Open AccessHigh‐level production and purification of a fully active recombinant dextransucrase from Leuconostoc mesenteroides NRRL B‐512F
Author(s) -
Moulis Claire,
Arcache Audrey,
Escalier PierreClaude,
Rinaudo Marguerite,
Monsan Pierre,
RemaudSimeon Magali,
PotockiVeronese Gabrielle
Publication year - 2006
Publication title -
fems microbiology letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.899
H-Index - 151
eISSN - 1574-6968
pISSN - 0378-1097
DOI - 10.1111/j.1574-6968.2006.00347.x
Subject(s) - leuconostoc mesenteroides , dextransucrase , recombinant dna , chemistry , food science , leuconostoc , microbiology and biotechnology , bacteria , biochemistry , biology , lactic acid , fermentation , lactobacillus , genetics , gene
Recombinant expression of the dextransucrase dsr S gene by Escherichia coli was optimized to produce 5850 U L −1 culture of DSR‐S, corresponding to a 30‐fold increase compared with previous studies. Rational deletions of the signal peptide, the beginning of the variable region and the last four repeats of the C‐terminal end caused no loss of activity. This new variant successfully purified was remarkably stable. With a k cat of 584 s −1 , it is the most efficient recombinant glucansucrase described to date. The synthesized polymer possesses more than 95% of α‐1,6 links, like the dextran produced by the native enzyme, and innovative gel properties were obtained.