Korean Journal of Microbiology (미생물학회지)

The Microbiological Society of Korea (한국미생물학회)
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Characterization of two β-mannanases from Cellulosimicrobium sp. YB-43

Cellulosimicrobium sp. YB-43에 의해 생산되는 2종류 β-mannanase의 특성분석

  • Yoon, Ki-Hong (Food Science and Biotechnology Major, Woosong University)
  • 윤기홍 (우송대학교 바이오식품과학전공)
  • Received : 2015.08.05
  • Accepted : 2015.09.04
  • Published : 2015.09.30
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Abstract

A bacterial strain producing extracellular mannanases was isolated from soil of chestnut tree farm located in Gongju city of Korea by enrichment culture using Avicel as a carbon source. 16S rDNA sequence of the isolate YB-43 was highly homologous to those of genus Cellulosimicrobium strains with sequence similarities of above 99.6%. Mannanase productivity was significantly increased when the Cellulosimicrobium sp. YB-43 was grown in the presence of locust bean gum (LBG) or konjac. The mannanases were partially purified to be mannanase A (ManA) and mannanase C (ManC) by DEAE-Sepharose column and Q-Sepharose column chromatography from the culture filtrate of Cellulosimicrobium sp. YB-43 grown in LB medium supplemented with 0.7% LBG for 24 h. The partially purified ManA showed the highest activity at $55^{\circ}C$ and pH 6.5, while ManC activity was optimal at $65^{\circ}C$ and pH 7.5. ManA was stable up to $40^{\circ}C$ for 1 h, but ManC activity decreased significantly even after 1 h at $20^{\circ}C$. ManA and ManC showed difference from each other according to their substrate specificities and predominant products resulting from the mannanase hydrolysis for mannooligosaccharides. As a result, Cellulosimicrobium sp. YB-43 was found to produce two different kinds of mannanases.

탄소원으로 Avicel이 첨가된 배지에서 공주에 소재한 밤 나무 농장의 토양 미생물을 증균 배양하여 mannanase를 생산하는 미생물을 분리하였다. 분리균 YB-43의 16S rDNA 서열은 Cellulosimicrobium 속 균주와 유사도가 99.6% 이상으로 가장 높게 나타났다. Locust bean gum (LBG)이나 konjac이 첨가된 배지에서 분리균의 mannanase 생산성이 크게 증가하였다. 0.7% LBG가 첨가된 LB 배지에서 Cellulosimicrobium sp. YB-43이 균체외로 생산한 mannanase를 DEAE-Sepharose와 Q-Sepharose 컬럼 크로마토그래피로 부분 정제하여 mannanase A (ManA)와 mannanase C (ManC)로 분획하였다. ManA는 $55^{\circ}C$와 pH 6.5, ManC는 $65^{\circ}C$ pH 7.5에서 최대활성을 보였으며, ManA는 $40^{\circ}C$ 이하에서 1시간 동안 실활되지 않았으나 ManC는 $20^{\circ}C$에서도 상당량 실활되었다. 또한 ManA와 ManC는 기질특이성과 mannooligosaccharides의 최종 분해산물에도 차이가 있는 것으로 나타났다. 이로 보아 Cellulosimicrobium sp. YB-43은 특성이 서로 다른 2종류 mannanases를 생산하는 것으로 판단된다.

Keywords

References

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