Microbiology and Biotechnology Letters (한국미생물·생명공학회지)

The Korean Society for Microbiology and Biotechnology (한국미생물·생명공학회)
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Purification and Characterization of Cyclodextrin Glucanotransferase from Bacillus sp. El

Bacillus sp. E1이 생성하는 Cyclodextrin Glucanotransferase의 정제 및 특성

  • Park, Cheon-Seok (Department of Food Science and Technology and Research Center for New Bio-materials in Agirculture, Seoul National University) ;
  • Woo, Eui-Jeon (Department of Food Science and Technology and Research Center for New Bio-materials in Agirculture, Seoul National University) ;
  • Kuk, Seung-Uk (Department of Food Science and Technology and Research Center for New Bio-materials in Agirculture, Seoul National University) ;
  • Seo, Byung-Cheol (Department of Food Science and Technology and Research Center for New Bio-materials in Agirculture, Seoul National University) ;
  • Park, Kwan-Hwa (Department of Food Science and Technology and Research Center for New Bio-materials in Agirculture, Seoul National University) ;
  • Lim, Hoon (Miwon Foods Co., R&D Div.)
  • 박천석 (서울대학교 농과대학 식품공학과 농업생물신소재연구센터) ;
  • 우의전 (서울대학교 농과대학 식품공학과 농업생물신소재연구센터) ;
  • 국승욱 (서울대학교 농과대학 식품공학과 농업생물신소재연구센터) ;
  • 서병철 (서울대학교 농과대학 식품공학과 농업생물신소재연구센터) ;
  • 박관화 (서울대학교 농과대학 식품공학과 농업생물신소재연구센터) ;
  • 임훈 (미원식품(주) 기술연구소)
  • Published : 1992.04.01
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Abstract

Bacillus sp. was isolated from soil for its strong activity of cyclodextrin glucanotransferase (CGTase, EC 2.4.1.19). The enzyme was purified by gel filtration and anion exchange column chromatography using FPLC. The purified enzyme exhibited its maximum CGTase activity in the pH range of 6~8 and the temperature range of 50~$70^{\circ}C$. The molecular weight was estimated as 114,000 by SDS-PAGE. The isoelectric point of the enzyme was 4.3. The CGTase of Bacillus sp. E l produced $\beta$-cyclodextrin mainly and did not produce a-cyclodextrin. The product ratio of $\beta$-cyclodextrin to $\gamma$-cyclodextrin was 7:l.

Cyclodextrin glucanotransferase 생산균주 선발배지를 이용하여 국내 토양으로부터 CGTase 활성이 우수한 Bacillus sp. E1균주를 분리하였다. FPLC를 이용하여 gel filtration과 anion exchange column chromatography를 한 결과 순수 정제된 단일 단백질을 얻을 수 있었으며, 정제된 효소의 최적 작용 pH 범위는 6에서 8까지 였고, 온도는 $60^{\circ}C$에서 최적을 나타냈다. 정제된 단백질의 분자량은 114,000, 등전점은 4.3이었다. 생성된 cyclodextrin은 $\beta$와 $\gamma$-cyclodextrin이 주였으며, 특이하게도 $\alpha$-cyclodextrin은 거의 생성되지 않았다. 작용 후 25시간 후 최대의 $\beta$-cyclodextrin이 생성되었으며, 이때 $\beta$-cyclodextrin과 $\gamma$-cyclodextrin의 생성비율은 7:1이었다.

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