Cloned (Comment) | Organism |
---|---|
expression of wild-type and mutant enzymes in Escherichia coli strain BL21(DE3) | Streptococcus gordonii |
Protein Variants | Comment | Organism |
---|---|---|
I259D | site-directed mutagenesis, almost inactive mutant | Streptococcus gordonii |
I259E | site-directed mutagenesis, almost inactive mutant | Streptococcus gordonii |
I259V | site-directed mutagenesis, the mutant shows 20% reduced activity compared to the wild-type enzyme | Streptococcus gordonii |
I260D | site-directed mutagenesis, the mutant shows 86% reduced activity compared to the wild-type enzyme | Streptococcus gordonii |
I260E | site-directed mutagenesis, the mutant shows 89% reduced activity compared to the wild-type enzyme | Streptococcus gordonii |
I260V | site-directed mutagenesis, the mutant shows 20% increased activity compared to the wild-type enzyme, the mutant is activable by NaF in contrast to the wild-type enzyme | Streptococcus gordonii |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.041 | - |
diphosphate | recombinant mutant I260D, pH 9.0, 25°C | Streptococcus gordonii | |
0.054 | - |
diphosphate | recombinant mutants I260V and I260E, pH 9.0, 25°C | Streptococcus gordonii | |
0.055 | - |
diphosphate | recombinant mutant I259V, pH 9.0, 25°C | Streptococcus gordonii | |
0.062 | - |
diphosphate | recombinant wild-type enzyme, pH 9.0, 25°C | Streptococcus gordonii | |
0.163 | - |
diphosphate | recombinant mutant I259D, pH 9.0, 25°C | Streptococcus gordonii | |
0.248 | - |
diphosphate | recombinant mutant I259E, pH 9.0, 25°C | Streptococcus gordonii |
Localization | Comment | Organism | GeneOntology No. | Textmining |
---|---|---|---|---|
soluble | - |
Streptococcus gordonii | - |
- |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Mn2+ | required, activates at 2 mM | Streptococcus gordonii |
Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|
27000 | - |
1 x * 27000 + 1 * 57000, recombinant mutant I259V, native PAGE in absence of Mn2+, 1 * 32000 + 1 x 62000, recombinant wild-type enzyme, native PAGE in absence of Mn2+ | Streptococcus gordonii |
32000 | - |
1 x * 27000 + 1 * 57000, recombinant mutant I259V, native PAGE in absence of Mn2+, 1 * 32000 + 1 x 62000, recombinant wild-type enzyme, native PAGE in absence of Mn2+ | Streptococcus gordonii |
50000 | - |
recombinant mutant I259D, native PAGE, in absence of Mn2+ | Streptococcus gordonii |
57000 | - |
1 x * 27000 + 1 * 57000, recombinant mutant I259V, native PAGE in absence of Mn2+, 1 * 32000 + 1 x 62000, recombinant wild-type enzyme, native PAGE in absence of Mn2+ | Streptococcus gordonii |
60000 | - |
recombinant mutant I260D, native PAGE, in presence or absence of Mn2+ | Streptococcus gordonii |
60000 | - |
recombinant mutants I260E and I259E, native PAGE, in absence of Mn2+ | Streptococcus gordonii |
61000 | - |
recombinant mutant I260V, native PAGE, in absence of Mn2+ | Streptococcus gordonii |
62000 | - |
recombinant mutant I260E, native PAGE, in presence of Mn2+ | Streptococcus gordonii |
64000 | - |
recombinant mutant I260V, native PAGE, in presence of Mn2+ | Streptococcus gordonii |
78000 | - |
recombinant wild-type enzyme, native PAGE, in presence of Mn2+ | Streptococcus gordonii |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Streptococcus gordonii | P95765 | - |
- |
Streptococcus gordonii DL1 | P95765 | - |
- |
Purification (Comment) | Organism |
---|---|
recombinant wild-type and mutant enzymes from Escherichia coli strain BL21(DE3) by anion exchange and hydrophobic interaction chromatography | Streptococcus gordonii |
Subunits | Comment | Organism |
---|---|---|
dimer | 1 x * 27000 + 1 * 57000, recombinant mutant I259V, native PAGE in absence of Mn2+, 1 * 32000 + 1 x 62000, recombinant wild-type enzyme, native PAGE in absence of Mn2+ | Streptococcus gordonii |
More | the hinge region plays an important role in opening and closing of the active site between the N- and C-terminal domains of PPase. Family II PPases are dimer under physiological conditions with Mn2+ present, and may dissociate into monomers at low enzyme concentration in the absence of Mn2+ | Streptococcus gordonii |
Synonyms | Comment | Organism |
---|---|---|
inorganic pyrophosphatase | - |
Streptococcus gordonii |
PPase | - |
Streptococcus gordonii |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
25 | - |
assay at | Streptococcus gordonii |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
11 | - |
diphosphate | recombinant mutant I259D, pH 9.0, 25°C | Streptococcus gordonii | |
48 | - |
diphosphate | recombinant mutant I259E, pH 9.0, 25°C | Streptococcus gordonii | |
560 | - |
diphosphate | recombinant mutant I260D, pH 9.0, 25°C | Streptococcus gordonii | |
570 | - |
diphosphate | recombinant mutant I260E, pH 9.0, 25°C | Streptococcus gordonii | |
4420 | - |
diphosphate | recombinant mutant I259V, pH 9.0, 25°C | Streptococcus gordonii | |
6290 | - |
diphosphate | recombinant wild-type enzyme, pH 9.0, 25°C | Streptococcus gordonii | |
6480 | - |
diphosphate | recombinant mutant I260V, pH 9.0, 25°C | Streptococcus gordonii |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
9 | - |
assay at | Streptococcus gordonii |
General Information | Comment | Organism |
---|---|---|
evolution | family II soluble inorganic pyrophosphatase | Streptococcus gordonii |
additional information | the hinge region plays an important role in opening and closing of the active site between the N- and C-terminal domains of PPase | Streptococcus gordonii |
kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
67.5 | - |
diphosphate | recombinant mutant I259D, pH 9.0, 25°C | Streptococcus gordonii | |
193.5 | - |
diphosphate | recombinant mutant I259E, pH 9.0, 25°C | Streptococcus gordonii | |
10560 | - |
diphosphate | recombinant mutant I260E, pH 9.0, 25°C | Streptococcus gordonii | |
13660 | - |
diphosphate | recombinant mutant I260D, pH 9.0, 25°C | Streptococcus gordonii | |
80360 | - |
diphosphate | recombinant mutant I259V, pH 9.0, 25°C | Streptococcus gordonii | |
101500 | - |
diphosphate | recombinant wild-type enzyme, pH 9.0, 25°C | Streptococcus gordonii | |
120000 | - |
diphosphate | recombinant mutant I260V, pH 9.0, 25°C | Streptococcus gordonii |