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URN etd-0202113-190603 Statistics This thesis had been viewed 1271 times. Download 7 times. Author Mio-Fen Cheng Author's Email Address No Public. Department Bioengineering Year 2012 Semester 1 Degree Master Type of Document Master's Thesis Language zh-TW.Big5 Chinese Page Count 53 Title Studies on Partial Purification of Fructosyltransferase from Xanthophyllomyces dendrorhous Keyword Xanthophyllomyces dendrorhous neofructooligosaccharides neofructooligosaccharides Xanthophyllomyces dendrorhous Abstract The intracellular neo-fructosyltransferase from the yeast Xanthophyllomyces dendrorhous TU-31 catalyzes the conversion of sucrose into neofructooligosaccharides (neo-FOS). Neo-FOS comprises neokestose and neonystose. Yeast cells obtained from a 5-L fermenter were disrupted by bead beater, thus the neo-fructosyltransferase was extracted. The crude neo-fructosyltransferase was concentrated by a hollow fiber module with a pore size of 100 k Da and proteins with the MW smaller than 100 k Da were removed. The increase of specific activity at 5.42 folds and an enzyme recovery of up to 97% were achieved. Precipitation of the enzyme by adding two volumes of acetone (–20oC) resulted in a purification folds of 4.26. However the recovery of the enzyme was as low as 5%. The acetone precipitation was not applicable for the purification of this enzyme. The concentrated enzyme was further purified by preparative isoelectrofocusing and it was found the enzyme activity peaked at around pH 3. The partially purified enzyme was active (> 97% relative activity) in the pH range of 5.4-7.4 and was most active at 50oC. Advisor Committee Dey-Chyi Sheu - advisor
Files Date of Defense 2013-01-29 Date of Submission 2013-02-04