Isolation and Characterization of Three Kinds of Lipopeptides Produced by Bacillus subtilis JKK238 from Jeot-Kal of Korean Traditional Fermented Fishes

한국 전통젓갈에서 분리한 Bacillus subtilis JKK238 균주 유래 세 종류 Lipopeptide의 분리 및 특성

About seven hundred bacterial strains were collected from Jeot-Kal, a Korean traditional fermented fishes, in various Korean districts. One of the strains designated JKK238 has its ability to antagonize in vitro the growth of a wide variety of plant pathogenic fungi responsible for diseases of economical importance. The JKK238 strain was isolated from Oh-Jeot, a kind of fermented shrimps, of Kangkyeung in Korea, and was identified as Bacillus subtilis based on its physiological characteristics, fatty acids compositions of cellular wall, and 16S rDNA sequence analysis. We isolated simply antimicrobial lipopeptides (AMLP) by $25\%$ ammonium sulfate precipitation of 3 days-old tryptic soy broth cultures of the JKK238 strain. Further analysis of AMLP revealed that B. subtilis JKK238 produces a wide variety of antifungal lipopeptide isomers from the iturin, fengycin and surfactin families simultaneously. Above results indicate that the JKK238 strain can be added to the limited number B. subtilis strains reported to co-produce the three kinds of lipopeptide families.

한국 전통 젓갈로부터 항균성 lipopeptide를 생산하는 JKK238 균주를 분리하여 16S rDNA및 세포벽의 지방산 조성 분석 등에 따라 Bacillus subtilis로 동정하였다. JKK238 균주를 3일간 배양한 TSB 액체배지의 $25\%$ ammonium sulfate 침전물이 대부분의 lipopeptide를 포함하고 이들이 다양한 식물병원성 곰팡이나 세균의 성장을 강력히 저해함을 알 수 있었다. 또한 FPLC에 의해 분리 및 정제된 각 lipopeptide의 Maldi-tof분석에서, 이 균주가 iturin A, fengycin, surfactin과 같은 세 종류의 lipopeptide의 isomer들을 동시에 생산하는 균주임을 밝혔다.