Purification and properties of beta-fructofuranosidase from Aspergillus oryzae ATCC 76080.

A fructooligosaccharide-producing beta-fructofuranosidase was purified from the crude extract of Aspergillus oryzae ATCC 76080 through successive steps of ultrafiltration, DEAE-Sepharose CL-6B ion-exchange chromatography, preparative isoelectric focusing electrophoresis and Sephacryl S-200 gel filtration. The purified enzyme had an optimal pH of 5-6, an optimal temperature of 50 degrees C, a Km value of 0.53 M for catalyzing selftransfer reaction from sucrose. The molecular weight was 87 kDa by gel filtration. Mercuric ion (0.25 mM), p-hydroxymercuribenzoate (0.25 mM) and N-bromosuccinimide (0.5 mM), significantly inhibited the enzyme activity. The enzyme showed both transfructosylation and hydrolytic action in 0.5 to 50% sucrose. The transfructosylation ratio increased as the sucrose concentration increased and it was 88.5% at 50% sucrose. The main fructooligosaccharides produced from sucrose were 1-kestose and nystose.