Microbiology and Biotechnology Letters (한국미생물·생명공학회지)

The Korean Society for Microbiology and Biotechnology (한국미생물·생명공학회)
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Mannanase Production by a Soybean Isolate, Bacillus subtilis WL-7.

된장 분리균 Bacillus subtilis WL-7에 의한 Mannanase 생산

  • 권민아 (우송대학교 식품생명과학부, 생물소재 응용연구센터) ;
  • 김현숙 (우송대학교 식품생명과학부) ;
  • 이미성 (씨티씨바이오 중앙연구소) ;
  • 최준호 (씨티씨바이오 중앙연구소) ;
  • 윤기홍 (우송대학교 식품생명과학부, 생물소재 응용연구센터)
  • Published : 2003.09.01
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Abstract

A bacterium producing the extracellular mannanase was isolated from Korean soybean paste. The isolate WL-7 has been identified as Bacillus subtiis on the basis on its 16S rRNA sequence, fatty acid composition, morphology and biochemical properties. The mannanase of culture supernatant was the most active around $55^{\circ}C$ and pH $6.0^{\circ}C$, and retained 90% of its maximum activity at range of pH 5.0∼7.5 and $50∼60^{\circ}C$. The additional carbohydrates including lactose, $\alpha$-cellulose, avicel, locust bean gum (LBG), wheat bran and konjak increased dramatically the mannanase productivity of strain WL-7. Especially, the maximum mannanase productivity was reached to 224 U/ml in LB medium supplemented with both 0.5% LBG and 0.5% konjak, which was approximately 200-folds more than that in LB medium. It was suggested that the increase of mannanase production was owing to induction of mannanase biosynthesis by both LBG and konjak hydrolysates transported following initial hydrolysis by extracellular mannanase during the cell growth.

전통 발효식품인 된장으로부터 mannanas의 생산균으로 분리된 WL-7균주는 형태적 특성, 생화학적 성질 및 16S rRNA의 염기서열에 근거하여 Bacillus subtilis로 동정되었다. B. subtilis WL-7의 배양상등액으로 제조한 조효소액을 이용하여 mannanase 활성을 측정한 결과, 55$^{\circ}C$와 pH 6.0에서 최대활성을 보였으며 , 반응온도 $50∼60^{\circ}C$와 반응 pH 5.0∼7.5에서 최대활성의 90% 이상의 활성을 나타냈다. 분리균WL-7의 mannanase생산성을 높이기 위해 LB배지에 소당류와 고분자 탄수화물을 첨가하여 배양한 결과, lactose, 밀기울, avicel, $\alpha$-cellulose, locust bean gum(LBG), konjak 등에 의해 mannanase생산성이 상당량 증가되었다. 특히 0.5% LBG와 0.5% konjak이 동시에 첨가된 LB 배지에서 10시간 배양하였을 때 배양상등액의 효소 활성이 224 U/ml로 최대 효소생산성을 보였으며, 이는 LBC와 konjak을 첨가하지 않은 배지에서 보다 효소 생산성이 200배 이상이 증가하였다. LBG와 konjak을 첨가하였을 때 B. subtilis WL-7의최대성장도가 약간 증가한 것에 비해 mannanase생산성은 현저히 증가된 것으로 보아 배양액 중의 LBG와 konjak의 가수분해 산물이 mannanas리 생합성을 유도한 것으로 판단된다.

Keywords

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