Korean Journal of Microbiology (미생물학회지)

The Microbiological Society of Korea (한국미생물학회)
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Characterization of a Chitinase Gene and Screening of Cold Active Chitinase from Polar Microorganisms

극지유래 저온활성 Chitinase 생산균주의 스크리닝과 Chitinase 유전자 클로닝

  • Park, Yu Kyung (Division of Life Sciences, Korea Polar Research Institute (KOPRI)) ;
  • Kim, Jung Eun (Division of Life Sciences, Korea Polar Research Institute (KOPRI)) ;
  • Lee, Hyoungseok (Division of Life Sciences, Korea Polar Research Institute (KOPRI)) ;
  • Kim, Ji Hyun (Division of Life Sciences, Korea Polar Research Institute (KOPRI)) ;
  • Park, Ha Ju (Division of Life Sciences, Korea Polar Research Institute (KOPRI)) ;
  • Kim, Dockyu (Division of Life Sciences, Korea Polar Research Institute (KOPRI)) ;
  • Park, Mira (Division of Life Sciences, Korea Polar Research Institute (KOPRI)) ;
  • Yim, Joung Han (Division of Life Sciences, Korea Polar Research Institute (KOPRI)) ;
  • Kim, Il-Chan (Division of Life Sciences, Korea Polar Research Institute (KOPRI))
  • 박유경 (극지연구소 생명과학연구부) ;
  • 김정은 (극지연구소 생명과학연구부) ;
  • 이형석 (극지연구소 생명과학연구부) ;
  • 김지현 (극지연구소 생명과학연구부) ;
  • 박하주 (극지연구소 생명과학연구부) ;
  • 김덕규 (극지연구소 생명과학연구부) ;
  • 박미라 (극지연구소 생명과학연구부) ;
  • 임정한 (극지연구소 생명과학연구부) ;
  • 김일찬 (극지연구소 생명과학연구부)
  • Received : 2012.07.27
  • Accepted : 2012.11.05
  • Published : 2012.12.31
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Abstract

Of the 169 strains of microorganisms stored in Polar and Alpine Microbial Collection of Korea Polar Research Institute, 27 strains were selected for their chitinase activity on ZoBell plates supplemented with 0.4% colloidal chitin. Among them, PAMC 21693 strain have shown the highest chitinolytic enzyme activity toward pNP-$(GlcNAc)_1$ at low temperature and the highest growth rate at $4^{\circ}C$. We cloned a full-length chitinase gene of 2,857 bp which contains an open reading frame of 2,169 bp encoding 872-amino acid polypeptide. Recombinant chitinase protein was expressed in E. coli and its molecular weight was confirmed 96 kDa. In this paper, we suggest the potential use of cold-active chitinase from polar microorganisms in the field of biotechnology.

극지의 다양한 환경으로부터 분리되어 극지연구소 PAMC(Polar and Alpine Microbial Collection)에 보관중인 169개 균주들을 0.4% colloidal chitin이 첨가된 ZoBell 고체배지에서 배양하여 chitinase 활성을 균주 27개를 선별하였다. 그 중 PAMC 21693 균주는 저온에서 pNP-$(GlcNAc)_1$를 기질로 사용했을 때 가장 큰 활성을 보였고, $4-37^{\circ}C$의 온도 범위 중 $4^{\circ}C$에서 가장 높은 개체수 증가율을 보였다. PAMC 21693의 chitinase 유전자를 클로닝한 결과 2,619 bp의 ORF를 포함하는 총 2,857 bp의 염기서열을 확보하였다. 대장균에서 chitinase 유전자의 재조합 단백질을 발현한 결과 분자량 96 kDa의 재조합 단백질을 확인할 수 있었다. 본 논문에서는 극지 미생물 유래 저온활성 chitinase들의 생물공학 분야에서의 이용가능성을 제시하였다.

Keywords

References

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