Microbiology and Biotechnology Letters (한국미생물·생명공학회지)

The Korean Society for Microbiology and Biotechnology (한국미생물·생명공학회)
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Optimization of \beta-mammanase Production from Bacillus subtilis JS-1.

\beta-Mannanase를 생산하는 Bacillus subtilis JS-1의 분리 및 효소 생산성

  • 임지수 (배재대학교 생명과학부) ;
  • 정진우 (배재대학교 생명과학부) ;
  • 이종수 (배재대학교 생명과학부) ;
  • 강대경 (이지바이오시스템 생물자원연구소) ;
  • 강하근 (이지바이오시스템 생물자원연구소)
  • Published : 2003.03.01
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Abstract

A bacteria strain producing extracellular $\beta$-mannanase was isolated from soil and was identified as Bacillus subtilis by 16S rRNA sequence comparison and biochemical determinations. The optimum pH and temperature for the $\beta$-mannanase activity were 5.0 and 5.5$^{\circ}C$, respectively. The zymogram technique revealed a single protein band exhibiting $\beta$-mannanase activity from the culture supernatant. The molecular mass of the enzyme was estimated at approximately 130 kDa. The addition of 0.5% lactose or 0.5% locust bean gum to the LB medium caused to Increase significantly the $\beta$-mannanase productivity from Bacillus subtilis JS-1. The cells grown on LB medium supplemented with lactose produced maximal enzyme activity at the stationary phase. In contrast to this, the $\beta$-mannanase was induced at the logarithmic phase from the cells grown on LB medium supplemented with locust bean gum. The discrepancy in induction times suggests that $\beta$-mannanase was induced by different induction mechanisms depending on the carbon sources in Bacillus subtilis JS-1 .

토양으로부터 $\beta$-mannanase활성이 우수한 균주를 분리하여 형태학적, 생화학적 동정과정을 거쳐 Bacillus subtilis JS-1으로 동정하였다. 분리균이 생산하는 $\beta$-mannanase 효소의 최적활성은 55$^{\circ}C$와 pH 5.0이었다. 탄소원이 다른 배지에서 배양한 분리 균주의 상등액을 전기영동하여 효소활성을 관찰한 결과 탄소원에 상관없이 분자량 130kDa에 해당하는 단일 단백질만이 효소 활성을 나타내었다 Bacillus subtilis JS-1은 탄소원으로 lactose와 locust bean gum이 존재할 때 $\beta$-mannanase 생산성이 크게 증가하는 것으로 나타났으며, lactose와 locust bean gum이 각각 0.5 % 존재할 때 배양 상등액의 $\beta$-mannanase 활성은 30U/ml과 45U/ml로 탄소원이 없는 대조구에 비해 최대 18배 정도 생산성이 증가하였다. 배지에 locust bean gum을 첨가하였을 때 효소 생산성 뿐만 아니라 균체의 성장도 함께 증가하는 것으로 보아 분리균주는 locust bean gum을 분해하여 에너지원으로 이용하는 것으로 판단된다

Keywords

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