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Purification and Characterization of Antibacterial Compound Produced by Bacillus subtilis MJP1  

Yim, Eun-Jung (Department of Food and Nutrition, Chosun University)
Yang, Eun-Ju (Department of Food and Nutrition, Chosun University)
Chang, Hae-Choon (Department of Food and Nutrition, Chosun University)
Publication Information
Microbiology and Biotechnology Letters / v.38, no.1, 2010 , pp. 84-92 More about this Journal
Abstract
Antibacterial compound from Bacillus subtilis MJP1 was purified using C18 Sep-Pak cartridge, ion exchange chromatography, and gel filtration chromatography. The purified antibacterial compound showed antibacterial activity against Listeria monocytogenes, Bacillus subtilis, Staphylococcus aureus subsp. aureus, and Enterococcus faecalis. The purified antibacterial compound was found to be stable at $100^{\circ}C$ for 5 min and in the pH range of 3.0~9.0, but it was unstable at pH 10.0. It was inactivated by proteinase K and pronase E, and heat treatment at $121^{\circ}C$ for 15 min, but it was stable with lipase and $\alpha$-amylase treatment, which indicated its proteineous nature. Ultra performance liquid chromatography and electrospray ionization tandem mass spectrometry analysis were used to identify the purified antibacterial compound and confirmed the existence of two peptides (3356.54 Da, 3400.5244 Da).
Keywords
Bacillus subtilis; antibacterial activity; peptide; purification;
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1 Babasaki, K., T. Takao, Y. Shimonishi, and K. Kurahashi. 1985. Subtilosin A, a new antibiotic peptide produced by Bacillus subtilis 168: isolation, structural analysis, and biogenesis. J. Biochem.(Tokyo). 98: 585-603.
2 Bradford, M. M. 1976. A rapid and sensitive method quantities of protein utilizing the principle of protein-dye binding. Anal. Biochem. 72: 248-254.   DOI   ScienceOn
3 Daeschel, M. A. 1989. Antimicrobial substances from lactic acid bacteria for use as food preservatives. Food Technol. 43: 164-167.
4 Smith, B. J. 1994. Chemical cleavage of proteins. Methods Mol. Biol. 32: 297-309.
5 Von Döhren, H. 1995. Peptides, pp. 129-171. In L. C. Vining and C. Stuttard. (eds.), Genetics and Biochemistry of Antibiotic Production. Butterworth-Heinemann, Newton, M.A.
6 Zheng, G., L. Z. Yan, J. C. Vederas, and P. Zuber. 1999. Genes of the sbo-alb locus of Bacillus subtilis are required for production of the antilisterial bacteriocin subtilosin. J. Bacteriol. 181: 7346-7355.
7 Hoover, D. G. and S. K. Harlander. 1993. Screening methods for detecting bacteriocin activity, pp. 23-39. In D. G. Hoover and L. R. Steenson. (eds.), Bacteriocins of Lactic Acid Bacteria. Academic Press, Inc., San Diego, U.S.A.
8 Oscariz, J. C., L. Cinatas, H. Holo, I. Lasa, I. F. Nes, and A. G. Pisabarro. 2006. Purification and sequencing of cerein 7B, a novel bacteriocin produced by Bacillus cereus Bc7. FEMS Microbiol. Lett. 254: 108-115.   DOI   ScienceOn
9 Paik, S. H., A. Chakicherla, and J. N. Hansen. 1998. Identification and characterization of the structural and transporter genes for, and the chemical and biological properties of, sublancin 168, a novel lantibiotic produced by Bacillus subtilis 168. J. Biol. Chem. 273: 23134-23142.   DOI
10 Jansen, E. F. and D. J. Hirschmann. 1944. Subtilin-an antibacterial product of Bacillus subtilis: culturing conditions and properties. Arch. Biochem. 4: 297-309.
11 Tagg, J. R. and A. R. Mcgiven. 1971. Assay system for bacteriocins. Appl. Microbiol. 21: 943.
12 Paik, H. D., S. S. Bae, S. H. Park, and J. G. Pan. 1997. Identification and partial characterization of tochicin, a bacteriocin produced by Bacillus thuringiensis subsp. tochigiensis. J. Ind. Microbiol. Biotechnol. 19: 294-298.   DOI   ScienceOn
13 Farries, T. C., A. Harris, A. D. Auffret, and A. Aitken. 1991. Removal of N-acetyl groups from blocked peptides with acylpeptide hydrolase: Stabilization of the enzyme and its application to protein sequencing. Eur. J. Biochem. 196: 679-685.   DOI   ScienceOn
14 Cherif, A., S. Chehimi, F. Limem, B. M. Hansen, N. B. Hendriksen, D. Daffonchio, and A boudabos. 2003. Detection and characterization of the novel bacteriocin entomocin 9, and safety evaluation of its producer, Bacillus thuringiensis ssp. entomocidus HD9. J. Appl. Microbiol. 95: 990-1000.   DOI   ScienceOn
15 Piard, J. C. and M. Desmazeaud. 1992. Inhibiting factors produced by lactic acid bacteria. 2. Bacteriocins and other antimicrobial factors. Dairy. Sci. Tech. 72: 113-142.   DOI
16 Uyangaa, T. 2007. Studies on the culture medium for large production of spores of Bacillus subtilis and purification of antibacterial activity of Bacillus sp. strain. (PhD dissertation). An seoung: Hangkyoung University.
17 Delves-Broughton, J. 1990. Nisin and its uses as a food preservative. Food Technol. 44: 100-117.
18 Schagger, H. and G. von Jagow. 1987. Tricine-sodium dodecyl sulfate-polyacrylamide gel electrophoresis for the separation of proteins in the range from 1 to 100 kDa. Anal. Biochem. 166: 368-379.   DOI   ScienceOn
19 Cherif, A., H. Ouzari, D. Daffonchio, H. Cherif, K. Ben Slama, A. Hassen, S. Jaoua, and A. Boudabous. 2001. Thuricin 7: a novel bacteriocin produced by Bacillus thuringiensis BMG1.7, a new strain isolated from soil. Lett. Appl. Microbiol. 32: 243-247.   DOI   ScienceOn
20 Jack R. W., J. R. Tagg, and B. Ray. 1995. Bacteriocins of gram-positive bacteria. Microbiol. Rev. 59: 171-200.
21 Schallmey, M., A. Singh, and O. P. Ward. 2004. Developments in the use of Bacillus species for industrial production. Can. J. Microbiol. 50: 1-17.   DOI   ScienceOn
22 Yang, E. J. and H. C. Chang. 2007. Characterization of bacteriocin-like substances produced by Bacillus subtilis MJP1. Kor. J. Microbiol. Biotechnol. 35: 339-346.
23 Tagg, J. R., A. S. Dajani, and L. W. Wannamarker. 1976. Bacteriocin of Gram-positive bacteria. Bacteriol. Rev. 40: 722-756.
24 Oscariz, J. C., I. Lasa, and A. G. Pisabarro. 1999. Detection and characterization of cerein 7, a new bacteriocin produced by Bacillus cereus with a broad spectrum of activity. FEMS Microbiol. Lett. 178: 337-341.
25 Bizani, D. and A. Brandelli. 2002. Characterization of a bacteriocin produced by a newly isolated Bacillus sp. strain 8A. J. Appl. Microbiol. 93: 512-519.   DOI   ScienceOn
26 Jiraphocakul, S., T. W. Sulivan, and K. M. Shahani. 1990. Influence of a dried Bacillus subtilis culture and antibiotics on performance and intestinal microflora in turkeys. Poult. Sci. 69: 1966-1973.   DOI   ScienceOn
27 Meyer, H. E., M. Heber, B. Eisermann, H. Korte, J. W. Metzger, and G. Jung. 1994. Sequence analysis of lantibiotics: chemical derivatization procedures allow a fast access to complete Edman degradation. Anal. Biochem. 223: 185-190.   DOI   ScienceOn
28 Niall. H. D. 1973. Automated Edman degradation: The protein sequenator. Methods Enzymol. 27: 942-1010.   DOI