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Literature summary extracted from

  • Zalatan, J.G.; Fenn, T.D.; Herschlag, D.
    Comparative enzymology in the alkaline phosphatase superfamily to determine the catalytic role of an active-site metal ion (2008), J. Mol. Biol., 384, 1174-1189.
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

EC Number Cloned (Comment) Organism
3.1.3.1 expressed in Escherichia coli as an N-terminal maltose binding protein (MBP) fusion construct (AP-MBP) Escherichia coli

Protein Variants

EC Number Protein Variants Comment Organism
3.1.3.1 E22Y kcat/KM (1/M*sec) (4-nitrophenyl phosphate): 7200, (3-nitrobenzyl phosphate): 31, (methyl phosphate): 1.6, (methyl 4-nitrophenyl phosphate): 35, (bis-4-nitrophenyl phosphate): 0.07, (4-nitrophenyl sulfate): 0.0000029 Escherichia coli
3.1.3.1 R166S kcat/KM (1/M*sec) (4-nitrophenyl phosphate): 100000, (3-nitrobenzyl phosphate): 2300, (methyl phosphate): 110, (methyl 4-nitrophenyl phosphate): 0.48, (bis-4-nitrophenyl phosphate): 0.05, (4-nitrophenyl sulfate): 0.000058 Escherichia coli
3.1.3.1 R166S/E322Y kcat/KM (1/M*sec) (4-nitrophenyl phosphate): 1.6, (3-nitrobenzyl phosphate): below 0.2, (methyl phosphate): not determined, (methyl 4-nitrophenyl phosphate): 0.24, (bis-4-nitrophenyl phosphate): 0.021, (4-nitrophenyl sulfate): below 0.000001 Escherichia coli
3.1.3.1 W322A kcat/KM (1/M*sec) (4-nitrophenyl phosphate): 8900, (3-nitrobenzyl phosphate): not determined, (methyl phosphate): not determined, (methyl 4-nitrophenyl phosphate): 18, (bis-4-nitrophenyl phosphate): 0.037, (4-nitrophenyl sulfate): not determined Escherichia coli

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
3.1.3.1 Mg2+ removal of a third metal ion site near the bimetallo site, containing Mg2+, suggests that the Mg2+ ion participates in general base catalysis. Mg2+ ion stabilizes the transferred phosphoryl group in the transition state, and this interaction is distinct from those mediated by the Zn2+ bimetallo site. Positioning of charged or polar groups to interact with all three nonbridging oxygen atoms of the transferred phosphoryl group is important for catalysis of phosphate monoester hydrolysis Escherichia coli
3.1.3.1 Zn2+ removal of a third metal ion site near the bimetallo site, containing Mg2+, suggests that the Mg2+ ion participates in general base catalysis. Mg2+ ion stabilizes the transferred phosphoryl group in the transition state, and this interaction is distinct from those mediated by the Zn2+ bimetallo site. Positioning of charged or polar groups to interact with all three nonbridging oxygen atoms of the transferred phosphoryl group is important for catalysis of phosphate monoester hydrolysis Escherichia coli

Organism

EC Number Organism UniProt Comment Textmining
3.1.3.1 Escherichia coli P00634
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
3.1.3.1 using affinity chromatography. Typical yields for a 6-L culture are 30-40 mg of pure protein Escherichia coli

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
3.1.3.1 3-nitrobenzyl phosphate + H2O
-
 Escherichia coli 3-nitrobenzoate + phosphate
-
 ?
3.1.3.1 4-nitrophenyl phosphate + H2O
-
 Escherichia coli 4-nitrophenol + phosphate
-
 ?
3.1.3.1 4-nitrophenyl sulfate + H2O
-
 Escherichia coli 4-nitrophenol + sulfate
-
 ?
3.1.3.1 bis-4-nitrophenyl phosphate + H2O
-
 Escherichia coli 4-nitrophenol + phosphate
-
 ?
3.1.3.1 methyl 4-nitrophenyl phosphate + H2O
-
 Escherichia coli methyl phosphate + 4-nitrophenol
-
 ?
3.1.3.1 methyl 4-nitrophenyl phosphorothioate + H2O only the R-enantiomer is detectably hydrolyzed by the enzyme Escherichia coli 4-nitrophenol + methyl phosphorothioate
-
 ?
3.1.3.1 methyl phosphate + H2O
-
 Escherichia coli methanol + phosphate
-
 ?

Synonyms

EC Number Synonyms Comment Organism
3.1.3.1 alkaline phosphatase
-
 Escherichia coli

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
3.1.3.1 25
-
 assay at Escherichia coli

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
3.1.3.1 additional information
-
 additional information wild-type enzyme: kcat/KM (1/M*sec) (4-nitrophenyl phosphate): 33000000, (3-nitrobenzyl phosphate): 18000000, (methyl phosphate): 1200000, (methyl 4-nitrophenyl phosphate): 18, (bis-4-nitrophenyl phosphate): 0.05, (4-nitrophenyl sulfate): 0.01 Escherichia coli

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
3.1.3.1 8
-
 assay at Escherichia coli