Purification and Characterization of Cyclodextrin Glucanotransferase from Bacillus stearothermophilus KJ16

Bacillus stearothermophilus KJ16이 생산하는 Cyclodextrin Glucanotransferase 의 정제와 효소특성

  • Kwon, Hyun-Ju ;
  • Nam, Soo-Wan ;
  • Kim, Kwang-Hyun ;
  • Song, Seong-Koo ;
  • Yun, Jong-Won ;
  • Kim, Byung-Woo
  • 권현주 (동의대학교 미생물학과) ;
  • 남수완 (동의대학교 미생물학과) ;
  • 김광현 (동의대학교 미생물학과) ;
  • 송승구 (부산대학교 화학공학과) ;
  • 윤종원 (대구대학교 생물공학과) ;
  • 김병우 (동의대학교 미생물학과)
  • Published : 1998.06.01


Cyclodextrin glucanotransferase from B. stearothermophilus KJ16 that can produce both cyclodextrin glucanotransferase and cyclodextrinase was purified by ammonium sulfate precipitation, DEAE-cellulose chromatography, Sephadex G-100 chromatography, and FPLC. The molecular weight of the purifice enzyme was about 65,000 dalton by SDS-PAGE. The optimal pH and temperature were 6.0 and $60^{\circ}C$, respectively. The enzyme was stable at $50^{\circ}C$ for 1 hr and in the pH range of 5.5 and 8.5. Mercaptoethanol and dithiothreitol inhibited the enzyme activity strongly. The enzyme produced 60% cyclodextrin(CD) from 5% soluble starch with the $^{\alpha}$, $^{\beta}$, $^{\gamma}$-CD ratio of 42:46:12. Amylopectin was the most suitable substrate with 67% conversion to CD.



  1. J. Am. Chem. Soc. v.61 The conversion of starch to crystalline dextrin by the action of a new type of amylase separated from cultures Aeromonal Tilden, E. B.;Hudson, C. S.
  2. J. Jpn. Soc. Starch Sci. v.33 Compatative studies of CGTases from Bacillus ohbensis, Bacilus macerans and Bacillus circulans and porduction of cyclodextrin using those CGTase Yagi, Y.;Sato, M.;Ishikura, T.
  3. Kor. J. Appl. Mirobiol. Bioeng. v.17 Purification and characterization of cyclodextrin glucanotransferase excreted from newly isolated alkalophilic Bacillus cirulans Shin, H. D.;Lee, S. H.;Lee, Y. H.
  4. J. Jpn. Soc. Starch Sci. v.29 Comparison of action of cyclodextrin glucanotransferase from B. megaterium, B. circulans, B. stearothermophilus and B. macerans Kitahata, S.;Okada, S.
  5. Kor. J. Appl. Micrbiol. Biotechnol. v.18 Cyclodextrin glucanotransferase from Bacillus stearothermophilus : Purification by affinity chromatography and its properties Ahn, J. H.;Hwang, J. B.;Kim, S. H.
  6. J. Ferment. Bioeng. v.71 Cyclomaltodextrin glucanotransferase producing moderate thermophile Akimaru, K.;Yagi, T.;Yamamoto. S.
  7. Kor. J. Appl. Microbiol. Biotechnol. v.21 Screening of new alkalophilic strain for overproduction of cyclodextrin glucanotransferase Do, E. J.;Park, J. B.;Lee, Y. H.
  8. Arch. Microbiol. v.111 Cyclodextrin glucanotansferase von K. pneumoniae Bender, H.
  9. Agric. Biol. Chem. v.40 Purification and properties of cyclodextrin glycosyltransferase of an alkalophilic Bacillus sp Nakamura, N.;Horikoshi, K.
  10. Kor. J. Appl. Microbiol. Bioeng. v.17 Isolation and chracterization cyclodextrin glycosyltransferase producing alkalophilic Bacillus sp Yu, J. H.;Chung, Y. J.;Lee, J. S.
  11. Kor. J. Appl. Microbiol. Biotehnol. v.20 Purification and characterization of cyclodextrin glucanotransferase from Bacillus sp. E1 Park, C. S.;Woo, E. J.;Kuk, S. U.;Seo, B. C.Park, K. H.;Lim, H.
  12. Kor. J. Appl. Microbiol. Biotehcnol. v.24 Isolation of a Bacillus sp. producing both cyclodextrin glucanotransferase and cyclodextrinase and characterization of the enzymes Kwon, H. J.;Nam, S. W.;Kim, K. H.;Kwak, Y. G.;Kim, B. W.
  13. Kor. J. Appl. Microbiol. Biotehcnol. v.24 Catabolite repression of cyclodextrin glucanotransferase and cyclodextrinase syntheses in Bacillus sp. KJ16 Kim, B. W.;Kwon, H. J.;Lee, K. H.
  14. Appl. Environ. Microbiol. v.58 Cyclization charcteristics of cyclodextrin glucanotransferase are conferred by the NH₂-terminal region of the enzyme Fujiwara, S.;Kakihara, H.;Kim, B. W.;Leujeune, A.;Kanemoto, M.;Sakaguchi, K.;Imanaka, T.
  15. Anal. Biochem. v.181 A spectrophotometric assay for the cyclization activity of cyclomaltohexaose glucanotransferase Lejeune, A.;Sakaguchi, K.;Imanaka, T.
  16. Anal. Biochem. v.150 Measurement of protein using bicinchoninic acid Smith, P. K.;Hrohn, R. I.;Hermanson, G. T.;Mallia, A. K.;Gartner, F. H.;Provenzano, M. D.;Fujimoto, E. K.;Goeke, N. M.;Olson, B. J.;Klenk, D. C.
  17. Nature v.277 Cleavage of structural proteins during the assembly of the head of bacteriophage T4 Laemmli, U. K.
  18. Kor. J. Appl. Microbiol. Biotehcnol. v.18 Production of cyclomaltodextrin from Bacillus stearomophilus Hwang, J. B.;Kim, S. H.;Lee, T. K.;Yang, H. C.