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.
Department of Food and Nutrition, Providence University, Shalu, Taiwan R.O.C.
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D001236:Aspergillus oryzae / Q000201:enzymology*; D004591:Electrophoresis, Polyacrylamide Gel; D004795:Enzyme Stability; D005632:Fructose / Q000378:metabolism; D005656:Fungal Proteins / Q000037:antagonists & inhibitors / Q000302:isolation & purification* / Q000378:metabolism; D006026:Glycoside Hydrolases / Q000037:antagonists & inhibitors / Q000302:isolation & purification* / Q000378:metabolism; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D008970:Molecular Weight; D009844:Oligosaccharides / Q000096:biosynthesis; D013379:Substrate Specificity; D013395:Sucrose / Q000378:metabolism; D013696:Temperature; D043324:beta-Fructofuranosidase
D005656:Fungal Proteins; D009844:Oligosaccharides; D005632:Fructose; D013395:Sucrose; D006026:Glycoside Hydrolases; D043324:beta-Fructofuranosidase