Korean Journal of Microbiology (미생물학회지)

The Microbiological Society of Korea (한국미생물학회)
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Characterization of Bacillus licheniformis B1 ${\beta}$-1,4-Glucanase Overproduced in Escherichia coli

대장균에서 과잉생산된 Bacillus licheniformis B1의 ${\beta}$-1,4-Glucanase 특성

  • Song, Hye-Jung (Department of Biotechnology, The Research Institute for Basic Sciences, Hoseo University) ;
  • Kim, Hwang-Yeon (Department of Biotechnology, The Research Institute for Basic Sciences, Hoseo University) ;
  • Hwang, Jae-Sung (Department of Biotechnology, The Research Institute for Basic Sciences, Hoseo University) ;
  • Kim, Han-Bok (Department of Biotechnology, The Research Institute for Basic Sciences, Hoseo University)
  • 송혜정 (호서대학교 자연과학대 생명공학과, 기초과학연구소) ;
  • 김황연 (호서대학교 자연과학대 생명공학과, 기초과학연구소) ;
  • 황재성 (호서대학교 자연과학대 생명공학과, 기초과학연구소) ;
  • 김한복 (호서대학교 자연과학대 생명공학과, 기초과학연구소)
  • Received : 2010.02.02
  • Accepted : 2010.03.04
  • Published : 2010.03.31
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Abstract

The ${\beta}$-1,4-glucanase gene of Bacillus licheniformis B1 was expressed in Esherichia coli BL21, and a protein with a mass of 50 kDa that was soluble was overproduced. A protein with a mass of 37 kDa was secreted from B. licheniformis. It seems that the ${\beta}$-1,4-glucanase produced in E. coli contained the leader peptide and unprocessed carboxy-terminal region, but its processing occurred in the carboxyterminal in Bacillus. The optimal temperature of ${\beta}$-1,4-glucanase was $40^{\circ}C$. The enzyme still had 76% maximal activity at $60^{\circ}C$. The optimal pH of the enzyme was 7. The enzyme retained considerable activities over the weak-acidic, neutral, and weak-basic pH range. Acidic fungal cellulases are used in food, detergent, pulp, paper, textile industries. However, studies about neutral and alkaline cellulase are not enough. The cellulase developed in this study may be useful for industrial applications in the fields of biofuel development.

Bacillus licheniformis B1의 ${\beta}$-1,4-glucanase 유전자는 Escherichia coli BL21에서 발현되어, 이로부터 수용성의 50 kDa 단백질이 과량생산되었다. 반면에 B. licheniformis에서는 37 kDa의 형태가 분비되었다. E. coli에서 발현된 ${\beta}$-1,4-glucanase는 leader peptide가 제거되지 않고 포함되어 있고 Bacillus에서는 효소의 carboxy 말단에서 processing이 일어난 것으로 보인다. E. coli에서 생산된 ${\beta}$-1,4-glucanase의 최적온도는 $40^{\circ}C$이었지만, $60^{\circ}C$에서도 최대치의 76% 활성을 유지하였다. 효소의 최적 pH는 7이었고, 효소의 활성은 전체적으로 보면 약산성, 중성 및 약알칼리 영역에 널리 걸쳐 있었다. Cellulase는 식품, 세제, 펄프, 제지, 섬유산업 등 다양한 분야에서 주로 산성의 곰팡이계 효소가 이용되고 있으나, 중성 및 알칼리성 cellulase 연구 및 개발은 미흡한 편이다. 본 연구에서 개발된 중성 cellulase가 바이오 연료 개발 등의 분야에서 활용되기를 기대해 본다.

Keywords

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