Cloned (Comment) | Organism |
---|---|
expression of His-tagged wild-type and mutant enzymes in Escherichia coli TOP10 cells | Limosilactobacillus reuteri |
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 | 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 | UniProt | Comment | Textmining |
---|---|---|---|
Limosilactobacillus reuteri | Q8GP32 | - |
- |
Limosilactobacillus reuteri 121 | Q8GP32 | - |
- |
Purification (Comment) | Organism |
---|---|
recombinant His-tagged wild-type and mutant enzymes from Escherichia coli TOP10 cells by nickel affinity chromatography | Limosilactobacillus reuteri |
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 | 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 | Comment | Organism |
---|---|---|
InuJ | - |
Limosilactobacillus reuteri |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
37 | - |
assay at | Limosilactobacillus reuteri |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
5.4 | - |
assay at | Limosilactobacillus reuteri |