Korean Journal of Food Science and Technology (한국식품과학회지)

Korean Society of Food Science and Technology (한국식품과학회)
권호 표지

Characterization of the Antagonistic Activity against Lactobacillus plantarum and Induction of Bacteriocin Production

김치로부터 Lactobacillus plantarum 생육저해 박테리오신 생산균주의 분리 및 박테리오신 생산의 유도효과

  • Yang, Eun-Ju (Department of Food and Nutrition, Chosun university) ;
  • Chang, Ji-Yoon (Department of Food and Nutrition, Chosun university) ;
  • Lee, Hyong-Joo (Department of Food Science and Technology, Seoul National University) ;
  • Kim, Jeong-Hwan (Department of Food Science and Technology, Gyeongsang National University) ;
  • Chung, Dae-Kyun (Department of Genetic Engineering, Kyung Hee University) ;
  • Lee, Jong-Hoon (Department of Food and Biotechnology, Kyunggi University) ;
  • Chang, Hae-Choon (Department of Food and Nutrition, Chosun university)
  • Published : 2002.04.01
Download PDF
⟨ Previous Next ⟩

Abstract

A new bacteriocin producing lactic acid bacteria having antagonistic activity against Lactobacillus plantarum, was isolated from Kimchi. It was identified as Leuconostoc mesenteroides, and designated as Leuconostoc mesenteroides B7. The bacteriocin from Leuconostoc mesenteroides B7 named as bacteriocin B7 was stable in the pH range $2.5{\sim}9.5$. Bacteriocin B7 was active over a wide temperature range from $4^{\circ}C$ to $120^{\circ}C$. It was inactivated by proteinase K, trypsin, ${\alpha}-chymotrypsin$, and protease treatments indicating its proteinous nature. Tricine-SDS-PAGE of the purified bacteriocin B7 showed the presence of a single band, having a molecular mass of about 3,500 dalton. Mixed culture of the producer and the indicator, Lb. plantarum KFRI 464 or Lb. delbruekii KFRI 347, increased production of bacteriocin B7. This result suggested the presence of bacteriocin inducing factor in the indicator strain. The inducing factor was localized in cell debris and intracellular faction of the indicator cell, Lb. plantarum KFRI 464. Treatment of the inducing factor with proteinase K destroyed inducing activity. This result strongly suggested that the inducing factor is a protein.

Lactobacillus plantarum에 대해 항균활성을 지니는 유산균을 김치로부터 분리하였다. 이 균은 Leuconostoc mesenteroides으로 동정되었으며 Leuconostoc mesenteroides B7으로 명명하였다. Leuconostoc mesenteroides B7이 생산하는 박테리오신은(박테리오신 B7) pH 및 열 안정성이 뛰어나 pH $2.5{\sim}9.5$ 그리고 $4^{\circ}C{\sim}120^{\circ}C$ 열처리에서도 항균활성을 안정하게 유지하였다. 박테리오신 B7은 proteinase K, trypsin, ${\alpha}-chymotrypsin$, 그리고 protease 처리에 의해 항균활성이 실활되므로 peptide나 단백질로 이루진 구조임을 알 수 있었다. 정제된 박테리오신 B7은 Tricine-SDS-PAGE를 통하여 분자량이 약 3,500 dalton임을 확인하였다. Leuconostoc mesenteroides B7과 이에 감수성인 균주, Lb. plantarum KFRI 464 혹은 Lb. delbruekii KFRI 347와의 혼합배양에 의하여 박테리오신 B7 생산이 증가됨을 확인하였고, 이는 박테리오신 생산 유도물질이(inducing factor) 감수성균주내에 존재함을 시사한다. 감수성균주의 세포분획을 통하여 inducing factor는 감수성균주의 세포벽과 세포내에 존재함을 알았다. 이 inducing factor가 proteinase K처리로 박테리오신 유도활성을 상실함으로써 이는 단백질성물질임을 시사하였다.

Keywords

References

  1. So, M.H. and Kim, Y.B. Identification of psychrotrophic lactic acid bacteria isolated from Kimchi. Korean J. Food Sci. Technol.27(4): 495-505 (1995)
  2. Kang, S.M., Yung, W.S., Kim, Y.C., Joung, E.Y. and Han, Y.G. Strain improvement of Leuconostoc mesenteroides for Kimchi fer-mentation and effect of starter. Korean J. Appl. Microbiol. Bio-technol. 23(4): 461-471 (1995)
  3. Lee, C.W., Ko, C.Y. and Ha, D.M. Microfloral of the lactic acid bacteria during Kimchi fermentation and identification of the iso-lates. Korean J. Appl. Microbiol. Biotechnol. 20(10): 102-109 (1992)
  4. Kim, O.M., Kim, M.K., Lee, K.R. and Kim, S.D. Selective serial antimicrobial effects of spice extracts against Lactobacillus plan-tarum and Leuconostoc mesenteroides isolated from Kimchi.Korean c 26(5): 373-378 (1998)
  5. Kim, M.H., Oh, S.W., Hong, S.P. and Yoon S.K. Microbiology/fermentation: antimicrobial characteristics of chitosan and chito-san oligosaccharides on the microorganism related to Kimchi. Korean J. Food Sci. Technol. 30(6): 1439-1447 (1998)
  6. Kim, S.I., Kim, I.C. and Chang, H.C. Isolation and identification of antimicrobial agent producing microorganisms and sensitive strain from soil. J. Korean Soc. Food Sci. Nutr. 28(3): 526-553 (1999)
  7. Tagg, J.R. and Mcgiven, A.R. Assay system for bacteriocin. Appl. Microbiol. 21:943 (1971)
  8. Schagger, H. and von Jagow, G. Tncin-sodium dodecyl sulfate-polyacryamide gel electrophoresis for the separation of proteins in the range from 1 to 100 kDa. Anal. Biochem. 166: 368-379(1987) https://doi.org/10.1016/0003-2697(87)90587-2
  9. Daba, H.S., Pandian, J.F., Gossenelin, R.E., Simard, J.H. and Lac-roix, C. Detection and activity of a bacteriocin produced by Leu-conostoc mesenteroides. Appl. Environ. Microbiol. 57: 3450-3455(1991)
  10. Daschel, M.A., Mckenney, M.C. and McDonald, L.C. Bacterio-cidal activity of Lactobacillus plantarum C-ll. Food Microbiol.7: 91-98 (1990) https://doi.org/10.1016/0740-0020(90)90014-9
  11. Mathieu, F, Suwandhi, I.S.N., Rekhif, J.F. and Lefebvre, G.Mesenterocin 52, a bacteriocin produced by Leuconostoc mesenteroides sp. mesenteroides FR52. J. Appl. Bacteriol. 74: 372-379(1993) https://doi.org/10.1111/j.1365-2672.1993.tb05141.x
  12. Liu, W. and Hansen, J.N. Some chemical and physical properties of nisin, a small-protein antibiotic produced by Lactococcus lac-tis. Appl. Environ. Microbiol. 56: 2551-2558(1990)
  13. Rammelsberg, M. and Radler, F. Antibacterial polypeptide of Lac-tobacillus species. J. Appl. Bacteriol. 69: 177-184 (1990) https://doi.org/10.1111/j.1365-2672.1990.tb01507.x
  14. Piard. J.C., Delorme, F, Giraffa, G., Commissaire, J. and Des-mazeud, M. Evidence for a bacteriocin produced by Lactococcus lactis CNRZ 481. Neth. Milk Dairy J. 44: 143-158 (1990)
  15. Quadri, L.E.N., Kleerebezeem, M., Kuipers, O.R, Roy, K.L.,Veras, J.C. and Stiles, M.E. Characterization of a locus from Car-nobacterium piscicola LV 1713 involved in bacteriocin production and immunity: evidence for global inducer-mediated transcrip-tional regulation. J. Bacteriol. 179: 6163-6179 (1997) https://doi.org/10.1128/jb.179.19.6163-6171.1997
  16. Axelsson, L. and Askild, 0. The genes involved in production of and immunity to sakacin A, a bacteriocin from Lactobacillus sake Lb 706. J. Bacteriol. 177: 2125-2137 (1995) https://doi.org/10.1128/jb.177.8.2125-2137.1995
  17. Diep, D.B., Havarstein, L.S. and Nes, I.F. A bacteriocin-like pep-tide induces bacteriocin synthesis in L. plantarum Cll. Mol. Microbiol. 18: 631-639 (1995) https://doi.org/10.1111/j.1365-2958.1995.mmi_18040631.x
  18. Eijsink, V.G.H., Bruberg, M.B., Middelhoven, P.H. and Nes, I.F.Induction of bacteriocin production in Lactobacillus sake by a secreted peptide. J. Bacteriol. 178: 2232-2237 (1996) https://doi.org/10.1128/jb.178.8.2232-2237.1996
  19. Barefoot, S.F, Chen, Y-R., Bodine, T. A., Shearer, M. Y. and Hughes, M. D. Identification and purification of a protein that induces production of the Lactobacillus acidophilus bactenocin Lactacin B. Appl. Microbiol. 60: 3522-3528 (1994)
  20. Saucier, L., Poon, A. and Stiles, M.E. Induction of bacteriocin in Cartobacterium piscicola LV17. J. Appl. Bacteriol. 78: 684-690(1995) https://doi.org/10.1111/j.1365-2672.1995.tb03116.x