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Literature summary for 2.4.1.9 extracted from

  • Anwar, M.A.; Leemhuis, H.; Pijning, T.; Kralj, S.; Dijkstra, B.W.; Dijkhuizen, L.
    The role of conserved inulosucrase residues in the reaction and product specificity of Lactobacillus reuteri inulosucrase (2012), FEBS J., 279, 3612-3621.
    View publication on PubMed
Search for more literature for 2.4.1.9

Cloned(Commentary)

Cloned (Comment) Organism
expression of His-tagged wild-type and mutant enzymes in Escherichia coli TOP10 cells Limosilactobacillus reuteri

Protein Variants

Protein Variants Comment Organism
A417N site-directed mutagenesis, unaltered activity compared to the wild-type enzyme Limosilactobacillus reuteri
A425P site-directed mutagenesis, slightly reduced activity compared to the wild-type enzyme Limosilactobacillus reuteri
A425P/A538S/N543S/D548R/W551T site-directed mutagenesis, the mutant shows 65% reduced activity compared to the wild-type enzyme Limosilactobacillus reuteri
A489G site-directed mutagenesis, slightly reduced activity compared to the wild-type enzyme Limosilactobacillus reuteri
A538S site-directed mutagenesis, the mutation, located behind the general acid/base, increases the enzyme activity two to threefold Limosilactobacillus reuteri
D548R site-directed mutagenesis, increased activity compared to the wild-type enzyme Limosilactobacillus reuteri
G416E site-directed mutagenesis, the mutation at the rim of the active site pocket in loop 415-423, increases the hydrolytic activity twofold, without significantly changing the transglycosylation activity Limosilactobacillus reuteri
K415R site-directed mutagenesis, unaltered activity compared to the wild-type enzyme Limosilactobacillus reuteri
additional information construction of 15 single and four multiple inulosucrase mutants that of residues conserved in inulosucrase enzymes, most of the inulosucrase mutants behave similarly to the wild-type enzyme. But some inulosucrase variants show higher transglycosylation specificity, higher catalytic rates, and different fructooligosaccharide size distributions, without changing the beta(2-1) linkage type in the product Limosilactobacillus reuteri
N365K site-directed mutagenesis, slightly increased activity compared to the wild-type enzyme Limosilactobacillus reuteri
N543S site-directed mutagenesis, slightly increased activity compared to the wild-type enzyme Limosilactobacillus reuteri
N543S site-directed mutagenesis, the mutation, located adjacent to the +1/+2 subsite residue R544, results in synthesis of a reduced variety of fructooligosaccharides compared to the wild-type enzyme Limosilactobacillus reuteri
P516L site-directed mutagenesis, unaltered activity compared to the wild-type enzyme Limosilactobacillus reuteri
S442N site-directed mutagenesis, slightly reduced activity compared to the wild-type enzyme Limosilactobacillus reuteri
T366L site-directed mutagenesis, slightly reduced activity compared to the wild-type enzyme Limosilactobacillus reuteri
T366L/A425P/N365K site-directed mutagenesis, the mutant shows 47% reduced activity compared to the wild-type enzyme Limosilactobacillus reuteri
T413K site-directed mutagenesis, slightly increased activity compared to the wild-type enzyme Limosilactobacillus reuteri
T413K/K415R/G416E/A425P/S442N/W486L/P516L site-directed mutagenesis, the mutant synthesizes 1-kestose only, but at low efficiency, it shows 94% reduced activity compared to the wild-type enzyme Limosilactobacillus reuteri
W486L site-directed mutagenesis, the mutant shows increased activity compared to the wild-type enzyme Limosilactobacillus reuteri
W551T site-directed mutagenesis, slightly increased activity compared to the wild-type enzyme Limosilactobacillus reuteri

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
additional information Limosilactobacillus reuteri wild-type inulosucrase synthesizes mostly fructooligosaccharides up to a degree of polymerization of 15 and relatively low amounts of inulin polymer ?
-
 ?
additional information Limosilactobacillus reuteri 121 wild-type inulosucrase synthesizes mostly fructooligosaccharides up to a degree of polymerization of 15 and relatively low amounts of inulin polymer ?
-
 ?
sucrose + [(2->1)-beta-D-fructosyl]n Limosilactobacillus reuteri
-
 glucose + [(2->1)-beta-D-fructosyl]n+1
-
 ?
sucrose + [(2->1)-beta-D-fructosyl]n Limosilactobacillus reuteri 121
-
 glucose + [(2->1)-beta-D-fructosyl]n+1
-
 ?

Organism

Organism UniProt Comment Textmining
Limosilactobacillus reuteri Q8GP32
-
-
Limosilactobacillus reuteri 121 Q8GP32
-
-

Purification (Commentary)

Purification (Comment) Organism
recombinant His-tagged wild-type and mutant enzymes from Escherichia coli TOP10 cells by nickel affinity chromatography Limosilactobacillus reuteri

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
additional information wild-type inulosucrase synthesizes mostly fructooligosaccharides up to a degree of polymerization of 15 and relatively low amounts of inulin polymer Limosilactobacillus reuteri ?
-
 ?
additional information wild-type inulosucrase synthesizes mostly fructooligosaccharides up to a degree of polymerization of 15 and relatively low amounts of inulin polymer Limosilactobacillus reuteri 121 ?
-
 ?
sucrose + [(2->1)-beta-D-fructosyl]n
-
 Limosilactobacillus reuteri glucose + [(2->1)-beta-D-fructosyl]n+1
-
 ?
sucrose + [(2->1)-beta-D-fructosyl]n via first transfructosylation product 1-kestose Limosilactobacillus reuteri glucose + [(2->1)-beta-D-fructosyl]n+1
-
 ?
sucrose + [(2->1)-beta-D-fructosyl]n
-
 Limosilactobacillus reuteri 121 glucose + [(2->1)-beta-D-fructosyl]n+1
-
 ?
sucrose + [(2->1)-beta-D-fructosyl]n via first transfructosylation product 1-kestose Limosilactobacillus reuteri 121 glucose + [(2->1)-beta-D-fructosyl]n+1
-
 ?

Subunits

Subunits Comment Organism
More three-dimensional structures of wild-type and mutant enzymes, comparison to the structure of levansucrase, EC 2.4.1.10, overview Limosilactobacillus reuteri

Synonyms

Synonyms Comment Organism
InuJ
-
 Limosilactobacillus reuteri

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
37
-
 assay at Limosilactobacillus reuteri

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
5.4
-
 assay at Limosilactobacillus reuteri