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김치유산균인 Lactococcus lactis subsp. lactis YH-10가 생산하는 박테리오신의 특성

Characterization of Bacteriocin, lacticin YH-10, Produced by Lactococcus lactis subsp. lactis YH-10 Isolated from Kimchi

  • Park, Eun-Min (Department of Biotechnology and Bioengineering, Pukyong National University) ;
  • Kim, Young-Hwa (Department of Biotechnology and Bioengineering, Pukyong National University) ;
  • Park, So-Jin (Department of Biotechnology and Bioengineering, Pukyong National University) ;
  • Kim, Yun-Im (Department of Biotechnology and Bioengineering, Pukyong National University) ;
  • Ha, Yu-Mi (Department of Biotechnology and Bioengineering, Pukyong National University) ;
  • Kim, Sung-Koo (Department of Biotechnology and Bioengineering, Pukyong National University)
  • 발행 : 2004.08.01

초록

API test를 통해서 김치에서 분리한 유산균이 Lactococcus lactis subsp. lactis의 한 종임 을 확인하고 Lactococcus lactis subsp. lactis YH-10이라 명명하고, 생산된 박테리오신을 lacticin YH-10으로 명명하였다. Lactococcus lactis subsp. lactis YH-10을 $25^{\circ}C$, 3$0^{\circ}C$와 37$^{\circ}C$에서 배양한 결과 $25^{\circ}C$에서 박테리오신을 생성하였다. Lacticin YH-10은 여러 가지 단백질 분해효소에 의해 항균활성을 소실하였으므로, 단백질로된 박테리오신임을 확인하였다. 또한 $\alpha$-amylase, $\beta$-amylase, glucoamylase의 처리에도 활성을 일부 소실하였으므로 박테리오신의 분류 중 classIV에 속함을 알 수 있었다 Ammonium sulfate정제한 후 SDS-PAGE를 통해 박테리오신의 분자량이 대략 14kDa임이 확인되었다. Lacticin YH-10은 단백질계 물질로서 인체 내에서 분해가 가능하기 때문에 일반 화학 보존제 보다 인체에 안정함을 알 수 있다. Lacticin YH-10은 낮은 pH와 높은 온도에서 항균활성을 가지고 있기 때문에 식품보존제로서 이용할 수 있다.

A bacteriocin-producing lactic acid bacteria was isolated from Kimchi on MRS selective media with the use of Lactobacillus delbrueckii subsp. delbrueckii as an indicator strain. The strain YH-10 was identified as Lactococcus lactis subsp. lactis through the API test. The crude bacteriocin (freeze-dried 50% ammonium sulfate precipitate of culture supernatant) produced by the strain was named as lacticin YH-10. Lacticin YH-10 showed the growth inhibitory activity against Gram positive pathogenic bacteria and other lactic acid bacteria. The bacteriocin was inactivated by proteases such as protamex and aroase AP-10 and partially inactivated by amylase, proteinase K, trypsin, and papain. The lacticin YH-10 remained its activity with the treatment of heat at 10$0^{\circ}C$ for 60 min or the changes of pH 2 to 11. However, the activity was lost at high pH combined with the exposure to 10$0^{\circ}C$. The bacteriocin production of the strain was started in the exponential phase and stopped in the stationary phase. The approximate molecular mass of the bacteriocin produced by the strain was approximate 14 kDa in the analysis on SDS-PAGE.

키워드

참고문헌

  1. Bogovic-Matiasic, B., I. Rogelj, I. F. Nes and H. Holo. 1998. Isolation and characterization of two bacteriocins of Lactobacillus acidophilus LF221. J. Microbiol. Biotechnol. 49, 606-612 https://doi.org/10.1007/s002530051221
  2. Cha, D. S. and D. M. Ha. 1996. Ioslation of Leuconostoc mesenteroides subsp. mesenteroises DU-0608 with antibacterial activity from kimchi and characterization of its bacteriocin. J. Microbiol. Biotechnol. 6, 270-277
  3. Chintas, L. M., P. Casaus, M. F. Fernandez and P. E. Hernandez. 1998. Comparative antimicrobial activity of enterocin L50, pediocin PA-1, nisin A and lactocin S against spoilage and foodborne pathogenic bacteria. J. Food Microbiol. 15, 289-298 https://doi.org/10.1006/fmic.1997.0160
  4. Choi, Y. O. and C. Ahn. 1997. Plasmid-associated bacteriocin produced by Leuconostoc sp. LAB145-3A isolated from Kimchi. J. Microbiol. Biotechnol. 7, 409-416
  5. Cleveland, J., T. J. Montville, I. F. Nes and M. L. Chikindas. 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
  6. Daba, H., S. Pandiann, J. F. Gosselin, R. E. Simard, J. Hung and C. Lacroix. 1991. Detection and activity of a bacteriocin produced by Leuconostoc mesenteroides. Appl. Environ. Microbiol. 57, 3450-3455
  7. I, von der Weid, Alviano, D. S., Santos, A. L. S, Soares, R. M. A., Alviano, C. S. and Seldin, L. 2003. Antimicrobial activity of Paenibacillus peoriae strain NRRL BD-62 against a broad spectrum of phytopathogenic bacteria and fungi. J. Appl. Microbiol. 95, 1143-1151 https://doi.org/10.1046/j.1365-2672.2003.02097.x
  8. Jack, R. W., J. R. Tagg and B. Ray. 1996. Bacteriocins of gram-positive bacteria. Microbiol. Rev. 59, 171-200
  9. Janzen, J. J. 1970. Economic losses resulting from mastitis. J. Dairy Sci. 5, 1151-1161
  10. Jo, Y. B., K. M. Bae, S. K. Kim and H. K. Jun. 1996. Evaluation of optimum conditions for bacteriocin production from Lactobacillus sp. JB-42 isolated from Kimchi. J. Microbiol. Biotechnol. 6, 63-67
  11. Kim, C. H., G. E. Ji and C. Ahn. 2000. Purification and molecular characterization of a bacteriocin from Pediococcus sp. KCA 1303-10 isolated fermented flatfish. Food Sci. Biotechnol. 9, 270-276
  12. Kim, H. J., J. H. Kim, J. H. Son, H. J. Seo, S. J. Park, N. S. Paek, and S. K. Kim. 2004. Characterization of Bacteriocin Produced by Lactobacillus bulgaricus. J. Microbiol. Biotechnol. 14, 503-508
  13. Kim, S. K., E. J. Lee, K. Y. Park and H. K. Jun. 1998. Bacteriocin produced by Lactobacillus curvatus SE1 Isolated from Kimchi. J. Microbiol. Biotechnol. 8, 588-594
  14. Kimura, H., H. Matsusaki, T. Sashihara, K. Sonomoto and A. Ishizaki. 1998. Purification and partial identification of bacteriocin ISK-1, a new lantibiotic produced by pediococcus sp. ISK-1. Biosci. Biotechnol. Biochem. 62, 2341-2345 https://doi.org/10.1271/bbb.62.2341
  15. Ko, S. H. and C. Ahn. 2000. Bacteriocin production by lactococcus lactis KCA2386 isolated from White Kimchi. Food Sci. Biotechnol. 9, 263-269
  16. Lee, H. J., Y. J. Joo, C. S. Park, S. H. Kim, I. K. Hwang, J. S. Ahn and T. I. Mheen. 1999. Identification and characterization of bacteriocin-producing lactic acid bacteria isolated from Kimchi. J. Microbiol. Biotechnol. 9, 282-291
  17. Matsusaki, H., K. Sonomoto and A. Ishizaki. 1997. Bacteriocins, growth inhibitory substances, of lactic acid bacteria. Microbiol. Rev. 15, 125-133
  18. Muriana P. M. and T. R. Klaenhammer. 1991. Cloning, phenotypic expression and DNA sequence of the gene for lactacin F, antimicrobial peptide produced by Lactobacillus spp. J. Bacteriol. 173, 1779-1788
  19. Ralph, W. J., J. R. Tagg. and N. B. Ray. 1995. Bacteriocins of Gram-positive bacteria. Microbiol. Rev. 59, 171-200
  20. Smelis, J., S. Rolfer and J. Metaxopoulos. 1994. Sakacin B, a bacteriocin produced by Lactobacillus sake isolated from Greek dry fermentation sausages. J. Appl. Bacteriol. 76, 475-486 https://doi.org/10.1111/j.1365-2672.1994.tb01105.x
  21. Tagg, J. R. and A. R. McGiven. 1971. Assay system for bacteriocins. Appl. Microbiol. 21, 943-945
  22. Tagg, J. R., A. S. Dajani and L. W. Wannamaker. 1976. Bacteriocins of gram positive bacteria. Bacteriol. Rev. 40, 722-756
  23. Yan, T. R. and C. S. Lee. 1997. Characterization of a partially purified bacteriocin, Fermenticin B, from Lactobacillus fermentum. Biotechnol. Lett. 8, 741-744 https://doi.org/10.1023/A:1018327907435

피인용 문헌

  1. Purification and Characterization of the Bacteriocin Produced by Lactococcus sp. KD 28 Isolated from Kimchi vol.25, pp.2, 2015, https://doi.org/10.5352/JLS.2015.25.2.180