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The Korean Society for Biotechnology and Bioengineering (한국생물공학회)
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Optimization for Production of Exo-β-1,3-glucanase (Laminarinase) from Aspergillus oryzae in Saccharomyces cerevisiae

Saccharomyces cerevisiae에서 Aspergillus oryzae 유래의 exo-β-1,3-glucanase (laminarinase)의 생산 최적화

  • Kim, Min-Jung (Department of Biomaterial Control (BK21 program)) ;
  • Nam, Soo-Wan (Department of Biomaterial Control (BK21 program)) ;
  • Tamano, Koichi (Research Institute for Cell Engineering, National Institute of Advanced Industrial Science and Technology (AIST)) ;
  • Machida, Masayuki (Research Institute for Cell Engineering, National Institute of Advanced Industrial Science and Technology (AIST)) ;
  • Kim, Sung-Koo (Department of Biotechnology and Bioengineering, Pukyong National University) ;
  • Kim, Yeon-Hee (Department of Biomaterial Control (BK21 program))
  • 김민정 (동의대학교 바이오물질제어학과) ;
  • 남수완 (동의대학교 바이오물질제어학과) ;
  • ;
  • ;
  • 김성구 (부경대학교 생물공학과) ;
  • 김연희 (동의대학교 바이오물질제어학과)
  • Received : 2011.07.28
  • Accepted : 2011.10.17
  • Published : 2011.10.31
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Abstract

In this study, a EXGA gene code for exo-β-1,3-glucanase from Aspergillus oryzae was overexpressed and secretory produced in Saccharomyces cerevisiae. To overexpress the β-1,3-glucanase, pGInu-exgA and pAInu-exgA plasmids having GAL10 and ADH1 promoter, respectively, and exoinulinase signal sequence (Inu s.s) were constructed and introduced in S. cerevisiae SEY2102 and 2805. The recombinant β-1,3-glucanase was successfully expressed and secreted into the medium and the β--1,3-glucanase activity in 2102/pGInu-exgA and 2102/pAInu-exgA strain were 5.01 unit/mL and 4.09 unit/mL, respectively. In the 2805/pGInu-exgA and 2805/pAInu-exgA strain, the β-1,3-glucanase activity showed 3.23 unit/mL and 3.22 unit/mL, respectively. Secretory efficiency in each strain reached 95% to 98%. Subsequently, the recombinant β1,3-glucanase was used for ethanol production. Ethanol productivity in 2102/pAInu-exgA strain was 0.83 g/L when pre-treated Laminaria japonica which has initial reducing sugar of 1.4 g/L was used as substrate. It is assumed that the polysaccharides of Laminaria japonica was effectively saccharified by recombinant β-1,3-glucanase, resulting in increase of ethanol productivity. These results suggested that recombinant β-1,3-glucanase was efficiently overexpressed and secreted in S. cerevisiae SEY2102 as host strain by using ADH1 promoter-Inu s.s system.

Keywords

References

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