Cloning and heterologous expression of the ftfCNC-2(1) gene from Weissella confusa MBFCNC-2(1) as an extracellular active fructansucrase in Bacillus subtilis

Amarila Malik, Maria Tyas Hapsari, Iwao Ohtsu, Shu Ishikawa, Hiroshi Takagi

Research output: Contribution to journalArticle

5 Citations (Scopus)

Abstract

Fructan-exopolysaccharides (fructan-EPS) (inulin and levan) and their oligosaccharides (fructooligosaccharides, FOS) have drawn considerable interest in the food and pharmaceutical industries. EPS-producing lactic acid bacteria have been reported to produce β-fructans (inulin and levan), as well as α-glucans, by the function of sucrase enzymes, i.e., fructansucrase and glucansucrase. A fructansucrase ftfCNC-2(1) gene from Weissella confusa strain MBFCNC-2(1) was previously cloned in Escherichia coli. In this study, we aimed to express the ftf[CNC-2(1)] gene in Bacillus subtilis to obtain the active form of the extracellular recombinant protein FTF[CNC-2(1)]. This cloning was achieved by inserting the gene in-fusion with the signal sequence of the B.subtilis subtilisin E. SDS-polyacrylamide gel electrophoresis analysis and in situ activity assay with Periodic Acid-Schiff staining revealed that the recombinant FTF[CNC-2(1)] was successfully expressed as an extracellular protein from B.subtilis DB403 in its active form, which was confirmed using sucrose and raffinose.

Original languageEnglish
Pages (from-to)515-520
Number of pages6
JournalJournal of Bioscience and Bioengineering
Volume119
Issue number5
DOIs
Publication statusPublished - 1 May 2015

Keywords

  • Bacillus subtilis
  • Fructansucrase
  • Fructosyltransferase
  • Ftf
  • Inulosucrase
  • Weissella confusa

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