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Information on EC 3.1.3.1 - alkaline phosphatase and Organism(s) Escherichia coli and UniProt Accession P00634
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3.1.3.1 alkaline phosphataseIUBMB Comments
Wide specificity. Also catalyses transphosphorylations. The human placental enzyme is a zinc protein. Some enzymes hydrolyse diphosphate (cf. EC 3.6.1.1 inorganic diphosphatase)
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Word Map
- 3.1.3.1
- osteoblast
- osteocalcin
-
osteogenic
- collagen
- aminotransferase
- bilirubin
- women
- mesenchymal
- albumin
- pancreatitis
- marrow
- parathyroid
- phosphorus
- urinary
- cholesterol
- osteoporosis
- creatinine
- resorption
- arterial
- fracture
- children
-
alt
- necrosis
- pain
- femoral
- spine
- transaminase
-
osteogenesis
-
lumbar
- biliary
-
morphogenetic
- osteopontin
- anterior
- bile
- femur
- periodontal
-
radiological
-
gamma-glutamyl
- calcification
- dental
-
postoperative
-
biocompatibility
- gg
-
preoperative
- transpeptidase
- ligament
-
admission
-
postmenopausal
- trabecular
-
hepatoprotective
- biotechnology
- agriculture
- medicine
- food industry
- environmental protection
- analysis
- molecular biology
- diagnostics
- synthesis
The taxonomic range for the selected organisms is: Escherichia coli
The enzyme appears in selected viruses and cellular organisms
The enzyme appears in selected viruses and cellular organisms
Synonyms
ap, alkaline phosphatase, bone alkaline phosphatase, placental alkaline phosphatase, alpase, apase, intestinal alkaline phosphatase, tnsalp, phosphomonoesterase, secreted alkaline phosphatase, 
more
topSYNONYM 
ORGANISM
UNIPROT
COMMENTARY 
LITERATURE
alkaline phenyl phosphatase
-
-
-
-
alkaline phosphohydrolase
-
-
-
-
alkaline phosphomonoesterase
-
-
-
-
ALP
AP-TNAP
-
-
-
-
APase
-
-
-
-
APASED
-
-
-
-
BC6
-
-
-
-
EAP
-
-
-
-
Germ-cell alkaline phosphatase
-
-
-
-
glycerophosphatase
-
-
-
-
H-AP
-
-
-
-
High molecular weight phosphatase
-
-
-
-
IAP
-
-
-
-
L-AP
-
-
-
-
Liver/bone/kidney isozyme
-
-
-
-
Low molecular weight phosphatase
-
-
-
-
M-ALP
-
-
-
-
Nagao isozyme
-
-
-
-
non-specific alkaline phosphatase
-
-
-
-
phosphatase, alkaline
-
-
-
-
phosphomonoesterase
-
-
-
-
PLAP-like
-
-
-
-
RAN1
-
-
-
-
Regan isozyme
-
-
-
-
TNSALP
-
-
-
-
ALP
-
-
-
-
REACTION
REACTION DIAGRAM
COMMENTARY
ORGANISM
UNIPROT
LITERATURE
a phosphate monoester + H2O = an alcohol + phosphate
double in-line displacement mechanism involving two-metal ion catalysis
a phosphate monoester + H2O = an alcohol + phosphate
Wide specificity. Also catalyses transphosphorylations. Some enzymes hydrolyse diphosphate
a phosphate monoester + H2O = an alcohol + phosphate
Arg166 contributes to catalysis through binding interactions and through additional transition state stabilization that may arise from complementarity of the guanidinum group to the geometry of the trigonal bipyramidal transition state
a phosphate monoester + H2O = an alcohol + phosphate
Wide specificity. Also catalyses transphosphorylations. Some enzymes hydrolyse diphosphate
-
a phosphate monoester + H2O = an alcohol + phosphate
Wide specificity. Also catalyses transphosphorylations. Some enzymes hydrolyse diphosphate, catalytically active His153 and His328
-
PATHWAY SOURCE
PATHWAYS
BRENDA
-
-
SYSTEMATIC NAME
IUBMB Comments
phosphate-monoester phosphohydrolase (alkaline optimum)
Wide specificity. Also catalyses transphosphorylations. The human placental enzyme is a zinc protein. Some enzymes hydrolyse diphosphate (cf. EC 3.6.1.1 inorganic diphosphatase)
CAS REGISTRY NUMBER
COMMENTARY
9001-78-9
-
SUBSTRATE
PRODUCT
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate)
(Substrate)
LITERATURE
(Substrate)
(Substrate)
COMMENTARY
(Product)
(Product)
LITERATURE
(Product)
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
r=reversible
ir=irreversible
?=not specified
methyl 3-nitrophenyl phosphate + H2O
methyl phosphate + 3-nitrophenol
-
-
-
?
methyl 4-nitrophenyl phosphate + H2O
methyl phosphate + 4-nitrophenol
-
-
-
?
methyl 4-nitrophenyl phosphorothioate + H2O
4-nitrophenol + methyl phosphorothioate
only the R-enantiomer is detectably hydrolyzed by the enzyme
-
-
?
2'-[2-benzthiazole]-6'-hydroxybenzthiazole phosphate + H2O
2'-[2-benzthiazole]-6'-hydroxybenzthiazole + phosphate
-
weakly fluorescent substrate
highly fluorescent product
?
4-methylumbelliferyl phosphate + H2O
4-methylumbelliferone + phosphate
-
-
-
-
?
methyl p-nitrophenyl phosphate + H2O
methanol + p-nitrophenol + phosphate
-
-
-
-
?
methyl p-nitrophenyl phosphorothioate + H2O
methanol + p-nitrophenol + phosphorothioate
-
-
-
-
?
N-acetylcysteamine S-phosphate + H2O
N-acetylcysteamine + phosphate
-
-
-
-
?
p-nitrophenyl phosphate + H2O
p-nitrophenol + phosphate
-
-
-
-
?
4-nitrophenyl phosphate + H2O
4-nitrophenol + phosphate
-
-
-
?
4-nitrophenyl phosphate + H2O
4-nitrophenol + phosphate
-
-
-
?
p-nitrophenyl phosphate + H2O
p-nitrophenol + phosphate
-
-
-
?
4-nitrophenyl phosphate + H2O
4-nitrophenol + phosphate
-
-
-
-
?
4-nitrophenyl phosphate + H2O
4-nitrophenol + phosphate
-
non-specific, phosphoseryl intermediate
-
?
4-nitrophenyl phosphate + H2O
4-nitrophenol + phosphate
-
strong support for a model in which electrostatic interactions between the bimetallo Zn2+ site and a nonbridging phosphate ester oxygen atom make a significant contribution to the large rate enhancement observed for alkaline phosphatase-catalyzed phosphate monoester hydrolysis
-
-
?
additional information
?
-
-
the enzyme also catalyses the transfer of the phosphoryl group to alcohols
-
?
additional information
?
-
-
the enzyme is involved in recovering of phosphate esters when free phosphate is depleted
-
?
additional information
?
-
-
the enzyme catalyzes the oxidation of phosphite to phosphate and molecular H2: H2PO3 + H2O = H3PO4 + H2
-
-
?
NATURAL SUBSTRATE
NATURAL PRODUCT
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate)
(Substrate)
LITERATURE
(Substrate)
(Substrate)
COMMENTARY
(Product)
(Product)
LITERATURE
(Product)
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
r=reversible
ir=irreversible
?=not specified
additional information
?
-
-
the enzyme is involved in recovering of phosphate esters when free phosphate is depleted
-
?
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
Mg2+
Zn2+
Mg2+
-
enzyme contains 1 Mg2+ ion per monomer, located near the active site, interacting with phosphate
Zn
Zn2+
additional information
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
Zn2+
metalloenzyme, mutant T59A contains sn amount similar to the wild-type enzyme, while mutant T59R has almost undetectable amounts of bound metal
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
Zn2+
-
enzyme contains 2 Zn2+ ions per monomer, located near the active site, interacting with phosphate
Zn2+
-
the enzyme employs a binuclear metallocenter of two Zn2+ ions approximately 4 A apart to facilitate catalysis of phosphate monoester hydrolysis. Alkaline phosphatase catalysis is ultrasensitive to charge sequestered between the active site zinc ions
Zn2+
-
strong support for a model in which electrostatic interactions between the bimetallo Zn2+ site and a nonbridging phosphate ester oxygen atom make a significant contribution to the large rate enhancement observed for alkaline phosphatase-catalyzed phosphate monoester hydrolysis
additional information
-
each subunit contains an active centre with three distinct metal binding sites and one phosphorylatable Ser
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
Urea
-
combined treatment with thioglycolic acid, 0.175%, and 6 M urea leads to reduction of disulfide bonds, loss of activity and separation into subunits
Zn2+
-
mutant H412Y in presence of a phosphoryl group acceptor and Zn2+ at 0.1 mM
thioglycolic acid
-
combined treatment with thioglycolic acid, 0.175%, and 6 M urea leads to reduction of disulfide bonds, loss of activity and separation into subunits
additional information
Zn2+ and Mg2+ do not affect the mutant T59R enzyme
-
additional information
-
Zn2+ and Mg2+ do not affect the mutant T59R enzyme
-
ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
Tris
-
activation at 0.2 M, increases with increasing concentration to a plateau
Zn2+
-
stimulation of mutant enzymes, in presence or absence of a phosphoryl group acceptor in the reaction, except for mutnat H412Y, whose activity is reduced in presence of phosphoryl group acceptor and 0.1 mM Zn2+
additional information
Zn2+ and Mg2+ do not affect the mutant T59R enzyme
-
additional information
-
Zn2+ and Mg2+ do not affect the mutant T59R enzyme
-
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
0.0005
4-nitrophenyl phosphate
mutant enzyme E332Y, at pH 8.0 and 25°C
0.0021
4-nitrophenyl phosphate
mutant enzyme R166S/E322Y/K328A, at pH 8.0 and 25°C
0.0022
4-nitrophenyl phosphate
mutant enzyme E322Y/K328A, at pH 8.0 and 25°C
0.0024
4-nitrophenyl phosphate
mutant enzyme D153A/K328A, at pH 8.0 and 25°C
0.0026
4-nitrophenyl phosphate
mutant enzyme D153A, at pH 8.0 and 25°C
0.0036
4-nitrophenyl phosphate
mutant enzyme D101A, at pH 8.0 and 25°C
0.0048
4-nitrophenyl phosphate
mutant enzyme D101A/R166S/D153A, at pH 8.0 and 25°C
0.005
4-nitrophenyl phosphate
mutant enzyme R166S, at pH 8.0 and 25°C
0.0053
4-nitrophenyl phosphate
mutant enzyme R166S/D153A, at pH 8.0 and 25°C
0.0054
4-nitrophenyl phosphate
mutant enzyme K328A, at pH 8.0 and 25°C
0.0062
4-nitrophenyl phosphate
mutant enzyme D101A/D153A, at pH 8.0 and 25°C
0.0084
4-nitrophenyl phosphate
mutant enzyme D101A/R166S, at pH 8.0 and 25°C
0.011
4-nitrophenyl phosphate
mutant enzyme D101A/D153A/K328A, at pH 8.0 and 25°C
0.012
4-nitrophenyl phosphate
mutant enzyme D101A/R166S/D153A/E322Y, at pH 8.0 and 25°C
0.0218
4-nitrophenyl phosphate
pH 8.0, 25°C, recombinant wild-type enzyme
0.027
4-nitrophenyl phosphate
mutant enzyme R166S/E322Y, at pH 8.0 and 25°C
0.02982
4-nitrophenyl phosphate
single-chain Fv antibody fusion protein
0.052
4-nitrophenyl phosphate
mutant enzyme D153A/E322Y, at pH 8.0 and 25°C
0.063
4-nitrophenyl phosphate
mutant enzyme R166S/K328A, at pH 8.0 and 25°C
0.078
4-nitrophenyl phosphate
mutant enzyme D101A/E322Y/K328A, at pH 8.0 and 25°C
0.14
4-nitrophenyl phosphate
mutant enzyme D101A/D153A/E322Y/K328A, at pH 8.0 and 25°C
0.14
4-nitrophenyl phosphate
mutant enzyme R166S/D153A/K328A, at pH 8.0 and 25°C
0.15
4-nitrophenyl phosphate
mutant enzyme D101A/E322Y, at pH 8.0 and 25°C
0.16
4-nitrophenyl phosphate
mutant enzyme D101A/R166S/D153A/E322Y/K328A, at pH 8.0 and 25°C
0.22
4-nitrophenyl phosphate
mutant enzyme R166S/D153A/E322Y/K328A, at pH 8.0 and 25°C
0.35
4-nitrophenyl phosphate
mutant enzyme D153A/E322Y/K328A, at pH 8.0 and 25°C
0.44
4-nitrophenyl phosphate
mutant enzyme R166S/D153A/E322Y, at pH 8.0 and 25°C
0.00000001
4-nitrophenyl phosphate
-
Vmax (mmol/min mg of protein): 1.8 (dimer), 0.42 (tetramer)
0.0035
4-nitrophenyl phosphate
-
mutant S105L, pH 8.0, 25°C, in absence of a phosphoryl group acceptor
0.005
4-nitrophenyl phosphate
-
mutant T155M, pH 8.0, 25°C, in absence of a phosphoryl group acceptor
0.0073
4-nitrophenyl phosphate
-
mutant H412Y, pH 8.0, 25°C, in absence of a phosphoryl group acceptor
0.0094
4-nitrophenyl phosphate
-
wild-type enzyme, pH 8.0, 25°C, in absence of a phosphoryl group acceptor
0.0095
4-nitrophenyl phosphate
-
mutant E341K, pH 8.0, 25°C, in absence of a phosphoryl group acceptor
0.0097
4-nitrophenyl phosphate
-
mutant S105L, pH 8.0, 25°C, in presence of a phosphoryl group acceptor
0.011
4-nitrophenyl phosphate
-
mutant E322K, pH 8.0, 25°C, in presence of a phosphoryl group acceptor
0.0176
4-nitrophenyl phosphate
-
mutant D369N, pH 8.0, 25°C, in absence of a phosphoryl group acceptor
0.0211
4-nitrophenyl phosphate
-
wild-type enzyme, pH 8.0, 25°C, in presence of a phosphoryl group acceptor
0.0221
4-nitrophenyl phosphate
-
mutant E322K, pH 8.0, 25°C, in absence of a phosphoryl group acceptor
0.0374
4-nitrophenyl phosphate
-
mutant E341K, pH 8.0, 25°C, in presence of a phosphoryl group acceptor
0.057
4-nitrophenyl phosphate
-
mutant T155M, pH 8.0, 25°C, in presence of a phosphoryl group acceptor
0.0597
4-nitrophenyl phosphate
-
mutant D369N, pH 8.0, 25°C, in presence of a phosphoryl group acceptor
0.0691
4-nitrophenyl phosphate
-
mutant D434E, pH 8.0, 25°C, in absence of a phosphoryl group acceptor
0.198
4-nitrophenyl phosphate
-
mutant H412Y, pH 8.0, 25°C, in presence of a phosphoryl group acceptor
0.29
4-nitrophenyl phosphate
-
fusion protein with ZZ protein, pH not specified in the publication, temperature not specified in the publication
0.29
4-nitrophenyl phosphate
-
native enzyme, pH not specified in the publication, temperature not specified in the publication
0.31
4-nitrophenyl phosphate
-
conjugate between enzyme and ZZ protein, pH not specified in the publication, temperature not specified in the publication
2.44
4-nitrophenyl phosphate
-
pH 10.5, 37°C, enzyme immobilized by phosphatase-polyresorcinol complex
0.0094
p-nitrophenyl phosphate
-
wild type enzyme, in absence of a phosphate acceptor
0.02124
p-nitrophenyl phosphate
-
wild type enzyme, in presence of a phosphate acceptor
0.05972
p-nitrophenyl phosphate
-
mutant enzyme D369N, in presence of a phosphate acceptor
0.176
p-nitrophenyl phosphate
-
mutant enzyme D369N, in absence of a phosphate acceptor
2.941
p-nitrophenyl phosphate
-
mutant enzyme D369A, in absence of a phosphate acceptor
additional information
additional information
-
kinetic characterization of the homodimeric complementation mutants, and heterodimeric hybrid formation
-
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
0.000029
4-nitrophenyl phosphate
mutant enzyme D101A/R166S/D153A/E322Y/K328A, at pH 8.0 and 25°C
0.000057
4-nitrophenyl phosphate
mutant enzyme R166S/E322Y/K328A, at pH 8.0 and 25°C
0.00021
4-nitrophenyl phosphate
pH 8.0, 25°C, recombinant mutant T59R
0.00024
4-nitrophenyl phosphate
mutant enzyme D101A/D153A/E322Y/K328A, at pH 8.0 and 25°C
0.00032
4-nitrophenyl phosphate
mutant enzyme D101A/R166S/D153A/E322Y, at pH 8.0 and 25°C
0.00073
4-nitrophenyl phosphate
mutant enzyme D153A/E322Y/K328A, at pH 8.0 and 25°C
0.00089
4-nitrophenyl phosphate
mutant enzyme D101A/E322Y/K328A, at pH 8.0 and 25°C
0.00096
4-nitrophenyl phosphate
mutant enzyme R166S/D153A/E322Y/K328A, at pH 8.0 and 25°C
0.0014
4-nitrophenyl phosphate
mutant enzyme D153A/E322Y, at pH 8.0 and 25°C
0.0026
4-nitrophenyl phosphate
mutant enzyme R166S/D153A/K328A, at pH 8.0 and 25°C
0.0034
4-nitrophenyl phosphate
mutant enzyme E322Y/K328A, at pH 8.0 and 25°C
0.0083
4-nitrophenyl phosphate
mutant enzyme R166S/D153A/E322Y, at pH 8.0 and 25°C
0.016
4-nitrophenyl phosphate
mutant enzyme R166S/K328A, at pH 8.0 and 25°C
0.061
4-nitrophenyl phosphate
mutant enzyme D101A/R166S/D153A/K328A, at pH 8.0 and 25°C
0.07
4-nitrophenyl phosphate
mutant enzyme R166S/D153A, at pH 8.0 and 25°C
0.083
4-nitrophenyl phosphate
mutant enzyme D101A/R166S/D153A, at pH 8.0 and 25°C
1.1
4-nitrophenyl phosphate
mutant enzyme D153A/K328A, at pH 8.0 and 25°C
1.6
4-nitrophenyl phosphate
mutant enzyme D101A/D153A/K328A, at pH 8.0 and 25°C
2.2
4-nitrophenyl phosphate
mutant enzyme D101A/D153A, at pH 8.0 and 25°C
2.3
4-nitrophenyl phosphate
pH 8.0, 25°C, recombinant wild-type enzyme
3 - 6
4-nitrophenyl phosphate
mutant enzyme D101A, at pH 8.0 and 25°C
4.2
4-nitrophenyl phosphate
mutant enzyme D101A/R166S, at pH 8.0 and 25°C
4.6
4-nitrophenyl phosphate
mutant enzyme D101A/E322Y, at pH 8.0 and 25°C
74.7
4-nitrophenyl phosphate
pH 8.0, 25°C, recombinant wild-type enzyme
113.6
4-nitrophenyl phosphate
single-chain Fv antibody fusion protein
0.016
4-nitrophenyl phosphate
-
mutant E322K, pH 8.0, 25°C, in absence of a phosphoryl group acceptor
0.027
4-nitrophenyl phosphate
-
mutant E322K, pH 8.0, 25°C, in presence of a phosphoryl group acceptor
0.042
4-nitrophenyl phosphate
-
mutant E341K, pH 8.0, 25°C, in absence of a phosphoryl group acceptor
0.052
4-nitrophenyl phosphate
-
mutant T155M, pH 8.0, 25°C, in absence of a phosphoryl group acceptor
0.059
4-nitrophenyl phosphate
-
mutant H412Y, pH 8.0, 25°C, in presence of a phosphoryl group acceptor
0.067
4-nitrophenyl phosphate
-
mutant H412Y, pH 8.0, 25°C, in absence of a phosphoryl group acceptor
0.1
4-nitrophenyl phosphate
-
mutant E341K, pH 8.0, 25°C, in presence of a phosphoryl group acceptor
0.32
4-nitrophenyl phosphate
-
mutant T155M, pH 8.0, 25°C, in presence of a phosphoryl group acceptor
0.39
4-nitrophenyl phosphate
-
mutant D369N, pH 8.0, 25°C, in absence of a phosphoryl group acceptor
2.4
4-nitrophenyl phosphate
-
mutant D369N, pH 8.0, 25°C, in presence of a phosphoryl group acceptor
3.8
4-nitrophenyl phosphate
-
mutant S105L, pH 8.0, 25°C, in absence of a phosphoryl group acceptor
5.8
4-nitrophenyl phosphate
-
mutant S105L, pH 8.0, 25°C, in presence of a phosphoryl group acceptor
44.4
4-nitrophenyl phosphate
-
wild-type enzyme, pH 8.0, 25°C, in absence of a phosphoryl group acceptor
65.6
4-nitrophenyl phosphate
-
mutant D434E, pH 8.0, 25°C, in absence of a phosphoryl group acceptor
80.5
4-nitrophenyl phosphate
-
wild-type enzyme, pH 8.0, 25°C, in presence of a phosphoryl group acceptor
285
4-nitrophenyl phosphate
-
conjugate between enzyme and ZZ protein, pH not specified in the publication, temperature not specified in the publication
370
4-nitrophenyl phosphate
-
fusion protein with ZZ protein, pH not specified in the publication, temperature not specified in the publication
378
4-nitrophenyl phosphate
-
native enzyme, pH not specified in the publication, temperature not specified in the publication
0.01
p-nitrophenyl phosphate
-
mutant enzyme D369A, in absence of a phosphate acceptor
0.4
p-nitrophenyl phosphate
-
mutant enzyme D369N, in absence of a phosphate acceptor
80.5
p-nitrophenyl phosphate
-
wild type enzyme, in presence of a phosphate acceptor
230
p-nitrophenyl phosphate
-
mutant enzyme D369N, in presence of a phosphate acceptor
additional information
additional information
kcat/KM (1/Msec) wild-type: 140000, mutant R166K: 20, mutant R166S: 24
-
additional information
additional information
-
kcat/KM (1/Msec) wild-type: 140000, mutant R166K: 20, mutant R166S: 24
-
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
-
kcat/KM VALUE [1/mMs-1]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
0.00002
4-nitrophenyl phosphate
mutant enzyme D101A/R166S/E322Y/K328A, at pH 8.0 and 25°C
0.00017
4-nitrophenyl phosphate
mutant enzyme D101A/R166S/D153A/E322Y/K328A, at pH 8.0 and 25°C
0.00039
4-nitrophenyl phosphate
mutant enzyme R166S/E322Y/K328A, at pH 8.0 and 25°C
0.00094
4-nitrophenyl phosphate
mutant enzyme D101A/R166S/D153A/E322Y, at pH 8.0 and 25°C
0.0011
4-nitrophenyl phosphate
mutant enzyme D101A/E322Y/K328A, at pH 8.0 and 25°C
0.0016
4-nitrophenyl phosphate
mutant enzyme R166S/E322Y, at pH 8.0 and 25°C
0.002
4-nitrophenyl phosphate
mutant enzyme D101A/D153A/E322Y/K328A, at pH 8.0 and 25°C
0.0031
4-nitrophenyl phosphate
mutant enzyme D101A/E322Y, at pH 8.0 and 25°C
0.0044
4-nitrophenyl phosphate
mutant enzyme R166S/D153A/E322Y/K328A, at pH 8.0 and 25°C
0.013
4-nitrophenyl phosphate
mutant enzyme D101A/D153A/E322Y, at pH 8.0 and 25°C
0.019
4-nitrophenyl phosphate
mutant enzyme R166S/D153A/E322Y, at pH 8.0 and 25°C
0.24
4-nitrophenyl phosphate
mutant enzyme R166S/K328A, at pH 8.0 and 25°C
0.28
4-nitrophenyl phosphate
mutant enzyme D101A/R166S/K328A, at pH 8.0 and 25°C
0.32
4-nitrophenyl phosphate
mutant enzyme D153A/E322Y/K328A, at pH 8.0 and 25°C
0.42
4-nitrophenyl phosphate
mutant enzyme D101A/R166S/E322Y, at pH 8.0 and 25°C
1.5
4-nitrophenyl phosphate
mutant enzyme E322Y/K328A, at pH 8.0 and 25°C
2.3
4-nitrophenyl phosphate
mutant enzyme D153A/E322Y, at pH 8.0 and 25°C
3.6
4-nitrophenyl phosphate
mutant enzyme R166S/D153A/K328A, at pH 8.0 and 25°C
5.5
4-nitrophenyl phosphate
mutant enzyme D101A/R166S/D153A/K328A, at pH 8.0 and 25°C
13
4-nitrophenyl phosphate
mutant enzyme R166S/D153A, at pH 8.0 and 25°C
20
4-nitrophenyl phosphate
mutant enzyme D101A/R166S/D153A, at pH 8.0 and 25°C
28
4-nitrophenyl phosphate
mutant enzyme D101A/K328A, at pH 8.0 and 25°C
58
4-nitrophenyl phosphate
mutant enzyme D101A/R166S, at pH 8.0 and 25°C
120
4-nitrophenyl phosphate
mutant enzyme D101A/D153A/K328A, at pH 8.0 and 25°C
330
4-nitrophenyl phosphate
mutant enzyme D101A/D153A, at pH 8.0 and 25°C
440
4-nitrophenyl phosphate
mutant enzyme D153A/K328A, at pH 8.0 and 25°C
Ki VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
0.423
phosphate
-
pH 10.5, 37°C, enzyme immobilized by phosphatase-polyresorcinol complex
IC50 VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
4
phosphate
Escherichia coli
-
pH 10.5, 37°C, enzyme immobilized by phosphatase-polyresorcinol complex
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
additional information
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
10.2
-
maximal activity of both free enzyme and enzyme immobilized by phosphatase-polyresorcinol complex
8.5
8.5
-
wild-type enzyme and alkaline phosphatase fused to a C-terminal region of Pseudomonas sp. MIS38 lipase
pH RANGE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
5 - 10
-
activity profile of free enzyme is comparable to immobilized enzyme
7.5 - 9.5
-
pH 7.5: about 60% of maximal activity, pH 9.5: about 65% of maximal activity, wild-type enzyme and alkaline phosphatase fused to a C-terminal region of Pseudomonas sp. MIS38 lipase
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
25
60 - 70
-
wild-type enzyme and alkaline phosphatase fused to a C-terminal region of Pseudomonas sp. MIS38 lipase
90
-
maximal activity of both free enzyme and enzyme immobilized by phosphatase-polyresorcinol complex
TEMPERATURE RANGE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
30 - 50
-
activity profile of free enzyme is comparable to immobilized enzyme
30 - 85
-
30°C: about 60% of maximal activity, 85°C: about 40% of maximal activity, wild-type enzyme and alkaline phosphatase fused to a C-terminal region of Pseudomonas sp. MIS38 lipase
70 - 90
-
at temperature values between 70 and 90°C, the immobilized phosphatase shows an activity about 10% higher than that of the free enzyme
ORGANISM
COMMENTARY
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
-
UniProt
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
SOURCE
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY
GeneOntology No.
LITERATURE
SOURCE
PDB
SCOP
CATH
UNIPROT
ORGANISM
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
44000
recombinant mutant T59R, sucrose density gradient sedimentation
87000
recombinant wild-type and mutant T59A enzymes, sucrose density gradient sedimentation
SUBUNIT
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
dimer
additional information
dimer
-
activity of the dimer is three or four times higher than that of the tetramer
additional information
Thr59 is involved in stabilization of the dimeric interface, structural analysis
additional information
-
Thr59 is involved in stabilization of the dimeric interface, structural analysis
additional information
-
heterodimer formation in vitro, overview, structural assembly of the enzyme into dimer induces cooperative interactions between the monomers necessary for the formation of the functional form of the holoenzyme
POSTTRANSLATIONAL MODIFICATION
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
CRYSTALLIZATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
a 2.1 A X-ray diffraction structure of Arg166Ser alkaline phosphatase is presented, which shows little difference in enzyme structure compared to the wild-type enzyme but shows a significant reorientation of the bound phosphate
crystal structure of the covalent phospho-enzyme intermediate of the H331Q mutant enzyme and of the transition state complex between the wild-type enzyme and vanadate
hanging drop vapour diffusion method, structures if the inactive wild-type enzyme with cobalt anf the structure of the active D153H/K328W enzyme with cobalt
mutant enzyme D101A/D153A, hanging drop vapor diffusion method, using 22% (w/v) PEG3350, 0.1 mM Bis-Tris, pH 5.0, 0.2 mM ammonium sulfate
PROTEIN VARIANTS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
D101A
the mutant shows 64fold decreased catalytic efficiency compared to the wild type enzyme
D101A/D153A
the mutant shows 190fold decreased catalytic efficiency compared to the wild type enzyme
D101A/D153A/E322Y
the mutant shows 48000000fold decreased catalytic efficiency compared to the wild type enzyme
D101A/D153A/E322Y/K328A
the mutant shows 320000000fold decreased catalytic efficiency compared to the wild type enzyme
D101A/D153A/K328A
the mutant shows 5300fold decreased catalytic efficiency compared to the wild type enzyme
D101A/E322Y
the mutant shows 20000000fold decreased catalytic efficiency compared to the wild type enzyme
D101A/E322Y/K328A
the mutant shows 570000000fold decreased catalytic efficiency compared to the wild type enzyme
D101A/K328A
the mutant shows 23000fold decreased catalytic efficiency compared to the wild type enzyme
D101A/R166S
the mutant shows 11000fold decreased catalytic efficiency compared to the wild type enzyme
D101A/R166S/D153A
the mutant shows 32000fold decreased catalytic efficiency compared to the wild type enzyme
D101A/R166S/D153A/E322Y
the mutant shows 670000000fold decreased catalytic efficiency compared to the wild type enzyme
D101A/R166S/D153A/E322Y/K328A
the mutant shows 3700000000fold decreased catalytic efficiency compared to the wild type enzyme
D101A/R166S/D153A/K328A
the mutant shows 120000fold decreased catalytic efficiency compared to the wild type enzyme
D101A/R166S/E322Y
the mutant shows 15000000000fold decreased catalytic efficiency compared to the wild type enzyme
D101A/R166S/E322Y/K328A
the mutant shows more than 30000000000fold decreased catalytic efficiency compared to the wild type enzyme
D101A/R166S/K328A
the mutant shows 2300000fold decreased catalytic efficiency compared to the wild type enzyme
D101S
weaker phosphate binding can be attributed to the increased flexibility of the Arg166 side chain, faster phosphate release is responsible for the 35fold higher activity
D153A
the mutant shows 230fold decreased catalytic efficiency compared to the wild type enzyme
D153A/E322Y
the mutant shows 270000fold decreased catalytic efficiency compared to the wild type enzyme
D153A/E322Y/K328A
the mutant shows 2000000fold decreased catalytic efficiency compared to the wild type enzyme
D153A/K328A
the mutant shows 1400fold decreased catalytic efficiency compared to the wild type enzyme
D153G
weaker phosphate binding can be attributed to the increased flexibility of the Arg166 side chain, faster phosphate release is responsible for the 5fold higher activity, reduced magnesium affinity, maximal activity is only achieved with the addition of exogenous magnesium
D153H
reduced magnesium affinity, maximual activity is only achieved with the addition of exogenous magnesium
D153H/K328A
reduced magnesium affinity, maximal activity is only achieved with the addition of exogenous magnesium
D153H/K328W
mutant enzyme containing Co2+ has higher catalytic efficiency than the wild-type enzyme containing cobalt
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
E322Y/K328A
the mutant shows 420000fold decreased catalytic efficiency compared to the wild type enzyme
E332Y
the mutant shows 88000fold decreased catalytic efficiency compared to the wild type enzyme
K328A
R166A
R166Q
mutation has very little effect on turnover number, in presence of phosphate acceptor the substrate binding decreases over 50fold
R166S
R166S/D153A
the mutant shows 49000fold decreased catalytic efficiency compared to the wild type enzyme
R166S/D153A/E322Y
the mutant shows 33000000fold decreased catalytic efficiency compared to the wild type enzyme
R166S/D153A/E322Y/K328A
the mutant shows 140000000fold decreased catalytic efficiency compared to the wild type enzyme
R166S/D153A/K328A
the mutant shows 180000fold decreased catalytic efficiency compared to the wild type enzyme
R166S/E322Y
R166S/E322Y/K328A
the mutant shows 1600000000fold decreased catalytic efficiency compared to the wild type enzyme
R166S/K328A
the mutant shows 2600000fold decreased catalytic efficiency compared to the wild type enzyme
S102A
activity is significantly less than that of the wild-type enzyme, but 100000-10000000fold greater than the non-enzymatic reaction,1000-10000fold reduction in the turnover number
S102C
activity is significantly less than that of the wild-type enzyme, but 100000-10000000fold greater than the non-enzymatic reaction, 100fold reduction in turnover number
S102G
activity is significantly less than that of the wild-type enzyme, but 100000-10000000fold greater than the non-enzymatic reaction, 1000-10000fold reduction in the turnover number
S102L
activity is significantly less than that of the wild-type enzyme, but 100000-10000000fold greater than the non-enzymatic reaction, 1000-10000fold reduction in the turnover number
T59A
site-directed mutagenesis, exists as a dimer, shows catalytic activity and metal content similar to the wild-type enzyme
T59R
site-directed mutagenesis, exists as a monomer, shows 10000fold reduced activity compared to the wild-type, highly reduced metal content, highly reduced thermal stability
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
D153G
-
the mutant has 5fold higher catalytic activity but no change in Km at pH 8.0 in 50 mM Tris-HCl. The mutation also affects Mg2+ binding, resulting in an enzyme with lower metal affinity. The mutation also affects the position of the water ligands of Mg2+ and the loop Gln152-Thr155 is shifted by 0.3 A away from the active site. The weaker Mg2+ binding of the mutant compared with the wild type is caused by an altered coordination sphere in the proximity of the Mg2+ ion and also by the loss of an electrostatic interaction, Mg2+/COO-Asp153, in the mutant
D369N
D434E
-
site-directed mutagenesis, reduced activity, increased kcat and Km compared to the wild-type enzyme
T81A
-
the T81A mutant displays a somewhat lower affinity than the wild-type enzyme for both substrate and phosphate, while kcat does not change significantly
additional information
K328A
the mutant shows 840fold decreased catalytic efficiency compared to the wild type enzyme
R166A
mutation has very little effect on turnover number, in presence of phosphate acceptor the substrate binding decreases over 50fold, phosphate inhibition is reduced 50fold
R166S
mutation has very little effect on turnover number, in presence of phosphate acceptor the substrate binding decreases over 50fold, phosphate inhibition is reduced 50fold
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
R166S
the mutant shows 6300fold decreased catalytic efficiency compared to the wild type enzyme
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
R166S/E322Y
the mutant shows 39000000fold decreased catalytic efficiency compared to the wild type enzyme
D369N
-
mutant enzyme shows reduced turnover rates and increased Km-value. The reaction mechanism of the mutant enzyme involves only 1 metal with the possible assistance of a His side chain
additional information
alkaline phosphatase is subject to circular permutation with its novel termini at the loops near the active site, and the original termini are linked by a flexible linker. While a permutant with the termini at original residues 407 and 408 is not active, a permutant with termini at residues 90 and 94 shows significant activity. Also the addition of a randomized residue at position 91 and 93 as well as outer peptide epitopes yields several mutants with specific activity comparable to the wild-type enzyme with similar outer peptides
additional information
-
alkaline phosphatase is subject to circular permutation with its novel termini at the loops near the active site, and the original termini are linked by a flexible linker. While a permutant with the termini at original residues 407 and 408 is not active, a permutant with termini at residues 90 and 94 shows significant activity. Also the addition of a randomized residue at position 91 and 93 as well as outer peptide epitopes yields several mutants with specific activity comparable to the wild-type enzyme with similar outer peptides
additional information
the results show that the role of the arginine side chain extends beyond its positive charge, as the Arg166Lys mutant is as compromised in activity as Arg166Ser. Through measurement of individual reaction steps, a free energy profile for the hydrolysis of the enzyme-phosphate intermediate is constructed. This analysis indicates that the arginine side chain strengthens binding by approximately 3 kcal/mol and provides an additional 1-2 kcal/mol stabilization of the chemical transition state
additional information
-
the results show that the role of the arginine side chain extends beyond its positive charge, as the Arg166Lys mutant is as compromised in activity as Arg166Ser. Through measurement of individual reaction steps, a free energy profile for the hydrolysis of the enzyme-phosphate intermediate is constructed. This analysis indicates that the arginine side chain strengthens binding by approximately 3 kcal/mol and provides an additional 1-2 kcal/mol stabilization of the chemical transition state
pH STABILITY
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
90
-
10 min, pH 8.5, containing 10 mM MgCl2 and 1 mM ZnSO4, about 50% loss of activity
95
additional information
95
-
10 min, pH 8.5, containing 10 mM MgCl2 and 1 mM ZnSO4, aout 80% loss of activity
additional information
-
the thermal stabilities of the immobilized phosphatase are higher than those of the native one
GENERAL STABILITY
ORGANISM
UNIPROT
LITERATURE
plays an indirect but important role in stabilizing the enzyme in its most active conformations
STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
the storage stabilities of the immobilized phosphatase are higher than those of the native one
-
PURIFICATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
using affinity chromatography. Typical yields for a 6-L culture are 30-40 mg of pure protein
CLONED (Commentary)
ORGANISM
UNIPROT
LITERATURE
circularly permutated alkaline phosphatase, expression in Escherichia coli
expressed in Escherichia coli as an N-terminal maltose binding protein (MBP) fusion construct (AP-MBP)
single-chain Fv antibody fusion protein expressed in Escherichia coli BL21(DE3)pLysS
Escherichia coli alkaline phosphatase is fused to a C-terminal region of Pseudomonas sp. MIS38 lipase (PML) and examined for secretion using the Escherichia coli cells carrying the heterologous type I secretion system. PML is one of the passenger proteins of TISS and contains 12 repetitive sequences and a secretion signal at the C-terminal region. The fusion protein is efficiently secreted to the extracellular medium, while alkaline phosphatase is not secreted at all, indicating that the secretion of alkaline phosphatase is promoted by a secretion signal of Pseudomonas sp. MIS38 lipase. The fusion protein purified from the culture supernatant existed as a homodimer, like AP, and is indistinguishable from alkaline phosphatase in enzymatic properties and stability
-
expression of wild-type and mutants in several Escherichia coli strains, intragenic complementation
-
RENATURED/Commentary
ORGANISM
UNIPROT
LITERATURE
APPLICATION
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
analysis
-
construction o a fusion proein between the ZZ protein, two specific Fc-fragment-binding domains derived from the B-domain of SpA, and alkaline phosphatase. The ZZ-AP fusion retains full parental activities and exhibits an approximately tenfold higher sensitivity than that of ZZ-AP conjugate in enzyme-linked immunosorbent assay. The ZZ-AP fusion is a promising immunoreagent for IgG detection and a potential biolinker between antibodies and reporter enzymes. Compared with the parents, the equilibrium dissociation constant of ZZ-AP conjugate is decreased by 32% and catalytic activity is decreased by 24%
biotechnology
-
alkaline phosphatase from Escherichia coli is immobilized by copolymerization with resorcinol. The phosphatase-polyresorcinol complex synthesized retains about 74% of the original enzymatic activity. On addition to soil, free enzyme is completely inactivated in 4 days, whereas the phosphatase-polyresorcinol complex is comparatively stable.Barley seed coated with the immobilized enzyme exhibits higher rhizosphere phosphatase activity. Under pot culture conditions, an increase in the soil inorganic phosphorus is detected when the seed is encapsulated with the phosphatase-polyresorcinol complex, and a positive influence on biomass and inorganic phosphorus concentration of shoot is observed
diagnostics
-
widely used enzyme, e.g. in ELISA, enzyme-linked immunosorbent assay
molecular biology
-
analytically widely used enzyme, e.g. in ELISA, enzyme-linked immunosorbent assay
REF.
AUTHORS
TITLE
JOURNAL
VOL.
PAGES
YEAR
ORGANISM (UNIPROT)
PUBMED ID
SOURCE
Dealwis, C.G.; Brennan, C.; Christianson, K.; Mandecki, W.; Abad-Zapatero C.
Crystallographic analysis of reversible metal binding observed in a mutant (Asp153-->Gly) of Escherichia coli alkaline phosphatase
Biochemistry
31
13967-13973
1995
Escherichia coli
Stinson, R.A.; Chan, J.R.A.
Alkaline phosphatase and its function as a protein phosphatase
Adv. Protein Phosphatases
4
127-151
1987
Bos taurus, Escherichia coli, Homo sapiens, Sus scrofa
-
Dealwis, C.G.; Chen, L.; Brennan, C.; Mandecki, W.; Abad-Zapatero, C.
3-D structure of the D153G mutant of Escherichia coli alkaline phosphatase: an enzyme with weaker magnesium binding and increased catalytic activity
Protein Eng.
8
865-871
1995
Escherichia coli
Wyckhoff, H.W.; Handschumacher, M.; Murthy, H.M.K.; Sowadski, J.M.
The three dimensional structure of alkaline phosphatase from E. coli
Adv. Enzymol. Relat. Areas Mol. Biol.
55
453-480
1983
Escherichia coli
Coleman, J.E.; Gettins, P.
Alkaline phosphatase, solution structure, and mechanism
Adv. Enzymol. Relat. Areas Mol. Biol.
55
381-452
1983
Bacillus subtilis, Bacillus licheniformis, Bos taurus, Escherichia coli, Homo sapiens, Micrococcus sodonensis, Rattus norvegicus
Reid, T.W.; Wilson, I.B.
E. coli alkaline phosphatase
The Enzymes, 3rd Ed. (Boyer, P. D. , ed. )
4
373-415
1971
Escherichia coli
-
Torriani, A.
Alkaline phosphatase of Escherichia coli
Methods Enzymol.
12
212-218
1968
Escherichia coli
-
Scutt, P.B.; Moss, D.W.
Reversible inactivation of alkaline phosphatase in acid solution
Enzymologia
35
157-167
1968
Escherichia coli, Homo sapiens
Stadtman, T.C.
Alkaline phosphatases
The Enzymes, 2nd Ed. (Boyer, P. D. , Lardy, H. , Myrbck, K. , eds. )
5
55-71
1961
Bos taurus, Escherichia coli, Sus scrofa
-
Atlan, D.; Portalier, R.
Purification of extracellular alkaline phosphatase released by Escherichia coli excretory mutants
Appl. Microbiol. Biotechnol.
26
318-322
1987
Escherichia coli
-
Coleman, J.E.; Gettins, P.
Alkaline phosphatase: an enzyme with multiple catalytic metal ions at each active center: 31P and 113Cd NMR in solution correlated with the crystal structure
Front. Bioinorg. Chem. (Lect. Int. Conf. Bioinorg. Chem. , 2nd Meeting Date 1985, Xavier, A. V. , ed. )
547-561
1986
Escherichia coli
-
Holtz, K.M.; Kantrowitz, E.R.
The mechanism of the alkaline phosphatase reaction: insight from NMR, crystallography and site-specific mutagenesis
FEBS Lett.
462
7-11
1999
Escherichia coli (P00634), Escherichia coli
Martinez, M.B.; Schendel, F.J.; Flickinger, M.C.; Nelsestuen, G.L.
Kinetic properties of enzyme populations in vivo: alkaline phosphatase of the Escherichia coli periplasm
Biochemistry
31
11500-11509
1992
Escherichia coli
Tibbitts, T.T.; Xu, X.; Kantrowitz, E.R.
Kinetics and crystal structure of a mutant Escherichia coli alkaline phosphatase (Asp-369->Asn): a mechanism involving one zinc per active site
Protein Sci.
3
2005-2014
1994
Escherichia coli
Malamy, M.; Horecker, B.L.
Alkaline phosphatase (crystalline)
Methods Enzymol.
9
639-642
1966
Escherichia coli, Escherichia coli C4F1
-
Polakowski, R.; Craig, D.B.; Skelley, A.; Dovichi, N.J.
Single molecules of highly purified bacterial alkaline phosphatase have identical activity
J. Am. Chem. Soc.
122
4853-4855
2000
Escherichia coli
-
Boulanger, R.R., Jr.; Kantrowitz, E.R.
Characterization of a monomeric Escherichia coli alkaline phosphatase formed upon a single amino acid substitution
J. Biol. Chem.
278
23497-23501
2003
Escherichia coli (P00634), Escherichia coli
Hehir, M.J.; Murphy, J.E.; Kantrowitz, E.R.
Characterization of heterodimeric alkaline phosphatases from Escherichia coli: An Investigation of Intragenic Complementation
J. Mol. Biol.
304
645-656
2000
Escherichia coli
Savchenko, A.; Wang, W.; Vieille, C.; Zeikus, J.G.
Alkaline phosphatase from Thermotoga neapolitana
Methods Enzymol.
331
298-305
2001
Escherichia coli, Thermotoga neapolitana
Wang, J.; Stieglitz, K.A.; Kantrowitz, E.R.
Metal specificity is correlated with two crucial active site residues in Escherichia coli alkaline phosphatase
Biochemistry
44
8378-8386
2005
Escherichia coli (P00634), Escherichia coli
Nikolic-Hughes, I.; O'Brien, P.J.; Herschlag, D.
Alkaline phosphatase catalysis is ultrasensitive to charge sequestered between the active site zinc ions
J. Am. Chem. Soc.
127
9314-9315
2005
Escherichia coli
Kojima, M.; Ayabe, K.; Ueda, H.
Importance of terminal residues on circularly permutated Escherichia coli alkaline phosphatase with high specific activity
J. Biosci. Bioeng.
100
197-202
2005
Escherichia coli (P00634), Escherichia coli
Yang, K.; Metcalf, W.W.
A new activity for an old enzyme: Escherichia coli bacterial alkaline phosphatase is a phosphite-dependent hydrogenase
Proc. Natl. Acad. Sci. USA
101
7919-7924
2004
Escherichia coli
Gudjonsdottir, K.; Asgeirsson, B.
Effects of replacing active site residues in a cold-active alkaline phosphatase with those found in its mesophilic counterpart from Escherichia coli
FEBS J.
275
117-127
2008
Vibrio sp., Escherichia coli (P00634), Escherichia coli
Orhanovic, S.; Bucevic-Popovic, V.; Pavela-Vrancic, M.; Vujaklija, D.; Gamulin, V.
Effect of a T81A mutation at the subunit interface on catalytic properties of alkaline phosphatase from Escherichia coli
Int. J. Biol. Macromol.
40
54-58
2006
Escherichia coli
Zalatan, J.G.; Catrina, I.; Mitchell, R.; Grzyska, P.K.; Obrien, P.J.; Herschlag, D.; Hengge, A.C.
Kinetic isotope effects for alkaline phosphatase reactions: implications for the role of active-site metal ions in catalysis
J. Am. Chem. Soc.
129
9789-9798
2007
Escherichia coli
Angkawidjaja, C.; Kuwahara, K.; Omori, K.; Koga, Y.; Takano, K.; Kanaya, S.
Extracellular secretion of Escherichia coli alkaline phosphatase with a C-terminal tag by type I secretion system: purification and biochemical characterization
Protein Eng. Des. Sel.
19
337-343
2006
Escherichia coli
OBrien, P.J.; Lassila, J.K.; Fenn, T.D.; Zalatan, J.G.; Herschlag, D.
Arginine coordination in enzymatic phosphoryl transfer: evaluation of the effect of Arg166 mutations in Escherichia coli alkaline phosphatase
Biochemistry
47
7663-7672
2008
Escherichia coli (P00634), Escherichia coli
Pilar, M.C.; Ortega, N.; Perez-Mateos, M.; Busto, M.D.
Alkaline phosphatase-polyresorcinol complex: characterization and application to seed coating
J. Agric. Food Chem.
57
1967-1974
2009
Escherichia coli
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
J. Mol. Biol.
384
1174-1189
2008
Escherichia coli (P00634)
Atyaksheva, L.F.; Chukhrai, E.S.; Poltorak, O.M.
The catalytic properties of alkaline phosphatases under various conditions
Russ. J. Phys. Chem. A
82
1947-1951
2008
Bos taurus, Escherichia coli, Gallus gallus (Q92058)
-
Liu, X.; Wang, H.; Liang, Y.; Yang, J.; Zhang, H.; Lei, H.; Shen, Y.; Sun, Y.
Production and characterization of a single-chain Fv antibody-alkaline phosphatase fusion protein specific for clenbuterol
Mol. Biotechnol.
45
56-64
2010
Escherichia coli (P00634), Escherichia coli
Tang, J.B.; Yang, H.M.; Liang, S.J.; Chen, Y.; Mu, Q.J.; Zhang, J.B.
Comparative characterization of recombinant ZZ protein-alkaline phosphatase and its application in enzyme immunoassays
Appl. Microbiol. Biotechnol.
97
153-158
2013
Escherichia coli
Liu, W.; Zhang, R.; Tian, N.; Xu, X.; Cao, Y.; Xian, M.; Liu, H.
Utilization of alkaline phosphatase PhoA in the bioproduction of geraniol by metabolically engineered Escherichia coli
Bioengineered
6
288-293
2015
Escherichia coli
Sunden, F.; Peck, A.; Salzman, J.; Ressl, S.; Herschlag, D.
Extensive site-directed mutagenesis reveals interconnected functional units in the alkaline phosphatase active site
eLife
2015
e06181
2015
Escherichia coli (P00634), Escherichia coli
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