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Literature summary for 3.1.3.1 extracted from

  • Hjoerleifsson, J.G.; Asgeirsson, B.
    Cold-active alkaline phosphatase is irreversibly transformed into an inactive dimer by low urea concentrations (2016), Biochim. Biophys. Acta, 1864, 755-765 .
    View publication on PubMed

Cloned(Commentary)

Cloned (Comment) Organism
expressed in Escherichia coli LMG194 cells Vibrio splendidus

Protein Variants

Protein Variants Comment Organism
F355W the mutant shows slightly reduced catalytic efficiency compared to the wild type enzyme Vibrio splendidus
R336L the mutant shows extreme thermo-instability and its activity drops by half at 0.1 M of urea within 1.5 h Vibrio splendidus
W155F the mutant shows reduced catalytic efficiency compared to the wild type enzyme Vibrio splendidus
W274F the mutant shows reduced catalytic efficiency compared to the wild type enzyme Vibrio splendidus
W301F the mutant shows reduced catalytic efficiency compared to the wild type enzyme Vibrio splendidus
W460F the mutant shows reduced catalytic efficiency compared to the wild type enzyme Vibrio splendidus
W460I inactive Vibrio splendidus
W460Y the mutant shows reduced catalytic efficiency compared to the wild type enzyme Vibrio splendidus
W475F the mutant shows reduced catalytic efficiency compared to the wild type enzyme Vibrio splendidus
W512F the mutant shows wild type activity Vibrio splendidus

General Stability

General Stability Organism
the enzyme is irreversibly transformed into an inactive dimer by low urea concentrations Vibrio splendidus

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.194
-
4-nitrophenyl phosphate mutant enzyme W475F, at pH 9.8 and 10°C Vibrio splendidus
0.194
-
4-nitrophenyl phosphate wild type enzyme, at pH 9.8 and 10°C Vibrio splendidus
0.212
-
4-nitrophenyl phosphate mutant enzyme W512F, at pH 9.8 and 10°C Vibrio splendidus
0.218
-
4-nitrophenyl phosphate mutant enzyme F355W, at pH 9.8 and 10°C Vibrio splendidus
0.218
-
4-nitrophenyl phosphate mutant enzyme W155F, at pH 9.8 and 10°C Vibrio splendidus
0.239
-
4-nitrophenyl phosphate mutant enzyme W301F, at pH 9.8 and 10°C Vibrio splendidus
0.252
-
4-nitrophenyl phosphate mutant enzyme W460Y, at pH 9.8 and 10°C Vibrio splendidus
0.354
-
4-nitrophenyl phosphate mutant enzyme W460F, at pH 9.8 and 10°C Vibrio splendidus
0.462
-
4-nitrophenyl phosphate mutant enzyme W274F, at pH 9.8 and 10°C Vibrio splendidus

Metals/Ions

Metals/Ions Comment Organism Structure
Mg2+ contains Mg2+ Vibrio splendidus

Organism

Organism UniProt Comment Textmining
Vibrio splendidus
-
-
-

Purification (Commentary)

Purification (Comment) Organism
Strep-Tactin affinity column chromatography, and Superdex 200 gel filtration Vibrio splendidus

Storage Stability

Storage Stability Organism
25°C, 25 mM MOPS buffer, pH 8.0 with 500 mM NaCl, 2 days, no loss of activity Vibrio splendidus

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
4-nitrophenyl phosphate + H2O
-
Vibrio splendidus 4-nitrophenol + phosphate
-
?

Subunits

Subunits Comment Organism
homodimer
-
Vibrio splendidus

Temperature Stability [°C]

Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
51
-
melting temperature Vibrio splendidus

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
44
-
4-nitrophenyl phosphate mutant enzyme W475F, at pH 9.8 and 10°C Vibrio splendidus
65
-
4-nitrophenyl phosphate mutant enzyme W460F, at pH 9.8 and 10°C Vibrio splendidus
125
-
4-nitrophenyl phosphate mutant enzyme W155F, at pH 9.8 and 10°C Vibrio splendidus
137
-
4-nitrophenyl phosphate mutant enzyme W301F, at pH 9.8 and 10°C Vibrio splendidus
255
-
4-nitrophenyl phosphate mutant enzyme F355W, at pH 9.8 and 10°C Vibrio splendidus
275
-
4-nitrophenyl phosphate mutant enzyme W460Y, at pH 9.8 and 10°C Vibrio splendidus
300
-
4-nitrophenyl phosphate mutant enzyme W274F, at pH 9.8 and 10°C Vibrio splendidus
302
-
4-nitrophenyl phosphate wild type enzyme, at pH 9.8 and 10°C Vibrio splendidus
313
-
4-nitrophenyl phosphate mutant enzyme W512F, at pH 9.8 and 10°C Vibrio splendidus

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
184
-
4-nitrophenyl phosphate mutant enzyme W460F, at pH 9.8 and 10°C Vibrio splendidus
230
-
4-nitrophenyl phosphate mutant enzyme W475F, at pH 9.8 and 10°C Vibrio splendidus
573
-
4-nitrophenyl phosphate mutant enzyme W155F, at pH 9.8 and 10°C Vibrio splendidus
573
-
4-nitrophenyl phosphate mutant enzyme W301F, at pH 9.8 and 10°C Vibrio splendidus
650
-
4-nitrophenyl phosphate mutant enzyme W274F, at pH 9.8 and 10°C Vibrio splendidus
869
-
4-nitrophenyl phosphate mutant enzyme W460Y, at pH 9.8 and 10°C Vibrio splendidus
1170
-
4-nitrophenyl phosphate mutant enzyme F355W, at pH 9.8 and 10°C Vibrio splendidus
1480
-
4-nitrophenyl phosphate mutant enzyme W512F, at pH 9.8 and 10°C Vibrio splendidus
1560
-
4-nitrophenyl phosphate wild type enzyme, at pH 9.8 and 10°C Vibrio splendidus