Characterization of a novel fructosyltransferase from Lactobacillus reuteri that synthesizes high-molecular-weight inulin and inulin oligosaccharides.
Fructosyltransferase (FTF) enzymes produce fructose polymers (fructans) from sucrose. Here, we report the isolation and characterization of an FTF-encoding gene from Lactobacillus reuteri strain 121. A C-terminally truncated version of the ftf gene was successfully expressed in Escherichia coli. When incubated with sucrose, the purified recombinant FTF enzyme produced large amounts of fructo-oligosaccharides (FOS) with beta-(2-->1)-linked fructosyl units, plus a high-molecular-weight fructan polymer (>10(7)) with beta-(2-->1) linkages (an inulin). FOS, but not inulin, was found in supernatants of L. reuteri strain 121 cultures grown on medium containing sucrose. Bacterial inulin production has been reported for only Streptococcus mutans strains. FOS production has been reported for a few bacterial strains. This paper reports the first-time isolation and molecular characterization of (i) a Lactobacillus ftf gene, (ii) an inulosucrase associated with a generally regarded as safe bacterium, (iii) an FTF enzyme synthesizing both a high molecular weight inulin and FOS, and (iv) an FTF protein containing a cell wall-anchoring LPXTG motif. The biological relevance and potential health benefits of an inulosucrase associated with an L. reuteri strain remain to be established.
van Hijum
S A F T
SA
Centre for Carbohydrate Bioengineering, TNO-RUG, Department of Microbiology, Groningen Biomolecular Sciences and Biotechnology Institute, University of Groningen, Haren, The Netherlands.
van Geel-Schutten
G H
GH
Rahaoui
H
H
van der Maarel
M J E C
MJ
Dijkhuizen
L
L
D000595:Amino Acid Sequence; D004926:Escherichia coli / Q000235:genetics; D006602:Hexosyltransferases / Q000235:genetics / Q000378:metabolism*; D007444:Inulin / Q000378:metabolism*; D007778:Lactobacillus / Q000201:enzymology* / Q000378:metabolism; D008969:Molecular Sequence Data; D008970:Molecular Weight; D009844:Oligosaccharides / Q000378:metabolism*; D011994:Recombinant Proteins / Q000378:metabolism; D017386:Sequence Homology, Amino Acid; D013395:Sucrose / Q000378:metabolism; D001426:Bacterial Proteins
D006602:Hexosyltransferases; D007444:Inulin; D009844:Oligosaccharides; D011994:Recombinant Proteins; D013395:Sucrose; C021451:inulosucrase; D001426:Bacterial Proteins