Purification and characterization of beta-fructofuranosidase from Bifidobacterium infantis.
beta-Fructofuranosidase activities of eight strains of Bifidobacteria, intestinal bacteria, were assayed and Bifidobacterium infantis was selected for purification of the enzyme. beta-Fructofuranosidase activity was recovered in the supernatant fraction after disruption of B. infantis cells with sonication and was purified to homogeneity by ammonium sulfate fractionation, and DEAE-cellulose, butyl-Toyopearl and Sephacryl S-300 column chromatographies. The enzyme (molecular weight (M.W.) 232000) was composed of three identical subunits (M.W. 75000) whose NH2-terminal amino acids were threonine. The enzyme was stable at pH 6-8, having the optimum activity at pH 6.0-6.2. The enzyme activity was stable under 40 degrees C and the optimal temperature was 55 degrees C. This enzyme catalyzed the hydrolysis of sucrose, 1-kestose, nystose, inulin and raffinose at the relative velocities of 100, 297, 365, 140 and 3.8, respectively, but did not catalyze the hydrolysis of maltose or cellobiose. These results indicated that this fructooligosaccharide hydrolyzing enzyme is a novel type of beta-fructofuranosidase.
Imamura
L
L
Faculty of Pharmaceutical Sciences, Toyama Medical and Pharmaceutical University, Japan.
Hisamitsu
K
K
Kobashi
K
K
D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000032:analysis; D001644:Bifidobacterium / Q000201:enzymology*; D002241:Carbohydrates / Q000737:chemistry; D002855:Chromatography, Thin Layer; D004591:Electrophoresis, Polyacrylamide Gel; D005632:Fructose / Q000032:analysis; D005947:Glucose / Q000032:analysis; D006026:Glycoside Hydrolases / Q000737:chemistry* / Q000302:isolation & purification; D006863:Hydrogen-Ion Concentration; D008969:Molecular Sequence Data; D008970:Molecular Weight; D013379:Substrate Specificity; D013696:Temperature; D043324:beta-Fructofuranosidase
D001426:Bacterial Proteins; D002241:Carbohydrates; D005632:Fructose; D006026:Glycoside Hydrolases; D043324:beta-Fructofuranosidase; D005947:Glucose