Food Science and Preservation (한국식품저장유통학회지)

The Korean Society of Food Preservation (한국식품저장유통학회)
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Isolation of Bacillus velezensis SSH100-10 with Antifungal Activity from Korean Traditional Soysauce and Characterization of Its Antifungal Compounds

전통재래 간장으로부터 항진균 활성 B. velezensis SSH100-10의 분리와 그 항진균 물질의 특성 구명

  • Chang, Mi (Department of Food and Nutrition, Kimchi Research Center, Chosun University) ;
  • Moon, Song Hee (Department of Food and Nutrition, Kimchi Research Center, Chosun University) ;
  • Chang, Hae Choon (Department of Food and Nutrition, Kimchi Research Center, Chosun University)
  • 장미 (조선대학교 식품영양학과.김치연구센터) ;
  • 문송희 (조선대학교 식품영양학과.김치연구센터) ;
  • 장해춘 (조선대학교 식품영양학과.김치연구센터)
  • Received : 2012.08.27
  • Accepted : 2012.10.05
  • Published : 2012.10.30
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Abstract

The SSH100-10 bacterial strain, which exhibits strong antifungal (anti-mold and anti-yeast) activity, was isolated from traditional korean soysauce aged 100 years. The strain was identified as Bacillus velezensis based on Gram-staining, the biochemical properties and 16S rRNA gene sequence determination. B. velezensis SSH100-10 showed strong proteinase activity and NaCl tolerance, but did not produce enterotoxin. Two-antifungal compounds from B. velezensis SSH100-10 were purified using SPE, preparative HPLC, and reverse phase-HPLC. The purified antifungal compounds were identified as $C_{14}$ and $C_{15}$ iturin through MALDI-TOF-MS and amino acid composition analysis. The stability characteristics of the antifungal compounds after temperature, pH, and enzyme treatments suggested that B. velezensis SSH100-10 produced more than two antifungal compounds; pH-stable $C_{14}$ iturin A and $C_{15}$ iturin A, and unidentified pH-unstable compounds. The results suggested that B. velezensis SSH100-10 can be used in soybean fermentation as a starter. Moreover it has potential as a biopreservative in the food and feed industry and as a biocontrol agent in the field of agriculture.

향미가 우수한 100년 묵은 재래 간장에서 부터 강력한 항진균 활성(antifungal)균주를 분리 동정하여 Bacillus velelzensis SSH100-10으로 명명하였다. B. velezensis SSH100-10은 식품 위해 및 식품 변패곰팡이에 대한 항곰팡이 활성과 장류에 이취를 주는 산막효모 등에 대한 항효모 활성이 동시에 있으며, 강력한 단백분해활성을 나타내었다. NaCl 내염성도 우수하여 12% NaCl 농도하에서도 생육 48시간에 $A_{600}$에서 3.51의 생육도를 나타내었다. 또한 최근 Bacillus subtilis group 중 일부 균주에서 설사형 독소인 enterotixon을 나타내는 균주들에 대한 보고가 증가함에 따라, 안전성검증의 일환으로 enterotoxin 생성 여부를 조사하였을 때, 본 분리 균주는 enterotoxin 검지반응에서 음성을 나타내었다. B. velezensis SSH100-10이 생산하는 항진균 활성물질을 SPE, preparative HPLC, reverse phase-HPLC로 정제 후, MALDI-TOF-MS와 아미노산 조성 분석을 통하여 그 항진균 원인 물질이 $C_{14}$ iturin A와 $C_{15}$ iturin A 임을 구명하였다. 또한 B. velezensis SSH100-10의 배양상징액과 분리 정제된 항진균 물질 $C_{14}$ iturin A와 $C_{15}$ iturin A와 대조구로 시판되는 $C_{14{\sim}15}$ iturin A의 pH, 온도, 효소 안정성 실험을 통하여 B. velezensis SSH100-10은 pH, 열, 효소처리에 매우 안정한 iturin A 이외에도 pH에 불안정한 또 다른 구명되지 않은 항진균 물질을 생산함을 보고하였다. 본 연구에서 보고된 강력한 항진균 활성을 지니는 B. velezensis SSH100-10은 맛있는 발효과정의 종균으로서의 작용과 더불어 콩발효식품에서 유해요소가 될 수 있는 바람직하지 못한 미생물을 제어함으로서 콩발효식품의 안전과 위생수준을 개선할 수 있는 우수한 종균으로 활용될 수 있을 것이다.

Keywords

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