Abstract
An antibacterial metabolite was isolated from Paenibacillus polymyxa HKA-15, a soybean bacterial endophyte. The purification of the crude metabolite from Paenibacillus polymyxa HKA-15 was done by column chromatography. In TLC, a spot with an R f value of 0.86 (±0.02) from the purified fraction showed bioactivity against Xanthomonas campestris pv. phaseoli M-5. In SDS-PAGE, the purified antibiotic was separated in the molecular weight range of 3.5 kDa. The exact molecular weight of the active compound was identified as 1,347.7 Da using MS-MS analysis. Infra red spectrum and 1H NMR analysis showed the presence of amino acids and fatty acids in the active compound. The characterization of the antibacterial compound revealed its lipopeptide nature. In an agar diffusion assay, the crude metabolite showed a broad spectrum of activity, being able to inhibit the growth of the fungal pathogen, Rhizoctonia bataticola, Macrophomina phaseolina and Fusarium udum. A stronger inhibition was observed against bacterial pathogens viz., X. campestris pv.phaseoli M-5, X. campestris pv. phaseoli CP-1-1, Xanthomonas oryzae, Ralstonia solanacearum and Micrococcus luteus.








Similar content being viewed by others
Explore related subjects
Discover the latest articles and news from researchers in related subjects, suggested using machine learning.References
Beatty PH, Jensen SE (2002) Paenibacillus polymyxa produces fusaricidin-type antifungal antibiotics active against Leptosphaeria maculans, the causative agent of blackleg disease of canola. Can J Microbiol 48:159–169
Bizani D, Dominguez APM, Brandelli A (2005a) Purification and partial chemical characterization of the antimicrobial peptide cerein 8A. Lett Appl Microbiol 41:269–273
Bizani D, Motta A, Morrissy JAC, Terra RMS, Souto AA, Brandelli A (2005b) Antibacterial activity of cerein 8A, a bacteriocin like peptide produced by Bacillus cereus. Int Microbiol 8:125–131
Fontoura R, Spada JC, Silveira ST, Tsai SM, Brandelli A (2009) Purification and characterization of an antimicrobial peptide produced by Pseudomonas sp. strain 4B. World J Microbiol Biotechnol 25:205–213
Hyun JW, Kim YH, Lee YS, Park WM (1999) Isolation and Evaluation of protective effect against Fusarium wilt of sesame plants of antibiotic substance from Bacillus polymyxa KB-8. Plant Pathol J 15:152–157
Kajimura Y, Sugiyama M, Kaneda M (1995) Bacillopeptins, new cyclic lipopeptide antibiotics from Bacillus subtilis FR-2. J Antibiot 48:1095–1103
Kaufman E, Geisler N, Weber K (1984) SDS-PAGE strongly overestimates the molecular masses of the neurofialmental proteins. FEBS Lett 170:81–84
Kim HS, Park J, Choi SW, Choi KH, Lee GP, Ban SJ, Lee CH, Kim CS (2003) Isolation and Characterization of Bacillus strains for biological control. J Microbiol 41:196–201
Laemmli LK (1970) Cleavage of structural proteins during the assembly of head of bacteriophage T4. Nature 227:680
Lee H, Churey JJ, Worobo RW (2008) Purification and structural characterization of bacillomycin F produced by a bacterial honey isolate active against Byssochlamys fulva H25. J Appl Microbiol 105:663–673
Makkar RS, Cameotra SS (1999) Structural characterization of a biosurfactant produced by Bacillus sutilis at 45°C. J Surf Det 2:367–372
Martin NL, Hu H, Moake M, Churey JJ, Whittal R, Worobo RW, Vederas JC (2003) Isolation, structural characterization, and properties of mattacin (Polymyxin M), a cyclic peptide antibiotic produced by Paenibacillus kobensis M. J Bio Chem 278:13124–13132
Mhammedi A, Peypoux F, Besson F, Michel G (1982) Bacillomycin F, a new antibiotic of iturin group; isolation and characterization. J Antibiot 35:306–311
Motta AS, Lorenzini DM, Brandelli A (2007) Purification and partial characterization of an antimicrobial peptide produced by a novel Bacillus sp. isolated from the amazon basin. Curr Microbiol 54:282–286
Pichard B, Larue JP, Thouvenot D (1995) Gavesrin and saltavalin, new peptide antibiotics produced by Bacillus polymyxa. FEMS Microbiol Lett 133:215–218
Pueyo MT, Bloch CJ, Carmona RAM, Masico P (2009) Lipopeptides produced by a soil Bacillus megaterium strain. Microb Ecol 57:367–378
Raza W, Yang W, Shen QR (2008) Paenibacillus polymyxa: Antibiotics, hydrolytic enzymes and hazard management. J Plant Pathol 90:419–430
Safdi N, Cherif M, Hajlaui MR, Boudabbous A, Belanger R (2002) Isolation and partial purification of antifungal metabolites produced by Bacillus cereus. Ann Microbiol 52:323–337
Selim S, Negrel J, Govaerts C, Gianinazzi S, Tuinen DV (2005) Isolation and partial characterization of antagonistic peptides produced by Paenibacillus sp. Strain B2 isolated from the sorghum mycorhizosphere. Appl Environ Microbiol 71:6501–6507
Senthilkumar M, Govindasamy V, Annapurna K (2007a) Role of antibiosis in suppression of charcoal rot disease by soybean endophyte Paenibacillus sp. HKA-15. Curr Microbiol 55:25–29
Senthilkumar M, Govindasamy V, Dureja P, Annapurna K (2007b) Purification and partial characterization of antifungal peptides from soybean endophyte-Paenibacillus sp strain HKA-15. J Plant Biochem Biotech 16:131–134
Shufie EHA, Ibrahim N, El-Sabour HA, Mostafa YA (2008) Purification and Characterization of bacteriocin produced by isolated strain of Lactococcus lactis. J Appl Sci Res 4:1315–1321
Tamehiro N, Okamoto HY, Okamoto S, Ubukata M, Hamada M, Naganawa H, Ochi K (2002) Bacilysocin, a Novel Phospholipid antibiotic produced by Bacillus subtilis 168. Antimicrob Agents Chemother 46:315–320
Tendulkar SR, Saikumar YK, Patel V, Raghotama S, Munshi TK, Balaram P, Chatoo BB (2007) Isolation, purification and characterization of an antifungal molecule produced by Bacillus licheniformis BC98, and its effect on phytopathogen Magnoporthe grisea. J Appl Microbiol 103:2331–2339
Weid VD, Alviano DS, Santos ALS, Soares RMA, Alviano CX, Seldin L (2003) Antimicrobial activity of Paenibacillus peoriae strain NRRL BD-62 against a broad spectrum of phytopathogenic bacteria and fungi. J Appl Microbiol 95:1143–1151
Wu S, Jia S, Sun D, Chen M, Chen X, Zhong J, Huan L (2005) Purification and characterization of two novel antimicrobial peptides subpeptin JM4-A and subpeptin JM4-B produced by Bacillus subtilis JM4. Curr Microbiol 51:292–296
Yoshida S, Hiradate S, Tsukamot T, Hatakeda K, Shirata A (2000) Antimicrobial activity of culture filtrate of Bacillus amyloliquefaciens RC-2 isolated from mulberry leaves. Biocontrol 91:181–187
Zanatta ZGCN, Moura AB, Maia LC, Santos AS (2007) Bioassay for selection of biocontroller bacteria against bean common blight (Xanthomonas axonopodis pv. phaseoli). Braz J Microbiol 38:511–515
Acknowledgments
The first author thanks the Jawaharlal Nehru Memorial Fund, New Delhi, India for the financial assistance in the form of PhD scholarship for this study.
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Mageshwaran, V., Walia, S. & Annapurna, K. Isolation and partial characterization of antibacterial lipopeptide produced by Paenibacillus polymyxa HKA-15 against phytopathogen Xanthomonas campestris pv. phaseoli M-5. World J Microbiol Biotechnol 28, 909–917 (2012). https://doi.org/10.1007/s11274-011-0888-y
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s11274-011-0888-y


