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

Abstract

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.

Keywords

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

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