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http://dx.doi.org/10.5352/JLS.2005.15.3.317

Expression of Paenibacillus macerans Cycloinulooligosaccharide Fructanotransferase in Saccharomyces cerevisiae  

Kim Hyun-Chul (Department of Biotechnology & Bioengineering, Dong-Eui University)
Kim Jeong-Hyun (Department of Biomaterial Control, Dong-Eui University)
Jeon Sung-Jong (Department of Biomaterial Control, Dong-Eui University)
Choi Woo-Bong (Department of Biomaterial Control, Dong-Eui University)
Nam Soo-Wan (Department of Biomaterial Control, Dong-Eui University)
Publication Information
Journal of Life Science / v.15, no.3, 2005 , pp. 317-322 More about this Journal
Abstract
The cycloinulooligosaccharide fructanotransferase (CFTase) gene (cft) from Paenibacillus macerans was subcloned into an E. coli-yeast shuttle vector, pYES2.0, resulting in pYGECFTN. The plasmid pYGECFTN (8.6 kb) was introduced into Saccharomyces cerevisiae SEY2102 cells and then the transformants were selected on the synthetic defined media lacking uracil. The cft gene expression in yeast transformant was demonstrated by the analyses cyclofructan (CF) spots on thin-layer chromatogram. The recombinant CFTase was not secreted into the medium and localized in the periplasmic space. The production of CF was observed after 5 min of the enzymatic reaction with inulin. The optimun pH and temperature for CF production were found to be at pH 8.0 and $45^{\circ}C$, respectively. Enzyme activity was stably maintained up to $55^{\circ}C$. The CF was produced from all inulin sources and was most efficiently produced from dahlia tubers and Jerusalem artichokes.
Keywords
Paenibacillus macerans; Saccharomyces cerevisiae; cycloinulooligosaccharide fructanotransferase; cyclofructan; inulin;
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