. Biotechnological potential of antimicrobial peptides from flowers. Peptides 2008; 29:1842-1851.

Finkina EI, Shramova EI, Tagaev AA, Ovchinnikova TV. A novel defensins from the lentil Lens culinaris seeds. Biochem Biophys Res Commun. 2008; 371: 860-865.

Wong JH, Ng, TB. Sesquin, a potent defensin-like antimicrobial peptide from ground beans with inhibitory activities toward tumor cells and HIV-1 reverse transcriptase. Peptides 2005; 26: 1120–1126.

Gao A, Hakimi SM, Mittanck CA, Wu Y, Woerner MB, Stark DM, et al. Fungal pathogen protection in potato by expression of a plant defensin peptide. Nat Biotechnol 2000; 18: 1307-1310.

Avato P, Bucci R, Tava A, Vitali C, Rosato A, Bialy B, et al. Antimicrobial activity of saponins from Medicago sp.: Structure-activity relationship. Phytother Res 2006; 20: 454-457.

Emmert EAB, Milner JL, Lee JC, Pulvermacher KL, Olivares HA, Clardy J, et al. Effect of canavanine from alfaalfa seeds on the population biology of Bacillus cereus. Appl Environ Microbiol 1988; 64: 4683-4688.


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