Journal of the Korean Society of Food Science and Nutrition (한국식품영양과학회지)

The Korean Society of Food Science and Nutrition (한국식품영양과학회)
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Isolation of a Bacteriocin - Producing Lactobacillus sakei Strain from Kimchi

김치에서 박테리오신을 분비하는 Lactobacillus sakei균주의 분리

  • 김한택 (경상대학교 대학원 응용생명과학부 농업생명과학연구소) ;
  • 박재용 (경상대학교 대학원 응용생명과학부 농업생명과학연구) ;
  • 이강권 (삼성 에버랜드(주) 식품연구) ;
  • 김정환 (경상대학교 대학원 응용생명과학부 농업생명과학연구소)
  • Published : 2004.03.01
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Abstract

Bacteriocin producing lactic acid bacteria (LAB) were isolated from Kimchi by using spot-on-the-lawn method. Listeria monocytogenes, Staphylococcus aureus, and Lactobacillus plantarum were used as indicators. One isolate (P3-l) produced a bacteriocin efficiently inhibiting the growth of Listeria monocytogenes. 16S rDNA sequence and sugar utilization test identified that P3-1 was a Lactobacillus sakei strain. Accordingly, the isolate was named as Lactobacillus sakei P3-1. L. sakei P3-1 produced a bacteriocin which efficiently inhibited the growth of Listeria monocytogenes but did not inhibit other Gram positive and negative organisms tested. The bacteriocin was stable against heat, organic solvent, and pH variation and it retained 50% of activity after 10 min heat treatment at 10$0^{\circ}C$. The molecular weight of Sakacin P3-1 was estimated to be 4 kDa by SDS-PAGE.

배추김치로부터 식품유해균인 Listeria monocytogenes를 저해하는 박테리오신을 생산하는 유산균, Lactobacillus sakei P3-1이 분리되었다. 형태학적, 생화학적 특성조사와 최종적으로 PCR로 증폭하여 얻은 16S rDNA 염기서열 결정을 통해서 L. sakei로 동정되었다. L. sakei P3-1이 분비하는 박테리오신은 여러 그람 양성 및 음성균들 중에서 단지 L. monocytogenes만을 저해하는 그래서 저해범위가 매우 좁은 박테리오신으로 확인되었다. 이온교환 크로마토그래피에 의해서 박테리오신은 부분 정제되었으며 박테리오신의 열처리 안정성을 조사한 결과 121$^{\circ}C$에서 15분간 그리고10$0^{\circ}C$에서 10분간 열처리 후에도 각각 12.5%와 50%의 역가가 잔존하여 상당한 열안정성을 지니고 있음을 알 수 있었다. MRS배지에서 배양중 배양온도가 박테리오신 역가에 미치는 영향을 조사한 결과 3$0^{\circ}C$에서 배양할 때 그리고 18시간 이상 배양에서 가장 높은 1,000 AU/mL 역가를 보였다. 한편 SDS-PAGE 및 activity staining에 의해 측정된 박테리오신의 분자량은 4,000이었다. L. monocytogenes 생육 억제능, 작은 분자량 및 높은 열안정성 등의 성질들을 종합적으로 고려할 때 L. sakei P3-1이 생산하는 박테리오신은 박테리오신들 중에서 class II-a에 속하는 것으로 추정된다.

Keywords

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