Cloned (Comment) | Organism |
---|---|
expression in Escherichia coli | Saccharolobus shibatae |
expression in Escherichia coli | Ferroplasma acidiphilum |
Crystallization (Comment) | Organism |
---|---|
protein crystals are obtained at 20°C by vapor diffusion, crystal structure is determined at 1.5 A resolution | Ferroplasma acidiphilum |
protein crystals of wild-type enzyme and mutant enzyme Q249P/G250E are obtained at 20°C by vapor diffusion, crystal structure is determined at 1.5 A resolution | Saccharolobus shibatae |
Protein Variants | Comment | Organism |
---|---|---|
A241N/Q249P/G250E/F276S | kcat and kcat/Km values for 4-nitrophenyl butyrate hydrolysis are significantly lower than the values of the wild-type enzyme. The mutant shows activations by heat treatment at either 40°C or 50°C | Saccharolobus shibatae |
N248A/P256Q/E257G/S283F | kcat and kcat/Km values for 4-nitrophenyl butyrate hydrolysis are significantly higher than the values of the wild-type enzyme. The pH-activity profile of the mutant for the hydrolysis of 4-nitrophenyl butyrate is distinct from that of the wild-type enzyme. The optimum pH for activity of the mutant (pH 8.0) is higher than that of the wild-type enzyme (pH 5.0) | Ferroplasma acidiphilum |
Q249P/G250E | kcat and kcat/Km values for 4-nitrophenyl butyrate hydrolysis are similar to the wild-type values and significantly higher than the values of the A241N/Q249P/G250E/F276S mutant enzyme | Saccharolobus shibatae |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.05 | - |
4-nitrophenyl butyrate | pH 6.0, 35°C, mutant enzyme A241N/Q249P/G250E/F276S | Saccharolobus shibatae | |
0.13 | - |
4-nitrophenyl hexanoate | pH 6.0, 35°C, mutant enzyme N248A/P256Q/E257G/S283F | Ferroplasma acidiphilum | |
0.13 | - |
4-nitrophenyl butyrate | pH 6.0, 35°C, wild-type enzyme | Saccharolobus shibatae | |
0.13 | - |
4-nitrophenyl hexanoate | pH 6.0, 35°C, wild-type enzyme | Ferroplasma acidiphilum | |
0.18 | - |
4-nitrophenyl butyrate | pH 6.0, 35°C, mutant enzyme Q249P/G250E | Saccharolobus shibatae | |
0.32 | - |
4-nitrophenyl butyrate | pH 6.0, 35°C, wild-type enzyme | Ferroplasma acidiphilum | |
0.37 | - |
4-nitrophenyl propionate | pH 6.0, 35°C, wild-type enzyme | Ferroplasma acidiphilum | |
0.48 | - |
4-nitrophenyl butyrate | pH 6.0, 35°C, mutant enzyme N248A/P256Q/E257G/S283F | Ferroplasma acidiphilum | |
0.7 | - |
4-nitrophenyl propionate | pH 6.0, 35°C, mutant enzyme N248A/P256Q/E257G/S283F | Ferroplasma acidiphilum | |
0.73 | - |
4-nitrophenyl acetate | pH 6.0, 35°C, wild-type enzyme | Ferroplasma acidiphilum | |
1.2 | - |
4-nitrophenyl acetate | pH 6.0, 35°C, mutant enzyme N248A/P256Q/E257G/S283F | Ferroplasma acidiphilum |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
additional information | the enzyme does not bind iron atoms | Ferroplasma acidiphilum |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Ferroplasma acidiphilum | Q2PCE5 | - |
- |
Saccharolobus shibatae | Q5NU42 | - |
- |
Saccharolobus shibatae DSM 5389 | Q5NU42 | - |
- |
Purification (Comment) | Organism |
---|---|
- |
Saccharolobus shibatae |
- |
Ferroplasma acidiphilum |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
4-nitrophenyl acetate + H2O | - |
Ferroplasma acidiphilum | 4-nitrophenol + acetate | - |
? | |
4-nitrophenyl butyrate + H2O | - |
Saccharolobus shibatae | 4-nitrophenol + butyrate | - |
? | |
4-nitrophenyl butyrate + H2O | - |
Ferroplasma acidiphilum | 4-nitrophenol + butyrate | - |
? | |
4-nitrophenyl butyrate + H2O | - |
Saccharolobus shibatae DSM 5389 | 4-nitrophenol + butyrate | - |
? | |
4-nitrophenyl hexanoate + H2O | - |
Ferroplasma acidiphilum | 4-nitrophenol + hexanoate | - |
? | |
4-nitrophenyl propionate + H2O | - |
Ferroplasma acidiphilum | 4-nitrophenol + propionate | - |
? |
Synonyms | Comment | Organism |
---|---|---|
EStFa | - |
Ferroplasma acidiphilum |
SshEstI | - |
Saccharolobus shibatae |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
35 | - |
assay at | Ferroplasma acidiphilum |
50 | - |
hydrolysis of 4-nitrophenol butyrate, wild-type enzyme and mutant enzyme exceeded 120 min | Ferroplasma acidiphilum |
Temperature Minimum [°C] | Temperature Maximum [°C] | Comment | Organism |
---|---|---|---|
20 | 55 | 20°C: about 40% of maximal activity, 55°C: about 55% of maximal activity, hydrolysis of 4-nitrophenol butyrate, wild-type enzyme | Ferroplasma acidiphilum |
Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
---|---|---|---|
50 | - |
the enzyme is inactivated at 50°C in biphasic kinetics where the t1/2 value of the fast phase of inactivation is less than 1 min. The half-life of mutant enzyme N248A/P256Q/E257G/S283F exceeds 120 min | Ferroplasma acidiphilum |
60 | - |
half-life of mutant enzyme A241N/Q249P/G250E/F276S is 30 min | Saccharolobus shibatae |
90 | - |
half-life of wild-type enzyme is greater than 60 min | Saccharolobus shibatae |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
2 | 8 | 4-nitrophenyl propionate | pH 6.0, 35°C, wild-type enzyme | Ferroplasma acidiphilum | |
6 | - |
4-nitrophenyl butyrate | pH 6.0, 35°C, mutant enzyme A241N/Q249P/G250E/F276S | Saccharolobus shibatae | |
11 | - |
4-nitrophenyl hexanoate | pH 6.0, 35°C, wild-type enzyme | Ferroplasma acidiphilum | |
13 | - |
4-nitrophenyl acetate | pH 6.0, 35°C, wild-type enzyme | Ferroplasma acidiphilum | |
31 | - |
4-nitrophenyl acetate | pH 6.0, 35°C, mutant enzyme N248A/P256Q/E257G/S283F | Ferroplasma acidiphilum | |
56 | - |
4-nitrophenyl butyrate | pH 6.0, 35°C, wild-type enzyme | Ferroplasma acidiphilum | |
59 | - |
4-nitrophenyl propionate | pH 6.0, 35°C, mutant enzyme N248A/P256Q/E257G/S283F | Ferroplasma acidiphilum | |
101 | - |
4-nitrophenyl butyrate | pH 6.0, 35°C, mutant enzyme N248A/P256Q/E257G/S283F | Ferroplasma acidiphilum | |
159 | - |
4-nitrophenyl hexanoate | pH 6.0, 35°C, mutant enzyme N248A/P256Q/E257G/S283F | Ferroplasma acidiphilum | |
859 | - |
4-nitrophenyl butyrate | pH 6.0, 35°C, wild-type enzyme | Saccharolobus shibatae | |
872 | - |
4-nitrophenyl butyrate | pH 6.0, 35°C, mutant enzyme Q249P/G250E | Saccharolobus shibatae |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
5 | - |
hydrolysis of 4-nitrophenol butyrate, wild-type enzyme | Ferroplasma acidiphilum |
6 | - |
assay at | Ferroplasma acidiphilum |
8 | - |
- |
Saccharolobus shibatae |
8 | - |
hydrolysis of 4-nitrophenol butyrate, mutant enzyme N248A/P256Q/E257G/S283F | Ferroplasma acidiphilum |
pH Minimum | pH Maximum | Comment | Organism |
---|---|---|---|
3 | 8 | pH 3.0: about 60% of maximal activity, pH 8.0: about 60% of maximal activity, hydrolysis of 4-nitrophenol butyrate, wild-type enzyme | Ferroplasma acidiphilum |
kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
17.8 | - |
4-nitrophenyl acetate | pH 6.0, 35°C, wild-type enzyme | Ferroplasma acidiphilum | |
25.8 | - |
4-nitrophenyl acetate | pH 6.0, 35°C, mutant enzyme N248A/P256Q/E257G/S283F | Ferroplasma acidiphilum | |
74.4 | - |
4-nitrophenyl propionate | pH 6.0, 35°C, wild-type enzyme | Ferroplasma acidiphilum | |
79.5 | - |
4-nitrophenyl hexanoate | pH 6.0, 35°C, wild-type enzyme | Ferroplasma acidiphilum | |
84.6 | - |
4-nitrophenyl propionate | pH 6.0, 35°C, mutant enzyme N248A/P256Q/E257G/S283F | Ferroplasma acidiphilum | |
119 | - |
4-nitrophenyl butyrate | pH 6.0, 35°C, mutant enzyme A241N/Q249P/G250E/F276S | Saccharolobus shibatae | |
173 | - |
4-nitrophenyl butyrate | pH 6.0, 35°C, wild-type enzyme | Ferroplasma acidiphilum | |
211 | - |
4-nitrophenyl butyrate | pH 6.0, 35°C, mutant enzyme N248A/P256Q/E257G/S283F | Ferroplasma acidiphilum | |
1275 | - |
4-nitrophenyl hexanoate | pH 6.0, 35°C, mutant enzyme N248A/P256Q/E257G/S283F | Ferroplasma acidiphilum | |
4807 | - |
4-nitrophenyl butyrate | pH 6.0, 35°C, mutant enzyme Q249P/G250E | Saccharolobus shibatae | |
6850 | - |
4-nitrophenyl butyrate | pH 6.0, 35°C, wild-type enzyme | Saccharolobus shibatae |