Kinetic properties of an inulosucrase from Lactobacillus reuteri 121.
نویسندگان
چکیده
Inulosucrases catalyze transfer of a fructose moiety from sucrose to a water molecule (hydrolysis) or to an acceptor molecule (transferase), yielding inulin. Bacterial inulin production is rare and a biochemical analysis of inulosucrase enzymes has not been reported. Here we report biochemical characteristics of a purified recombinant inulosucrase enzyme from Lactobacillus reuteri. It displayed Michaelis-Menten type of kinetics with substrate inhibition for the hydrolysis reaction. Kinetics of the transferase reaction is best described by the Hill equation, not reported before for these enzymes. A C-terminal deletion of 100 amino acids did not appear to affect enzyme activity or product formation. This truncated form of the enzyme was used for biochemical characterization.
منابع مشابه
Single amino acid residue changes in subsite -1 of inulosucrase from Lactobacillus reuteri 121 strongly influence the size of products synthesized.
Bacterial fructansucrase enzymes belong to glycoside hydrolase family 68 and catalyze transglycosylation reactions with sucrose, resulting in the synthesis of fructooligosaccharides and/or a fructan polymer. Significant differences in fructansucrase enzyme product specificities can be observed, i.e. in the type of polymer (levan or inulin) synthesized, and in the ratio of polymer versus fructoo...
متن کاملPuri¢cation of a novel fructosyltransferase from Lactobacillus reuteri strain 121 and characterization of the levan produced
Fructosyltransferase (FTF) enzymes have been characterized from various Gram-positive bacteria, but not from Lactobacillus sp. In a screening of 182 lactobacilli for polysaccharide production only one strain, Lactobacillus reuteri strain 121, was found to produce a fructan being a levan. Here we report the first-time identification and biochemical characterization of a Lactobacillus FTF enzyme....
متن کاملPurification of a novel fructosyltransferase from Lactobacillus reuteri strain 121 and characterization of the levan produced.
Fructosyltransferase (FTF) enzymes have been characterized from various Gram-positive bacteria, but not from Lactobacillus sp. In a screening of 182 lactobacilli for polysaccharide production only one strain, Lactobacillus reuteri strain 121, was found to produce a fructan being a levan. Here we report the first-time identification and biochemical characterization of a Lactobacillus FTF enzyme....
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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-ol...
متن کاملBiochemical and molecular characterization of a levansucrase from Lactobacillus reuteri.
Lactobacillus reuteri strain 121 employs a fructosyltransferase (FTF) to synthesize a fructose polymer [a fructan of the levan type, with beta(2-->6) linkages] from sucrose or raffinose. Purification of this FTF (a levansucrase), and identification of peptide amino acid sequences, allowed isolation of the first Lactobacillus levansucrase gene (lev), encoding a protein (Lev) consisting of 804 am...
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ورودعنوان ژورنال:
- FEBS letters
دوره 534 1-3 شماره
صفحات -
تاریخ انتشار 2003