title: Purification and characterization of 2,6-beta-D-fructan 6-levanbiohydrolase from Streptomyces exfoliatus F3-2.
abstract: Streptomyces exfoliatus F3-2 produced an extracellular enzyme that converted levan, a beta-2,6-linked fructan, into levanbiose. The enzyme was purified 50-fold from culture supernatant to give a single band on sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). The molecular weights of this enzyme were 54,000 by SDS-PAGE and 60,000 by gel filtration, suggesting the monomeric structure of the enzyme. The isoelectric point of the enzyme was determined to be 4.7. The optimal pH and temperature of the enzyme for levan degradation were pH 5.5 and 60 degrees C, respectively. The enzyme was stable in the pH range 3.5 to 8.0 and also up to 50 degrees C. The enzyme gave levanbiose as a major degradation product from levan in an exo-acting manner. It was also found that this enzyme catalyzed hydrolysis of such fructooligosaccharides as 1-kestose, nystose, and 1-fructosylnystose by liberating fructose. Thus, this enzyme appeared to hydrolyze not only beta-2,6-linkage of levan, but also beta-2,1-linkage of fructooligosaccharides. From these data, the enzyme from S. exfoliatus F3-2 was identified as a novel 2,6-beta-D-fructan 6-levanbiohydrolase (EC 3.2.1.64).
authors.affiliation: Laboratory of Applied Microbiology, Research Group of Molecular Bioscience, Division of Applied Bioscience, Graduate School of Agriculture, Hokkaido University, Sapporo 060-8589, Japan.
authors.forename: K
authors.initials: K
authors.lastname: Saito
authors.forename: K
authors.initials: K
authors.lastname: Kondo
authors.forename: I
authors.initials: I
authors.lastname: Kojima
authors.forename: A
authors.initials: A
authors.lastname: Yokota
authors.forename: F
authors.initials: F
authors.lastname: Tomita
mesh_terms: D004187:Disaccharides / Q000378:metabolism*; D005630:Fructans / Q000378:metabolism*; D006026:Glycoside Hydrolases / Q000302:isolation & purification* / Q000378:metabolism*; D006863:Hydrogen-Ion Concentration; D008970:Molecular Weight; D013302:Streptomyces / Q000201:enzymology*; D013379:Substrate Specificity; D013696:Temperature
chemical_list: D004187:Disaccharides; D005630:Fructans; C077708:levanbiose; C072599:levan; D006026:Glycoside Hydrolases; C403663:2,6-beta-fructan 6-levanbiohydrolase