Journal of Chitin and Chitosan (한국키틴키토산학회지)

The Korean Society for Chitin and Chitosan (한국키틴키토산학회)
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Characterization of Amylase Produced by Bacillus subtilis KMKW4 Isolated from Kimchi

김치로부터 분리된 Bacillus subtilis KMKW4의 amylase 생산 및 효소활성

  • Yang, Su-Jin (Department of Food Science and Technology, Catholic University of Daegu) ;
  • Kang, Eun-Ji (Department of Food Science and Technology, Catholic University of Daegu) ;
  • Lee, Rae-Hyun (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) ;
  • 강은지 (대구가톨릭대학교 식품공학전공) ;
  • 이래현 (대구가톨릭대학교 식품공학전공) ;
  • 정희경 ((재)대구테크노파크 바이오헬스융합센터) ;
  • 박창수 (대구가톨릭대학교 식품공학전공) ;
  • 홍주헌 (대구가톨릭대학교 식품공학전공)
  • Received : 2014.02.11
  • Accepted : 2014.03.06
  • Published : 2014.03.31
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Abstract

In this study, one GRAS strain was screened from Kimchi, a Korean traditional fermented food, as a microorganism producing amylase due to formation of 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, therefore was named with Bacillus subtilis KMKW4. When the nutrient broth medium was prepared with 3% NaCl, 5% glucose and the initial medium pH 6.0, the B. subtilis KMKW4 was showed the maximum growth. Among soluble starch, corn starch, maize amylopectin and wheat starch, corn starch was the most effective carbon source in the production of amylase by B. subtilis KMKW4. The amylase from B. subtilis KMKW4 showed the highest activities at pH 6.0 and $50^{\circ}C$ conditions, and corn starch was the most proper substrate on the enzyme activity. When the corn starch was used as substrate, the production of sugars by enzyme activity was increased for 24 hr and then the enzyme activity was maintained constantly.

본 연구는 곡류 효소식품 제조에 있어 starter로서 활용 가능한 amylase의 발굴을 위하여 배추김치로부터 amylase를 생산하는 균주를 분리하였고, 분리한 미생물들의 동정을 통하여 GRAS 미생물로부터 생산되는 amylase의 효소활성을 조사하였다. Amylase 활성을 검토한 결과 가장 높은 효소활성을 보인 균주는 경상남도 마산시에서 제조된 배추김치였으며, 16S ribosomal RNA 유전자 염기서열 분석에서 Bacillus subtilis로 동정되었으며 본 균주를 B. subtilis KMKW4로 명명하였다. B. subtilis KMKW4 균주의 생육에 대한 배지 성분의 특성을 검토한 결과 pH 6.0, NaCl 3%, glucose 5%에서 가장 높은 균생육 특성을 나타내었다. 다양한 탄소원의 영향은 corn starch를 영양원으로 이용하였을 때 가장 높은 amylase 효소활성을 나타내었으며, 효소활성에 미치는 생육 조건을 검토한 결과 pH 6.0, $50^{\circ}C$에서 가장 높은 효소활성을 나타내었다. 기질 특이성은 corn starch > soluble starch > wheat starch > maize amylopectin 순으로 효소활성을 나타내었으며 기질에 따른 배양시간별 효소활성은 corn starch가 180분에서 187 U/mL로 가장 우수하였다. B. subtilis가 생산하는 amylase와 corn starch에 대한 전분분해능 및 총당 생성율 변화는 24시간에서 가장 높은 총당 생성 및 전분분해를 보여주었다.

Keywords

Acknowledgement

Supported by : 한국연구재단

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