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Production of cyclofructan by cycloinulooligosaccharide fructanotransferase expressed in Saccharomyces cerevisiae


, : Production of cyclofructan by cycloinulooligosaccharide fructanotransferase expressed in Saccharomyces cerevisiae. Korean Journal of Microbiology & Biotechnology 32(1): 60-66, March

The cycloinulooligosaccharide fructanotransferase (CFTase) gene (cft) from Paenibacillus polymyxa was subcloned into the E. coli-yeast shuttle vector, pYES2.0 (GAL1 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 cft gene under the control of GAL I 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 3 h of enzymatic reaction with inulin. The pH and temperature optimum for the CF production from inulin was pH 8.0 and 45degreeC, 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.

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