Molecular cloning of the gene encoding the di-D-fructofuranose 1,2': 2,3' dianhydride hydrolysis enzyme (DFA IIIase) from Arthrobacter sp. H65-7
2003
Saito, K. (National Agricultural Research Center for Hokkaido Region, Sapporo (Japan)) | Sumita, Y. | Nagasaka, Y. | Tomita, F. | Yokota, A.
The gene encoding an intracellular enzyme hydrolyzing di-D-fructofuranose 1,2':2,3' dianhydride (DFA 3) (DFA 3ase) was cloned from the genomic DNA of Arthrobacter sp. H65-7 for the first time. The single open reading frame (ORF) of the DFA 3ase gene consisted of 1368-bp encoding 455 amino acids. DFA 3ase showed a phylogenetically distinct position from other inulindegrading enzymes and showed similarity only with inulin fructotransferases (depolymerizing) (inulase 2, EC 2.4.1.93) from Arthrobacter globiformis C11-1, Arthrobacter sp. A-6, and Arthrobacter sp. H65-7 (48.7-50.3%), and inulin fructotransferase (DFA I-producing) (EC 2.4.1.200) from A. globiformis S14-3 (44.4%). An Escherichia coli transformant harboring a recombinant plasmid, pINB2, in which the DFA 3ase gene was fused with the v-galactosidase of pUC19 and under the control of the lac promoter, expressed DFA 3ase and the cloned enzyme produced inulobiose from DFA 3 similarly to the DFA 3ase of the wild-type strain, Arthrobacter sp. H65-7.
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