Journal of Microbiology and Biotechnology

The Korean Society for Microbiology and Biotechnology (한국미생물·생명공학회)
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Cloning of Dextransucrase Gene from Leuconostoc citreum HJ-P4 and Its High-Level Expression in E. coli by Low Temperature Induction

  • Yi, Ah-Rum (Department of Food Science and Technology, Chungbuk National University) ;
  • Lee, So-Ra (Department of Food Science and Technology, Chungbuk National University) ;
  • Jang, Myoung-Uoon (Department of Food Science and Technology, Chungbuk National University) ;
  • Park, Jung-Mi (Department of Food Science and Technology, Chungbuk National University) ;
  • Eom, Hyun-Ju (Department of Food Science and Technology, Chungbuk National University) ;
  • Han, Nam-Soo (Department of Food Science and Technology, Chungbuk National University) ;
  • Kim, Tae-Jip (Department of Food Science and Technology, Chungbuk National University)
  • Published : 2009.08.31
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Abstract

A dextransucrase (LcDS) gene from Leuconostoc citreum HJ-P4 has been amplified and cloned in E. coli. The LcDS gene consists of 4,431 nucleotides encoding 1,477 amino acid residues sharing 63-98% of amino acid sequence identities with other known dextransucrases from Leuc. mesenteroides. Interestingly, 0.1 mM of IPTG induction at $15^{\circ}C$ remarkably increased the LcDS productivity to 19,187 U/I culture broth, which was over 330-fold higher than that induced at $37^{\circ}C$. Optimal reaction temperature and pH of LcDS were determined as $35^{\circ}C$ and pH 5.5 in 20 mM sodium acetate buffer, respectively. Meanwhile, 0.1 mM $CaCl_2$ increased its activity to the maximum of 686 U/mg, which was 2.1-fold higher than that in the absence of calcium ion. Similar to the native Leuconostoc dextransucrase, recombinant LcDS could successfully produce a series of isomaltooligosaccharides from sucrose and maltose, on the basis of its transglycosylation activity.

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References

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