Chromatographic separation and kinetic properties of fructosyltransferase from Aureobasidium pullulans.
Efficient chromatographic separation of fructosyltransferase from Aureobasidium pullulans was achieved on a preparative scale using a weak anion-exchanger Sepabeads FP-DA. The recovery yield was about 70% and the purification factor reached a value of 28. The molecular weight of the enzyme determined by size-exclusion chromatography was 570,000. The enzyme exhibited both hydrolytic and transferase activity when the latter was higher in the whole concentration range and completely dominating at higher sucrose concentrations. It was found that sucrose was the only donor of fructosyl moiety used in the transfer reaction. The initial rate method was used for the investigation of the kinetics of the action of fructosyltransferase on sucrose in the concentration range 30-2,430 mM. The initial transfructosylation rate was well fitted with a linear function of the sucrose activity where the activity coefficient was an exponentially decreasing function of the sucrose concentration.
Antosová
Monika
M
Department of Chemical and Biochemical Engineering, Institute of Chemical and Environmental Engineering, Faculty of Chemical and Food Technology, Slovak University of Technology, Radlinského 9, 812 37 Bratislava, Slovakia.
Illeová
Viera
V
Vandáková
Marcela
M
Druzkovská
Alexandra
A
Polakovic
Milan
M
D001203:Ascomycota / Q000145:classification* / Q000201:enzymology*; D002852:Chromatography, Ion Exchange / Q000379:methods*; D004789:Enzyme Activation; D004795:Enzyme Stability; D006602:Hexosyltransferases / Q000737:chemistry* / Q000302:isolation & purification*; D007700:Kinetics; D013045:Species Specificity; D001426:Bacterial Proteins
D006602:Hexosyltransferases; C021451:inulosucrase; D001426:Bacterial Proteins