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Literature summary for 3.1.3.1 extracted from

  • Koutsioulis, D.; Wang, E.; Tzanodaskalaki, M.; Nikiforaki, D.; Deli, A.; Feller, G.; Heikinheimo, P.; Bouriotis, V.
    Directed evolution on the cold adapted properties of TAB5 alkaline phosphatase (2008), Protein Eng. Des. Sel., 21, 319-327.
    View publication on PubMed

Cloned(Commentary)

Cloned (Comment) Organism
expressed in Escherichia coli Antarctic bacterium TAB5

Protein Variants

Protein Variants Comment Organism
G149D mutant shows increased activity after thermal inactivation at 60°C, 120 min: 11.4%, kcat (1/sec) (4-nitrophenyl phosphate): 117 Antarctic bacterium TAB5
G87A mutant shows increased activity after thermal inactivation at 60°C, 120 min: 11.4%, kcat (1/sec) (4-nitrophenyl phosphate): 100 Antarctic bacterium TAB5
H135E double mutant shows decreased activity after thermal inactivation at 60°C, 120 min: 0.4%, kcat (1/sec) (4-nitrophenyl phosphate): 303 Antarctic bacterium TAB5
H135E/G149D double mutant shows decreased activity after thermal inactivation at 60°C, 120 min: 1%, kcat (1/sec) (4-nitrophenyl phosphate): 361 Antarctic bacterium TAB5
S338T double mutant shows decreased activity after thermal inactivation at 60°C, 120 min: 0.6%, kcat (1/sec) (4-nitrophenyl phosphate): 114 Antarctic bacterium TAB5
S42G double mutant shows decreased activity after thermal inactivation at 60°C, 120 min: 0%, kcat (1/sec) (4-nitrophenyl phosphate): 133 Antarctic bacterium TAB5
S42G/H135E double mutant shows decreased activity after thermal inactivation at 60°C, 120 min: 0%, kcat (1/sec) (4-nitrophenyl phosphate): 70 Antarctic bacterium TAB5
S42G/S338T double mutant shows decreased activity after thermal inactivation at 60°C, 120 min: 0%, kcat (1/sec) (4-nitrophenyl phosphate): 71 Antarctic bacterium TAB5
S86A mutant shows increased activity after thermal inactivation at 60°C, 120 min: 15.6%, kcat (1/sec) (4-nitrophenyl phosphate): 203 Antarctic bacterium TAB5
S86A/G87A double mutant shows increased activity after thermal inactivation at 60°C, 120 min: 39.4%, kcat (1/sec) (4-nitrophenyl phosphate): 30 Antarctic bacterium TAB5

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
35860
-
deduced from cDNA Antarctic bacterium TAB5

Organism

Organism UniProt Comment Textmining
Antarctic bacterium TAB5 Q9KWY4
-
-

Purification (Commentary)

Purification (Comment) Organism
using a Q-Sepharose fast flow column and a Sephacryl S200 high-resolution gel-filtration column Antarctic bacterium TAB5

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
4-nitrophenyl phosphate + H2O
-
Antarctic bacterium TAB5 4-nitrophenol + phosphate
-
?

Synonyms

Synonyms Comment Organism
alkaline phosphatase
-
Antarctic bacterium TAB5
TAB5 AP
-
Antarctic bacterium TAB5

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
15
-
assay at Antarctic bacterium TAB5

Temperature Stability [°C]

Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
additional information
-
residual activity after after thermal inactivation at 60°C, 120 min: 2.4% Antarctic bacterium TAB5
10 30
-
Antarctic bacterium TAB5

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
30
-
4-nitrophenyl phosphate pH 10.0, 15°C, mutant S86A/G87A Antarctic bacterium TAB5
70
-
4-nitrophenyl phosphate pH 10.0, 15°C, mutant S42G/H135E Antarctic bacterium TAB5
71
-
4-nitrophenyl phosphate pH 10.0, 15°C, mutant S42G/S338T Antarctic bacterium TAB5
100
-
4-nitrophenyl phosphate pH 10.0, 15°C, mutant G87A Antarctic bacterium TAB5
114
-
4-nitrophenyl phosphate pH 10.0, 15°C, mutant S338T Antarctic bacterium TAB5
117
-
4-nitrophenyl phosphate pH 10.0, 15°C, mutant G149D Antarctic bacterium TAB5
133
-
4-nitrophenyl phosphate pH 10.0, 15°C, mutant S42G Antarctic bacterium TAB5
203
-
4-nitrophenyl phosphate pH 10.0, 15°C, mutant S86A Antarctic bacterium TAB5
236
-
4-nitrophenyl phosphate pH 10.0, 15°C, wild-type Antarctic bacterium TAB5
303
-
4-nitrophenyl phosphate pH 10.0, 15°C, mutant H135E Antarctic bacterium TAB5
361
-
4-nitrophenyl phosphate pH 10.0, 15°C, mutant H135E/G149D Antarctic bacterium TAB5

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
10
-
assay at Antarctic bacterium TAB5