Journal of Korean Society of Forest Science (한국산림과학회지)

Korean Society of Forest Science (한국산림과학회)
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Characterization of Cellulase Gene (MsGHF45) from Monochamus saltuarius Expressed in Yeast

효모에 발현된 북방수염하늘소(Monochamus saltuarius) Cellulase gene MsGHF45의 특성에 관한 연구

  • Ko, Hyeon-Jin (Department of Biomedical Sciences, Kangwon National University) ;
  • Ko, Hyunjun (Department of Biomedical Sciences, Kangwon National University) ;
  • Hong, Soon-Kwan (Department of Biomedical Sciences, Kangwon National University) ;
  • Park, Yong Chul (Department of Biomedical Sciences, Kangwon National University)
  • 고현진 (강원대학교 의생명과학과) ;
  • 고현준 (강원대학교 의생명과학과) ;
  • 홍순관 (강원대학교 의생명과학과) ;
  • 박용철 (강원대학교 의생명과학과)
  • Received : 2016.02.15
  • Accepted : 2016.05.03
  • Published : 2016.06.30
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Abstract

In this study, the cellulase of Monochamus saltuarius (MsGHF45) gene was introduced in Kluyveromyces lactis, successfully. The molecular weight of recombinant enzyme was determined by SDS-PAGE and western blotting. The enzymatic activity was confirmed by native-PAGE containing carboxymethyl cellulose as a substrate. The optimul pH and temperature of recombinant MsGHF45 was pH5 and $40^{\circ}C$. The barium ($Ba^{2+}$) and ferrous ($Fe^{2+}$) enhanced enzyme activity, and the mercuty ($Hg^{2+}$) inhibited its activity.

화석연료의 고갈 문제로 인하여 다양한 대체에너지가 개발 중이며 본 연구에서는 바이오에탄올 연구의 초석으로 사용하기 위해 곤충의 효소를 연구하였다. 북방수염하늘소(Monochamus saltuarius Gebler)가 지닌 cellulase (MsGHF45) 유전자를 Kluyveromyces lactis에 형질전환 시켰다. 형질전환 된 효모는 활성이 있는 cellulase 효소를 성공적으로 생산하였다. 재조합 MsGHF45의 분자량은 SDS-PAGE와 western blot을 통하여 확인하였다. 효소의 활성은 기질로써 carboxymethyl cellulose를 첨가한 native-PAGE를 통해서 검증하였다. 효소 활성의 최적조건을 알아본 결과, pH는 5에서, 온도는 $40^{\circ}C$에서 나타났다. 바륨이온($Ba^{2+}$)과 철이온($Fe^{2+}$)은 효소활성을 저해하지 않았으나, 칼슘이온($Ca^{2+}$), 코발트이온($Co^{2+}$), 구리이온($Cu^{2+}$), 수은이온($Hg^{2+}$), 마그네슘이온($Mg^{2+}$), 망간이온($Mn^{2+}$) 그리고 아연이온($Zn^{2+}$)은 활성을 저해하였다. 특히, 수은이온은 효소활성을 66.5% 감소시켰다. 본 연구는 다양한 효소를 이용한 biofuel 연구에 참고자료로 사용될 것이다.

Keywords

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