Cloning and sequencing of a gene coding for an extracellular dextransucrase (DSRB) from Leuconostoc mesenteroides NRRL B-1299 synthesizing only a alpha (1-6) glucan.
The coding region for a novel Leuconostoc mesenteroides NRRL B-1299 dextransucrase gene (dsrB) was isolated and sequenced. Using degenerate primers homologous to a conserved region present in dextransucrases from Streptococcus (GTFs) and L. mesenteroides NRRL B-512F (DSRS) and conserved amino acid sequences located in the N-terminal catalytic region of these enzymes, about 60% of the DSRB encoding gene was isolated. Two sites, BamHI and HindIII, were identified which allowed one 0.5-kbp probe to be obtained to isolate the 5' and the 3' ends of dsrB. The nucleotide sequence of the dsrB gene was determined and found to consist of an open reading frame (ORF) of 4521 base pairs (bp) coding for a 1507-amino acid protein with an M1 of 168,511. The amino acid sequence is very close to that of DSRS. Like DSRS, the dextran produced appeared to be composed of only alpha (1-6) glucosidic bonds, and the oligosaccharides synthesized in the presence of acceptor maltose were also composed of alpha (1-6) linked glucosyl residues in addition to the maltosyl residue. DSRB thus appears to be a novel dextransucrase from L. mesenteroides NRRL B-1299. DSRB produces a dextran different from the typical dextran containing alpha (1-6) and alpha (1-2) linkages produced when this strain is grown in the presence of sucrose.
Monchois
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Centre de Bioingénierie Gilbert Durand, UMR CNRS 5504, LA INRA, INSA, Toulouse, France.
Remaud-Simeon
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Monsan
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Willemot
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D000595:Amino Acid Sequence; D003001:Cloning, Molecular; D004926:Escherichia coli / Q000235:genetics; D005798:Genes, Bacterial; D005936:Glucans / Q000096:biosynthesis*; D005964:Glucosyltransferases / Q000235:genetics*; D007934:Leuconostoc / Q000201:enzymology / Q000235:genetics*; D008969:Molecular Sequence Data
D005936:Glucans; D005964:Glucosyltransferases; C020025:dextransucrase