As. Pac. J. Mol. Biol. & Biotech., January 2012 Vol. 20, issue 4, 33-42
Molecular cloning and in silico characterization of fructansucrase gene from Weissella confusa MBFCNC-2(1) isolated from local beverage
Amarila Malik
Laboratory of Microbiology and Biotechnology, Faculty of Pharmacy, Universitas Indonesia, Kampus UI Depok, Depok 16424, Indonesia.
* Author for correspondence: Dr Amarila Malik
Laboratory of Microbiology and Biotechnology, Faculty of Pharmacy, Universitas Indonesia, Kampus UI Depok, Depok 16424, Indonesia.
Tel: 62-21-7270031 ext. 304, Fax: 62-21-7863433, Email: This email address is being protected from spambots. You need JavaScript enabled to view it. or This email address is being protected from spambots. You need JavaScript enabled to view it.
Abstract.
Fructan-type exopolysaccharides (EPS) (inulin and levan) and their oligosaccharides (fructooligosaccharides/FOS) are of great interest to the food and pharmaceutical industries. EPS producing-lactic acid bacteria (LAB) have been reported to produce beta fructans (both inulin and levan) as well as α-glucans through the action of sucrase enzymes fructansucrase (FS) and glucansucrase (GS). The Weissella confusa strain MBFCNC-2(1), isolated from a sample of a local beverage, and was shown to possess the open reading frame (ORF) of the FS coding gene (ftf) with high similarity to a putative inulosucrase of L. reuteri, which was designated ftfCNC-2(1). In total, 2,137 bp of full length ftfCNC-2(1) DNA was successfully cloned and sequenced, and a putative FTF protein of 900 amino acids was analyzed. A core region was identified as an active domain belonging to glycoside hydrolase 68. The N-terminal region contained a signal sequence followed by a sequence similar to the N-terminus of gtfKg15 (putatively identified as glucansucrase); a putative substrate binding domain was interestingly shown to be Serine Threonin rich. The C-terminus region contained an LPXTG motif, a cell wall anchoring domain, which was also Threonin rich. For further study, the ftfCNC-2(1) gene was successfully subcloned into a shuttle vector for future expression in either E. coli or B. subtilis.