Food Science and Preservation (한국식품저장유통학회지)

The Korean Society of Food Preservation (한국식품저장유통학회)
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Characterization of alkaline cellulase from Bacillus subtilis 4-1 isolated from Korean traditional soybean paste

전통 장류에서 분리된 알칼리성 Cellulase 생성 Bacillus subtilis 4-1 균주의 효소학적 특성

  • Baek, Seong Yeol (Fermented Food Division, Department of Agro-food Resource, NAAS, RDA) ;
  • Lee, You Jung (Fermented Food Division, Department of Agro-food Resource, NAAS, RDA) ;
  • Yun, Hye Ju (Fermented Food Division, Department of Agro-food Resource, NAAS, RDA) ;
  • Park, Hye Young (Fermented Food Division, Department of Agro-food Resource, NAAS, RDA) ;
  • Yeo, Soo-Hwan (Fermented Food Division, Department of Agro-food Resource, NAAS, RDA)
  • 백성열 (농촌진흥청 국립농업과학원 농식품자원부 발효식품과) ;
  • 이유정 (농촌진흥청 국립농업과학원 농식품자원부 발효식품과) ;
  • 윤혜주 (농촌진흥청 국립농업과학원 농식품자원부 발효식품과) ;
  • 박혜영 (농촌진흥청 국립농업과학원 농식품자원부 발효식품과) ;
  • 여수환 (농촌진흥청 국립농업과학원 농식품자원부 발효식품과)
  • Received : 2014.04.22
  • Accepted : 2014.05.27
  • Published : 2014.06.30
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Abstract

In this study, we isolated a cellulase-producing bacterium isolated from traditional Korean fermented soybean paste and investigated the effect of culture conditions on the production of cellulase. This bacterium, which was identified as Bacillus subtilis 4-1 through 16S rRNA gene sequence analysis, showed the highest cellulase activity when the cells were grown at $45^{\circ}C$ for 24 hours in the CMC medium supplemented with 1.0% of soluble starch and 0.1% yeast extract. The initial optimum pH of the medium was observed in the range of 5.0~9.0. The optimal pH and temperature for the production of cellulase from B. subtilis 4-1 were pH 9.0 and $60^{\circ}C$ respectively. In addition, the enzyme showed significant activity in the temperature range of $20{\sim}90^{\circ}C$, which indicates that B. subtilis 4-1 cellulase is an alkaline-resistance and thermo-stable enzyme. This enzyme showed higher activity with CMC as the substrate for endo-type cellulase than avicel or pNPG as the exo-type substrates for exo-type cellulase and ${\beta}$-glucosidase. These results suggest that the cellulase produced from B. subtilis 4-1 is a complex enzyme rather than a mono-enzyme.

수집한 장류 시료에서 분리한 균주들을 CMC를 함유하는 배지에 접종하여 섬유소 분해 활성이 우수한 4-1 균주를 선발하였다. 4-1 균주의 16S rRNA 염기서열을 분석한 결과, B. subtilis로 동정되었다. B. subtilis 4-1의 효소생산을 위한 최적 배양조건은 탄소원으로 1.0% soluble starch와 질소원으로 0.1% yeast extract를 첨가하여 $45^{\circ}C$에서 24시간 배양하였을 때로 나타났다. 최적 배양 pH를 조사한 결과, pH 5.0~9.0에서 cellulase 효소활성이 높았다. B. subtilis 4-1의 조효소 특성은 효소반응의 최적 pH가 pH 9.0, 반응온도는 $60^{\circ}C$에서 효소활성이 가장 높았으며, $20{\sim}90^{\circ}C$ 온도에서 60분간 열처리시 효소 활성이 80%이상 유지되었다. 따라서 B. subtilis 4-1에 의해 생산되는 cellulase는 내알칼리성 효소로 추정되며, 높은 열에도 안정한 것으로 나타났다. B. subtilis 4-1이 생산하는 cellulase는 CMC에 가장 높은 효소활성을 나타내었으며 avicel과 pNPG에서도 활성을 보여 복합효소로 생각된다.

Keywords

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