Functional, Genetic, and Bioinformatic Characterization of Dextransucrase (DSRBCB4) Gene in Leuconostoc mesenteroides B-1299CB4

  • Kang, Hee-Kyoung (School of Biological Sciences and Technology, Chonnam National University) ;
  • Kim, Young-Min (School of Biological Sciences and Technology, Chonnam National University) ;
  • Kim, Do-Man (School of Biological Sciences and Technology, Chonnam National University)
  • Published : 2008.06.30

Abstract

A gene encoding a dextransucrase (dsrBCB4) that synthesizes only ${\alpha}$-1,6-linked dextran was cloned from Leuconostoc mesenteroides B-1299CB4. The coding region consisted of an open reading frame (ORF) of 4,395 bp that coded a 1,465-amino-acids protein with a molecular mass of 163,581 Da. The expressed recombinant DSRBCB4 (rDSRBCB4) synthesized oligosaccharides in the presence of maltose or isomaltose as an acceptor, plus the products included ${\alpha}$-1,6-linked glucosyl residues in addition to the maltosyl or isomaltosyl residue. Alignments of the amino acid sequence of DSRBCB4 with glucansucrases from Streptococcus and Leuconostoc identified conserved amino acid residues in the catalytic core that are critical for enzyme activity. The mutants D530N, E568Q, and D641N displayed a 98- to 10,000-fold reduction of total enzyme activity.

Keywords

References

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