Cloned (Comment) | Organism |
---|---|
expressed in Escherichia coli as His-tagged fusion protein | Acetivibrio thermocellus |
Protein Variants | Comment | Organism |
---|---|---|
D236A | decreased activity compared to the wild type enzyme | Acetivibrio thermocellus |
D236E | decreased activity compared to the wild type enzyme | Acetivibrio thermocellus |
D236N | similar activity compared to the wild type enzyme | Acetivibrio thermocellus |
D392E | decreased activity compared to the wild type enzyme | Acetivibrio thermocellus |
D392N | decreased activity compared to the wild type enzyme | Acetivibrio thermocellus |
H189A | decreased activity compared to the wild type enzyme | Acetivibrio thermocellus |
H189D | decreased activity compared to the wild type enzyme | Acetivibrio thermocellus |
H189E | decreased activity compared to the wild type enzyme | Acetivibrio thermocellus |
H189Q | decreased activity compared to the wild type enzyme | Acetivibrio thermocellus |
H264A | decreased activity compared to the wild type enzyme | Acetivibrio thermocellus |
H264N | increased activity compared to the wild type enzyme | Acetivibrio thermocellus |
H264Q | decreased activity compared to the wild type enzyme | Acetivibrio thermocellus |
H376D | decreased activity compared to the wild type enzyme | Acetivibrio thermocellus |
H376N | decreased activity compared to the wild type enzyme | Acetivibrio thermocellus |
R237A | decreased activity compared to the wild type enzyme | Acetivibrio thermocellus |
R237K | increased activity compared to the wild type enzyme | Acetivibrio thermocellus |
R237Q | decreased activity compared to the wild type enzyme | Acetivibrio thermocellus |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
1 | - |
p-nitrophenyl phosphate | mutant enzyme H376N, at 45°C, in 50 mM Tris-HCl (pH 7.5), 0.5 mM MnCl2 | Acetivibrio thermocellus | |
1.3 | - |
p-nitrophenyl phosphate | mutant enzyme D392E, at 45°C, in 50 mM Tris-HCl (pH 7.5), 0.5 mM MnCl2 | Acetivibrio thermocellus | |
1.6 | - |
bis-p-nitrophenyl phosphate | mutant enzyme D236A, at 45°C, in 50 mM Tris-HCl (pH 8.0) 0.5 mM NiCl2 | Acetivibrio thermocellus | |
2.2 | - |
bis-p-nitrophenyl phosphate | mutant enzyme D236N, at 45°C, in 50 mM Tris-HCl (pH 8.0) 0.5 mM NiCl2 | Acetivibrio thermocellus | |
3.4 | - |
bis-p-nitrophenyl phosphate | mutant enzyme H264A, at 45°C, in 50 mM Tris-HCl (pH 8.0), 0.5 mM NiCl2 | Acetivibrio thermocellus | |
3.6 | - |
bis-p-nitrophenyl phosphate | mutant enzyme H189D, at 45°C, in 50 mM Tris-HCl (pH 8.0), 0.5 mM MnCl2 | Acetivibrio thermocellus | |
4.9 | - |
p-nitrophenyl phosphate | mutant enzyme D236A, at 45°C, in 50 mM Tris-HCl (pH 7.5), 0.5 mM MnCl2 | Acetivibrio thermocellus | |
5 | - |
p-nitrophenyl phosphate | mutant enzyme D236N, at 45°C, in 50 mM Tris-HCl (pH 7.5), 0.5 mM MnCl2 | Acetivibrio thermocellus | |
5 | - |
bis-p-nitrophenyl phosphate | mutant enzyme D236N, at 45°C, in 50 mM Tris-HCl (pH 8.0), 0.5 mM MnCl2 | Acetivibrio thermocellus | |
5 | - |
bis-p-nitrophenyl phosphate | mutant enzyme D392E, at 45°C, in 50 mM Tris-HCl (pH 8.0), 0.5 mM MnCl2 | Acetivibrio thermocellus | |
5 | - |
bis-p-nitrophenyl phosphate | mutant enzyme H376D, at 45°C, in 50 mM Tris-HCl (pH 8.0) 0.5 mM MnCl2 | Acetivibrio thermocellus | |
6.3 | - |
bis-p-nitrophenyl phosphate | wild type enzyme, at 45°C, in 50 mM Tris-HCl (pH 8.0), 0.5 mM NiCl2 | Acetivibrio thermocellus | |
6.6 | - |
p-nitrophenyl phosphate | mutant enzyme D236E, at 45°C, in 50 mM Tris-HCl (pH 7.5), 0.5 mM MnCl2 | Acetivibrio thermocellus | |
6.6 | - |
bis-p-nitrophenyl phosphate | mutant enzyme H189A, at 45°C, in 50 mM Tris-HCl (pH 8.0) 0.5 mM MnCl2 | Acetivibrio thermocellus | |
7.2 | - |
bis-p-nitrophenyl phosphate | mutant enzyme H376N, at 45°C, in 50 mM Tris-HCl (pH 8.0), 0.5 mM MnCl2 | Acetivibrio thermocellus | |
7.4 | - |
bis-p-nitrophenyl phosphate | mutant enzyme D392N, at 45°C, in 50 mM Tris-HCl (pH 8.0), 0.5 mM MnCl2 | Acetivibrio thermocellus | |
9.6 | - |
bis-p-nitrophenyl phosphate | mutant enzyme H264N, at 45°C, in 50 mM Tris-HCl (pH 8.0), 0.5 mM NiCl2 | Acetivibrio thermocellus | |
11 | - |
bis-p-nitrophenyl phosphate | mutant enzyme H264Q, at 45°C, in 50 mM Tris-HCl (pH 8.0), 0.5 mM NiCl2 | Acetivibrio thermocellus | |
12 | - |
bis-p-nitrophenyl phosphate | mutant enzyme D236A, at 45°C, in 50 mM Tris-HCl (pH 8.0), 0.5 mM MnCl2 | Acetivibrio thermocellus | |
12 | - |
bis-p-nitrophenyl phosphate | mutant enzyme R237K, at 45°C, in 50 mM Tris-HCl (pH 8.0), 0.5 mM NiCl2 | Acetivibrio thermocellus | |
12 | - |
p-nitrophenyl phosphate | wild type enzyme, at 45°C, in 50 mM Tris-HCl (pH 7.5), 0.5 mM MnCl2 | Acetivibrio thermocellus | |
13 | - |
p-nitrophenyl phosphate | mutant enzyme R237K, at 45°C, in 50 mM Tris-HCl (pH 7.5), 0.5 mM MnCl2 | Acetivibrio thermocellus | |
18 | - |
bis-p-nitrophenyl phosphate | mutant enzyme R237A, at 45°C, in 50 mM Tris-HCl (pH 8.0), 0.5 mM NiCl2 | Acetivibrio thermocellus | |
18 | - |
bis-p-nitrophenyl phosphate | mutant enzyme R237Q, at 45°C, in 50 mM Tris-HCl (pH 8.0), 0.5 mM NiCl2 | Acetivibrio thermocellus | |
24 | - |
bis-p-nitrophenyl phosphate | mutant enzyme H264A, at 45°C, in 50 mM Tris-HCl (pH 8.0), 0.5 mM MnCl2 | Acetivibrio thermocellus | |
26 | - |
bis-p-nitrophenyl phosphate | mutant enzyme H189Q, at 45°C, in 50 mM Tris-HCl (pH 8.0), 0.5 mM MnCl2 | Acetivibrio thermocellus | |
38 | - |
bis-p-nitrophenyl phosphate | mutant enzyme H264Q, at 45°C, in 50 mM Tris-HCl (pH 8.0), 0.5 mM MnCl2 | Acetivibrio thermocellus | |
58 | - |
bis-p-nitrophenyl phosphate | mutant enzyme H189E, at 45°C, in 50 mM Tris-HCl (pH 8.0), 0.5 mM MnCl2 | Acetivibrio thermocellus | |
75 | - |
p-nitrophenyl phosphate | mutant enzyme H264N, at 45°C, in 50 mM Tris-HCl (pH 7.5), 0.5 mM MnCl2 | Acetivibrio thermocellus | |
88 | - |
bis-p-nitrophenyl phosphate | wild type enzyme, at 45°C, in 50 mM Tris-HCl (pH 8.0), 0.5 mM MnCl2 | Acetivibrio thermocellus |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Mn2+ | required for hydrolysis of bis-p-nitrophenyl phosphate | Acetivibrio thermocellus | |
Ni2+ | required for hydrolysis of bis-p-nitrophenyl phosphate | Acetivibrio thermocellus |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Acetivibrio thermocellus | - |
- |
- |
Purification (Comment) | Organism |
---|---|
nickel-agarose chromatography | Acetivibrio thermocellus |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
bis-p-nitrophenyl phosphate + H2O | - |
Acetivibrio thermocellus | p-nitrophenyl phosphate + phosphate | - |
? | |
p-nitrophenyl phosphate + H2O | - |
Acetivibrio thermocellus | p-nitrophenol + phosphate | - |
? |
Synonyms | Comment | Organism |
---|---|---|
PNKP | belongs to the dinuclear metallophosphoesterase superfamily | Acetivibrio thermocellus |
polynucleotide kinase/phosphatase | multifunctional enzyme that catalyzes the phosphorylation of 5'-OH termini of DNA or RNA polynucleotides and the dephosphorylation of 3'-phosphate and 2'-phosphate ribonucleotides | Acetivibrio thermocellus |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.26 | - |
bis-p-nitrophenyl phosphate | mutant enzyme H264Q, at 45°C, in 50 mM Tris-HCl (pH 8.0), 0.5 mM MnCl2 | Acetivibrio thermocellus | |
0.52 | - |
bis-p-nitrophenyl phosphate | mutant enzyme D392N, at 45°C, in 50 mM Tris-HCl (pH 8.0), 0.5 mM MnCl2 | Acetivibrio thermocellus | |
0.92 | - |
bis-p-nitrophenyl phosphate | mutant enzyme H264Q, at 45°C, in 50 mM Tris-HCl (pH 8.0), 0.5 mM NiCl2 | Acetivibrio thermocellus | |
1.1 | - |
bis-p-nitrophenyl phosphate | mutant enzyme R237A, at 45°C, in 50 mM Tris-HCl (pH 8.0), 0.5 mM NiCl2 | Acetivibrio thermocellus | |
1.23 | - |
bis-p-nitrophenyl phosphate | mutant enzyme H264A, at 45°C, in 50 mM Tris-HCl (pH 8.0), 0.5 mM MnCl2 | Acetivibrio thermocellus | |
1.7 | - |
bis-p-nitrophenyl phosphate | wild type enzyme, at 45°C, in 50 mM Tris-HCl (pH 8.0), 0.5 mM NiCl2 | Acetivibrio thermocellus | |
1.88 | - |
p-nitrophenyl phosphate | mutant enzyme D236E, at 45°C, in 50 mM Tris-HCl (pH 7.5), 0.5 mM MnCl2 | Acetivibrio thermocellus | |
2.13 | - |
bis-p-nitrophenyl phosphate | mutant enzyme H376D, at 45°C, in 50 mM Tris-HCl (pH 8.0) 0.5 mM MnCl2 | Acetivibrio thermocellus | |
2.5 | - |
p-nitrophenyl phosphate | mutant enzyme R237K, at 45°C, in 50 mM Tris-HCl (pH 7.5), 0.5 mM MnCl2 | Acetivibrio thermocellus | |
2.53 | - |
bis-p-nitrophenyl phosphate | mutant enzyme D236A, at 45°C, in 50 mM Tris-HCl (pH 8.0) 0.5 mM NiCl2 | Acetivibrio thermocellus | |
3 | 6 | bis-p-nitrophenyl phosphate | mutant enzyme H189D, at 45°C, in 50 mM Tris-HCl (pH 8.0), 0.5 mM MnCl2 | Acetivibrio thermocellus | |
3.13 | - |
bis-p-nitrophenyl phosphate | mutant enzyme H376N, at 45°C, in 50 mM Tris-HCl (pH 8.0),0.5 mM MnCl2 | Acetivibrio thermocellus | |
3.33 | - |
bis-p-nitrophenyl phosphate | mutant enzyme H189Q, at 45°C, in 50 mM Tris-HCl (pH 8.0), 0.5 mM MnCl2 | Acetivibrio thermocellus | |
3.42 | - |
p-nitrophenyl phosphate | mutant enzyme D392E, at 45°C, in 50 mM Tris-HCl (pH 7.5), 0.5 mM MnCl2 | Acetivibrio thermocellus | |
3.63 | - |
bis-p-nitrophenyl phosphate | mutant enzyme R237K, at 45°C, in 50 mM Tris-HCl (pH 8.0), 0.5 mM NiCl2 | Acetivibrio thermocellus | |
3.72 | - |
bis-p-nitrophenyl phosphate | mutant enzyme H189A, at 45°C, in 50 mM Tris-HCl (pH 8.0), 0.5 mM MnCl2 | Acetivibrio thermocellus | |
4.05 | - |
bis-p-nitrophenyl phosphate | mutant enzyme H376N, at 45°C, in 50 mM Tris-HCl (pH 8.0),0.5 mM MnCl2 | Acetivibrio thermocellus | |
5.25 | - |
p-nitrophenyl phosphate | mutant enzyme H376N, at 45°C, in 50 mM Tris-HCl (pH 7.5), 0.5 mM MnCl2 | Acetivibrio thermocellus | |
8.8 | - |
bis-p-nitrophenyl phosphate | mutant enzyme D392E, at 45°C, in 50 mM Tris-HCl (pH 8.0), 0.5 mM MnCl2 | Acetivibrio thermocellus | |
9.33 | - |
bis-p-nitrophenyl phosphate | mutant enzyme D236N, at 45°C, in 50 mM Tris-HCl (pH 8.0) 0.5 mM NiCl2 | Acetivibrio thermocellus | |
9.58 | - |
p-nitrophenyl phosphate | mutant enzyme D236A, at 45°C, in 50 mM Tris-HCl (pH 7.5), 0.5 mM MnCl2 | Acetivibrio thermocellus | |
12 | - |
bis-p-nitrophenyl phosphate | mutant enzyme H264N, at 45°C, in 50 mM Tris-HCl (pH 8.0), 0.5 mM NiCl2 | Acetivibrio thermocellus | |
13.77 | - |
bis-p-nitrophenyl phosphate | mutant enzyme D392N, at 45°C, in 50 mM Tris-HCl (pH 8.0), 0.5 mM MnCl2 | Acetivibrio thermocellus | |
15 | - |
p-nitrophenyl phosphate | wild type enzyme, at 45°C, in 50 mM Tris-HCl (pH 7.5), 0.5 mM MnCl2 | Acetivibrio thermocellus | |
16.9 | - |
bis-p-nitrophenyl phosphate | wild type enzyme, at 45°C, in 50 mM Tris-HCl (pH 8.0), 0.5 mM MnCl2 | Acetivibrio thermocellus | |
18 | - |
bis-p-nitrophenyl phosphate | mutant enzyme H264A, at 45°C, in 50 mM Tris-HCl (pH 8.0), 0.5 mM NiCl2 | Acetivibrio thermocellus | |
19.33 | - |
bis-p-nitrophenyl phosphate | mutant enzyme R237A, at 45°C, in 50 mM Tris-HCl (pH 8.0), 0.5 mM NiCl2 | Acetivibrio thermocellus | |
23.17 | - |
bis-p-nitrophenyl phosphate | wild type enzyme, at 45°C, in 50 mM Tris-HCl (pH 8.0), 0.5 mM MnCl2 | Acetivibrio thermocellus | |
30.83 | - |
p-nitrophenyl phosphate | wild type enzyme, at 45°C, in 50 mM Tris-HCl (pH 7.5), 0.5 mM MnCl2 | Acetivibrio thermocellus | |
33 | - |
bis-p-nitrophenyl phosphate | mutant enzyme R237Q, at 45°C, in 50 mM Tris-HCl (pH 8.0), 0.5 mM NiCl2 | Acetivibrio thermocellus | |
38.33 | - |
p-nitrophenyl phosphate | mutant enzyme D236N, at 45°C, in 50 mM Tris-HCl (pH 7.5), 0.5 mM MnCl2 | Acetivibrio thermocellus | |
43.5 | - |
bis-p-nitrophenyl phosphate | mutant enzyme D236A, at 45°C, in 50 mM Tris-HCl (pH 8.0), 0.5 mM MnCl2 | Acetivibrio thermocellus | |
52.33 | - |
bis-p-nitrophenyl phosphate | mutant enzyme H264Q, at 45°C, in 50 mM Tris-HCl (pH 8.0), 0.5 mM MnCl2 | Acetivibrio thermocellus | |
65.83 | - |
bis-p-nitrophenyl phosphate | mutant enzyme D236N, at 45°C, in 50 mM Tris-HCl (pH 8.0), 0.5 mM MnCl2 | Acetivibrio thermocellus | |
117.5 | - |
bis-p-nitrophenyl phosphate | mutant enzyme H189E, at 45°C, in 50 mM Tris-HCl (pH 8.0), 0.5 mM MnCl2 | Acetivibrio thermocellus | |
146.7 | - |
p-nitrophenyl phosphate | mutant enzyme H264N, at 45°C, in 50 mM Tris-HCl (pH 7.5), 0.5 mM MnCl2 | Acetivibrio thermocellus | |
253.3 | - |
bis-p-nitrophenyl phosphate | mutant enzyme H264A, at 45°C, in 50 mM Tris-HCl (pH 8.0), 0.5 mM MnCl2 | Acetivibrio thermocellus |