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학술저널
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한국미생물생명공학회 한국미생물·생명공학회지 한국미생물·생명공학회지 제46권 제2호
발행연도
2018.1
수록면
102 - 110 (9page)

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A bacterial strain capable of metabolizing myo-inositol (MI) and converting to other substances was isolated from soil of orchard. The isolate, named YB-46, was grown on minimal medium supplemented with MI as the sole carbon source and was presumed to belonging to genus Enterobacter according to the 16S rDNA sequence. Escherichia coli transformant converting MI into unknown metabolites was selected from a metagenomic library prepared with fosmid pCC1FOS vector. Plasmid was isolated from the transformant, and the inserted gene was partially sequenced. From the nucleotide sequence, an iolG gene was identified to encode myo-inositol dehydrogenase (IolG) consisting of 336 amino residues. The IolG showed amino acid sequence similarity of about 50% with IolG of Enterobacter aerogenes and Bacillus subtilis. The His-tagged IolG (HtIolG) fused with hexahistidine at C-terminus was produced and purified from cell extract of recombinant E. coli. The purified HtIolG showed maximal activity at 45℃ and pH 10.5 with the highest activity for MI and D-glucose, and more than 90% of maximal activity for D-chiro-inositol, D-mannitol and D-xylose. Km and Vmax values of the HtIolG for MI were 1.83 mM and 0.724 μmol/min/mg under the optimal reaction condition, respectively. The activity of HtIolG was increased 1.7 folds by Zn2+, but was significantly inhibited by Co2+ and SDS.

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