Skip to main content
PMC home page
Food Science and Biotechnology logoLink to Food Science and Biotechnology
. 2016 Apr 30;25(2):525–532. doi: 10.1007/s10068-016-0073-z

Antimicrobial activity of Bacillus amyloliquefaciens EMD17 isolated from Cheonggukjang and potential use as a starter for fermented soy foods

Jae Yong Lee 1, Jae Min Shim 1, Zhuang Yao 1, Xiaoming Liu 1, Kang Wook Lee 1, Hyun-Jin Kim 1,3, Kyung-Sik Ham 2, Jeong Hwan Kim 1,3,✉
PMCID: PMC6049179  PMID: 30263301

Abstract

A Bacillus strain with antimicrobial activity was isolated from Cheonggukjang and identified as Bacillus amyloliquefaciens. Filtered culture supernatants of B. amyloliquefaciens EMD17 strongly inhibited growth of Bacillus cereus ATCC14579, Listeria monocytogenes ATCC19111, and an ochratoxin (OTA) producing Penicillium spp. The antimicrobial activity was not decreased by heat, pH, and proteases treatment, indicating a non-proteinous nature of the antimicrobial substance. A B. amyloliquefaciens EMD17 culture added to a B. cereus ATCC14579 culture killed B. cereus cells completely in 6 h, showing the bacteriocidal effect. The srfAA gene encoding surfactin synthetase A was detected using PCR, indicating that surfactin might be the responsible agent. Genes encoding fengycin and iturin were also detected. MS indicated production of lipopeptides, including surfactin. Cheonggukjang and Doenjang were prepared using B. amyloliquefaciens EMD17 as a starter and growth of spiked B. cereus cells was completely inhibited.

Keywords: Bacillus amyloliquefaciens, antimicrobial activity, lipopeptide, surfactin, starter

References

  • 1.Lee C-H, Youn Y, Song G-S, Kim YS. Immunostimulatory effects of traditional Doenjang. J. Korean Soc. Food Sci. Nutr. 2011;40:1227–1234. doi: 10.3746/jkfn.2011.40.9.1227. [DOI] [Google Scholar]
  • 2.Oh Y-S, Park J-E, Oh H-J, Kim JH, Oh M-C, Oh C-K, Oh Y-J, Lim S-B. Isolation and characteristics of microorganisms producing extracellular enzymes from Jeju traditional fermented soybean paste (Doenjang) J. Korean Soc. Food Sci. Nutr. 2010;39:47–53. doi: 10.3746/jkfn.2010.39.1.047. [DOI] [Google Scholar]
  • 3.Yoon KH, Shin HY. Medium optimization for the protease production by Bacillus licheniformis isolated from Cheonggukjang. Microbiol. Biotechnol. Lett. 2010;38:385–390. [Google Scholar]
  • 4.Gálvez A, López RL, Abriouel H. Application of bacteriocins in the control of foodborne pathogenic and spoilage bacteria. Crit. Rev. Biotechnol. 2008;28:125–152. doi: 10.1080/07388550802107202. [DOI] [PubMed] [Google Scholar]
  • 5.Settanni L, Corsetti A. Application of bacteriocins in vegetable food biopreservation. Int. J. Food Microbiol. 2008;121:123–138. doi: 10.1016/j.ijfoodmicro.2007.09.001. [DOI] [PubMed] [Google Scholar]
  • 6.Cha HA, Chung D, Hong S, Chung KS. Purification and characterization of an antimicrobial substance from Bacillus subtilis HH28 antagonistic to Bacillus cereus. Microbiol. Biotechnol. Lett. 2014;42:393–401. [Google Scholar]
  • 7.Kim YS, Yun SH, Jeong DY, Hahn K U-B. Isolation of Bacillus licheniformis producing antimicrobial agentsagainst Bacillus cereus and its properties. Korean J. Microbiol. 2010;46:270–277. [Google Scholar]
  • 8.Salum K, Lee HA, Kim JH. Properties of Bac W42, a bacteriocin produced by Bacillus subtilis W42 isolated from Cheonggukjang. J. Microbiol. Biotechnol. 2012;22:1092–1100. doi: 10.4014/jmb.1110.10002. [DOI] [PubMed] [Google Scholar]
  • 9.Yang EJ, Chang HC. Characterization of bacteriocin-like substances produced by Bacillus subtilis MJP1. Microbiol. Biotechnol. Lett. 2007;35:339–346. [Google Scholar]
  • 10.Heo K, Cho KM, Lee CK, Kim GM, Shin JH, Kim JS, Kim JH. Characterization of a fibrinolytic enzyme secreted by Bacillus amyloliquefaciens CB1 and its gene cloning. J. Microbiol. Biotechnol. 2013;23:974–983. doi: 10.4014/jmb.1302.02065. [DOI] [PubMed] [Google Scholar]
  • 11.Kim GM, Lee AR, Lee KW, Park JY, Chun J, Cha J, Song Y-S, Kim JH. Characterization of a 27 kDa fibrinolytic enzyme from Bacillus amyloliquefaciens CH51 isolated from Cheonggukjang. J. Microbiol. Biotechnol. 2009;19:997–1004. doi: 10.4014/jmb.0811.600. [DOI] [PubMed] [Google Scholar]
  • 12.Meerak J, Iida H, Watanabe Y, Miyashita M, Sato H, Nakagawa Y, Tahara Y. Phylogeny of g-polyglutamic acid-producing Bacillus strains isolated from fermented soybean foods manufactured in Asian countries. J. Gen. Appl. Microbiol. 2007;53:315–323. doi: 10.2323/jgam.53.315. [DOI] [PubMed] [Google Scholar]
  • 13.Park C, Choi Y-H, Shin H-J, Poo H, Song JJ, Kim C-J, Sung M-H. Effect of high molecular-weight poly-g-glutamic acid from Bacillus subtilis (Cheonggukjang) on Ca solubility and intestinal absorption. J. Microbiol. Biotechnol. 2005;15:855–858. [Google Scholar]
  • 14.Hoover DG, Harlander SK. Screening methods for detecting bacteriocin activity. In: Hoover DG, Steenson LR, editors. Bacteriocins of Lactic Acid Bacteria. San Diego, CA, USA: Academic Press; 1993. pp. 23–39. [Google Scholar]
  • 15.Kwon GH, Lee HA, Park JY, Kim JS, Lim J, Park CS, Kwon DY, Kim YS, Kim JH. Development of a RAPD-PCR method for identification of Bacillus species isolated from Chunggukjang. Int. J. Food Microbiol. 2009;129:282–287. doi: 10.1016/j.ijfoodmicro.2008.12.013. [DOI] [PubMed] [Google Scholar]
  • 16.Lee HA, Kim JH. Isolation of Bacillus amyloliquefaciens strains with antifungal activities from Meju. Prev. Nutr. Food. Sci. 2012;17:64–70. doi: 10.3746/pnf.2012.17.1.064. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 17.Salum K, Lee HA, Heo K, Kim JH. Properties of a bacteriocin from Bacillus subtilis H27 isolated from Cheonggukjang. Food Sci. Biotechnol. 2012;21:1745–1751. doi: 10.1007/s10068-012-0232-9. [DOI] [Google Scholar]
  • 18.Cho MJ, Lee JY, Lee KW, Cho KM, Lee CK, Kim GM, Shin JH, Kim JS, Kim HJ, Kim JH. Properties of Doenjang (Soybean Paste) fermented with multiple starters. J. Agric. Life Sci. 2014;48:291–300. doi: 10.14397/jals.2014.48.6.291. [DOI] [Google Scholar]
  • 19.Lee AR, Kim GM, Park JY, Jo HD, Cha J, Song YS, Chun J, Kim JH. Characterization of a 27 kDa fibrinolytic enzyme from Bacillus amyloliquefaciens CH86-1. J. Korean Soc. Appl. Biol. Chem. 2010;53:56–61. doi: 10.3839/jksabc.2010.010. [DOI] [Google Scholar]
  • 20.Yamamoto S, Shiraishi S, Suzuki S. Are cyclic lipopeptides produced by Bacillus amyloliquefaciens S13-3 responsible for the plant defence response in strawberry against Colletotrichum gloeosporioides? Lett. Appl. Microbiol. 2015;60:379–386. doi: 10.1111/lam.12382. [DOI] [PubMed] [Google Scholar]
  • 21.Meena KR, Kanwar SS. Lipopeptides as the antifungal and antibacterial agents: Applications in food safety and therapeutics. 2015. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 22.Stein T. Bacillus subtilis antibiotics: Structures, syntheses and specific functions. Mol. Microbiol. 2005;56:845–857. doi: 10.1111/j.1365-2958.2005.04587.x. [DOI] [PubMed] [Google Scholar]
  • 23.Mora I, Cabrefiga J, Montesinos E. Antimicrobial peptide genes in Bacillus strains from plant environments. Int. Microbiol. 2011;14:213–223. doi: 10.2436/20.1501.01.151. [DOI] [PubMed] [Google Scholar]
  • 24.Tapi A, Chollet-Imbert M, Scherens B, Jacques P. New approach for the detection of non-ribosomal peptide synthetase genes in Bacillus strains by polymerase chain reaction. Appl. Microbiol. Biotech. 2010;85:1521–1531. doi: 10.1007/s00253-009-2176-4. [DOI] [PubMed] [Google Scholar]
  • 25.Chung SH, Kong HS, Buyer JS, Lakshman DK, Lydon J, Kim SD, Roberts DP. Isolation and partial characterization of Bacillus subtilis ME488 for suppression of soilborne pathogens of cucumber and pepper. Appl. Microbiol. Biotechnol. 2008;80:115–123. doi: 10.1007/s00253-008-1520-4. [DOI] [PubMed] [Google Scholar]
  • 26.Mikkola R, Kolari M, Andersson MA, Helin J, Salkinoja-Salonen MS. Toxic lactonic lipopeptide from food poisoning isolates of Bacillus licheniformis. Eur. J. Biochem. 2000;267:4068–4074. doi: 10.1046/j.1432-1033.2000.01467.x. [DOI] [PubMed] [Google Scholar]
  • 27.Cho MJ, Lee JY, Kim JH. Microbial and physiochemical properties of Cheonggukjang fermented using Bacillus strains with antibacterial or antifungal activities. Food Sci. Biotechnol. 2014;23:1525–1532. doi: 10.1007/s10068-014-0208-z. [DOI] [Google Scholar]

Articles from Food Science and Biotechnology are provided here courtesy of Springer

RESOURCES