Activating Compound | Comment | Organism | Structure |
---|---|---|---|
L-cysteine | 20 mM used in assay conditions | Tabernaemontana divaricata |
Cloned (Comment) | Organism |
---|---|
expressed in Escherichia coli BL21(DE3) cells | Tabernaemontana divaricata |
Protein Variants | Comment | Organism |
---|---|---|
A67Y | the mutation enhances the catalytic efficiency of the enzyme about 8fold while the thermostability of the mutant enzyme remains unchanged. The specific activity against azo-casein is almost 2.4 times higher than wild type | Tabernaemontana divaricata |
S32T | the mutation enhances the catalytic efficiency of the enzyme about 8fold while the thermostability of the mutant enzyme remains unchanged | Tabernaemontana divaricata |
S32T/A67Y | the mutations enhance the catalytic efficiency of the enzyme about 8fold while the thermostability of the mutant enzyme remains unchanged | Tabernaemontana divaricata |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
E-64 | - |
Tabernaemontana divaricata |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.09687 | - |
N-benzoyl-Phe-Val-Arg-4-nitroanilide | mutant enzyme S32T/A67Y, in 50 mM Na-acetate buffer pH 5.0, at 60°C | Tabernaemontana divaricata | |
0.1273 | - |
N-benzoyl-Phe-Val-Arg-4-nitroanilide | wild type enzyme, in 50 mM Na-acetate buffer pH 5.0, at 60°C | Tabernaemontana divaricata | |
0.1499 | - |
N-benzoyl-Phe-Val-Arg-4-nitroanilide | mutant enzyme S32T, in 50 mM Na-acetate buffer pH 5.0, at 60°C | Tabernaemontana divaricata | |
0.1769 | - |
N-benzoyl-Phe-Val-Arg-4-nitroanilide | mutant enzyme A67Y, in 50 mM Na-acetate buffer pH 5.0, at 60°C | Tabernaemontana divaricata |
Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|
40000 | - |
x * 40000, SDS-PAGE | Tabernaemontana divaricata |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Tabernaemontana divaricata | P83654 | - |
- |
Purification (Comment) | Organism |
---|---|
Ni-NTA column chromatography | Tabernaemontana divaricata |
Renatured (Comment) | Organism |
---|---|
dialysis | Tabernaemontana divaricata |
Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
---|---|---|---|
30.78 | - |
wild type enzyme, with azocasein as substrate, in 50 mM Na-acetate buffer pH 5.0, at 60°C | Tabernaemontana divaricata |
63.11 | - |
mutant enzyme S32T/A67Y, with azocasein as substrate, in 50 mM Na-acetate buffer pH 5.0, at 60°C | Tabernaemontana divaricata |
73.63 | - |
mutant enzyme S32T, with azocasein as substrate, in 50 mM Na-acetate buffer pH 5.0, at 60°C | Tabernaemontana divaricata |
86.95 | - |
mutant enzyme A67Y, with azocasein as substrate, in 50 mM Na-acetate buffer pH 5.0, at 60°C | Tabernaemontana divaricata |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
azocasein + H2O | - |
Tabernaemontana divaricata | ? | - |
? | |
Gelatin + H2O | - |
Tabernaemontana divaricata | ? | - |
? | |
N-benzoyl-Phe-Val-Arg-4-nitroanilide + H2O | - |
Tabernaemontana divaricata | N-benzoyl-Phe-Val-Arg + 4-nitroaniline | - |
? |
Subunits | Comment | Organism |
---|---|---|
? | x * 40000, SDS-PAGE | Tabernaemontana divaricata |
Synonyms | Comment | Organism |
---|---|---|
Erv-C | - |
Tabernaemontana divaricata |
ervatamin-C | - |
Tabernaemontana divaricata |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
70 | - |
- |
Tabernaemontana divaricata |
Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
---|---|---|---|
40 | 80 | after 10 incubation, the enzyme shows 100% activity between 40 and 55°C, about 97% activity at 60°C, about 65% activity at 70°C, 50% activity at 72°C, about 25% activity at 75°C, and no activity at 80°C | Tabernaemontana divaricata |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.23 | - |
N-benzoyl-Phe-Val-Arg-4-nitroanilide | wild type enzyme, in 50 mM Na-acetate buffer pH 5.0, at 60°C | Tabernaemontana divaricata | |
1.43 | - |
N-benzoyl-Phe-Val-Arg-4-nitroanilide | mutant enzyme S32T/A67Y, in 50 mM Na-acetate buffer pH 5.0, at 60°C | Tabernaemontana divaricata | |
2.23 | - |
N-benzoyl-Phe-Val-Arg-4-nitroanilide | mutant enzyme S32T, in 50 mM Na-acetate buffer pH 5.0, at 60°C | Tabernaemontana divaricata | |
2.65 | - |
N-benzoyl-Phe-Val-Arg-4-nitroanilide | mutant enzyme A67Y, in 50 mM Na-acetate buffer pH 5.0, at 60°C | Tabernaemontana divaricata |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
6.5 | 7.5 | - |
Tabernaemontana divaricata |
pH Minimum | pH Maximum | Comment | Organism |
---|---|---|---|
4.5 | 8 | more than 50% activity between pH 4.5 and 8.0 | Tabernaemontana divaricata |
IC50 Value | IC50 Value Maximum | Comment | Organism | Inhibitor | Structure |
---|---|---|---|---|---|
0.0001712 | - |
mutant enzyme S32T/A67Y, in 50 mM Na-acetate buffer pH 5.0, at 60°C | Tabernaemontana divaricata | E-64 | |
0.0002137 | - |
mutant enzyme S32T, in 50 mM Na-acetate buffer pH 5.0, at 60°C | Tabernaemontana divaricata | E-64 | |
0.0002507 | - |
mutant enzyme A67Y, in 50 mM Na-acetate buffer pH 5.0, at 60°C | Tabernaemontana divaricata | E-64 | |
0.000349 | - |
wild type enzyme, in 50 mM Na-acetate buffer pH 5.0, at 60°C | Tabernaemontana divaricata | E-64 |
kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
1.8 | - |
N-benzoyl-Phe-Val-Arg-4-nitroanilide | wild type enzyme, in 50 mM Na-acetate buffer pH 5.0, at 60°C | Tabernaemontana divaricata | |
14.75 | - |
N-benzoyl-Phe-Val-Arg-4-nitroanilide | mutant enzyme A67Y, in 50 mM Na-acetate buffer pH 5.0, at 60°C | Tabernaemontana divaricata | |
14.76 | - |
N-benzoyl-Phe-Val-Arg-4-nitroanilide | mutant enzyme S32T/A67Y, in 50 mM Na-acetate buffer pH 5.0, at 60°C | Tabernaemontana divaricata | |
14.87 | - |
N-benzoyl-Phe-Val-Arg-4-nitroanilide | mutant enzyme S32T, in 50 mM Na-acetate buffer pH 5.0, at 60°C | Tabernaemontana divaricata |