Characterization of ${\beta}$-1,4-Glucanase Activity of Bacillus licheniformis B1 in Chungkookjang

청국장 발효균주 Bacillus licheniformis B1의 ${\beta}$-1,4-glucanase 특성

  • Hwang, Jae-Sung (Department of Biotechnology, The Research Institute for Basic Scinences, Hoseo University) ;
  • Yoo, Hyung-Jae (Department of Biotechnology, The Research Institute for Basic Scinences, Hoseo University) ;
  • Kim, Sung-Jo (Department of Biotechnology, The Research Institute for Basic Scinences, Hoseo University) ;
  • Kim, Han-Bok (Department of Biotechnology, The Research Institute for Basic Scinences, Hoseo University)
  • 황재성 (호서대학교 자연과학대 생명공학과, 호서대 기초과학연구소) ;
  • 유형재 (호서대학교 자연과학대 생명공학과, 호서대 기초과학연구소) ;
  • 김성조 (호서대학교 자연과학대 생명공학과, 호서대 기초과학연구소) ;
  • 김한복 (호서대학교 자연과학대 생명공학과, 호서대 기초과학연구소)
  • Published : 2008.03.31

Abstract

Fermented soybeans contain microorganisms, diverse enzymes, and bioactive compounds. Few studies on cellulase in Chungkookjang exist. Oligosaccharides play diverse roles of bioactivity. Through Congo red test and activity staining, it was confirmed that the enzyme solution contained cellulase. Optimal pH and temperature of the cellulase produced by Bacillus licheniformis B1 were 10 and $40^{\circ}C$, respectively. Through TLC analysis, it was demonstrated that the enzyme solution degraded carboxymethyl cellulose (CMC), whose main products contained dimer and trimer oligosaccharides. Cellulase activity of barley-Chungkookjnag fermented by the strain increased, compared with that of Chungkookjang. The cellulase was found to be a ${\beta}$-1,4-glucanase through the analysis of the cloned gene, showing polymorphism at 32 amino acid sites in the coding range of amino acid 10 and 460.

대두가 발효된 청국장에는 미생물, 다양한 효소와 생리활성물질이 존재한다. 청국장의 탄수화물을 분해하는 cellulase에 대한 연구는 많지 않다. Oligosaccharide는다양한 생리활성을 지니고 있다. Congo red test 및 활성염색을 통해, 효소액이 cellulase를 포함하는 것을 확인했다. 청국장 발효 균주 Bacillus licheniformis B1이 분비하는 cellulase활성의 최적 pH와 온도는 각각 10과 $40^{\circ}C$이었다. TLC분석을 통해 효소액은 carboxymethyl cellulose (CMC)를 분해하여 2탄당 이상을 생성함을 확인하였다. 본 균주를 이용해서 제조한 보리 청국장에서는 효소의 활성이 증가되었다. 본 균주의 cellulase 유전자를 클로닝하여 분석한 결과, 본 효소는 ${\beta}$-1,4-glucanase였으며, 전체 coding 영역 $10{\sim}460$번째 아미노산 영역 중, 32군데서 polymorphism을 보였다.

Keywords

References

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