Production of Cyclofructan by Cycloinulooligosaccharide Fructanotransferase Expressed in Saccharomyces cerevisiae.

Saccharomyces cerevisiae에서 발현된 Cycloinulooligosaccharide Fructanotransferase을 이용한 Cyclofructan의 생산

  • 임채권 (동의대학교 미생물학과) ;
  • 김현철 (동의대학교 바이오물질제어학과) ;
  • 김광현 (동의대학교 미생물학과) ;
  • 김병우 (동의대학교 미생물학과) ;
  • 남수완 (동의대학교 생명공학과)
  • Published : 2004.03.01


The cycloinulooligosaccharide fructanotransferase(CFTase) gene (cft) from Paenibacillus polymyxa was subcloned into the E. coli-yeast shuttle vector, pYES2.0 (GALI promoter). The constructed plasmid, pYGCFT (9.9 kb) was introduced into S. cerevisiae SEY2102 cell and then the yeast transformant was selected on the synthetic defined media lacking uracil Based on the cyclofructan(CF) spots on thin-layer chromatogram, the gene under the control of GALI promoter was successfully expressed in the yeast transformant. The recombinant CFTase was not secreted into the medium and was predominantly localized in the periplasmic space. CF was started to be produced after 3h of enzymatic reaction with inulin. The pH and temperature optimum for the CF production from inulin was pH 8.0 and 45$^{\circ}C$, respectively. Enzyme activity was stably maintained up to the pH of 10.0. The examination of the inulin sources revealed that a dahlia tuber and Jerusalem artichoke were the best for the production of CF.


Cyclofructan;cycloinulooligosaccharide fructanotransferase;expression;inulin;Paenibacillus polymyxa;Saccharomyces cerevisiae


  1. Anal. Chem. v.55 Use of dinitrosalicylic acid teagent for determination of reducing sugar Miller,G.L.
  2. Carbohydr. Res. v.241 Complexing of cycloinulo-oligosaccharides with metalions Uchiyama,T.;M.Kawamura;T.U.ragami;H.Okuno
  3. Carbohydr. Res. v.217 The crystal structure of cycloinulohexaose produced from inulin by cycloinulooligosaccharide fructanotransferase Sawada,M.;T.Tanaka;Y.Takai;T.Hanafrsa;T.Taniguchi;M.Kawamura;T.Uchiyama
  4. Proc. Natl. Acad. Sci. v.80 An Ma$\'{a}$l-SUC2 (α-factor-invertase) gene fusion for study of protein localization and gene expression in yeast Emr,S.D.;R.Schekman;M.C.Flessel;J.Thorner
  5. J. Microbiol. Biotechnol v.12 Cloning and characterization of cycloinulooligosaccharide fructanotransferase(CFTase) from Bacillus polymyxa MGL21 Jeon,S.J.;d.J.You;H.J.Kwon;s.Kanaya;N.Kunihiro;K.H.Kim;Y.H.Kim;B.W.Kim
  6. Trends Biotechnol. v.7 Cyclodextrin glycosyltransferase production;yield enhancement by overexpression of cloned genes Schmid,G.
  7. Biotechnol. Lett. v.15 Secretion and localization of invertase and inulinase in recombinant Saccharomyces cerevisiae Nam,S.W.;K.Yoda;M.Yamasaki
  8. Biotechnol. Lett. v.23 Expression of Bacillus macerans cyclodextrin glucanotransferase gene in Saccharomyces cerevisiae Nam,S.W.;H.Y.Park;J.H.Kim;J.H.Seo;N.S.Han;B.W.Kim
  9. Annu. rev. Biochem v.54 Assembly of asparaginelinked oligosaccharides Kornfeld,R.;s.Komfeld
  10. Yeast v.12 The targeting of Bacillus subtillis levansucrase in yeast is correlated to both the hydrophobicity of the signal peptide and the net charge of the N-terminus mature part Sotti,P.A.;M.Praestegaard,R.Chambert;M.R.PetitGlatron
  11. Carbohydr. Res. v.241 Complexing of cycloinulo-oligosaccharides with metal ions Uchiyama,T.;M.Kawamura;T.Uragami;H.Okuno
  12. Enz. Microbial. Technol. v.13 Cultivation and utilization of jerusalem artichkoke for ethanol, single cell protein, and high fructose syrup production Bajpai,P.K.;P.Bajpai
  13. J. Microbiol. Biotechnol v.4 Secretion of a Bacillus endoglucanase in Saccharomyces cerevisiae by its own signal sequence Han,Y.J.;D.O.Kang;S.C.Lee;B.Y.Kim;H.H.Suh;J.M.Kim;T.I.Mheen;J.S.Ahn
  14. Appl. Environ. Microbiol v.67 Molecular characterization of cycloinulooligosaccharide fructanotransferase from Bacillus macerans Kim.H.Y.;Y.J.Choi
  15. J. Chem. Soc v.72 Studies on fructosans Hirst,E.L.;D.J.Mcgilvary;E.G.V.Percival
  16. Biosci. Biotechnol. Biochem. v.57 Reaction catalyzed by cycloinulooligosaccharide fructanotransferase Kawamura,M.;T.Uchiyama
  17. Carbohydr. Res. v.260 Purification and some properties of cycloinulooligosaccharide fructanotransferase from Bacillus circulans OKUMZ31B Kawamura.M.;T.Uchiyama
  18. Biosic. Biotechnol. Biochem v.58 Purification and catalyztion of cycloinulooligosaccharide fructanotransferase (CFTase) Kushibe,S.;K,Mitsui;M.Yamagishi;K.Yamada;Y.Morimoto
  19. J. Bacteriol. v.153 Transformation of intact yeast cells treated with alkali cations Ito,H.;Y.Fukuda;K.Murata;A.Kimura
  20. Carbohydr. Res. v.192 Formation of cycloinulo-oligosaccharide from inulin by an extracclular enzyme of Bacillus circulans OKUMZ31B Kawamura,M.;T.Uchiyama;K.Takashi;T.Yukiyoshi;M.Kenji
  21. Appl. Environ. Microbiol. v.63 Recombinant thermostable cycloinulo-oligosaccharide fructanotransferase produced by Ssccharomyces cerevisiae Kanai,T.;N.Ueki;T.Kawaguchi;Y.Teranishi;H.Atomi;C.Tomorbaatar;M.Ueda;A.Tanaka
  22. J. Chem. Soc. Chem. Commun. v.1 a permethylated cyclic fructo-oligosaccharide host that can bind cation in solution Takai,Y.;Y.Okumura;S.Takahashi;M.Sawada;M.Kawamura;t.Uchiyama