생명과학회지 (Journal of Life Science)

  • 제23권7호
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  • Pages.863-868
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  • 2013
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  • 1225-9918(pISSN)
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  • 2287-3406(eISSN)
한국생명과학회 (Korean Society of Life Science)
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출아효모(Saccharomyces cerevisiae)에서 Bacillus sp. HY-20균주의 재조합 endoxylanase의 효율적 분비 발현

Efficient Secretory Expression of Recombinant Endoxylanase from Bacillus sp. HY-20 in Saccharomyces cerevisiae

  • 김민지 (동의대학교 생명공학과) ;
  • 김보현 (동의대학교 생명공학과) ;
  • 남수완 (동의대학교 생명공학과) ;
  • 최의성 (한국생명공학연구원 바이오화학연구센터) ;
  • 신동하 ((주)인섹트 바이오텍) ;
  • 조한영 (한국생명공학연구원 산업바이오소재연구센터) ;
  • 손광희 (한국생명공학연구원 산업바이오소재연구센터) ;
  • 박호용 (한국생명공학연구원 산업바이오소재연구센터) ;
  • 김연희 (동의대학교 생명공학과)
  • Kim, Min-Ji (Department of Biotechnology and Bioengineering, Dong-Eui University) ;
  • Kim, Bo-Hyun (Department of Biotechnology and Bioengineering, Dong-Eui University) ;
  • Nam, Soo-Wan (Department of Biotechnology and Bioengineering, Dong-Eui University) ;
  • Choi, Eui-Sung (Biochemicals and Synthetic Biology Research Center, Korea Research Institute of Bioscience and Biotechnology (KRIBB)) ;
  • Shin, Dong-Ha (Insect Biotech Co. Ltd.) ;
  • Cho, Han-Young (Industrial Biomaterials Research Center, Korea Research Institute of Bioscience and Biotechnology (KRIBB)) ;
  • Son, Kwang-Hee (Industrial Biomaterials Research Center, Korea Research Institute of Bioscience and Biotechnology (KRIBB)) ;
  • Park, Ho-Yong (Industrial Biomaterials Research Center, Korea Research Institute of Bioscience and Biotechnology (KRIBB)) ;
  • Kim, Yeon-Hee (Department of Biotechnology and Bioengineering, Dong-Eui University)
  • 투고 : 2013.05.30
  • 심사 : 2013.07.01
  • 발행 : 2013.07.30
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초록

Bacillus sp. HY-20균주 유래 endoxylanase를 코드하는 XylP 유전자를 효모에서 발현시키기 위해 두 개의 발현 플라스미드 pG-xylP와 pGMF-xylP를 구축하였다. 이들 플라스미드는 endoxylanase의 분비발현을 위해 각각 다른 분비서열인 XylP 유전자의 자체 분비서열(XylP s.s)과 최적화된 $MF{\alpha}$ 분비서열($MF{\alpha}_{opt}$ s.s)을 가지고 있으며, S. cerevisiae SEY2102와 FY833균주에 형질전환되어 그 분비활성이 비교 조사되었다. 재조합 endoxylanase는 분비발현시스템과 숙주세포에 따라 23.7~70.1 unit/ml의 활성으로 효모 세포에서 성공적으로 발현되었고, 그 중 SEY2102/pGMF-xylP 형질전환주를 이용해 baffled-flask 배양을 실시한 결과 최대 88.1 unit/ml의 endoxylanase 활성을 보임을 확인하였다. 대부분의 재조합 endoxylanase는 세포 외 분획에 효율적으로 분비 생산되었으며, $MF{\alpha}_{opt}$ 분비서열이 XylP 유전자의 자체 분비서열보다 endoxylanase를 더 효율적으로 분비시킴을 확인할 수 있었다. 그러므로 본 연구에서 개발된 발현시스템은 효모를 숙주세포로 하여 많은 양의 세포 외 endoxylanase의 생산을 가능하게 하고, 바이오에탄올 생산 및 산업적 응용에도 유용하게 사용 될 수 있으리라 기대된다.

The XylP gene, which encodes endoxylanase in Bacillus sp. HY-20, was subcloned, and two expression plasmids, pG-xylP and pGMF-xylP were constructed. These plasmids, which contain different signal sequences, XylP s.s and $MF{\alpha}_{opt}$ s.s, respectively, for the secretory expression of endoxylanase, were transformed into Saccharomyces cerevisiae SEY2102 and FY833, respectively. The recombinant endoxylanases were successfully expressed, with a total activity range of 23.7-70.1 unit/ml according to the expression system and host strain. The endoxylanase activity in SEY2102/pGMF-xylP reached a maximum of 88.1 unit/ml in baffled flask culture. Most of the recombinant endoxylanase was efficiently secreted in the extracellular fraction, and the $MF{\alpha}_{opt}$ s.s was more efficient for secreting endoxylanase in yeast than the XylP s.s. Therefore, the expression system developed in this study produces large extracellular amounts of endoxylanase using S. cerevisiae as the host strain, and it could be used in bioethanol production and industrial applications.

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참고문헌

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