Purification of \alpha-Cyclodextrin Glucanotransferase Excreted from Themophilic Geobacillus thermosac-chalytycus and Characterization of Transglycosylation Reaction of Glucosides.

호열성 Geobacillus thermosacchalytycus가 생산하는 \alpha-Cyclodextrin Glucanotransferase의 분리정제와 당전이 반응 특성

  • 이미숙 (경북대학교 자연과학대학 유전공학과) ;
  • 신현동 (이엔지바이오㈜) ;
  • 김태권 (경북대학교 자연과학대학 유전공학과) ;
  • 이용현 (경북대학교 자연과학대학 유전공학과, 이엔지바이오㈜)
  • Published : 2004.03.01

Abstract

$\alpha$-Cyclodextrin glucanotransferase excreted from a newly isolated Geobacillus thermosacchalytycus was purified through the ultrafiltraion, hydrophobic Sepharose CD-4B affinity chromatography, and gel filtration on Sephadex G-75, respectively. The molecular weight of the purified CGTase was 69 kDa and its N-terminal amino acid sequence was determined to be Asn-Leu-Asn-Lys-Val-Asn-Phe-Val-Ser-Asp-Val-Val-Val-Gln-Ile. The optimum pH and temperature were pH 6.0 and$ 60^{\circ}C$, respectively, and stably at the pH range of 6.0-8.0 and $60^{\circ}C$ in the presence of $Ca^{++}$. The excreted CGTase from the thermophilic G. thermosacchalytycus was $\alpha$-type showing a high coupling activity for the transglycosylation on various glucosides. The coupling reaction was carried out according to the random ternary complex mechanism.m.

Keywords

Geobacillus thermosacchalytycus;\alpha-cyclodextrin glucanotransferase;thermophiles;disproportionation reaction;glucosides

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