Cloned (Comment) | Organism |
---|---|
expressed in Escherichia coli BL21(DE3) cells | Aspergillus niger |
Crystallization (Comment) | Organism |
---|---|
DMML in complex with Mg2+ and in complex with Mg2+ and 3,3-difluorooxalacetate, hanging drop vapor diffusion method, using 0.2 M potassium thiocyanate, 0.1 M bis-Tris propane (pH 7.5), and 20k polyethylene glycol 3350, or using 14k polyethylene glycol 6000 and 0.1 M MES (pH 6.5) | Aspergillus niger |
Protein Variants | Comment | Organism |
---|---|---|
C124A | the mutant shows decreased kcat compared to the wild type enzyme | Aspergillus niger |
C124S | the mutant shows decreased kcat compared to the wild type enzyme | Aspergillus niger |
D59A | the mutant shows decreased kcat compared to the wild type enzyme | Aspergillus niger |
D59S | the mutant shows decreased kcat compared to the wild type enzyme | Aspergillus niger |
P240S | the mutant shows decreased kcat compared to the wild type enzyme | Aspergillus niger |
P240T | the mutant shows decreased kcat compared to the wild type enzyme | Aspergillus niger |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
3,3-difluoroxaloacetate | tight binding competitive inhibitor | Aspergillus niger | |
additional information | (2R,3S)-isocitrate, (R)-malate, (S)-malate, and carboxyphosphoenolpyruvate are not inhibitors | Aspergillus niger | |
oxalate | weak binding competitive inhibitor | Aspergillus niger | |
Phosphonopyruvate | - |
Aspergillus niger |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.027 | - |
oxaloacetate | mutant enzyme C124S, in 50 mM K+-HEPES (pH 7.5 and 25°C) | Aspergillus niger | |
0.22 | - |
oxaloacetate | wild type enzyme, in 50 mM K+-HEPES (pH 7.5 and 25°C) | Aspergillus niger | |
0.234 | - |
oxaloacetate | wild type enzyme, in 0.1 M imidazole (pH 7.6 and 25°C) | Aspergillus niger | |
1.11 | - |
oxaloacetate | mutant enzyme P240S, in 50 mM K+-HEPES (pH 7.5 and 25°C) | Aspergillus niger | |
1.22 | - |
oxaloacetate | mutant enzyme P240T, in 50 mM K+-HEPES (pH 7.5 and 25°C) | Aspergillus niger | |
20.14 | - |
oxaloacetate | mutant enzyme C124A, in 50 mM K+-HEPES (pH 7.5 and 25°C) | Aspergillus niger |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Mg2+ | required for activity (5 mM) | Aspergillus niger |
Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|
31986 | - |
4 * 31986, mass spectrometry | Aspergillus niger |
32000 | - |
4 * 32000, SDS-PAGE | Aspergillus niger |
32117 | - |
4 * 32117, calculated from amino acid sequence | Aspergillus niger |
110000 | - |
gel filtration | Aspergillus niger |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Aspergillus niger | - |
- |
- |
Purification (Comment) | Organism |
---|---|
Q-Sepharose column chromatography, ammonium sulfate precipitation, butyl Sepharose column chromatography, and phenyl Sepharose column chromatography | Aspergillus niger |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
oxaloacetate + H2O | DMML also possesses significant oxaloacetate acetyl hydrolase activity | Aspergillus niger | oxalate + acetate | - |
? |
Subunits | Comment | Organism |
---|---|---|
homotetramer | 4 * 32000, SDS-PAGE | Aspergillus niger |
homotetramer | 4 * 31986, mass spectrometry | Aspergillus niger |
homotetramer | 4 * 32117, calculated from amino acid sequence | Aspergillus niger |
Synonyms | Comment | Organism |
---|---|---|
(2R,3S)-dimethylmalate lyase | - |
Aspergillus niger |
An07g08390 | - |
Aspergillus niger |
DMML | - |
Aspergillus niger |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.00001 | - |
oxaloacetate | mutant enzyme Y44F, in 50 mM K+-HEPES (pH 7.5 and 25°C) | Aspergillus niger | |
0.081 | - |
oxaloacetate | mutant enzyme C124S, in 50 mM K+-HEPES (pH 7.5 and 25°C) | Aspergillus niger | |
0.082 | - |
oxaloacetate | mutant enzyme P240T, in 50 mM K+-HEPES (pH 7.5 and 25°C) | Aspergillus niger | |
0.125 | - |
oxaloacetate | mutant enzyme C124A, in 50 mM K+-HEPES (pH 7.5 and 25°C) | Aspergillus niger | |
0.357 | - |
oxaloacetate | mutant enzyme P240S, in 50 mM K+-HEPES (pH 7.5 and 25°C) | Aspergillus niger | |
0.48 | - |
oxaloacetate | wild type enzyme, in 50 mM K+-HEPES (pH 7.5 and 25°C) | Aspergillus niger | |
0.505 | - |
oxaloacetate | wild type enzyme, in 0.1 M imidazole (pH 7.6 and 25°C) | Aspergillus niger |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7.5 | - |
- |
Aspergillus niger |
pH Minimum | pH Maximum | Comment | Organism |
---|---|---|---|
5 | 8 | - |
Aspergillus niger |
Ki Value [mM] | Ki Value maximum [mM] | Inhibitor | Comment | Organism | Structure |
---|---|---|---|---|---|
0.0024 | - |
Phosphonopyruvate | in 50 mM K+-HEPES (pH 7.5 and 25°C) | Aspergillus niger | |
0.0025 | - |
3,3-difluoroxaloacetate | in 50 mM K+-HEPES (pH 7.5 and 25°C) | Aspergillus niger | |
1.6 | - |
oxalate | in 50 mM K+-HEPES (pH 7.5 and 25°C) | Aspergillus niger |
General Information | Comment | Organism |
---|---|---|
metabolism | DMML is a key enzyme in bacterial nicotinate catabolism | Aspergillus niger |
kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
2 | - |
oxaloacetate | wild type enzyme, in 0.1 M imidazole (pH 7.6 and 25°C) | Aspergillus niger | |
2 | - |
oxaloacetate | wild type enzyme, in 50 mM K+-HEPES (pH7.5 and 25°C) | Aspergillus niger | |
70000 | - |
oxaloacetate | mutant enzyme P240T, in 50 mM K+-HEPES (pH 7.5 and 25°C) | Aspergillus niger | |
300000 | - |
oxaloacetate | mutant enzyme P240S, in 50 mM K+-HEPES (pH 7.5 and 25°C) | Aspergillus niger | |
900000 | - |
oxaloacetate | mutant enzyme C124A, in 50 mM K+-HEPES (pH 7.5 and 25°C) | Aspergillus niger | |
3000000 | - |
oxaloacetate | mutant enzyme C124S, in 50 mM K+-HEPES (pH 7.5 and 25°C) | Aspergillus niger |