Food Science and Preservation (한국식품저장유통학회지)

The Korean Society of Food Preservation (한국식품저장유통학회)
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Amylase activity and characterization of Bacillus subtilis CBD2 isolated from Doenjang

된장으로부터 분리된 Bacillus subtilis CBD2의 생육특성 및 amylase 활성

  • Yang, Su-Jin (Department of Food Science and Technology, Catholic University of Daegu) ;
  • Lee, Dae-Hoon (Department of Food Science and Technology, Catholic University of Daegu) ;
  • Park, Hye-Mi (Department of Food Science and Technology, Catholic University of Daegu) ;
  • Jung, Hee Kyoung (Biohealth Convergence Center, Daegu Technopark) ;
  • Park, Chang-Su (Department of Food Science and Technology, Catholic University of Daegu) ;
  • Hong, Joo-Heon (Department of Food Science and Technology, Catholic University of Daegu)
  • 양수진 (대구가톨릭대학교 식품공학전공) ;
  • 이대훈 (대구가톨릭대학교 식품공학전공) ;
  • 박혜미 (대구가톨릭대학교 식품공학전공) ;
  • 정희경 ((재)대구테크노파크 바이오헬스융합센터) ;
  • 박창수 (대구가톨릭대학교 식품공학전공) ;
  • 홍주헌 (대구가톨릭대학교 식품공학전공)
  • Received : 2014.02.06
  • Accepted : 2014.04.03
  • Published : 2014.04.30
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Abstract

In this study, one GRAS strain was screened from doenjang, a traditional Korean fermented food, as a microorganism producing amylase due to the formation of a clear zone on the medium including soluble starch. From the analysis of the gene sequence of 16S ribosomal RNA, the strain was identified as Bacillus subtilis and was therefore named Bacillus subtilis CBD2. When the nutrient broth medium was prepared with 3% NaCl, 5% glucose, and the initial medium pH 7.0, the B. subtilis CBD2 showed maximum growth. Among soluble starch, corn starch, maize amylopectin, and wheat starch, soluble starch was the most effective carbon source in the production of amylase by B. subtilis CBD2. The amylase from B. subtilis CBD2 showed the highest activities at pH 8.0 and $50^{\circ}C$, and corn starch was the most proper substrate for the enzyme activity. When corn starch was used as a substrate, the production of sugars through enzyme activity increased for 24 h, and then the enzyme activity became constant.

본 연구에서는 식품산업에 적용 가능한 amylase의 발굴을 위하여 된장으로부터 amylase를 생산하는 균주를 분리하였고, 분리된 미생물들의 동정을 통하여 GRAS 미생물로 부터 생산되는 amylase에 대한 효소 특성을 규명하였다. 그 결과 균주 유래 16S ribosomal RNA 유전자 염기서열에서 B. subtilis로 동정되어 본 균주를B. subtilis CBD2로 명명하였다. B. subtilis CBD2 균주의 생육에 미치는 배지 성분에 대한 특성을 검토한 결과 본 균주는 pH 7.0, NaCl 3%, glucose 10%에서 가장 높은 균 생육 특성을 나타내었다. 그리고 본 균주의 amylase 생산에 미치는 탄소원의 영향을 검토한 결과, 본 균주는 soluble starch, corn starch, maize amylopectin 및 wheat starch 중에서 soluble starch를 탄소원으로 사용하였을 때 가장 높은 amylase 생산성을 나타내었다. B. subtilis CBD2유래 amylase의 활성에 미치는 반응조건을 검토한 결과 본 균주 유래 amylase는 pH 8.0 그리고 $50^{\circ}C$에서 가장 높은 효소 활성을 나타내었으며 기질특이성에 대한 검토에서는 corn starch>wheat starch>soluble starch>maize amylopectin 순으로 효소 활성을 나타내었다. B. subtilis CBD2가 생성하는 amylase 효소와 corn starch 기질 반응에 대한 전분 분해능 및 당 생성율 변화에서는 24시간에서 가장 높은 전분 분해능 및 당 생성율을 나타내어 corn starch를 이용한 효소 반응은 24시간이 가장 적절함을 확인할 수 있었다. 따라서 본 연구에서 분리된 균주는 GRAS 미생물인B. subtilis이었으며 본 균주 유래의 amylase 특성을 활용한다면 향 후 식품산업에 있어서 전분 분해 관련 분야에 유용하게 이용될 것으로 기대되어진다.

Keywords

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