Mi-Ran KI, Bashistha KANTH, Ki Ha MIN, Eui Kyoung JANG, Ki Baek YEO, Seung Pil PACK
언어
영어(ENG)
URL
https://www.earticle.net/Article/A174428
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원문정보
초록
영어
An unusual, internally duplicated, 60 kDa alpha-type carbonic anhydrase of Dunaliella tertiolecta, a unicellular green alga, (Dt-aCA), has been expressed successfully in E. coli system. Here, we demonstrated to express the N- terminal and C-terminal carbonic anhydrase domains of Dt-aCA as distinct proteins using a pET system to increase the expression levels more than one expressed as a duplicated form. The N- and C-terminal domain were expressed using a vector containing an octa-histidine (8 ⨯ His) tag at the C terminus. The purified N- terminal CA domain (residues 1–279) had an apparent molecular mass of; 35 kDa, and retained no activity to hydrolyte p-nitrophenyl acetate (p-NA) into p- nitrophenol and acetate and CO2 hydration. The purified C-terminal CA domain (residues 280-557) had an apparent molecular mass of; 31kDa and retained the ability to catalyze p-NA hydrolysis and CO2 hydration, which were enhanced by the presence of N-terminal domain. The expressions of both domains were increased as compared to that of duplicated dimeric form as one protein. The CA activity was successfully reconstituted by mixing the two domains, N- and C- terminal domains, although N-terminal domain did not show catalytic activity as distinct protein. These results demonstrate that the two domains of CA that has a duplicated dimeric structure can be expressed separately and functionally reconstituted in vitro.
한국생물공학회 [The Korean Society for Biotechnology and Bioengineering]
설립연도
1984
분야
공학>생물공학
소개
이 법인은 생물 공학의 발전과 보급에 이바지하고, 회원 상호 간의 연구 협력과 친목을 도모함을 목적으로 한다
1. 생물공학 분야의 발전을 위한 연구 협력
2. 생물공학의 실용화를 촉진시키기 위한 산학 협동
3. 학술연구 발표회, 강연회, 연수회 등 학술활동의 개최
4. 국,영문 학술지,소식지,학술회의 Proceedings 및 학술도서의 발간
5. 생물공학 발전을 위한 정책 건의
6. 기타 국제 교류 등 본 학회의 목적 달성을 위한 제반 활동