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Applied and Environmental Microbiology, January 2000, p. 252-256, Vol. 66, No. 1
0099-2240/0/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.

Purification and Characterization of 2,6-beta -D-Fructan 6-Levanbiohydrolase from Streptomyces exfoliatus F3-2

Katsuichi Saito,1 Kazuya Kondo,1 Ichiro Kojima,2 Atsushi Yokota,3,* and Fusao Tomita1

Laboratory of Applied Microbiology1 and Laboratory of Microbial Resources and Ecology,3 Research Group of Molecular Bioscience, Division of Applied Bioscience, Graduate School of Agriculture, Hokkaido University, Sapporo 060-8589, and Central Technical Research Laboratory, Nippon Mitsubishi Oil Corporation, Yokohama 231-0815,2 Japan

Received 8 July 1999/Accepted 27 October 1999

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°C, respectively. The enzyme was stable in the pH range 3.5 to 8.0 and also up to 50°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).


* Corresponding author. Mailing address: Laboratory of Microbial Resources and Ecology, Research Group of Molecular Bioscience, Division of Applied Bioscience, Graduate School of Agriculture, Hokkaido University, Kita-9 Nishi-9, Kita-ku, Sapporo 060-8589, Japan. Phone: (81)-11-706-2501. Fax: (81)-11-706-4961. E-mail: yokota{at}chem.agr.hokudai.ac.jp.


Applied and Environmental Microbiology, January 2000, p. 252-256, Vol. 66, No. 1
0099-2240/0/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.



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