Cloned (Comment) | Organism |
---|---|
expressed in Escherichia coli BL21(DE3) cells | Mycobacterium tuberculosis |
Protein Variants | Comment | Organism |
---|---|---|
G151A | the mutant shows wild type activity | Mycobacterium tuberculosis |
G151D | the mutant shows 1.5 times the activity of the wild type enzyme against N-formyl-Met-Ala-Ser and deformylates N-formyl-Met-Leu-Phe with higher efficiency than the wild type enzyme. The mutation increases deformylase thermostability (2fold higher activity at 50°C compared with wild type activity at 30°C) and completely loses its activity upon incubating with 200 mM H2O2 | Mycobacterium tuberculosis |
G49C | the mutant retains nearly 36.1% activity compared to the wild type enzyme | Mycobacterium tuberculosis |
G49P | the mutant is almost completely inactive | Mycobacterium tuberculosis |
L107E | the mutant retains less than 10% activity compared to the wild type enzyme | Mycobacterium tuberculosis |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
actinonin | complete inhibition at 0.005 mM | Mycobacterium tuberculosis |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
2.5 | - |
N-formyl-Met-Ala-Ser | mutant enzyme G151D, in 20 mM phosphate buffer, pH 7.4, temperature not specified in the publication | Mycobacterium tuberculosis | |
4.3 | - |
N-formyl-Met-Ala-Ser | wild type enzyme, in 20 mM phosphate buffer, pH 7.4, temperature not specified in the publication | Mycobacterium tuberculosis | |
4.4 | - |
N-formyl-Met-Ala-Ser | mutant enzyme G151A, in 20 mM phosphate buffer, pH 7.4, temperature not specified in the publication | Mycobacterium tuberculosis | |
7.8 | - |
N-formyl-Met-Ala-Ser | mutant enzyme G49C, in 20 mM phosphate buffer, pH 7.4, temperature not specified in the publication | Mycobacterium tuberculosis |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Iron | iron-containing enzyme | Mycobacterium tuberculosis |
Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|
22500 | - |
x * 22500, calculated from amino acid sequence | Mycobacterium tuberculosis |
29000 | - |
x * 29000, SDS-PAGE | Mycobacterium tuberculosis |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Mycobacterium tuberculosis | - |
- |
- |
Mycobacterium tuberculosis H37Rv | - |
- |
- |
Oxidation Stability | Organism |
---|---|
the presence of 500 mM H2O2 for 30 min does not affect the deformylase wild type activity | Mycobacterium tuberculosis |
Purification (Comment) | Organism |
---|---|
Ni-NTA column chromatography, gel filtration | Mycobacterium tuberculosis |
Renatured (Comment) | Organism |
---|---|
refolding by dialysing against 20 mM phosphate buffer, pH 7.4 | Mycobacterium tuberculosis |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
N-formyl-L-methionine + H2O | worst substrate | Mycobacterium tuberculosis | formate + L-methionine | - |
? | |
N-formyl-L-methionine + H2O | worst substrate | Mycobacterium tuberculosis H37Rv | formate + L-methionine | - |
? | |
N-formyl-Met-Ala-Ser + H2O | best substrate | Mycobacterium tuberculosis | formate + Met-Ala-Ser | - |
? | |
N-formyl-Met-Ala-Ser + H2O | best substrate | Mycobacterium tuberculosis H37Rv | formate + Met-Ala-Ser | - |
? | |
N-formyl-Met-Leu-Phe + H2O | - |
Mycobacterium tuberculosis | formate + Met-Leu-Phe | - |
? | |
N-formyl-Met-Leu-Phe + H2O | - |
Mycobacterium tuberculosis H37Rv | formate + Met-Leu-Phe | - |
? |
Subunits | Comment | Organism |
---|---|---|
? | x * 29000, SDS-PAGE | Mycobacterium tuberculosis |
? | x * 22500, calculated from amino acid sequence | Mycobacterium tuberculosis |
Synonyms | Comment | Organism |
---|---|---|
- |
Mycobacterium tuberculosis |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
20 | 30 | - |
Mycobacterium tuberculosis |
Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
---|---|---|---|
30 | 50 | the wild type enzyme is stable at 30°C with a half-life close to 4.5 h. At 40°C the half-life is reduced to 90 min and at 50°C to 40 min | Mycobacterium tuberculosis |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
2 | - |
N-formyl-Met-Ala-Ser | mutant enzyme G49C, in 20 mM phosphate buffer, pH 7.4, temperature not specified in the publication | Mycobacterium tuberculosis | |
3.5 | - |
N-formyl-Met-Ala-Ser | mutant enzyme G151A, in 20 mM phosphate buffer, pH 7.4, temperature not specified in the publication | Mycobacterium tuberculosis | |
3.6 | - |
N-formyl-Met-Ala-Ser | wild type enzyme, in 20 mM phosphate buffer, pH 7.4, temperature not specified in the publication | Mycobacterium tuberculosis | |
4.5 | - |
N-formyl-Met-Ala-Ser | mutant enzyme G151D, in 20 mM phosphate buffer, pH 7.4, temperature not specified in the publication | Mycobacterium tuberculosis |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7.4 | - |
- |
Mycobacterium tuberculosis |
IC50 Value | IC50 Value Maximum | Comment | Organism | Inhibitor | Structure |
---|---|---|---|---|---|
0.00012 | - |
wild type enzyme, in 20 mM phosphate buffer, pH 7.4, temperature not specified in the publication | Mycobacterium tuberculosis | actinonin | |
0.0008 | - |
mutant enzyme G151D, in 20 mM phosphate buffer, pH 7.4, temperature not specified in the publication | Mycobacterium tuberculosis | actinonin |
kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.26 | - |
N-formyl-Met-Ala-Ser | mutant enzyme G49C, in 20 mM phosphate buffer, pH 7.4, temperature not specified in the publication | Mycobacterium tuberculosis | |
0.795 | - |
N-formyl-Met-Ala-Ser | mutant enzyme G151A, in 20 mM phosphate buffer, pH 7.4, temperature not specified in the publication | Mycobacterium tuberculosis | |
0.842 | - |
N-formyl-Met-Ala-Ser | wild type enzyme, in 20 mM phosphate buffer, pH 7.4, temperature not specified in the publication | Mycobacterium tuberculosis | |
1.786 | - |
N-formyl-Met-Ala-Ser | mutant enzyme G151D, in 20 mM phosphate buffer, pH 7.4, temperature not specified in the publication | Mycobacterium tuberculosis |