Any feedback?
Please rate this page
(literature.php)
(0/150)

BRENDA support

Literature summary for 3.1.3.1 extracted from

  • Sunden, F.; Peck, A.; Salzman, J.; Ressl, S.; Herschlag, D.
    Extensive site-directed mutagenesis reveals interconnected functional units in the alkaline phosphatase active site (2015), eLife, 2015, e06181 .
    View publication on PubMedView publication on EuropePMC
Search for more literature for 3.1.3.1

Cloned(Commentary)

Cloned (Comment) Organism
expressed in Escherichia coli SM547(DE3) cells Escherichia coli

Crystallization (Commentary)

Crystallization (Comment) Organism
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 Escherichia coli

Protein Variants

Protein Variants Comment Organism
D101A the mutant shows 64fold decreased catalytic efficiency compared to the wild type enzyme Escherichia coli
D101A/D153A the mutant shows 190fold decreased catalytic efficiency compared to the wild type enzyme Escherichia coli
D101A/D153A/E322Y the mutant shows 48000000fold decreased catalytic efficiency compared to the wild type enzyme Escherichia coli
D101A/D153A/E322Y/K328A the mutant shows 320000000fold decreased catalytic efficiency compared to the wild type enzyme Escherichia coli
D101A/D153A/K328A the mutant shows 5300fold decreased catalytic efficiency compared to the wild type enzyme Escherichia coli
D101A/E322Y the mutant shows 20000000fold decreased catalytic efficiency compared to the wild type enzyme Escherichia coli
D101A/E322Y/K328A the mutant shows 570000000fold decreased catalytic efficiency compared to the wild type enzyme Escherichia coli
D101A/K328A the mutant shows 23000fold decreased catalytic efficiency compared to the wild type enzyme Escherichia coli
D101A/R166S the mutant shows 11000fold decreased catalytic efficiency compared to the wild type enzyme Escherichia coli
D101A/R166S/D153A the mutant shows 32000fold decreased catalytic efficiency compared to the wild type enzyme Escherichia coli
D101A/R166S/D153A/E322Y the mutant shows 670000000fold decreased catalytic efficiency compared to the wild type enzyme Escherichia coli
D101A/R166S/D153A/E322Y/K328A the mutant shows 3700000000fold decreased catalytic efficiency compared to the wild type enzyme Escherichia coli
D101A/R166S/D153A/K328A the mutant shows 120000fold decreased catalytic efficiency compared to the wild type enzyme Escherichia coli
D101A/R166S/E322Y the mutant shows 15000000000fold decreased catalytic efficiency compared to the wild type enzyme Escherichia coli
D101A/R166S/E322Y/K328A the mutant shows more than 30000000000fold decreased catalytic efficiency compared to the wild type enzyme Escherichia coli
D101A/R166S/K328A the mutant shows 2300000fold decreased catalytic efficiency compared to the wild type enzyme Escherichia coli
D153A the mutant shows 230fold decreased catalytic efficiency compared to the wild type enzyme Escherichia coli
D153A/E322Y the mutant shows 270000fold decreased catalytic efficiency compared to the wild type enzyme Escherichia coli
D153A/E322Y/K328A the mutant shows 2000000fold decreased catalytic efficiency compared to the wild type enzyme Escherichia coli
D153A/K328A the mutant shows 1400fold decreased catalytic efficiency compared to the wild type enzyme Escherichia coli
E322Y/K328A the mutant shows 420000fold decreased catalytic efficiency compared to the wild type enzyme Escherichia coli
E332Y the mutant shows 88000fold decreased catalytic efficiency compared to the wild type enzyme Escherichia coli
K328A the mutant shows 840fold decreased catalytic efficiency compared to the wild type enzyme Escherichia coli
R166S the mutant shows 6300fold decreased catalytic efficiency compared to the wild type enzyme Escherichia coli
R166S/D153A the mutant shows 49000fold decreased catalytic efficiency compared to the wild type enzyme Escherichia coli
R166S/D153A/E322Y the mutant shows 33000000fold decreased catalytic efficiency compared to the wild type enzyme Escherichia coli
R166S/D153A/E322Y/K328A the mutant shows 140000000fold decreased catalytic efficiency compared to the wild type enzyme Escherichia coli
R166S/D153A/K328A the mutant shows 180000fold decreased catalytic efficiency compared to the wild type enzyme Escherichia coli
R166S/E322Y the mutant shows 39000000fold decreased catalytic efficiency compared to the wild type enzyme Escherichia coli
R166S/E322Y/K328A the mutant shows 1600000000fold decreased catalytic efficiency compared to the wild type enzyme Escherichia coli
R166S/K328A the mutant shows 2600000fold decreased catalytic efficiency compared to the wild type enzyme Escherichia coli
S102G/D101A/D153A/R166S/E322Y/K328A inactive Escherichia coli

Inhibitors

Inhibitors Comment Organism Structure
phosphate
-
 Escherichia coli

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.00036
-
 4-nitrophenyl phosphate wild type enzyme, at pH 8.0 and 25°C Escherichia coli
0.0005
-
 4-nitrophenyl phosphate mutant enzyme E332Y, at pH 8.0 and 25°C Escherichia coli
0.0021
-
 4-nitrophenyl phosphate mutant enzyme R166S/E322Y/K328A, at pH 8.0 and 25°C Escherichia coli
0.0022
-
 4-nitrophenyl phosphate mutant enzyme E322Y/K328A, at pH 8.0 and 25°C Escherichia coli
0.0024
-
 4-nitrophenyl phosphate mutant enzyme D153A/K328A, at pH 8.0 and 25°C Escherichia coli
0.0026
-
 4-nitrophenyl phosphate mutant enzyme D153A, at pH 8.0 and 25°C Escherichia coli
0.0036
-
 4-nitrophenyl phosphate mutant enzyme D101A, at pH 8.0 and 25°C Escherichia coli
0.0048
-
 4-nitrophenyl phosphate mutant enzyme D101A/R166S/D153A, at pH 8.0 and 25°C Escherichia coli
0.005
-
 4-nitrophenyl phosphate mutant enzyme R166S, at pH 8.0 and 25°C Escherichia coli
0.0053
-
 4-nitrophenyl phosphate mutant enzyme R166S/D153A, at pH 8.0 and 25°C Escherichia coli
0.0054
-
 4-nitrophenyl phosphate mutant enzyme K328A, at pH 8.0 and 25°C Escherichia coli
0.0062
-
 4-nitrophenyl phosphate mutant enzyme D101A/D153A, at pH 8.0 and 25°C Escherichia coli
0.0084
-
 4-nitrophenyl phosphate mutant enzyme D101A/R166S, at pH 8.0 and 25°C Escherichia coli
0.011
-
 4-nitrophenyl phosphate mutant enzyme D101A/D153A/K328A, at pH 8.0 and 25°C Escherichia coli
0.012
-
 4-nitrophenyl phosphate mutant enzyme D101A/R166S/D153A/E322Y, at pH 8.0 and 25°C Escherichia coli
0.027
-
 4-nitrophenyl phosphate mutant enzyme R166S/E322Y, at pH 8.0 and 25°C Escherichia coli
0.052
-
 4-nitrophenyl phosphate mutant enzyme D153A/E322Y, at pH 8.0 and 25°C Escherichia coli
0.063
-
 4-nitrophenyl phosphate mutant enzyme R166S/K328A, at pH 8.0 and 25°C Escherichia coli
0.078
-
 4-nitrophenyl phosphate mutant enzyme D101A/E322Y/K328A, at pH 8.0 and 25°C Escherichia coli
0.14
-
 4-nitrophenyl phosphate mutant enzyme D101A/D153A/E322Y/K328A, at pH 8.0 and 25°C Escherichia coli
0.14
-
 4-nitrophenyl phosphate mutant enzyme R166S/D153A/K328A, at pH 8.0 and 25°C Escherichia coli
0.15
-
 4-nitrophenyl phosphate mutant enzyme D101A/E322Y, at pH 8.0 and 25°C Escherichia coli
0.16
-
 4-nitrophenyl phosphate mutant enzyme D101A/R166S/D153A/E322Y/K328A, at pH 8.0 and 25°C Escherichia coli
0.22
-
 4-nitrophenyl phosphate mutant enzyme R166S/D153A/E322Y/K328A, at pH 8.0 and 25°C Escherichia coli
0.35
-
 4-nitrophenyl phosphate mutant enzyme D153A/E322Y/K328A, at pH 8.0 and 25°C Escherichia coli
0.44
-
 4-nitrophenyl phosphate mutant enzyme R166S/D153A/E322Y, at pH 8.0 and 25°C Escherichia coli

Metals/Ions

Metals/Ions Comment Organism Structure
Mg2+ required for activity Escherichia coli
Zn2+ required for activity Escherichia coli

Organism

Organism UniProt Comment Textmining
Escherichia coli P00634
-
-

Purification (Commentary)

Purification (Comment) Organism
amylose resin column chromatography Escherichia coli

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
4-nitrophenyl phosphate + H2O
-
 Escherichia coli 4-nitrophenol + phosphate
-
 ?
methyl phosphate + H2O
-
 Escherichia coli methanol + phosphate
-
 ?

Subunits

Subunits Comment Organism
homodimer
-
 Escherichia coli

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
0.000029
-
 4-nitrophenyl phosphate mutant enzyme D101A/R166S/D153A/E322Y/K328A, at pH 8.0 and 25°C Escherichia coli
0.000057
-
 4-nitrophenyl phosphate mutant enzyme R166S/E322Y/K328A, at pH 8.0 and 25°C Escherichia coli
0.00024
-
 4-nitrophenyl phosphate mutant enzyme D101A/D153A/E322Y/K328A, at pH 8.0 and 25°C Escherichia coli
0.00032
-
 4-nitrophenyl phosphate mutant enzyme D101A/R166S/D153A/E322Y, at pH 8.0 and 25°C Escherichia coli
0.00073
-
 4-nitrophenyl phosphate mutant enzyme D153A/E322Y/K328A, at pH 8.0 and 25°C Escherichia coli
0.00089
-
 4-nitrophenyl phosphate mutant enzyme D101A/E322Y/K328A, at pH 8.0 and 25°C Escherichia coli
0.00096
-
 4-nitrophenyl phosphate mutant enzyme R166S/D153A/E322Y/K328A, at pH 8.0 and 25°C Escherichia coli
0.0014
-
 4-nitrophenyl phosphate mutant enzyme D153A/E322Y, at pH 8.0 and 25°C Escherichia coli
0.0026
-
 4-nitrophenyl phosphate mutant enzyme R166S/D153A/K328A, at pH 8.0 and 25°C Escherichia coli
0.0034
-
 4-nitrophenyl phosphate mutant enzyme E322Y/K328A, at pH 8.0 and 25°C Escherichia coli
0.0083
-
 4-nitrophenyl phosphate mutant enzyme R166S/D153A/E322Y, at pH 8.0 and 25°C Escherichia coli
0.016
-
 4-nitrophenyl phosphate mutant enzyme R166S/K328A, at pH 8.0 and 25°C Escherichia coli
0.061
-
 4-nitrophenyl phosphate mutant enzyme D101A/R166S/D153A/K328A, at pH 8.0 and 25°C Escherichia coli
0.07
-
 4-nitrophenyl phosphate mutant enzyme R166S/D153A, at pH 8.0 and 25°C Escherichia coli
0.083
-
 4-nitrophenyl phosphate mutant enzyme D101A/R166S/D153A, at pH 8.0 and 25°C Escherichia coli
0.5
-
 4-nitrophenyl phosphate mutant enzyme R166S, at pH 8.0 and 25°C Escherichia coli
1.1
-
 4-nitrophenyl phosphate mutant enzyme D153A/K328A, at pH 8.0 and 25°C Escherichia coli
1.6
-
 4-nitrophenyl phosphate mutant enzyme D101A/D153A/K328A, at pH 8.0 and 25°C Escherichia coli
2.2
-
 4-nitrophenyl phosphate mutant enzyme D101A/D153A, at pH 8.0 and 25°C Escherichia coli
3 6 4-nitrophenyl phosphate mutant enzyme D101A, at pH 8.0 and 25°C Escherichia coli
3.4
-
 4-nitrophenyl phosphate mutant enzyme K328A, at pH 8.0 and 25°C Escherichia coli
4.2
-
 4-nitrophenyl phosphate mutant enzyme D101A/R166S, at pH 8.0 and 25°C Escherichia coli
4.6
-
 4-nitrophenyl phosphate mutant enzyme D101A/E322Y, at pH 8.0 and 25°C Escherichia coli
7.6
-
 4-nitrophenyl phosphate mutant enzyme D153A, at pH 8.0 and 25°C Escherichia coli
12
-
 4-nitrophenyl phosphate wild type enzyme, at pH 8.0 and 25°C Escherichia coli

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
0.00002
-
 4-nitrophenyl phosphate mutant enzyme D101A/R166S/E322Y/K328A, at pH 8.0 and 25°C Escherichia coli
0.00017
-
 4-nitrophenyl phosphate mutant enzyme D101A/R166S/D153A/E322Y/K328A, at pH 8.0 and 25°C Escherichia coli
0.00039
-
 4-nitrophenyl phosphate mutant enzyme R166S/E322Y/K328A, at pH 8.0 and 25°C Escherichia coli
0.00094
-
 4-nitrophenyl phosphate mutant enzyme D101A/R166S/D153A/E322Y, at pH 8.0 and 25°C Escherichia coli
0.0011
-
 4-nitrophenyl phosphate mutant enzyme D101A/E322Y/K328A, at pH 8.0 and 25°C Escherichia coli
0.0016
-
 methyl phosphate mutant enzyme E322Y, at pH 8.0 and 25°C Escherichia coli
0.0016
-
 4-nitrophenyl phosphate mutant enzyme R166S/E322Y, at pH 8.0 and 25°C Escherichia coli
0.002
-
 4-nitrophenyl phosphate mutant enzyme D101A/D153A/E322Y/K328A, at pH 8.0 and 25°C Escherichia coli
0.0031
-
 4-nitrophenyl phosphate mutant enzyme D101A/E322Y, at pH 8.0 and 25°C Escherichia coli
0.0044
-
 4-nitrophenyl phosphate mutant enzyme R166S/D153A/E322Y/K328A, at pH 8.0 and 25°C Escherichia coli
0.011
-
 methyl phosphate mutant enzyme R166S, at pH 8.0 and 25°C Escherichia coli
0.013
-
 4-nitrophenyl phosphate mutant enzyme D101A/D153A/E322Y, at pH 8.0 and 25°C Escherichia coli
0.019
-
 4-nitrophenyl phosphate mutant enzyme R166S/D153A/E322Y, at pH 8.0 and 25°C Escherichia coli
0.061
-
 methyl phosphate mutant enzyme D101A/D153A, at pH 8.0 and 25°C Escherichia coli
0.24
-
 4-nitrophenyl phosphate mutant enzyme R166S/K328A, at pH 8.0 and 25°C Escherichia coli
0.28
-
 4-nitrophenyl phosphate mutant enzyme D101A/R166S/K328A, at pH 8.0 and 25°C Escherichia coli
0.32
-
 4-nitrophenyl phosphate mutant enzyme D153A/E322Y/K328A, at pH 8.0 and 25°C Escherichia coli
0.42
-
 4-nitrophenyl phosphate mutant enzyme D101A/R166S/E322Y, at pH 8.0 and 25°C Escherichia coli
1.1
-
 methyl phosphate mutant enzyme D153A, at pH 8.0 and 25°C Escherichia coli
1.5
-
 4-nitrophenyl phosphate mutant enzyme E322Y/K328A, at pH 8.0 and 25°C Escherichia coli
2.3
-
 4-nitrophenyl phosphate mutant enzyme D153A/E322Y, at pH 8.0 and 25°C Escherichia coli
2.7
-
 methyl phosphate mutant enzyme D101A, at pH 8.0 and 25°C Escherichia coli
3.6
-
 4-nitrophenyl phosphate mutant enzyme R166S/D153A/K328A, at pH 8.0 and 25°C Escherichia coli
5.5
-
 4-nitrophenyl phosphate mutant enzyme D101A/R166S/D153A/K328A, at pH 8.0 and 25°C Escherichia coli
7.2
-
 4-nitrophenyl phosphate mutant enzyme E332Y, at pH 8.0 and 25°C Escherichia coli
13
-
 4-nitrophenyl phosphate mutant enzyme R166S/D153A, at pH 8.0 and 25°C Escherichia coli
20
-
 4-nitrophenyl phosphate mutant enzyme D101A/R166S/D153A, at pH 8.0 and 25°C Escherichia coli
28
-
 4-nitrophenyl phosphate mutant enzyme D101A/K328A, at pH 8.0 and 25°C Escherichia coli
58
-
 4-nitrophenyl phosphate mutant enzyme D101A/R166S, at pH 8.0 and 25°C Escherichia coli
100
-
 4-nitrophenyl phosphate mutant enzyme R166S, at pH 8.0 and 25°C Escherichia coli
120
-
 4-nitrophenyl phosphate mutant enzyme D101A/D153A/K328A, at pH 8.0 and 25°C Escherichia coli
330
-
 4-nitrophenyl phosphate mutant enzyme D101A/D153A, at pH 8.0 and 25°C Escherichia coli
440
-
 4-nitrophenyl phosphate mutant enzyme D153A/K328A, at pH 8.0 and 25°C Escherichia coli
750
-
 4-nitrophenyl phosphate mutant enzyme K328A, at pH 8.0 and 25°C Escherichia coli
1200
-
 methyl phosphate wild type enzyme, at pH 8.0 and 25°C Escherichia coli
2800
-
 4-nitrophenyl phosphate mutant enzyme D153A, at pH 8.0 and 25°C Escherichia coli
9900
-
 4-nitrophenyl phosphate mutant enzyme D101A, at pH 8.0 and 25°C Escherichia coli
33000
-
 4-nitrophenyl phosphate wild type enzyme, at pH 8.0 and 25°C Escherichia coli