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Literature summary for 3.1.3.1 extracted from
- Alkaline phosphatase from Thermotoga neapolitana (2001), Methods Enzymol., 331, 298-305.
Activating Compound
| Activating Compound | Comment | Organism | Structure |
|---|---|---|---|
| Tris | activation at 0.2 M, increases with increasing concentration to a plateau | Escherichia coli |  |
| Tris | activation at 0.2 M, increases with increasing concentration to a plateau | Thermotoga neapolitana | |
Application
| Application | Comment | Organism |
|---|---|---|
| diagnostics | widely used enzyme, e.g. in ELISA, enzyme-linked immunosorbent assay | Escherichia coli |
| molecular biology | analytically widely used enzyme, e.g. in ELISA, enzyme-linked immunosorbent assay | Escherichia coli |
Cloned(Commentary)
| Cloned (Comment) | Organism |
|---|---|
| overexpression in Escherichia coli BL21(DE3) as C-terminally His-tagged enzyme | Thermotoga neapolitana |
Inhibitors
| Inhibitors | Comment | Organism | Structure |
|---|---|---|---|
| EDTA | inactivation of the native, but not of the recombinant enzyme, inhibition of the native enzyme is reversible by divalent metal ions, best effect with Co2+ | Thermotoga neapolitana | |
Metals/Ions
| Metals/Ions | Comment | Organism | Structure |
|---|---|---|---|
| Co2+ | required | Thermotoga neapolitana | |
| Mg2+ | enzyme contains 1 Mg2+ ion per monomer, located near the active site, interacting with phosphate | Escherichia coli | |
| Zn2+ | can partly substitute for Co2+ | Thermotoga neapolitana | |
| Zn2+ | enzyme contains 2 Zn2+ ions per monomer, located near the active site, interacting with phosphate | Escherichia coli | |
Molecular Weight [Da]
| Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
|---|---|---|---|
| 40000 | - |
2 * 40000, about, native enzyme, SDS-PAGE | Thermotoga neapolitana |
| 87000 | - |
native enzyme, gel filtration | Thermotoga neapolitana |
Natural Substrates/ Products (Substrates)
| Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| additional information | Escherichia coli | the enzyme is involved in recovering of phosphate esters when free phosphate is depleted | ? | - |
? | |
Organism
| Organism | UniProt | Comment | Textmining |
|---|---|---|---|
| Escherichia coli | - |
- |
- |
| Thermotoga neapolitana | - |
strain DSM 5068 | - |
Purification (Commentary)
| Purification (Comment) | Organism |
|---|---|
| native enzyme 2882.6fold and recombinant His-tagged enzyme expressed in Escherichia coli | Thermotoga neapolitana |
Reaction
| Reaction | Comment | Organism | Reaction ID |
|---|---|---|---|
| a phosphate monoester + H2O = an alcohol + phosphate | Wide specificity. Also catalyses transphosphorylations. Some enzymes hydrolyse diphosphate | Thermotoga neapolitana | |
| a phosphate monoester + H2O = an alcohol + phosphate | Wide specificity. Also catalyses transphosphorylations. Some enzymes hydrolyse diphosphate, catalytically active His153 and His328 | Escherichia coli |
Specific Activity [micromol/min/mg]
| Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
|---|---|---|---|
| 1309 | - |
purified enzyme, substrate 4-nitrophenyl phosphate | Thermotoga neapolitana |
Substrates and Products (Substrate)
| Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| 4-nitrophenyl phosphate + H2O | non-specific, phosphoseryl intermediate | Escherichia coli | 4-nitrophenol + phosphate | - |
? | |
| 4-nitrophenyl phosphate + H2O | non-specific, via phosphoseryl intermediate | Thermotoga neapolitana | 4-nitrophenol + phosphate | - |
? | |
| additional information | the enzyme also catalyses the transfer of the phosphoryl group to alcohols | Escherichia coli | ? | - |
? | |
| additional information | the enzyme is involved in recovering of phosphate esters when free phosphate is depleted | Escherichia coli | ? | - |
? | |
Subunits
| Subunits | Comment | Organism |
|---|---|---|
| dimer | 2 * 40000, about, native enzyme, SDS-PAGE | Thermotoga neapolitana |
| dimer | homodimer with alpha/beta structure | Escherichia coli |
Synonyms
| Synonyms | Comment | Organism |
|---|---|---|
| alkaline phosphatase | - |
Escherichia coli |
| alkaline phosphatase | - |
Thermotoga neapolitana |
| AP | - |
Escherichia coli |
| orthophosphoric-mono phosphohydrolase | - |
Escherichia coli |
| TNAP | - |
Thermotoga neapolitana |
Temperature Optimum [°C]
| Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
|---|---|---|---|
| 90 | - |
- |
Thermotoga neapolitana |
Temperature Range [°C]
| Temperature Minimum [°C] | Temperature Maximum [°C] | Comment | Organism |
|---|---|---|---|
| 20 | 90 | exponential increase in activity, optimum at 90°C | Thermotoga neapolitana |
Temperature Stability [°C]
| Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
|---|---|---|---|
| additional information | - |
Co2+ and Zn2+ increase the thernal stability by 30 and 23fold at 90°C | Thermotoga neapolitana |
| 87 | - |
half-life of the recombinant enzyme is 38 min | Thermotoga neapolitana |
| 90 | - |
half-life of the native enzyme is 4 h in presence of 2 mM Co2+ | Thermotoga neapolitana |
pH Optimum
| pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
|---|---|---|---|
| 9.9 | - |
- |
Thermotoga neapolitana |
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