Cloning, Expression, and Purification of Exoinulinase from Bacillus sp. snu-7

A gene encoding inulin-degrading enzyme of Bacillus sp. snu-7 with ORF of 1536 nucleotides was cloned. And it was overexpressed as His-tagged protein in E. coli BL21(DE3) pLysS using pRSET B vector containing mature enzyme sequence. Maximum enzyme production was achieved by IPTG (0.1 mM) induction a...

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Veröffentlicht in:	Journal of microbiology and biotechnology 2004-04, Vol.14 (2), p.344-349
Hauptverfasser:	Kim, K.Y. (Seoul National University, Seoul, Republic of Korea), Koo, B.S. (BioNgene Co., Ltd., Seoul, Republic of Korea), Jo, D.H. (Ajou University, Suwon, Republic of Korea), Kim, S.I. (Seoul National University, Seoul, Republic of Korea), E-mail: sikim@plaza.snu.ac.kr
Format:	Artikel
Sprache:	eng
Schlagworte:	BACILLUS Bacillus sp. snu-7 Biological and medical sciences Biotechnology CHROMATOGRAPHIE CHROMATOGRAPHY CROMATOGRAFIA exoinulinase Fundamental and applied biological sciences. Psychology immobilized metal affinity chromatography
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Zusammenfassung:	A gene encoding inulin-degrading enzyme of Bacillus sp. snu-7 with ORF of 1536 nucleotides was cloned. And it was overexpressed as His-tagged protein in E. coli BL21(DE3) pLysS using pRSET B vector containing mature enzyme sequence. Maximum enzyme production was achieved by IPTG (0.1 mM) induction at OD∧600 1.2 and 30℃ followed by 6 h incubation. The expressed protein purified through immobilized metal affinity chromatography showed molecular mass of 60 kDa on SDS-PAGE.
ISSN:	1017-7825