Biochemical characterization of an intracellular 6G-fructofuranosidase from Xanthophyllomyces dendrorhous and its use in production of neo-fructooligosaccharides (neo-FOSs).
An intracellular 6G-fructofuranosidase (endo-type enzyme) extracted from Xanthophyllomyces dendrorhous 269 efficiently hydrolyzes fructosyl-β-(2→1)-linked sucrose to produce neo-kestose as a main transglycosylation product. The enzyme with a molecular weight of 33 kDa was purified by DEAE-52 cellulose chromatography. Thirty-fivefold purification and a 13.4% enzyme activity recovery were achieved. Optimum enzyme activity occurred at pH 6.4 and 45 °C and the enzyme was stable at pH 4-7 and at 45 °C. Using sucrose as a substrate, the Km and Vmax values were, respectively, 511 mmol/l and 233 μmol/(min mg) for transfer activity and 62 mmol/l and 164 μmol/(min mg) for hydrolytic activity. Under optimum conditions, a maximum concentration (73.9 g/l) of neo-fructooligosaccharides catalyzed by the endo-enzyme was obtained. These findings suggest that the purified endo-enzyme exhibits a high transfructosylation activity and it has potential for the industrial production of neo-FOSs.
Chen
Jing
J
The State Key Laboratory of Food Science and Technology, School of Food Science and Technology, Jiangnan University, 1800 Lihu Road, Wuxi 214122, China.
Chen
Xiaoming
X
Xu
Xueming
X
Ning
Yawei
Y
Jin
Zhengyu
Z
Tian
Yaoqi
Y
D001487:Basidiomycota / Q000187:drug effects / Q000201:enzymology*; D002852:Chromatography, Ion Exchange; D004591:Electrophoresis, Polyacrylamide Gel; D004795:Enzyme Stability / Q000187:drug effects; D006863:Hydrogen-Ion Concentration / Q000187:drug effects; D007202:Indicators and Reagents / Q000494:pharmacology; D042541:Intracellular Space / Q000187:drug effects / Q000201:enzymology*; D007477:Ions; D007700:Kinetics; D008670:Metals / Q000494:pharmacology; D009844:Oligosaccharides / Q000096:biosynthesis*; D013395:Sucrose / Q000378:metabolism; D013696:Temperature; D043324:beta-Fructofuranosidase / Q000302:isolation & purification / Q000378:metabolism*
D007202:Indicators and Reagents; D007477:Ions; D008670:Metals; D009844:Oligosaccharides; C116580:fructooligosaccharide; D013395:Sucrose; D043324:beta-Fructofuranosidase