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Literature summary for 3.1.3.75 extracted from

  • Beassoni, P.R.; Otero, L.H.; Lisa, A.T.; Domenech, C.E.
    Using a molecular model and kinetic experiments in the presence of divalent cations to study the active site and catalysis of Pseudomonas aeruginosa phosphorylcholine phosphatase (2008), Biochim. Biophys. Acta, 1784, 2038-2044.
    View publication on PubMed
Search for more literature for 3.1.3.75

Cloned(Commentary)

Cloned (Comment) Organism
expressed in Escherichia coli ER2566 cells Pseudomonas aeruginosa

Protein Variants

Protein Variants Comment Organism
D262E mutant shows activity less than 2% with respect to native recombinant enzyme Pseudomonas aeruginosa
D267E mutant shows activity less than 2% with respect to native recombinant enzyme and is not activated by Cu2+ Pseudomonas aeruginosa
S166T mutant shows activity less than 2% with respect to native recombinant enzyme Pseudomonas aeruginosa
T35S mutant shows activity less than 2% with respect to native recombinant enzyme Pseudomonas aeruginosa

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
1.4
-
 p-nitrophenyl phosphate recombinant mutant enzyme D262E, at pH 5.0 and 37°C, in the presence of 0.5 mM Zn2+ Pseudomonas aeruginosa
2.5
-
 p-nitrophenyl phosphate recombinant mutant enzyme S166T, at pH 5.0 and 37°C, in the presence of 2 mM Mg2+ Pseudomonas aeruginosa
2.6
-
 p-nitrophenyl phosphate recombinant mutant enzyme T35S, at pH 5.0 and 37°C, in the presence of 0.5 mM Zn2+ Pseudomonas aeruginosa
3.2
-
 p-nitrophenyl phosphate recombinant wild type enzyme, at pH 5.0 and 37°C, in the presence of 2 mM Mg2+ Pseudomonas aeruginosa
3.5
-
 p-nitrophenyl phosphate recombinant wild type enzyme, at pH 5.0 and 37°C, in the presence of 0.5 mM Zn2+ Pseudomonas aeruginosa
4.3
-
 p-nitrophenyl phosphate recombinant mutant enzyme T35S, at pH 5.0 and 37°C, in the presence of 2 mM Mg2+ Pseudomonas aeruginosa
5.1
-
 p-nitrophenyl phosphate recombinant mutant enzyme D267E, at pH 5.0 and 37°C, in the presence of 0.5 mM Zn2+ Pseudomonas aeruginosa
5.4
-
 p-nitrophenyl phosphate recombinant mutant enzyme S166T, at pH 5.0 and 37°C, in the presence of 0.5 mM Zn2+ Pseudomonas aeruginosa
7.6
-
 p-nitrophenyl phosphate recombinant mutant enzyme D262E, at pH 5.0 and 37°C, in the presence of 2 mM Mg2+ Pseudomonas aeruginosa
11.5
-
 p-nitrophenyl phosphate recombinant mutant enzyme D267E, at pH 5.0 and 37°C, in the presence of 2 mM Mg2+ Pseudomonas aeruginosa
12.2
-
 p-nitrophenyl phosphate recombinant wild type enzyme, at pH 5.0 and 37°C, in the presence of 0.3 mM Cu2+ Pseudomonas aeruginosa
13.1
-
 p-nitrophenyl phosphate recombinant mutant enzyme T35S, at pH 5.0 and 37°C, in the presence of 0.3 mM Cu2+ Pseudomonas aeruginosa
18.7
-
 p-nitrophenyl phosphate recombinant mutant enzyme S166T, at pH 5.0 and 37°C, in the presence of 0.3 mM Cu2+ Pseudomonas aeruginosa
22.2
-
 p-nitrophenyl phosphate recombinant mutant enzyme D262E, at pH 5.0 and 37°C, in the presence of 0.3 mM Cu2+ Pseudomonas aeruginosa

Localization

Localization Comment Organism GeneOntology No. Textmining
periplasm
-
 Pseudomonas aeruginosa
-
-

Metals/Ions

Metals/Ions Comment Organism Structure
Cu2+ activator, the wild type recombinant phosphorylcholine phosphatase has higher affinity for Zn2+ and Cu2+ than for Mg2+, Cu2+ is able to diminish almost 3times the affinity of the enzyme for p-nitrophenyl with respect to the addition of Zn2+ or Mg2+ Pseudomonas aeruginosa
Mg2+ activator, the wild type recombinant phosphorylcholine phosphatase has higher affinity for Zn2+ and Cu2+ than for Mg2+ Pseudomonas aeruginosa
Zn2+ activator, the wild type recombinant phosphorylcholine phosphatase has higher affinity for Zn2+ and Cu2+ than for Mg2+ Pseudomonas aeruginosa

Organism

Organism UniProt Comment Textmining
Pseudomonas aeruginosa
-
-
-

Purification (Commentary)

Purification (Comment) Organism
-
 Pseudomonas aeruginosa

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
75
-
 wild type enzyme, at pH 5.0 and 37°C, in the presence of 2 mM Mg2+ Pseudomonas aeruginosa
114
-
 wild type enzyme, at pH 5.0 and 37°C, in the presence of 0.3 mM Cu2+ Pseudomonas aeruginosa
142
-
 wild type enzyme, at pH 5.0 and 37°C, in the presence of 0.5 mM Zn2+ Pseudomonas aeruginosa

Storage Stability

Storage Stability Organism
-20°C, 10 mM Tris-HCl, pH 8, containing 30% glycerol, more than 1 year, no loss of activity Pseudomonas aeruginosa

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
O-phosphocholine + H2O
-
 Pseudomonas aeruginosa choline + phosphate
-
 ?
p-nitrophenyl phosphate + H2O
-
 Pseudomonas aeruginosa 4-nitrophenol + phosphate
-
 ?

Synonyms

Synonyms Comment Organism
PChP
-
 Pseudomonas aeruginosa
phosphorylcholine phosphatase
-
 Pseudomonas aeruginosa

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
0.2
-
 p-nitrophenyl phosphate recombinant mutant enzyme D267E, at pH 5.0 and 37°C, in the presence of 2 mM Mg2+ Pseudomonas aeruginosa
0.7
-
 p-nitrophenyl phosphate recombinant mutant enzyme D267E, at pH 5.0 and 37°C, in the presence of 0.5 mM Zn2+ Pseudomonas aeruginosa
1100
-
 p-nitrophenyl phosphate recombinant mutant enzyme D262E, at pH 5.0 and 37°C, in the presence of 2 mM Mg2+ Pseudomonas aeruginosa
1400
-
 p-nitrophenyl phosphate recombinant mutant enzyme D262E, at pH 5.0 and 37°C, in the presence of 0.5 mM Zn2+ Pseudomonas aeruginosa
19000
-
 p-nitrophenyl phosphate recombinant mutant enzyme D262E, at pH 5.0 and 37°C, in the presence of 0.3 mM Cu2+ Pseudomonas aeruginosa
21000
-
 p-nitrophenyl phosphate recombinant mutant enzyme S166T, at pH 5.0 and 37°C, in the presence of 2 mM Mg2+ Pseudomonas aeruginosa
22000
-
 p-nitrophenyl phosphate recombinant mutant enzyme S166T, at pH 5.0 and 37°C, in the presence of 0.3 mM Cu2+ Pseudomonas aeruginosa
29000
-
 p-nitrophenyl phosphate recombinant mutant enzyme S166T, at pH 5.0 and 37°C, in the presence of 0.5 mM Zn2+ Pseudomonas aeruginosa
120000
-
 p-nitrophenyl phosphate recombinant mutant enzyme T35S, at pH 5.0 and 37°C, in the presence of 2 mM Mg2+ Pseudomonas aeruginosa
650000
-
 p-nitrophenyl phosphate recombinant mutant enzyme T35S, at pH 5.0 and 37°C, in the presence of 0.5 mM Zn2+ Pseudomonas aeruginosa
1600000
-
 p-nitrophenyl phosphate recombinant wild type enzyme, at pH 5.0 and 37°C, in the presence of 2 mM Mg2+ Pseudomonas aeruginosa
2500000
-
 p-nitrophenyl phosphate recombinant mutant enzyme T35S, at pH 5.0 and 37°C, in the presence of 0.3 mM Cu2+ Pseudomonas aeruginosa
5500000
-
 p-nitrophenyl phosphate recombinant wild type enzyme, at pH 5.0 and 37°C, in the presence of 0.5 mM Zn2+ Pseudomonas aeruginosa
7900000
-
 p-nitrophenyl phosphate recombinant wild type enzyme, at pH 5.0 and 37°C, in the presence of 0.3 mM Cu2+ Pseudomonas aeruginosa