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Isolation and Characterization of a Bacteriocin-Producing Lactobacillus sakei B16 from Kimchi

김치에서 박테리오신을 생산하는 Lactobacillus sakei B16의 분리 및 특성 분석

  • Ahn, Ji-Eun (Dept. of Food Science & Technology, Chungbuk National University) ;
  • Kim, Jin-Kyoung (Dept. of Food Science & Technology, Chungbuk National University) ;
  • Lee, Hyeong-Rho (Dept. of Agricultural Biotechnology, Seoul National University) ;
  • Eom, Hyun-Ju (Dept. of Food Science & Technology, Chungbuk National University) ;
  • Han, Nam-Soo (Dept. of Food Science & Technology, Chungbuk National University)
  • Received : 2012.02.09
  • Accepted : 2012.03.05
  • Published : 2012.05.31

Abstract

Lactic acid bacteria (LAB) are able to secrete antimicrobial peptides called bacteriocins, which inhibit other bacteria such as pathogenic microorganisms. Therefore, bacteriocin-producing starters can be used as natural biopreservatives for various foods. The objective of this study was to screen and characterize bacteriocin-producing LAB from Kimchi and to investigate their applicability as a starter in Kimchi fermentation. To screen bacteriocin-producing LAB, gram-positive and gram-negative bacteria were used as indicators. To measure the antimicrobial activities of isolates, agar well diffusion assay method was used. According to the results, bacteriocin produced by $Lb.$ $sakei$ B16 showed antimicrobial activity against $Listeria$ $monocytogenes$ ATCC 19115, $Escherichia$ $coli$ KCTC 1467, and$Lactobacillus$ $plantarum$ KTCT 3104. Furthermore, bacteriocin was very stable after treatment with high temperature and high and low pH, but its effects were inhibited by treatment with proteolytic enzymes such as trypsin, proteinase K, and ${\alpha}$-chymotrypsin, revealing their bacteriocin-like protein- based structure. These results suggest that $Lb.$ $sakei$ B16 and its bacteriocin are good candidates as a functional probiotic and natural biopreservative, respectively, in fermented foods.

본 연구는 김치 내에서 균들의 생육을 조절하고 김치의 저장성과 안전성을 향상시키고자 김치로부터 항균활성이 우수한 박테리오신 생산 유산균을 동정 및 박테리오신의 특성을 확인하고자 하였다. 그 결과 항균활성이 뛰어난 $Lb.$ $sakei$ B16을 선발하였고 이 균주에서 생산되는 박테리오신은 pH와 열에 안정하고 일반적으로 박테리오신이 그람 양성균을 저해하는 것에 비해 본 연구에서 분리한 박테리오신은 그람 음성 유해균을 저해하여 넓은 저해 범위를 갖는 장점이 있다. 반면, trypsin, proteinase K, ${\alpha}$-chymotrypsin을 처리하였을 때 항균 활성이 저하되는 것을 확인하였다. 따라서 본 연구에서 분리한 박테리오신은 다양한 환경 변화에서 안정하고 체내에 섭취되었을 때 단백질 가수분해 효소에 의해 분해됨으로써 인체에 영향을 주지 않을 것으로 생각된다. 또한 sakacin P cluster 염기서열 정보를 이용하여 $Lb.$ $sakei$ B16에서 박테리오신 유전자의 존재를 확인하였다. 본 연구를 통해 유산균이 생산하는 박테리오신을 이용하여 김치 제조 산업에서 스타터로 개발하여 김치 내에서 미생물의 생육을 조절하고 김치의 저장성과 품질을 향상시킬 수 있을 것이라 기대된다.

Keywords

References

  1. Kim HK, Lee KH, Kim JH. 2004. Bacteriocins produced by lactic acid bacteria isolated from Kimchi. J Agric Life Sci 38: 15-24.
  2. Zendo T, Yoneyama F, Sonomoto K. 2010. Lactococcal membrane-permeabilizing antimicrobial peptides. Appl Microbiol Biotechnol 88: 1-9. https://doi.org/10.1007/s00253-010-2764-3
  3. Nissen-Meyer J, Oppegard C, Rogne P, Haugen HS, Kristiansen PE. 2010. Structure and mode-of-action of the two-peptide (Class-IIb) bacteriocins. Probiotics Antimicro Prot 2: 52-60. https://doi.org/10.1007/s12602-009-9021-z
  4. Cleveland J, Montville TJ, Nes IF, Chikindas ML. 2001. Bacteriocins: safe, natural antimicrobials for food preservation. Int J Food Microbiol 71: 1-20. https://doi.org/10.1016/S0168-1605(01)00560-8
  5. Drider D, Fimland G, Hechard Y, McMullen LM, Prevost H. 2006. The continuing story of class IIa bacteriocins. Microbiol Mol Biol Rev 70: 564-582. https://doi.org/10.1128/MMBR.00016-05
  6. Hata T, Tanaka R, Ohmomo S. 2010. Isolation and characterization of plantaricin ASM1: a new bacteriocin produced by Lactobacillus plantarum A-1. Int J Food Microbiol 137: 94-99. https://doi.org/10.1016/j.ijfoodmicro.2009.10.021
  7. Alverez-Cisneros YM, Fernandez FJ, Wacher-Rodarte C, Aguilar MB, Sainz Espunes Tdel R, Ponce-Alquicira E. 2010. Biochemical characterization of a bacteriocin-like inhibitory substance produced by Enterococcus faecium MXVK29, isolated from mexican traditional sausage. J Sci Food Agric 14: 2475-2481.
  8. Devi SM, Halami PM. 2011. Detection and characterization of pediocin PA-1/AcH like bacteriocin producing lactic acid bacteria. Curr Microcbiol 63: 181-185. https://doi.org/10.1007/s00284-011-9963-8
  9. Kim JH. 1995. Inhibition of Listeria monosytogenes by bacteriocin(s) from lactic acid bacteria isolated from Kimchi. Agric Chem Biotechnol 38: 302-307.
  10. Chang JY, Lee HJ, Chang HC. 2007. Identification of the agent from Lactobacillus plantarum KFRI464 that enhances bacteriocin production by Leuconostoc citreum GJ7. J Appl Microbiol 103: 2504-2515. https://doi.org/10.1111/j.1365-2672.2007.03543.x
  11. Lim, SM, Im DS. 2009. Screening and characterization of probiotic lactic acid bacteria isolated from Korean fermented foods. J Microbiol Biotechnol 19: 178-186. https://doi.org/10.4014/jmb.0804.269
  12. Eom HJ, Park JM, Seo MJ, Kim MD, Han NS. 2008. Monitoring of Leuconostoc mesenteroides DRC starter in fermented vegetable by random integration of chloramphenicol acetyltransferase gene. J Ind Microbiol Biotechnol 35: 935-939.
  13. Chang JY, Chang HC. 2010. Improvements in the quality and shelf life of Kimchi by fermentation with the induced bacteriocin-producing strain Leuconostoc citreum GJ7 as a starter. J Food Sci 75: 103-110.
  14. Eom HJ, Seo DM, Han NS. 2007. Selection of psychrotrophic Leuconostoc spp. producing highly active dextransucrase from lactate fermented vegetables. Int J Food Microbiol 10: 61-67.
  15. Shin MS, Han SK, Ryu JS, Lee WK. 2008. Isolation and partial characterization of a bacteriocin produced by Pediococcus pentosaceus K23-2 isolated from Kimchi. J Appl Mircobiol 105: 331-339.
  16. Urso R, Rantsiou K, Cantoni C, Comi G, Cocolin L. 2006. Sequencing and experssion analysis of the sakacin P bacteriocin produced by a Lactobacillus sakei strain isolated from naturally fermented sausages. Appl Microbiol Biotechnol 71: 480-485. https://doi.org/10.1007/s00253-005-0172-x
  17. Xiraphi N, Georgalaki M, Rantsiou K, Cocolin L, Tsakalidou E, Drosinos EH. 2008. Purification and characterization of a bacteriocin produced by Leuconostoc mesenteroides E131. Meat Sci 80: 194-203. https://doi.org/10.1016/j.meatsci.2007.11.020

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