Microbiology and Biotechnology Letters (한국미생물·생명공학회지)

The Korean Society for Microbiology and Biotechnology (한국미생물·생명공학회)
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Simultaneous Overpexpression of Genes Encoding Cellulose- and Xylan-Degrading Enzymes through High Density Culture of a Recombinant Yeast Cell

재조합 효모 세포의 고농도배양을 통한 섬유소와 자일란 분해효소 유전자의 동시 과발현

  • Kim, Yeon-Hee (Biomedical Engineering and Biotechnology Major, Division of Applied Bioengineering, College of Engineering, Dong-Eui University) ;
  • Heo, Sun-Yeon (Microbial Biotechnology Research Center, Jeonbuk Branch Institute, Korea Research Institute of Bioscience and Biotechnology (KRIBB)) ;
  • Kim, Gun-Do (Department of Microbiology, College of Natural Sciences, Pukyong National University) ;
  • Nam, Soo-Wan (Biomedical Engineering and Biotechnology Major, Division of Applied Bioengineering, College of Engineering, Dong-Eui University)
  • 김연희 (동의대학교 공과대학 바이오응용공학부 의생명공학전공) ;
  • 허선연 (한국생명공학연구원 전북분원 미생물기능연구센터) ;
  • 김군도 (부경대학교 미생물학과) ;
  • 남수완 (동의대학교 공과대학 바이오응용공학부 의생명공학전공)
  • Received : 2018.10.16
  • Accepted : 2018.11.12
  • Published : 2018.12.28
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Abstract

For the coexpression of endoxylanase and endoglucanase genes in yeast Saccharomyces cerevisiae, the genes were separately inserted downstream of the yeast ADH1 promoters, resulting the plasmid pAGX3 (9.83 kb). In the batch culture on YPD medium of the yeast transformant, S. cerevisiae SEY2102/pAGX3, the total activities of the enzymes reached about 7.91 units/ml for endoxylanase and 0.43 units/ml for endoglucanase. In the fed-batch culture with intermittent feeding of yeast extract and glucose, the total activities of 24.9 units/ml for endoxylanase and 0.84 units/ml for endoglucanase were produced which were about 3.1-fold and 2.0-fold increased levels, respectively, compared to those of the batch culture. Most of endoxylanase and endoglucanase activities were found in the extracellular media. This recombinant yeast could be useful for the development of simultaneous saccharification bioprocess of the cellulose and xylan mixture.

Endoxylanase와 endoglucanase 유전자가 ADH1 프로모터 하류에 따로따로 삽입된 pAGX3 플라스미드를 함유한 Saccharomyces cerevisiae에서 endoxylanase와 endoglucanase 유전자는 성공적으로 발현되었으며, YPD 배지에서의 회분배양 결과, endoxylanase는 7.91 units/ml, endoglucanase는 0.43 units/ml에 달하는 총활성을 보였다. Yeast extract와 포도당을 간헐적으로 공급하는 유가배양에서 endoxylanase와 endoglucanase의 총활성은 24.9 units/ml과 0.84 units/ml을 각각 보였으며, 이는 회분배양에서 발현된 각각 활성의 3.1배와 2배에 해당되었다. 또한, 대부분의 endoxylanase와 endoglucanase 활성은 세포밖 배지에서 측정되어, 향후 이 재조합 효모는 섬유소(cellulose)와 xylan 혼합물의 동시당화 바이오공정 개발에 활용될 가능성이 높다 하겠다.

Keywords

References

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