Cloned (Comment) | Organism |
---|---|
expressed in Escherichia coli BL21(DE3)pLysS cells | Mycobacterium tuberculosis |
Protein Variants | Comment | Organism |
---|---|---|
D81A | the mutant shows strongly decreased activity compared to the wild type enzyme | Mycobacterium tuberculosis |
D81E | the mutant shows strongly decreased activity compared to the wild type enzyme | Mycobacterium tuberculosis |
D81E/N321I | the mutant shows strongly decreased activity compared to the wild type enzyme | Mycobacterium tuberculosis |
N321A | the mutant shows strongly decreased activity compared to the wild type enzyme | Mycobacterium tuberculosis |
N321I | the mutant shows strongly decreased activity compared to the wild type enzyme | Mycobacterium tuberculosis |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
Mg2+ | hydrolysis activity of the wild type enzyme is inhibited by increasing concentrations of Mg2+ with IC50 values in the low millimolar range | Mycobacterium tuberculosis | |
additional information | not inhibited by EDTA | Mycobacterium tuberculosis |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.097 | - |
acetyl-CoA | mutant enzyme D81A, at pH 7.5 and 25°C | Mycobacterium tuberculosis | |
0.123 | - |
acetyl-CoA | mutant enzyme N321A, at pH 7.5 and 25°C | Mycobacterium tuberculosis | |
0.163 | - |
acetyl-CoA | mutant enzyme D81E/N321I, at pH 7.5 and 25°C | Mycobacterium tuberculosis | |
0.233 | - |
acetyl-CoA | mutant enzyme D81E, at pH 7.5 and 25°C | Mycobacterium tuberculosis | |
0.253 | - |
acetyl-CoA | wild type enzyme, at pH 7.5 and 25°C | Mycobacterium tuberculosis | |
0.262 | - |
acetyl-CoA | mutant enzyme N321I, at pH 7.5 and 25°C | Mycobacterium tuberculosis |
Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|
72000 | - |
2 * 72000, SDS-PAGE | Mycobacterium tuberculosis |
171000 | - |
gel filtration | Mycobacterium tuberculosis |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
acetyl-CoA + 3-methyl-2-oxobutanoate + H2O | Mycobacterium tuberculosis | - |
(2S)-2-isopropylmalate + CoA | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Mycobacterium tuberculosis | - |
- |
- |
Purification (Comment) | Organism |
---|---|
HisTrap column chromatography | Mycobacterium tuberculosis |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
acetyl-CoA + 3-methyl-2-oxobutanoate + H2O | - |
Mycobacterium tuberculosis | (2S)-2-isopropylmalate + CoA | - |
? |
Subunits | Comment | Organism |
---|---|---|
homodimer | 2 * 72000, SDS-PAGE | Mycobacterium tuberculosis |
Synonyms | Comment | Organism |
---|---|---|
alpha-IPMS | - |
Mycobacterium tuberculosis |
Alpha-isopropylmalate synthase | - |
Mycobacterium tuberculosis |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.05 | - |
acetyl-CoA | mutant enzyme D81A, at pH 7.5 and 25°C | Mycobacterium tuberculosis | |
0.12 | - |
acetyl-CoA | mutant enzyme D81E, at pH 7.5 and 25°C | Mycobacterium tuberculosis | |
0.13 | - |
acetyl-CoA | mutant enzyme D81E/N321I, at pH 7.5 and 25°C | Mycobacterium tuberculosis | |
0.15 | - |
acetyl-CoA | mutant enzyme N321A, at pH 7.5 and 25°C | Mycobacterium tuberculosis | |
0.15 | - |
acetyl-CoA | mutant enzyme N321I, at pH 7.5 and 25°C | Mycobacterium tuberculosis | |
0.32 | - |
acetyl-CoA | wild type enzyme, at pH 7.5 and 25°C | Mycobacterium tuberculosis |