산업기술연구 (Journal of Industrial Technology)

  • 제36권
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  • Pages.33-37
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  • 2016
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  • 1229-9588(pISSN)
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  • 1598-1371(eISSN)
강원대학교 산업기술연구소 (Kangwon National University, Institute of Industrial Technology)
권호 표지

Intein을 이용한 대장균에서의 Trichoderma reesei 유래의 Cellobiohydrolase I 섬유소 결합 도메인의 발현

Intein-mediated expression of Trichoderma reesei Cellobiohydrolase I Cellulose Binding Domain in E. coli

  • 최신건 (강원대학교 생물공학과)
  • 발행 : 2016.02.29
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초록

Cellulose binding domains (CBDs) of cellulases are thought to assist in the hydrolysis of insoluble crystalline cellulose. To gain sufficient amount of CBDs, the self-cleavable intein tag was used for expression and purification of Trichoderma reesei cellobiohydrolase I CBD in E. coli. Synthetic CBD genes, CBD or linker-CBD were cloned into expression vector pTYB11. Recombinant CBDs were successfully purified by intein mediated purification with an affinity chitin-binding domain. The final yields of recombinant CBD and linker-CBD were 3.2 mg/L and 1.4 mg/L, respectively. The functional bindings of recombinant CBDs were confirmed by Avicel binding experiments. The simple and easy purification method using self-cleavable intein tag can be further used in pretreatment of crystalline cellulose or characterization of engineered CBDs.

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참고문헌

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