한국생물공학회:학술대회논문집

The Korean Society for Biotechnology and Bioengineering (한국생물공학회)
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The development of modified cellulase with higher activity by directed evolution

  • Kang, Whan-Koo (Department of Chemical Engineering, Hannam University) ;
  • Son, Jeong-Il (Department of Chemical Engineering, Hannam University) ;
  • Hwang, Sun-Duk (Department of Chemical Engineering, Hannam University) ;
  • Kim, Bum-Chang (Department of Chemical Engineering, Hannam University) ;
  • Kim, Hyoung-Sik (Department of Chemical Engineering, Hannam University) ;
  • Lee, Byung-Ryul (Department of Chemical Engineering, Hannam University) ;
  • Lee, Chul-Woo (Department of Chemical Engineering, Hannam University)
  • Published : 2003.10.22
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Abstract

In the study, we have obtained modified cellulase with higher cellulose degradation activity by molecular evolution method. Cellobiohydrolase(CBH I ) gene of Trichorderma reerri has been used. Cellulase mutant 228-G2 was selected and the activity of cellulase mutant 228-G2 was increased by 300% compared to original CBH I The 17 among 1542bases were found to be modified with mutant 228-G2.

본 연구에서는 Trichorderma reerri의 cellobiohydrolase (CBH1) gene을 이용하여 분자진화 방법을 이용한 cellulose분해 활성이 증가된 cellulase 변이체를 선별하고자 하였다. 재조합된 벡터가 도입된 균주의 발현을 SDS-PAGE로 확인한 후, 분자 진화에 의한 cellulase 변이체를 trypan blue staining법으로 cellulase 변이체 228-G2를 선별하였으며, cellulose분해 활성을 DNS법으로도 다시 확인 할 수 있었다. Cellulase 변이체 228-G2의 분해활성 증가율은 original CBH I에 비해 약 300%증가 된 것을 확인하였고, DNA sequence를 확인할 결과 1542bp 중 17개의 염기서열이 바뀐 것을 알 수 있었다.

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