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PMID:21229247

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Citation

Fraga Vidal, R, Moulis, C, Escalier, P, Remaud-Siméon, M and Monsan, P (2011) Isolation of a gene from Leuconostoc citreum B/110-1-2 encoding a novel dextransucrase enzyme. Curr. Microbiol. 62:1260-6

Abstract

The amplicon encoding dextransucrase DSR-F from Leuconostoc citreum B/110-1-2, a novel sucrose glucosyltransferase (GTF)-specific for α-1,6 and α-1,3 glucosidic bond synthesis, with α-1,4 branching was cloned, sequenced, and expressed into Escherichia coli JM109. Recombinant enzyme catalyzed oligosaccharides synthesis from sucrose as donor and maltose acceptor. The dsrF gene encodes for a protein (DSR-F) of 1,528 amino acids, with a theoretical molecular mass of 170447.72 Da (~170 kDa). From amino acid sequence comparison, it appears that DSR-F possesses the same domains as those described for GTFs. However, the variable region is longer than in other GTFs (by 100 amino acids) and two APY repeats (a 79 residue long motif with a high number of conserved glycine and aromatic residues, characterized by the presence of the three consecutive residues Ala, Pro, and Tyr) were identified in the glucan binding domain. The DSR-F catalytic domain possesses the catalytic triad involved in the glucosyl enzyme formation. The amino acid sequence of this domain shares a 56% identity with catalytic domain of the alternansucrase ASR from L. citreum NRRL B-1355 and with the catalytic domain of a putative alternansucrase sequence found in the genome of L. citreum KM20. A truncated active variant DSR-F-∆SP-∆GBD of 1,251 amino acids, with a molecular mass of 145 544 Da (~145 kDa), was obtained.

Links

PubMed Online version:10.1007/s00284-010-9851-7

Keywords

Amino Acid Sequence; Bacterial Proteins/chemistry; Bacterial Proteins/genetics; Bacterial Proteins/metabolism; Glucosyltransferases/chemistry; Glucosyltransferases/genetics; Glucosyltransferases/metabolism; Leuconostoc/chemistry; Leuconostoc/enzymology; Leuconostoc/genetics; Molecular Sequence Data; Sequence Alignment; Substrate Specificity

Significance

Annotations

Gene product Qualifier GO Term Evidence Code with/from Aspect Extension Notes Status

LEUCI:D5FS20

GO:0047849: dextransucrase activity

ECO:0000314:

F

Figure 2: Highperformance anion-exchange chromatography with pulsed amperometric detection was used to compare profiles. L. citreum B/110-1-2 (WT dsrF), DSR-F-DSP-DGBD (recombinant dextransucrase),DSR-S from mesenteroides NRRLB-512F(DSR-S is a dextransucrase). L. citreum B/110-1-2 produces a series of isomaltodextrins analogs with retention times that were very similar to the ones produced by the dextransucrase DSR-S from L. mesenteroides NRRLB-512F.

complete
CACAO 4573

LEUCI:D5FS20

enables

GO:0047849: dextransucrase activity

ECO:0000314: direct assay evidence used in manual assertion

F

Seeded From UniProt

complete


See also

References

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