Cloning of inulin fructotransferase (DFA III-producing) gene from Arthrobacter sp. L68-1

A gene of inulin fructotransferase (DFA III-producing) [EC 2.4.1.93] from Arthrobacter sp. L68-1 was cloned and the nucleotide was sequenced. The gene encoded a signal peptide (32 amino acid residues) for a secretion, and the mature enzyme protein was estimated to be consisted with 410 amino acid residues. The molecular mass of the native enzyme was calculated as 43.7 kDa by the sequence data. The deduced amino acid sequence of the enzyme had 79.0% homology with that of the Arthrobacter globiformis C11-1, and had 77.4% homology with that of the Arthrobacter sp. H65-7. It also had 43.7% homology with that of inulin fructotransferase (DFA I-producing) [EC 2.4.1.200] from A. globiformis S14-3. The cloned enzyme was immobilized using Chitopearl BCW 3510 as a carrier. The immobilized enzyme was able to use 10 times without a significant loss of the enzyme activity.