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Functional Expression and Characterization of Recombinant Chitinase from Antarctic Psychrophilic Bacterium Sanguibacter antarcticus KCTC 13143

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Title
Functional Expression and Characterization of Recombinant Chitinase from Antarctic Psychrophilic Bacterium Sanguibacter antarcticus KCTC 13143
Other Titles
남극 호냉성 세균 Sanguibacter antarcticus KCTC 13143의 재조합 키틴분해효소의 발현 및 특성연구
Authors
이창은
Kim, Il-Chan
김주영
Yim, Joung Han
Kim, Dockyu
Park, Ha Ju
Keywords
Cold-active; Endochitinase; Psychrophilic; Sanguibacter
Issue Date
2008
Citation
이창은, et al. 2008. Functional Expression and Characterization of Recombinant Chitinase from Antarctic Psychrophilic Bacterium Sanguibacter antarcticus KCTC 13143. 극지연구소. 극지연구소. 2008.09.25~.
Abstract
A psychrophilic Sanguibacter antarcticus KCTC 13143 has been previously isolated from sea sand on King Sejong Station in Antarctica. An cold-active endochitinase gene, designated chi21702, from KCTC 13143 was PCR-amplified, cloned into pEXP5-CT/TOPO vector, and overexpressed in Eschrichia coli BL21(DE3) without its signal-peptide and with an C-terminal His-tag. The recombinant chitinase (R-Chi21702) was approximately 56 kDa in size, most of which was converted to inclusion body having no chitinae activity against oligomers of N-acetylglucosamine. In next step, refolding experiment for the inert chitinase was carried out using optimized refolding conditions. The refolded R-Chi21702 was then purified using a Ni-NTA column, which restored its chitinase specific activity (5.0 U/mg). The optimal reaction temperature and pH of R-chitinase were determined to be 40 ℃ and pH 8.0, respectively.
URI
http://repository.kopri.re.kr/handle/201206/7873
Conference Name
극지연구소
Conference Place
극지연구소
Conference Date
2008.09.25~
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