한국생물공학회:학술대회논문집
- 2003.10a
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- Pages.753-757
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- 2003
Development of Enhanced Yeast Expression System for GAP Promoter by Directed Evolution
- Kang, Whan-Koo (Department of Chemical Engineering, Hannam University, Gene to Protein, Inc.) ;
- Hwang, Sun-Duk (Department of Chemical Engineering, Hannam University) ;
- Kim, Bum-Chang (Department of Chemical Engineering, Hannam University) ;
- Lee, Chul-Woo (Department of Chemical Engineering, Hannam University) ;
- Son, Jeong-Il (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, Bheong-Uk (Department of Biological Science, Kosin University, Gene to Protein, Inc.)
- Published : 2003.10.22
Abstract
Escherichia coli and Saccharomyces cerevisiae have been used as host for production of recombinant proteins. It is known that S. cerevisiae has advantages such as good folding and secretion capability, and safety as host over E. coli. But S. cerevisiae has shortcomings of low expression level which is just 20% of that of E. coli. To solve this problem, directed evolution method was tried to enhance the GAP promoter strength of S. cerevisiae in this study. As result, modified GAP promoter that has increased expression level of about 360% compared to that of wild type was selected.
외래 단백질 생산을 위한 숙주로는 Escherichia coli 및 Saccharomyces cerevisiae를 포함한 효모 균주들이 이용되어져 왔다. S. cerevisiae는 대장균에 비해 단백질의 접힘과 분비 능력이 우수하며, 안전한 균주(GRAS)로 알려져 있다. 그러나 S. cerevisiae의 단점은 대장균의 외래 단백질 발현양의 20% 이하 수준의 낮은 외래 단백질 발현율에 있다. 본 연구에서는 S. cerevisiae에서 외래 단백질의 발현을 향상시키기 위해 GAP promoter를 대상으로 분자진화를 수행하였으며, 이 결과 발현율이 360% 증가된 GAP promoter 변이체를 획득하였다.
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