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Cloning, expression and functional validation of a β-fructofuranosidase from Lactobacillus plantarum

Paper ID Volume ID Publish Year Pages File Format Full-Text
34517 45032 2014 10 PDF Available
Title
Cloning, expression and functional validation of a β-fructofuranosidase from Lactobacillus plantarum
Abstract

•This is the first report on the characterization of a β-fructofuranosidase from Lactobacillus plantarum.•The SacA enzyme was predicted to have an additional α-helix at the N-terminal end of the molecule.•The SacA enzyme can hydrolyze various fructans and FOS is the most preferred substrate.•Expression of sacA gene in Lactobacillus rhamnosus GG enabled the recombinant strain to metabolize FOS.

Fructooligosaccharides (FOS) are prebiotics that selectively stimulate the growth and activity of lactobacilli and bifidobacteria. These strains metabolize FOS with endogenous β-fructofuranosidase. In this study, a β-fructofuranosidase gene from Lactobacillus plantarum ST-III designated sacA was cloned into Escherichia coli, and the properties of the recombinant protein (SacA) were examined. The sacA gene encodes a peptide of 501 amino acids with a predicted molecular weight of 56.7 kDa. Sequence alignment revealed the presence of three highly conserved motifs, NDPNG, RDP and EC, indicating that the enzyme belongs to glycoside hydrolase family 32. The predicted three-dimensional structure of the SacA enzyme was similar to β-fructofuranosidases of bifidobacteria, such that it contained a five-blade β-propeller module and a β-sandwich domain with one additional N-terminal α-helix. The optimal reaction temperature and pH of the enzyme were 37 °C and 6.0, respectively. Substrate hydrolysis and kinetic parameters demonstrated that β-fructofuranosidase from L. plantarum ST-III liberated fructosyl residues from the non-reducing terminus of fructans, such as sucrose, FOS, levan or inulin, and FOS was the preferred substrate. The expression of the sacA gene in a non-FOS-fermenting strain, Lactobacillus rhamnosus GG, enabled the recombinant strain to metabolize FOS and sucrose.

Keywords
β-Fructofuranosidase; Lactobacillus plantarum; Fructooligosaccharides; Glycoside hydrolase family 32
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Cloning, expression and functional validation of a β-fructofuranosidase from Lactobacillus plantarum
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Publisher
Database: Elsevier - ScienceDirect
Journal: Process Biochemistry - Volume 49, Issue 5, May 2014, Pages 758–767
Authors
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Subjects
Physical Sciences and Engineering Chemical Engineering Bioengineering
Get Full-Text Now
Don't Miss Today's Special Offer
Price was $35.95
You save - $31
Price after discount Only $4.95
100% Money Back Guarantee
Full-text PDF Download
Online Support
Any Questions? feel free to contact us