Literature summary for 3.1.3.75 extracted from

Beassoni, P.R.; Otero, L.H.; Massimelli, M.J.; Lisa, A.T.; Domenech, C.E.
Critical active-site residues identified by site-directed mutagenesis in Pseudomonas aeruginosa phosphorylcholine phosphatase, a new member of the haloacid dehalogenases hydrolase superfamily (2006), Curr. Microbiol., 53, 534-539.

Activating Compound

Activating Compound	Comment	Organism	Structure
additional information	signal peptide alters the low-affinity site of recombinant PChP expressed in Escherichia coli	Pseudomonas aeruginosa

Cloned(Commentary)

Cloned (Comment)	Organism
overexpression of the wild-type and mutant enzymes in Escherichia coli ER2566, transformed with pTY-pchP327, pTYpchP349, or the mutated expression plasmids	Pseudomonas aeruginosa

Protein Variants

Protein Variants	Comment	Organism
D262E	low PChP activity	Pseudomonas aeruginosa
D265E	enzyme activity reduced to 30-35% of wild-type activity	Pseudomonas aeruginosa
D267E	low PChP activity	Pseudomonas aeruginosa
D31E	protein expression, but no enzyme activity	Pseudomonas aeruginosa
D33E	protein expression, but no enzyme activity	Pseudomonas aeruginosa
G261A	reduced activity as compared to wild-type	Pseudomonas aeruginosa
K242Q	is inactive	Pseudomonas aeruginosa
K242R	functional enzyme	Pseudomonas aeruginosa
S166T	reduced PChP activity	Pseudomonas aeruginosa
T35S	protein expression, low PChP activity	Pseudomonas aeruginosa

KM Value [mM]

KM Value [mM]	KM Value Maximum [mM]	Substrate	Comment	Organism
0.03	-	phosphocholine	at pH 5 and pH 7.4, without signal peptide	Pseudomonas aeruginosa
0.03	-	phosphocholine	mutant T35S	Pseudomonas aeruginosa
0.05	-	phosphocholine	at pH 5 and pH 7.4, with signal peptide	Pseudomonas aeruginosa
0.5	-	phosphocholine	at pH 5 and pH 7.4, without signal peptide	Pseudomonas aeruginosa
0.5	-	phosphocholine	mutant T35S	Pseudomonas aeruginosa
2	-	p-nitrophenyl phosphate	at pH 5, with and without signal peptide	Pseudomonas aeruginosa
2.1	-	p-nitrophenyl phosphate	mutant T35S	Pseudomonas aeruginosa
3.5	-	phosphocholine	at pH 7.4, with signal peptide	Pseudomonas aeruginosa
3.6	-	phosphocholine	at pH 5, with signal peptide	Pseudomonas aeruginosa

Metals/Ions

Metals/Ions	Comment	Organism	Structure
Mg2+	-	Pseudomonas aeruginosa

Organism

Organism	UniProt	Comment	Textmining
Pseudomonas aeruginosa	-	PAO1	-

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
1.3	-	mutant D262E, at pH 5, with p-nitrophenyl phosphate as substrate	Pseudomonas aeruginosa
1.5	-	mutant D267E, at pH 5, with p-nitrophenyl phosphate as substrate	Pseudomonas aeruginosa
2.4	-	mutant D267E, at pH 5, with phosphocholine as substrate	Pseudomonas aeruginosa
2.5	-	mutant D267E, at pH 7.4, with phosphocholine as substrate	Pseudomonas aeruginosa
3	-	mutant T35S, at pH 5, with phosphocholine as substrate	Pseudomonas aeruginosa
3.6	-	mutant T35S, at pH 7.4, with phosphocholine as substrate	Pseudomonas aeruginosa
5.9	-	mutant T35S, at pH 5, with p-nitrophenyl phosphate as substrate	Pseudomonas aeruginosa
6.5	-	mutant D262E, at pH 5, with phosphocholine as substrate	Pseudomonas aeruginosa
7	-	mutant S166T, at pH 5, with p-nitrophenyl phosphate as substrate	Pseudomonas aeruginosa
11.1	-	mutant D262E, at pH 7.4, with phosphocholine as substrate	Pseudomonas aeruginosa
20.3	-	mutant S166T, at pH 5, with phosphocholine as substrate	Pseudomonas aeruginosa
26.2	-	mutant S166T, at pH 7.4, with phosphocholine as substrate	Pseudomonas aeruginosa
38.8	-	mutant G261A, at pH 5, with phosphocholine as substrate	Pseudomonas aeruginosa
56.9	-	mutant K242R, at pH 5, with phosphocholine as substrate	Pseudomonas aeruginosa
63.4	-	mutant G261A, at pH 5, with p-nitrophenyl phosphate as substrate	Pseudomonas aeruginosa
64	-	mutant D265E, at pH 5, with p-nitrophenyl phosphate as substrate	Pseudomonas aeruginosa
64.8	-	mutant G261A, at pH 7.4, with phosphocholine as substrate	Pseudomonas aeruginosa
70	-	mutant D265E, at pH 7.4, with phosphocholine as substrate	Pseudomonas aeruginosa
71	-	mutant D265E, at pH 5, with phosphocholine as substrate	Pseudomonas aeruginosa
74.5	-	wild-type with motif 31DMDNT35, at pH 5, with p-nitrophenyl phosphate as substrate	Pseudomonas aeruginosa
74.6	-	mutant K242R, at pH 7.4, with phosphocholine as substrate	Pseudomonas aeruginosa
86.9	-	wild-type with motif 31DMDNT35, at pH 7.4, with phosphocholine as substrate	Pseudomonas aeruginosa
88.4	-	wild-type with motif 31DMDNT35, at pH 5, with phosphocholine as substrate	Pseudomonas aeruginosa
100	-	wild-type with motif 166S, motif 242K or motif 161GDTPDSD267, at pH 5, with p-nitrophenyl phosphate as substrate and at pH 5 and pH 7.4 with phosphocholine as substrate	Pseudomonas aeruginosa
106.7	-	mutant K242R, at pH 5, with p-nitrophenyl phosphate as substrate	Pseudomonas aeruginosa

Storage Stability

Storage Stability	Organism
-10°C, 10 mM Tris-HCl, pH 8, 30% glycerol	Pseudomonas aeruginosa

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.	Reac.
p-nitrophenyl phosphate + H2O	-	Pseudomonas aeruginosa	4-nitrophenol + phosphate	-	?
phosphocholine + H2O	D31, D33, D262, D267, S166 or K242 are important residues for catalysis	Pseudomonas aeruginosa	choline + phosphate	-	?

Synonyms

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