• Microbiology and Biotechnology Letters
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• v.27 no.6
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• pp.471-476
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• 1999

Inulin fructotransferase(depolymerizing)(EC 2.4.1.93)(inulaseII) which converts inulin into di-D-fructofuranose-1,2':2,3'-dianhydride (DFAIII) was purified from Arthrobacter ureafaciens KCTC 3387 using column chromatography on DEAE-Toyopearl 650M and gel filtration of Sephadex G-200. The enzyme was purified 7-fold with a yield of 11% from a culture supernatant. The purified enzyme gave a single band on polyacrylamide gel electrophoresis, and the molecular weight of the enzyme was estimated to be 45,000 by SDS-polyacrylamide gel electrophoresis. The optimum pH and temperature for the enzyme reaction were pH6.5~7.0 and $55{\circ}C$, respectively. The enzyme was stable within a pH range of 5.0 to 10.6 and up to $60^{\circ}C$. The Km of this enzyme for DFAIII production was 11.9mM. The enzyme was inactivated by $Hg^{2+}$ and after exhaustive digestion of inulin by this enzyme, 1-kestose and nystose were produced in addition of DFAIII.

• KSBB Journal
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• v.14 no.5
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• pp.539-542
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• 1999

For the mass production of DFAIII and for the development of techniques of separation and purification of it, the methods of production of DFAIII and its recovery was investigated by fermentation with the strain of Arthrobacter ureafaciens KCTC 3387 and by enzyme reaction. In the first method, DFAIII was produced by fermentation with the strain of Arthrobacter ureafaciens KCTC 3387 and recovered from culture supernatant with silica gel gy filtration, in the second method, it was produced by enzyme reaction and recoverd with the same method of the first, and in third method it was produced by fermentation and recovered by addition of ethanol to the culture supernatnat.Against 25g/L of initial concentration of inulin, 1.57, 4.40, 0.34 g/L of powder of DFAIII was recovered respectively and the rate of recovery was 6.3, 17.6 1.4% and the purity was estimated at 81, 97, 87% respectively. For the production of DFAIII and its recovery, enzyme reaction method was the highest in the rate of recovery and its purity. By fermentation method, DFAIII was produced with 50% fo initial concentration of substrate but th rate of recovery was lower than enzyme reaction method and purity was lowest among the three methods. Ethanol pricipitation method showed the lowest rate of recovery.