Cloned (Comment) | Organism |
---|---|
expressed in Escherichia coli AD494 (DE3) cells | Shewanella sp. |
Crystallization (Comment) | Organism |
---|---|
hanging drop vapour diffusion method, using a reservoir solution containing 30% (w/v) polyethylene glycol 4000, 0.2 M ammonium acetate, and 0.1 M Tris-HCl (pH 8.5) | Shewanella sp. |
Protein Variants | Comment | Organism |
---|---|---|
I115A | no activity | Shewanella sp. |
I115F | the mutant enzyme possesses a higher kcat at low temperatures compared to the wild type enzyme, the thermostability of the I115F mutant enzyme exhibits higher catalytic efficiency than the wild type enzyme | Shewanella sp. |
I115L | the mutant enzyme possesses a kcat at low temperatures similar to the wild type enzyme | Shewanella sp. |
I115M | the mutant enzyme possesses a higher kcat at low temperatures compared to the wild type enzyme, the thermostability of the I115M mutant enzyme exhibits higher catalytic efficiency than the wild type enzyme | Shewanella sp. |
I115V | the mutant enzyme possesses a kcat at low temperatures similar to the wild type enzyme | Shewanella sp. |
I307A | the mutant enzyme possesses a lower kcat at low temperatures compared to the wild type enzyme | Shewanella sp. |
I307F | the mutant enzyme possesses a lower kcat at low temperatures compared to the wild type enzyme | Shewanella sp. |
I307L | the mutant enzyme possesses a lower kcat at low temperatures compared to the wild type enzyme | Shewanella sp. |
I307M | the mutant enzyme possesses a lower kcat at low temperatures compared to the wild type enzyme | Shewanella sp. |
L145A | the mutant enzyme possesses a lower kcat at low temperatures compared to the wild type enzyme | Shewanella sp. |
L145F | the mutant enzyme possesses a lower kcat at low temperatures compared to the wild type enzyme | Shewanella sp. |
L145M | the mutant enzyme possesses a lower kcat at low temperatures compared to the wild type enzyme | Shewanella sp. |
L145V | the mutant enzyme possesses a lower kcat at low temperatures compared to the wild type enzyme | Shewanella sp. |
V79A | the mutant enzyme possesses a lower kcat at low temperatures compared to the wild type enzyme | Shewanella sp. |
V79I | the mutant enzyme possesses a lower kcat at low temperatures compared to the wild type enzyme | Shewanella sp. |
V79L | the mutant enzyme possesses a lower kcat at low temperatures compared to the wild type enzyme | Shewanella sp. |
V79M | no activity | Shewanella sp. |
V85A | no activity | Shewanella sp. |
V85F | the mutant enzyme possesses a lower kcat at low temperatures compared to the wild type enzyme | Shewanella sp. |
V85I | the mutant enzyme possesses a lower kcat at low temperatures compared to the wild type enzyme | Shewanella sp. |
V85L | the mutant enzyme possesses a lower kcat at low temperatures compared to the wild type enzyme | Shewanella sp. |
V85M | the mutant enzyme possesses a lower kcat at low temperatures compared to the wild type enzyme | Shewanella sp. |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.45 | - |
4-nitrophenyl phosphate | mutant enzyme I115F, in 50 mM MES-NaOH (pH 6.0), 2.5 mM MgCl2, at 5°C | Shewanella sp. | |
0.46 | - |
4-nitrophenyl phosphate | mutant enzyme I115L, in 50 mM MES-NaOH (pH 6.0), 2.5 mM MgCl2, at 5°C | Shewanella sp. | |
0.46 | - |
4-nitrophenyl phosphate | mutant enzyme I115M, in 50 mM MES-NaOH (pH 6.0), 2.5 mM MgCl2, at 5°C | Shewanella sp. | |
0.48 | - |
4-nitrophenyl phosphate | mutant enzyme V79I, in 50 mM MES-NaOH (pH 6.0), 2.5 mM MgCl2, at 5°C | Shewanella sp. | |
0.51 | - |
4-nitrophenyl phosphate | mutant enzyme L145A, in 50 mM MES-NaOH (pH 6.0), 2.5 mM MgCl2, at 5°C | Shewanella sp. | |
0.55 | - |
4-nitrophenyl phosphate | mutant enzyme V85F, in 50 mM MES-NaOH (pH 6.0), 2.5 mM MgCl2, at 5°C | Shewanella sp. | |
0.59 | - |
4-nitrophenyl phosphate | mutant enzyme L145V, in 50 mM MES-NaOH (pH 6.0), 2.5 mM MgCl2, at 5°C | Shewanella sp. | |
0.6 | - |
4-nitrophenyl phosphate | mutant enzyme I115V, in 50 mM MES-NaOH (pH 6.0), 2.5 mM MgCl2, at 5°C | Shewanella sp. | |
0.6 | - |
4-nitrophenyl phosphate | mutant enzyme V85M, in 50 mM MES-NaOH (pH 6.0), 2.5 mM MgCl2, at 5°C | Shewanella sp. | |
0.62 | - |
4-nitrophenyl phosphate | wild type enzyme, in 50 mM MES-NaOH (pH 6.0), 2.5 mM MgCl2, at 5°C | Shewanella sp. | |
0.66 | - |
4-nitrophenyl phosphate | mutant enzyme V79L, in 50 mM MES-NaOH (pH 6.0), 2.5 mM MgCl2, at 5°C | Shewanella sp. | |
0.71 | - |
4-nitrophenyl phosphate | mutant enzyme L145F, in 50 mM MES-NaOH (pH 6.0), 2.5 mM MgCl2, at 5°C | Shewanella sp. | |
0.8 | - |
4-nitrophenyl phosphate | mutant enzyme V85L, in 50 mM MES-NaOH (pH 6.0), 2.5 mM MgCl2, at 5°C | Shewanella sp. | |
0.83 | - |
4-nitrophenyl phosphate | mutant enzyme L145M, in 50 mM MES-NaOH (pH 6.0), 2.5 mM MgCl2, at 5°C | Shewanella sp. | |
2.4 | - |
4-nitrophenyl phosphate | mutant enzyme I307L, in 50 mM MES-NaOH (pH 6.0), 2.5 mM MgCl2, at 5°C | Shewanella sp. | |
4.01 | - |
4-nitrophenyl phosphate | mutant enzyme I307A, in 50 mM MES-NaOH (pH 6.0), 2.5 mM MgCl2, at 5°C | Shewanella sp. | |
5.03 | - |
4-nitrophenyl phosphate | mutant enzyme V79A, in 50 mM MES-NaOH (pH 6.0), 2.5 mM MgCl2, at 5°C | Shewanella sp. | |
6 | - |
4-nitrophenyl phosphate | mutant enzyme I307F, in 50 mM MES-NaOH (pH 6.0), 2.5 mM MgCl2, at 5°C | Shewanella sp. | |
6.3 | - |
4-nitrophenyl phosphate | mutant enzyme I307M, in 50 mM MES-NaOH (pH 6.0), 2.5 mM MgCl2, at 5°C | Shewanella sp. |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Zn2+ | contains 1.79 mol of Zn2+ per mole | Shewanella sp. |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Shewanella sp. | Q9S427 | - |
- |
Purification (Comment) | Organism |
---|---|
Q-Sepharose column chromatography, cellulofine-HAP column chromatography, and Mono-Q column chromatography | Shewanella sp. |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
4-nitrophenyl phosphate + H2O | - |
Shewanella sp. | 4-nitrophenol + phosphate | - |
? |
Synonyms | Comment | Organism |
---|---|---|
CAPTPase | - |
Shewanella sp. |
cold-active protein tyrosine phosphatase | - |
Shewanella sp. |
Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
---|---|---|---|
30 | 40 | during a 60 min incubation at 30°C, the activity of the I115M mutant enzyme gradually decreases to 62%, whereas the activity of the I115F mutant decreases to 38%, at 40°C the activities decrease to 48% for I115M and 10% for I115F, the wild type enzyme maintains 80% and 75% activity after incubation at 30°C and 40°C for 60 min | Shewanella sp. |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
3.33 | - |
4-nitrophenyl phosphate | mutant enzyme V79A, in 50 mM MES-NaOH (pH 6.0), 2.5 mM MgCl2, at 5°C | Shewanella sp. | |
7.1 | - |
4-nitrophenyl phosphate | mutant enzyme I307A, in 50 mM MES-NaOH (pH 6.0), 2.5 mM MgCl2, at 5°C | Shewanella sp. | |
11 | - |
4-nitrophenyl phosphate | mutant enzyme I307F, in 50 mM MES-NaOH (pH 6.0), 2.5 mM MgCl2, at 5°C | Shewanella sp. | |
11.4 | - |
4-nitrophenyl phosphate | mutant enzyme L145V, in 50 mM MES-NaOH (pH 6.0), 2.5 mM MgCl2, at 5°C | Shewanella sp. | |
21.9 | - |
4-nitrophenyl phosphate | mutant enzyme L145A, in 50 mM MES-NaOH (pH 6.0), 2.5 mM MgCl2, at 5°C | Shewanella sp. | |
26 | - |
4-nitrophenyl phosphate | mutant enzyme I307L, in 50 mM MES-NaOH (pH 6.0), 2.5 mM MgCl2, at 5°C | Shewanella sp. | |
36.9 | - |
4-nitrophenyl phosphate | mutant enzyme I307M, in 50 mM MES-NaOH (pH 6.0), 2.5 mM MgCl2, at 5°C | Shewanella sp. | |
38.3 | - |
4-nitrophenyl phosphate | mutant enzyme V85F, in 50 mM MES-NaOH (pH 6.0), 2.5 mM MgCl2, at 5°C | Shewanella sp. | |
40.7 | - |
4-nitrophenyl phosphate | mutant enzyme V79L, in 50 mM MES-NaOH (pH 6.0), 2.5 mM MgCl2, at 5°C | Shewanella sp. | |
62.3 | - |
4-nitrophenyl phosphate | mutant enzyme V85M, in 50 mM MES-NaOH (pH 6.0), 2.5 mM MgCl2, at 5°C | Shewanella sp. | |
66.2 | - |
4-nitrophenyl phosphate | mutant enzyme V79I, in 50 mM MES-NaOH (pH 6.0), 2.5 mM MgCl2, at 5°C | Shewanella sp. | |
129 | - |
4-nitrophenyl phosphate | mutant enzyme L145F, in 50 mM MES-NaOH (pH 6.0), 2.5 mM MgCl2, at 5°C | Shewanella sp. | |
129.7 | - |
4-nitrophenyl phosphate | mutant enzyme V85L, in 50 mM MES-NaOH (pH 6.0), 2.5 mM MgCl2, at 5°C | Shewanella sp. | |
167.1 | - |
4-nitrophenyl phosphate | mutant enzyme L145M, in 50 mM MES-NaOH (pH 6.0), 2.5 mM MgCl2, at 5°C | Shewanella sp. | |
184.1 | - |
4-nitrophenyl phosphate | mutant enzyme I115L, in 50 mM MES-NaOH (pH 6.0), 2.5 mM MgCl2, at 5°C | Shewanella sp. | |
184.8 | - |
4-nitrophenyl phosphate | mutant enzyme I115V, in 50 mM MES-NaOH (pH 6.0), 2.5 mM MgCl2, at 5°C | Shewanella sp. | |
184.9 | - |
4-nitrophenyl phosphate | wild type enzyme, in 50 mM MES-NaOH (pH 6.0), 2.5 mM MgCl2, at 5°C | Shewanella sp. | |
223.7 | - |
4-nitrophenyl phosphate | mutant enzyme I115M, in 50 mM MES-NaOH (pH 6.0), 2.5 mM MgCl2, at 5°C | Shewanella sp. | |
262 | - |
4-nitrophenyl phosphate | mutant enzyme I115F, in 50 mM MES-NaOH (pH 6.0), 2.5 mM MgCl2, at 5°C | Shewanella sp. |