Korean Journal of Microbiology (미생물학회지)

The Microbiological Society of Korea (한국미생물학회)
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Isolation and Identification of Halotolerant Bacillus sp. SJ-10 and Characterization of Its Extracellular Protease

세포외 Protease를 생산하는 내염성 Bacillus sp. SJ-10 균주의 분리 동정 및 효소 특성

  • Kim, Eun-Young (Department of Biotechnology, Pukyong National University) ;
  • Kim, Dong-Gyun (Department of Biotechnology, Pukyong National University) ;
  • Kim, Yu-Ri (Department of Biotechnology, Pukyong National University) ;
  • Choi, Sun-Young (Department of Biological Analysis, Chemical Defense Research Institute) ;
  • Kong, In-Soo (Department of Biotechnology, Pukyong National University)
  • 김은영 (부경대학교 생물공학과) ;
  • 김동균 (부경대학교 생물공학과) ;
  • 김유리 (부경대학교 생물공학과) ;
  • 최선영 (국군화생방방호사령부 화학방어 연구소 생물분석과) ;
  • 공인수 (부경대학교 생물공학과)
  • Received : 2009.04.27
  • Accepted : 2009.06.16
  • Published : 2009.06.30
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Abstract

A bacterium producing the halotolerant extracellular protease was isolated from squid jeotgal, and was identified as Bacillus sp. SJ-10 based on morphological, physiological and biochemical characteristics, as well as phylogenetic analysis using 16S rRNA gene sequence. The strain grew at $20^{\circ}C\sim55^{\circ}C$, pH 5~8, and 0%~14% NaCl and optimal growth conditions were $35{\pm}5^{\circ}C$, pH 7, and 5% NaCl. The major cellular fatty acids were anteiso-$C_{15:0}$, anteiso-$C_{17:0}$, and $C_{16:0}$ DNA G+C content was 50.58 mol% and menaquinone consisted of MK-7 Phylogenic analysis based on the 16S rRNA gene sequence indicated that SJ-10T belongs to the genus Bacillus. About 40 kDa of the salt-tolerant protease was purified by 40% ammonium sulfate saturation and Mono Q column chromatography. The optimal activity of the protease was pH 8 and stable at pH 5~10. The optimum temperature and NaCl concentration were $35{\pm}5^{\circ}C$ and $5{\pm}1%$, respectively.

오징어 젓갈로부터 호염성 균주를 분리하여 형태적, 생리 생화학적 특성 및 분자계통학적 분석을 통하여 동정하였고, 이 균주가 세포외로 생산하는 호염성 protease를 정제하여 그 특성을 분석하였다. 본 균주는 NaCl 0~14%, $20\sim55^{\circ}C$ 및 pH 5~8에서 성장하였으며, $35{\pm}5^{\circ}C$, pH 7 및 5% NaCl에서 최적으로 성장하였다. 주요 지방산 조성은 anteiso-$C_{15:0}$ (44.70%), anteiso-$C_{17:0}$ (15.76%) 및 $C_{16:0}$ (13.91%)이었고, menaquinone은 MK-7이었으며, DNA G+C 함량은 50.58 mol%였다. 16S rRNA 유전자 염기서열을 통한 계통분석 결과 Bacillus이었으며 Bacillus sp. SJ-10으로 명명하였다. SJ-10 균주 배양액으로부터 40% 황산암모늄 침전과 Mono Q 컬럼크로마토그래피 과정을 거쳐 collagen과 gelatin을 특이적으로 분해하는 약 40 kDa의 세포외 protease를 정제하였다. 이 효소는 $35{\pm}5^{\circ}C$, NaCl $5{\pm}1%$ 및 pH 8에서 최적 활성을 나타냈으며, pH 5~10 범위에서 안정하였다.

Keywords

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