한국균학회지 (The Korean Journal of Mycology)

한국균학회 (The Korean Society of Mycology)
권호 표지

Bacillus subtilis KS1이 생산하는 항진균물질의 정제 및 특성

Purification and characterization of antifungal compounds produced by Bacillus subtilis KS1

  • 류승우 (충남대학교 미생물학과) ;
  • 맹학영 (중부대학 교양학부 (화학)) ;
  • 맹필재 (충남대학교 미생물학과)
  • Ryoo, Sung-Woo (Department of Microbiology, Chungnam National University) ;
  • Maeng, Hack-Young (Department of Basic Education(Chemistry), Joongbu University) ;
  • Maeng, Pil-Jae (Department of Microbiology, Chungnam National University)
  • 발행 : 1996.12.30
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초록

인삼포 토양으로부터 강력한 항진균력을 보이는 KS1 균주를 분리하고 Bacillus subtilis로 동정하였다. 온상시험에서 B. subtilis KS1의 배양액은 오이잿빛곰팡이병, 밀붉은녹병 등의 진균성 농작물병에 대하여 탁월한 방제효과를 나타내었다. 또한, B. subtilis KS1 배양액의 butanol 분획은 Botrytis maydis, Chytridium lagenarium, Candida albicans 등 식물 또는 사람의 병원성 진균을 비롯한 몇몇 진균류에 대하여 억제효과를 나타내었다. 이 균주가 생성하는 항진균물질을 pep-RPC reverse phase column과 ${\mu}$ Bondapak $C_{18}$ reverse phase column을 이용하여 분리 정제하였다. 정제된 항진균물질의 열안정성 및 pH 안정성을 조사한 결과, $-20-121^{\circ}C$와 pH 4.0-10.0의 범위에서 안정하였다. 이 물질의 조성 및 구조적 특징을 HPLC와 $^1H-,\;^1H-^1H-$, COSY, NOESY, COSY-NOESY 및 HOHAHA NMR spectroscopy를 통하여 각각 분석한 결과, B. subtilis가 생산하는 고리형 peptide 중 iturin A에 속하는 물질로 확인되었다. 그러나, 지금까지 알려진 iturin A계 물질들의 ${\beta}$-아미노산에 붙어 있는 탄화수소 사슬이 곁가지가 없거나 terminus 또는 subterminus에 1개의 $CH_3$ 곁가지를 갖는 것과는 달리, SW1의 ${\beta}$-아미노산은 2개의 $-CH_3$ 곁가지가 붙어 있는 탄화수소 사슬을 가짐이 확인되었다.

A bacterial strain, KSl, possessing strong antifungal activity was isolated from soil samples of ginseng fields and identified as Bacillus subtilis. In greenhouse test, the culture filtrate of B. subtilis KS1 showed strong protective effect against several fungal diseases of agricultural plants such as cucumber gray mold and wheat leaf rust. In addition, the crude butanol fraction of the culture filtrate exhibited antagonistic effect against several fungi including plant or human pathogens, such as Botrytis maydis, Chytridium lagenarium and Candida albicans. The antifungal compound, SW1, produced by B. subtilis KS1 was purified through consecutive chromatographic separations on a pep-RPC column and a ${\mu}$ Bondapak $C_{18}$ reverse phase column. Temperature and pH showed little effect on the stability of the compound in the ranges $-20-121^{\circ}C$ and pH 4.0-10.0, respectively. The composition and structural characteristics of SW1 were analysed by HPLC and by $^1H-,\;^1H-^1H-COSY$, NOESY, COSY-NOESY and HOHAHA NMR spectroscopy, respectively, which revealed that the compound belongs to iturin A, a typical cyclic antifungal compound produced by B. subtilis. In contrast to the previously reported iturin A compounds which have one or no $-CH_3$ side chain in the hydrophobic hydrocarbon chain of ${\beta}-amino$ acids, SW1 was shown to have a ${\beta}-amino$ acid containing 12-carbon skeleton with two $-CH_3$ side chains.

키워드

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