Mutational analysis of the role of calcium ions in the Lactobacillus reuteri strain 121 fructosyltransferase (levansucrase and inulosucrase) enzymes

Bacterial fructosyltransferase enzymes belonging to glycoside hydrolase family 68 (GH68) are not known to require a metal cofactor. Here, we show that Ca2+ ions play an important structural role in the Lactobacillus reuteri 121 levansucrase (Lev) and inulosucrase (Inu) enzymes. Analysis of the Bacil...

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Bibliographic Details
Published in:FEBS letters 2005-02, Vol.579 (5), p.1124-1128
Main Authors: Ozimek, L.K., Euverink, G.J.W., van der Maarel, M.J.E.C., Dijkhuizen, L.
Format: Article
Language:English
Subjects:
Amino Acid Sequence
Bacillus subtilis
Binding Sites
Calcium - metabolism
Calcium - pharmacology
Calcium ion
Edetic Acid - pharmacology
Enzyme Stability - drug effects
Fructosyltransferase
Hexosyltransferases - chemistry
Hexosyltransferases - classification
Hexosyltransferases - genetics
Hexosyltransferases - metabolism
Inulosucrase
Ion Transport
Lactobacillus - enzymology
Lactobacillus - genetics
Lactobacillus reuteri
Levansucrase
Molecular Sequence Data
Mutagenesis
Mutation - genetics
Protein Denaturation - drug effects
Sequence Alignment
Temperature
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Summary:Bacterial fructosyltransferase enzymes belonging to glycoside hydrolase family 68 (GH68) are not known to require a metal cofactor. Here, we show that Ca2+ ions play an important structural role in the Lactobacillus reuteri 121 levansucrase (Lev) and inulosucrase (Inu) enzymes. Analysis of the Bacillus subtilis Lev 3D structure [Meng, G. and Futterer, K. (2003) Nat. Struct. Biol. 10, 935–941] has provided evidence for the presence of a bound metal ion, most likely Ca2+. Characterization of site-directed mutants in the putative Ca2+ ion-binding sites of Lb. reuteri Lev and Inu revealed that the Inu Asp520 and Lev Asp500 residues play an important role in Ca2+ binding. Sequence alignments of family GH68 proteins showed that this Ca2+ ion-binding site is (largely) present only in proteins of Gram-positive origin.
ISSN:0014-5793
1873-3468
DOI:10.1016/j.febslet.2004.11.113