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

BRENDA support

Literature summary for 3.1.3.1 extracted from

  • 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 (2008), FEBS J., 275, 117-127.
    View publication on PubMed
Search for more literature for 3.1.3.1

Protein Variants

Protein Variants Comment Organism
H116D slightly less heat-stable than the wild-type enzyme. Mutant enzyme shows much reduced catalytic efficiencies as compared to the wild-type enzyme. kcat/KM is 29.7fold lower than wild-type value Vibrio sp.
H116D/W274K more heat-tolerant than the wild-type enzyme. Mutant enzyme shows much reduced catalytic efficiencies as compared to the wild-type enzyme. kcat/KM is 17.1fold lower than wild-type value Vibrio sp.
W274K more heat-tolerant than the wild-type enzyme. Mutant enzyme shows much reduced catalytic efficiencies as compared to the wild-type enzyme. kcat/KM is 30.6fold lower than wild-type value Vibrio sp.

General Stability

General Stability Organism
loss of activity occurrs at urea concentrations below 1 M. The global structure is unfolded at urea concentrations in the range 1-3 M Vibrio sp.

Inhibitors

Inhibitors Comment Organism Structure
phosphate
-
 Escherichia coli
phosphate
-
 Vibrio sp.

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.021
-
 p-nitrophenyl phosphate 25°C, pH 8.0, wild-type enzyme Escherichia coli
0.08
-
 p-nitrophenyl phosphate 25°C, pH 9.8, wild-type enzyme Vibrio sp.
0.19
-
 p-nitrophenyl phosphate 25°C, pH 9.8, mutant enzyme W274K Vibrio sp.
1.25
-
 p-nitrophenyl phosphate 25°C, pH 9.8, mutant enzyme H116D Vibrio sp.
1.44
-
 p-nitrophenyl phosphate 25°C, pH 9.8, mutant enzyme H116D/W274K Vibrio sp.

Metals/Ions

Metals/Ions Comment Organism Structure
Mg2+ wild-type enzyme most likely has one Mg2+ in the active site in addition to three Mg2+ that bind elsewhere. At 18°C with 10 mM Mg2+, the enzyme activity is completely stable over 160 min. With no Mg2+ in solution, the activity drops to 20% of the initial activity after 160 min Vibrio sp.
Zn2+ wild-type enzyme most likely has two Zn2+. Zn2+ release from the active site coincides with total protein structure unfolding Vibrio sp.

Organism

Organism UniProt Comment Textmining
Escherichia coli P00634
-
-
Vibrio sp.
-
-
-

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
p-nitrophenyl phosphate + H2O
-
 Vibrio sp. p-nitrophenol + phosphate
-
 ?
p-nitrophenyl phosphate + H2O
-
 Escherichia coli p-nitrophenol + phosphate
-
 ?

Temperature Stability [°C]

Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
25
-
 t1/2: 94 min for wild-type value Vibrio sp.
30
-
 t1/2: 79 min for wild-type enzyme, 2600 min for mutant enzyme h116D Vibrio sp.
35
-
 t1/2: 31 min for wild-type enzyme, 63 min for mutant enzyme H116D, 2200 min for mutant enzyme H116S/W274K Vibrio sp.
40
-
 t1/2: 12 min for wild-type enzyme, 231 min for mutant enzyme W274K, 30 min for mutant enzyme H116D, 217 min for mutant enzyme H116D/W274K Vibrio sp.
45
-
 t1/2: 2.3 min for wild-type enzyme, 26 min for mutant enzyme W274K, 1.1 min for mutant enzyme H116D, 96 min for mutant enzyme H116D/W274K Vibrio sp.
50
-
 t1/2: 0.7 min for wild-type enzyme, 6 min for mutant enzyme W274K, 0.8 min for mutant enzyme H116D, 10 min for mutant enzyme H116D/W274K Vibrio sp.
55
-
 t1/2: 1.2 min for mutant enzyme W274K, 0.2 min for mutant enzyme H116D, 5 min for mutant enzyme H116D/W274K Vibrio sp.
55
-
 Tm-value for mutant enzyme H116D Vibrio sp.
57
-
 Tm-value for mutant enzymeW274K is 57.2°C Vibrio sp.
57
-
 Tm-value for wild-type enzyme is 56.5°C Vibrio sp.
59
-
 Tm-value for mutant enzyme H116D/W274K is 58.6°C Vibrio sp.
60
-
 t1/2: 0.5 min for mutant enzyme H116D/W274K Vibrio sp.

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
38
-
 p-nitrophenyl phosphate 25°C, pH 8.0, wild-type enzyme Escherichia coli
220
-
 p-nitrophenyl phosphate 25°C, pH 9.8, mutant enzyme W274K Vibrio sp.
830
-
 p-nitrophenyl phosphate 25°C, pH 9.8, mutant enzyme H116D Vibrio sp.
930
-
 p-nitrophenyl phosphate 25°C, pH 9.8, mutant enzyme H116D/W274K Vibrio sp.
1580
-
 p-nitrophenyl phosphate 25°C, pH 9.8, wild-type enzyme Vibrio sp.

Ki Value [mM]

Ki Value [mM] Ki Value maximum [mM] Inhibitor Comment Organism Structure
0.045
-
 phosphate 25°C, pH 8.0, wild-type enzyme Escherichia coli
0.39
-
 phosphate 25°C, pH 9.8, wild-type enzyme Vibrio sp.
0.77
-
 phosphate 25°C, pH 9.8, mutant enzyme W274K Vibrio sp.
3.45
-
 phosphate 25°C, pH 9.8, mutant enzyme H116D/W274K Vibrio sp.
5.25
-
 phosphate 25°C, pH 9.8, mutant enzyme H116D Vibrio sp.