Properties of a Bacteriocin Produced by Bacillus subtilis EMD4 Isolated from Ganjang (Soy Sauce)
4
조회
0
다운로드
4
인용
0
참고문헌
키워드
초록
A Bacillus species, EMD4, with strong antibacterial activity was isolated from ganjang (soy sauce) and identified as B. subtilis. B. subtilis EMD4 strongly inhibited the growth of B. cereus ATCC14579 and B. thuringiensis ATCC33679. The antibacterial activity was stable at pH 3-9 but inactive at pH 10 and above. The activity was fully retained after 15 min at 80°C but reduced by 50% after 15 min at 90°C. The activity was completely destroyed by proteinase K and protease treatment, indicating its proteinaceous nature. The bacteriocin (BacEMD4) was partially purified from culture supernatant by ammonium sulfate precipitation, and QSepharose and Sephadex G-50 column chromatographies. The specific activity was increased from 769.2 AU/mg protein to 8,347.8 AU/mg protein and the final yield was 12.6%. The size of BacEMD4 was determined to be 3.5 kDa by Tricine SDS-PAGE. The N-terminal amino acid sequence was similar with that of Subtilosin A. Nucleotide sequencing of the cloned gene confirmed that BacEMD4 was Subtilosin A. BacEMD4 showed bactericidal activity against B. cereus ATCC14579.
Abstract
A Bacillus species, EMD4, with strong antibacterial activity was isolated from ganjang (soy sauce) and identified as B. subtilis. B. subtilis EMD4 strongly inhibited the growth of B. cereus ATCC14579 and B. thuringiensis ATCC33679. The antibacterial activity was stable at pH 3-9 but inactive at pH 10 and above. The activity was fully retained after 15 min at 80°C but reduced by 50% after 15 min at 90°C. The activity was completely destroyed by proteinase K and protease treatment, indicating its proteinaceous nature. The bacteriocin (BacEMD4) was partially purified from culture supernatant by ammonium sulfate precipitation, and QSepharose and Sephadex G-50 column chromatographies. The specific activity was increased from 769.2 AU/mg protein to 8,347.8 AU/mg protein and the final yield was 12.6%. The size of BacEMD4 was determined to be 3.5 kDa by Tricine SDS-PAGE. The N-terminal amino acid sequence was similar with that of Subtilosin A. Nucleotide sequencing of the cloned gene confirmed that BacEMD4 was Subtilosin A. BacEMD4 showed bactericidal activity against B. cereus ATCC14579.
참고문헌 (28)
- Abdel-Mohsein H, "Isolation of bacteriocin-like substances producing bacteria from finished cattle-manure compost and activity evaluation against some food-borne pathogenic and spoilage bacteria", J. Gen. Appl. Microbiol, 56, pp.151~161 (2010)
- Babasaki K, "Subtilosin A, a new antibiotic peptide produced by Bacillus subtilis 168: isolation, structural analysis, and biogenesis", J. Biochem, 98, pp.585~603 (1985)
- Bizani D, "Characterization of a bacteriocin produced by a newly isolated Bacillus sp. strain 8 A", J. Appl. Microbiol, 93, pp.512~519 (2002)
- Bradford MM, "Rapid and sensitive methods for the quantification of microgram quantities of protein utilizing the principle of protein-dye binding", Anal. Biochem, 72, pp.248~254 (1976)
- Cherif A, "Characterization and partial purification of entomocin 110, a newly identified bacteriocin from Bacillus thuringiensis subsp. Entomocidus HD110", Microbiol. Res, 163, pp.684~692 (2008)
- Cintas LM, "Comparative antimicrobial activity of enterocin L50, pediocin PA-1, nisin A and lactocin S against spoilage and foodborne pathogenic bacteria", Food Microbiol, 15, pp.289~298 (1998)
- Cleveland J, "Bacteriocins: safe, natural antimicrobials for food preservation", Int. J. Food Microbiol, 71, pp.1~20 (2001)
- Gálvez A, "Application of bacteriocins in the control of foodborne pathogenic and spoilage bacteria", Crit. Rev. Biotechnol, 28, pp.125~152 (2008)
- Garg N, "Lantibiotics from Geobacillus thermodenitrificans", Proc. Natl. Acad. Sci, 109, pp.5241~5246
- Gray EJ, "A novel bacteriocin, thuricin 17, produced by plant growth promoting rhizobacteria strain Bacillus thuringiensis NEB17: isolation and classification", J. Appl. Microbiol, 100, pp.545~554 (2006)
- Hyronimus B, "Coagulin, a bacteriocin-like inhibitory substance produced by Bacillus coagulans I4", J. Appl. Microbiol, 85, pp.42~50 (1998)
- Kawulka KE, "Structure of subtilosin A, a cyclic antimicrobial peptide from Bacillus subtilis with unusual sulfur to R-carbon cross-links: formation and reduction of R-thio-R-amino acid derivatives", Biochemistry, 43, pp.3385~3395 (2004)
- Kamoun F, "Purification, amino acid sequence and characterization of Bacthuricin F4, a new bacteriocin produced by Bacillus thuringiensis", J. Appl. Microbiol, 98, pp.881~888 (2005)
- Salum Kindoli, "Properties of a Bacteriocin from Bacillus subtilis H27 Isolated from Cheonggukjang", Food Science and Biotechnology, 21(6), pp.1745~1751 (2012), 한국식품과학회[DOI]
- Kindoli, Salum, "Properties of Bac W42, a Bacteriocin Produced by Bacillus subtilis W42 Isolated from Cheonggukjang", Journal of Microbiology and Biotechnology, 22(8), pp.1092~1100 (2012), 한국미생물·생명공학회[DOI]
- Klaenhammer TR, "Genetics of bacteriocins produced by lactic acid bacteria", FEMS Microbiol. Rev, 12, pp.39~85 (1993)
- Oscáriz JC, "Detec tion and characterization of cerein 7, a new bacteriocin produced by Bacillus cereus with a broad spectrum of activity", FEMS Microbiol. Lett, 178, pp.337~341 (1999)
- O’Sullivan L, "Generation of food-grade lactococcal starters which produce the lantibiotics lacticin 3147 and lacticin 481", Appl. Environ. Microbiol, 69, pp.3681~3685 (2003)
- Riley MA, "Bacteriocins: evolution, ecology and application", Annu. Rev. Microbiol, 56, pp.117~137 (2002)
- Sabaté DC, "Inhibitory activity of surfactin, produced by different Bacillus subtilis subsp. subtilis strains, against Listeria monocytogenes sensitive and bacteriocinresistant strains", Microbiol. Res, 168, pp.125~129 (2013)
- Sambrook J, "Molecular Cloning: A Laboratory Manual" (2001), Cold Spring Harbor Laboratory Press
- Schägger H, "Tricine-sodium dodecyl sulphate polyacrylamide gel electrophoresis for the separation of protein in the range from 1 to 100 kDa", Anal. Biochem, 166, pp.368~379 (1987)
- Senbagam D, "Physical chemical and biological characterization of a new bacteriocin produced by Bacillus cereus NS02", Asian Pac. J. Trop. Med, 6, pp.934~941 (2013)
- Settanni L, "Application of bacteriocins in vegetable food biopreservation", Int. J. Food Microbiol, 121, pp.123~138 (2008)
- Sobrino-López A, "Use of nisin and other bacteriocins for preservation of dairy products", Int. Dairy J, 18, pp.329~343 (2007)
- Stein T., "Bacillus subtilis antibiotics: structures, syntheses and specific functions", Mol. Microbiol, 56, pp.845~857 (2005)
- Stein T, "Subtilosin production by two Bacillus subtilis subspecies and variance of the sbo-alb cluster", Appl. Environ. Microbiol, 70, pp.2349~2353 (2004)
- Vuyst LD, "Bacteriocins of Lactic Acid Bacteria. Microbiology, Genetics and Applications", pp.151~221 (1994), Chapman & Hall