Enzymic hydrolysis of di-D-fructofuranose 1, 2'; 2, 3' dianhydride with Arthrobacter ureafaciens

Enzymic hydrolysis of di-D-fructofuranose 1, 2'; 2, 3' dianhydride with the bacteria Arthrobacter ureafaciens was studied to elucidate its mechanism. Hydrolysis of the difructose dianhydride to D-fructose, which did not occur with yeast invertase [EC 3.2.1.26], was found to occur on incubation with an enzyme preparation from an autolysate of the above bacteria. However, incubation with enzyme which had been treated at 60 degrees for 30 min yielded an intermediate hydrolysis product. The product isolated was found to be inulobiose and to be hydrolyzed to D-fructose by the original enzyme, as well as by yeast invertase. It was thus shown that the hydrolysis of the difructose dianhydride to D-fructose with the crude enzyme took place not in a single step but in two separate steps at 2, 3' and 1, 2' linkages. It was not determined whether the entire process is mediated by one and the same beta-fructofuranosidase or by different enzymes.