Purification and properties of inulaseII from arthrobacter ureafaciens KCTC 3387 = Arthrobacter ureafaciens KCTC 3387이 생산하는 inulase II의 정제 및 특성

Abstract

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 650 M and gel filtration on 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 pH 6.5-7.0 and 55°C, respectively. The enzyme was stable within a pH range of 5.0 to 10.6 and up to 60°C. The K(m) of this enzyme for DFAIII production was 11.9 mM. The enzyme was inactivated by Hg2+ and after exhaustive digestion of inulin by this enzyme, 1-kestose and nystose were produced in addition to DFAIII.