Cloned (Comment) | Organism |
---|---|
expressed in Escherichia coli | Aspergillus niger |
expressed in Escherichia coli BL21(DE3) cells | Aspergillus niger |
into the vector pGEM-T Easy and subsequently into pET3c for expression in Escherichia coli BL21DE3 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 |
in complex with Mg2+, hanging drop vapour diffusion method | Aspergillus niger |
the crystal structure of DMML in complex with Mg2+ and in complex with Mg2+ and a substrate analog, the gem-diol of 3,3-difluoro-oxaloacetate, is determined | Aspergillus niger |
Protein Variants | Comment | Organism |
---|---|---|
C124A | reduced activity | Aspergillus niger |
C124A | site-directed mutant for testing the contributions made by key active-site residues | Aspergillus niger |
C124A | the mutant shows decreased kcat compared to the wild type enzyme | Aspergillus niger |
C124S | reduced activity | Aspergillus niger |
C124S | site-directed mutant for testing the contributions made by key active-site residues | Aspergillus niger |
C124S | the mutant shows decreased kcat compared to the wild type enzyme | Aspergillus niger |
D59A | reduced activity | Aspergillus niger |
D59A | site-directed mutant for testing the contributions made by key active-site residues | Aspergillus niger |
D59A | the mutant shows decreased kcat compared to the wild type enzyme | Aspergillus niger |
D59S | reduced activity | Aspergillus niger |
D59S | site-directed mutant for testing the contributions made by key active-site residues | Aspergillus niger |
D59S | the mutant shows decreased kcat compared to the wild type enzyme | Aspergillus niger |
P240S | reduced activity | Aspergillus niger |
P240S | site-directed mutant for testing the contributions made by key active-site residues | Aspergillus niger |
P240S | the mutant shows decreased kcat compared to the wild type enzyme | Aspergillus niger |
P240T | reduced activity | Aspergillus niger |
P240T | site-directed mutant for testing the contributions made by key active-site residues | Aspergillus niger |
P240T | the mutant shows decreased kcat compared to the wild type enzyme | Aspergillus niger |
Y44F | inactive | Aspergillus niger |
Y44F | site-directed mutant for testing the contributions made by key active-site residues | Aspergillus niger |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
(2R,3S)-isocitrate | - |
Aspergillus niger | |
(R)-malate | - |
Aspergillus niger | |
(S)-malate | - |
Aspergillus niger | |
3,3-difluoroxaloacetate | tight binding competitive inhibitor | Aspergillus niger | |
carboxyphosphoenolpyruvate | - |
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 C124S | Aspergillus niger | |
0.027 | - |
oxaloacetate | mutant enzyme C124S, at pH 7.5 and 25°C | Aspergillus niger | |
0.036 | - |
(2R,3S)-dimethylmalate | mutant C124A | Aspergillus niger | |
0.036 | - |
(2R,3S)-dimethylmalate | mutant enzyme C124A, at pH 7.5 and 25°C | Aspergillus niger | |
0.036 | - |
(2R,3S)-dimethylmalate | mutant enzyme C124A, in 50 mM K+-HEPES (pH 7.5 and 25°C) | Aspergillus niger | |
0.142 | - |
oxaloacetate | mutant C124A | Aspergillus niger | |
0.142 | - |
oxaloacetate | mutant enzyme C124A, at pH 7.5 and 25°C | Aspergillus niger | |
0.22 | - |
(2R,3S)-dimethylmalate | - |
Aspergillus niger | |
0.22 | - |
oxaloacetate | wild type | Aspergillus niger | |
0.22 | - |
(2R,3S)-dimethylmalate | wild type | Aspergillus niger | |
0.22 | - |
oxaloacetate | direct optical method, K+-Hepes as buffer | Aspergillus niger | |
0.22 | - |
oxaloacetate | wild type enzyme, at pH 7.5 and 25°C | Aspergillus niger | |
0.22 | - |
(2R,3S)-dimethylmalate | wild type enzyme, at pH 7.5 and 25°C | Aspergillus niger | |
0.22 | - |
(2R,3S)-dimethylmalate | wild type enzyme, in 50 mM K+-HEPES (pH 7.5 and 25°C) | Aspergillus niger | |
0.234 | - |
oxaloacetate | direct optical method, imidazole as buffer | Aspergillus niger | |
1.11 | - |
oxaloacetate | mutant enzyme P240S, at pH 7.5 and 25°C | Aspergillus niger | |
1.11 | - |
oxaloacetate | mutant P240S | Aspergillus niger | |
1.17 | - |
(2R)-ethyl-(3S)-methylmalate | - |
Aspergillus niger | |
1.17 | - |
(2R)-ethyl-(3S)-methylmalate | wild type enzyme, at pH 7.5 and 25°C | Aspergillus niger | |
1.17 | - |
(2R)-ethyl-(3S)-methylmalate | wild type enzyme, in 50 mM K+-HEPES (pH 7.5 and 25°C) | Aspergillus niger | |
1.22 | - |
oxaloacetate | mutant enzyme P240T, at pH 7.5 and 25°C | Aspergillus niger | |
1.22 | - |
oxaloacetate | mutant P240T | Aspergillus niger | |
2.2 | - |
(2R)-methylmalate | - |
Aspergillus niger | |
2.2 | - |
(2R)-methylmalate | wild type enzyme, at pH 7.5 and 25°C | Aspergillus niger | |
2.2 | - |
(2R)-methylmalate | wild type enzyme, in 50 mM K+-HEPES (pH 7.5 and 25°C) | Aspergillus niger | |
2.7 | - |
(2R)-propyl-(3S)-methylmalate | - |
Aspergillus niger | |
2.7 | - |
(2R)-propyl-(3S)-methylmalate | wild type enzyme, at pH 7.5 and 25°C | Aspergillus niger | |
2.7 | - |
(2R)-propyl-(3S)-methylmalate | wild type enzyme, in 50 mM K+-HEPES (pH 7.5 and 25°C) | Aspergillus niger | |
3.6 | - |
(2R,3S)-dimethylmalate | mutant C124S | Aspergillus niger | |
3.6 | - |
(2R,3S)-dimethylmalate | mutant enzyme C124S, at pH 7.5 and 25°C | Aspergillus niger | |
3.6 | - |
(2R,3S)-dimethylmalate | mutant enzyme C124S, in 50 mM K+-HEPES (pH 7.5 and 25°C) | Aspergillus niger | |
4.1 | - |
(2R,3S)-dimethylmalate | mutant enzyme P240T, at pH 7.5 and 25°C | Aspergillus niger | |
4.1 | - |
(2R,3S)-dimethylmalate | mutant P240T | Aspergillus niger | |
4.1 | - |
(2R,3S)-dimethylmalate | mutant enzyme P240T, in 50 mM K+-HEPES (pH 7.5 and 25°C) | Aspergillus niger | |
5.9 | - |
(2R,3S)-dimethylmalate | mutant enzyme P240S, at pH 7.5 and 25°C | Aspergillus niger | |
5.9 | - |
(2R,3S)-dimethylmalate | mutant P240S | Aspergillus niger | |
5.9 | - |
(2R,3S)-dimethylmalate | mutant enzyme P240S, in 50 mM K+-HEPES (pH 7.5 and 25°C) | Aspergillus niger | |
6 | - |
(2R,3S)-dimethylmalate | mutant D59S | Aspergillus niger | |
6 | - |
(2R,3S)-dimethylmalate | mutant enzyme D59S, at pH 7.5 and 25°C | Aspergillus niger | |
6 | - |
(2R,3S)-dimethylmalate | mutant enzyme D59S, in 50 mM K+-HEPES (pH 7.5 and 25°C) | Aspergillus niger | |
7.4 | - |
(2R,3S)-dimethylmalate | mutant D59A | Aspergillus niger | |
7.4 | - |
(2R,3S)-dimethylmalate | mutant enzyme D59A, at pH 7.5 and 25°C | Aspergillus niger | |
7.4 | - |
(2R,3S)-dimethylmalate | mutant enzyme D59A, in 50 mM K+-HEPES (pH 7.5 and 25°C) | Aspergillus niger | |
12 | - |
(2R)-ethylmalate | - |
Aspergillus niger | |
12 | - |
(2S,3R)-2-methylisocitrate | - |
Aspergillus niger | |
12 | - |
(2R,3S)-methylisocitrate | - |
Aspergillus niger | |
12 | - |
(2R,3S)-2-methylisocitrate | wild type enzyme, at pH 7.5 and 25°C | Aspergillus niger | |
12 | - |
(2R)-ethylmalate | wild type enzyme, at pH 7.5 and 25°C | Aspergillus niger | |
12 | - |
(2S,3R)-2-methylisocitrate | wild type enzyme, at pH 7.5 and 25°C | Aspergillus niger | |
12 | - |
(2R,3S)-2-methylisocitrate | wild type enzyme, in 50 mM K+-HEPES (pH 7.5 and 25°C) | Aspergillus niger | |
12 | - |
(2R)-ethylmalate | wild type enzyme, in 50 mM K+-HEPES (pH 7.5 and 25°C) | Aspergillus niger | |
12 | - |
(2S,3R)-2-methylisocitrate | wild type enzyme, in 50 mM K+-HEPES (pH 7.5 and 25°C) | Aspergillus niger |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Mg2+ | - |
Aspergillus niger | |
Mg2+ | required for activity | Aspergillus niger | |
Mg2+ | required for activity (5 mM) | Aspergillus niger | |
Mn2+ | - |
Aspergillus niger |
Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|
31986 | - |
4 * 31986, mass spectrometry | Aspergillus niger |
31990 | - |
mass spectrometry | Aspergillus niger |
31990 | - |
determined by mass spectrometry | Aspergillus niger |
32000 | - |
SDS-PAGE | Aspergillus niger |
32000 | - |
determined by SDS-PAGE | Aspergillus niger |
32117 | - |
4 * 32117, calculated from amino acid sequence | Aspergillus niger |
32120 | - |
calculated from amino acid sequence | Aspergillus niger |
32120 | - |
theoretical | Aspergillus niger |
110000 | - |
gel filtration | Aspergillus niger |
110000 | - |
determined by gel-filtration chromatography | Aspergillus niger |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
(2R,3S)-2,3-dimethylmalate | Aspergillus niger | - |
propanoate + pyruvate | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Aspergillus niger | Q2L887 | - |
- |
Purification (Comment) | Organism |
---|---|
by using a column chromatography-based protocol | Aspergillus niger |
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. |
---|---|---|---|---|---|---|
(2R)-ethyl-(3S)-methylmalate | - |
Aspergillus niger | ? | - |
? | |
(2R)-ethylmalate | - |
Aspergillus niger | ? | - |
? | |
(2R)-methylmalate | - |
Aspergillus niger | ? | - |
? | |
(2R)-propyl-(3S)-methylmalate | - |
Aspergillus niger | ? | - |
? | |
(2R,3S)-2,3-dimethylmalate | - |
Aspergillus niger | propanoate + pyruvate | - |
? | |
(2R,3S)-2-methylisocitrate | - |
Aspergillus niger | ? | - |
? | |
(2R,3S)-dimethylmalate | - |
Aspergillus niger | propanoate + pyruvate | - |
? | |
(2R,3S)-isocitrate | - |
Aspergillus niger | ? | - |
? | |
(2R,3S)-methylisocitrate | - |
Aspergillus niger | ? | - |
? | |
(2S)-methylmalate | - |
Aspergillus niger | ? | - |
? | |
(2S,3R)-2-methylisocitrate | - |
Aspergillus niger | ? | - |
? | |
(3S)-isopropylmalate | - |
Aspergillus niger | ? | - |
? | |
(R)-malate | - |
Aspergillus niger | ? | - |
? | |
(S)-malate | - |
Aspergillus niger | ? | - |
? | |
3,3-difluoroxaloacetate | - |
Aspergillus niger | ? | - |
? | |
3-butylmalate | - |
Aspergillus niger | ? | - |
? | |
carboxyphosphoenolpyruvate | - |
Aspergillus niger | ? | - |
? | |
additional information | (2R,3S)-isocitrate, (R)-malate, (S)-malate, (2S)-methylmalate, (3S)-isopropylmalate, 3-butylmalate, phosphonopyruvate, carboxyphenolpyruvate, 3,3-difluoroxaloacetate, oxalate are no substrates | Aspergillus niger | ? | - |
? | |
additional information | (2R,3S)-isocitrate, (R)-malate, (S)-malated, (2S)-methylmalate, (3S)-isopropylmalate, 3-butylmalate, phosphonopyruvate, carboxyphosphoenolpyruvate, 3,3-difluoroxaloacetate, and oxalate are no substrates | Aspergillus niger | ? | - |
? | |
additional information | DMML also possesses significant oxaloacetate acetyl hydrolase activity | Aspergillus niger | ? | - |
? | |
oxaloacetate | - |
Aspergillus niger | ? | - |
? | |
oxaloacetate | - |
Aspergillus niger | acetate + oxalate | - |
? | |
phosphonopyruvate | - |
Aspergillus niger | ? | - |
? |
Subunits | Comment | Organism |
---|---|---|
homotetramer | - |
Aspergillus niger |
homotetramer | 4 * 32000, SDS-PAGE | Aspergillus niger |
homotetramer | 4 * 31986, mass spectrometry | Aspergillus niger |
homotetramer | 4 * 32117, calculated from amino acid sequence | Aspergillus niger |
tetramer | x-ray crystallography | Aspergillus niger |
Synonyms | Comment | Organism |
---|---|---|
(2R,3S)-dimethylmalate lyase | - |
Aspergillus niger |
2,3-dimethylmalate lyase | - |
Aspergillus niger |
An07g08390 | - |
Aspergillus niger |
DMML | - |
Aspergillus niger |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
25 | - |
activity assay | Aspergillus niger |
Temperature Minimum [°C] | Temperature Maximum [°C] | Comment | Organism |
---|---|---|---|
4.5 | 7.5 | activity drops at higher pH | Aspergillus niger |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.00001 | - |
oxaloacetate | mutant Y44F, smaller than 0.00001 s-1 | Aspergillus niger | |
0.00001 | - |
(2R,3S)-dimethylmalate | mutant Y44F, smaller than 0.00001 s-1 | Aspergillus niger | |
0.00001 | - |
(S)-malate | smaller than 0.00001 s-1 | Aspergillus niger | |
0.00001 | - |
Phosphonopyruvate | smaller than 0.00001 s-1 | Aspergillus niger | |
0.00001 | - |
(R)-malate | smaller than 0.00001 s-1 | Aspergillus niger | |
0.00001 | - |
(2R,3S)-isocitrate | smaller than 0.00001 s-1 | Aspergillus niger | |
0.00001 | - |
carboxyphosphoenolpyruvate | smaller than 0.00001 s-1 | Aspergillus niger | |
0.00001 | - |
3,3-difluoroxaloacetate | smaller than 0.00001 s-1 | Aspergillus niger | |
0.00001 | - |
3-butylmalate | smaller than 0.00001 s-1 | Aspergillus niger | |
0.00001 | - |
(2S)-methylmalate | smaller than 0.00001 s-1 | Aspergillus niger | |
0.00001 | - |
(3S)-isopropylmalate | smaller than 0.00001 s-1 | Aspergillus niger | |
0.00001 | - |
(2R,3S)-dimethylmalate | mutant enzyme Y44F, in 50 mM K+-HEPES (pH 7.5 and 25°C) | Aspergillus niger | |
0.000121 | - |
(2R,3S)-dimethylmalate | mutant C124S | Aspergillus niger | |
0.000121 | - |
(2R,3S)-dimethylmalate | mutant enzyme C124S, at pH 7.5 and 25°C | Aspergillus niger | |
0.000121 | - |
(2R,3S)-dimethylmalate | mutant enzyme C124S, in 50 mM K+-HEPES (pH 7.5 and 25°C) | Aspergillus niger | |
0.00014 | - |
(2R,3S)-dimethylmalate | mutant D59S | Aspergillus niger | |
0.00014 | - |
(2R,3S)-dimethylmalate | mutant enzyme D59S, at pH 7.5 and 25°C | Aspergillus niger | |
0.00014 | - |
(2R,3S)-dimethylmalate | mutant enzyme D59S, in 50 mM K+-HEPES (pH 7.5 and 25°C) | Aspergillus niger | |
0.000178 | - |
(2R,3S)-dimethylmalate | mutant C124A | Aspergillus niger | |
0.000178 | - |
(2R,3S)-dimethylmalate | mutant enzyme C124A, at pH 7.5 and 25°C | Aspergillus niger | |
0.000178 | - |
(2R,3S)-dimethylmalate | mutant enzyme C124A, in 50 mM K+-HEPES (pH 7.5 and 25°C) | Aspergillus niger | |
0.000427 | - |
(2R,3S)-dimethylmalate | mutant D59A | Aspergillus niger | |
0.000427 | - |
(2R,3S)-dimethylmalate | mutant enzyme D59A, at pH 7.5 and 25°C | Aspergillus niger | |
0.000427 | - |
(2R,3S)-dimethylmalate | mutant enzyme D59A, in 50 mM K+-HEPES (pH 7.5 and 25°C) | Aspergillus niger | |
0.081 | - |
oxaloacetate | mutant C124S | Aspergillus niger | |
0.081 | - |
oxaloacetate | mutant enzyme C124S, at pH 7.5 and 25°C | Aspergillus niger | |
0.082 | - |
oxaloacetate | mutant enzyme P240T, at pH 7.5 and 25°C | Aspergillus niger | |
0.082 | - |
oxaloacetate | mutant P240T | Aspergillus niger | |
0.125 | - |
oxaloacetate | mutant C124A | Aspergillus niger | |
0.125 | - |
oxaloacetate | mutant enzyme C124A, at pH 7.5 and 25°C | Aspergillus niger | |
0.13 | - |
(2S,3R)-2-methylisocitrate | - |
Aspergillus niger | |
0.13 | - |
(2R,3S)-methylisocitrate | - |
Aspergillus niger | |
0.13 | - |
(2R,3S)-2-methylisocitrate | wild type enzyme, at pH 7.5 and 25°C | Aspergillus niger | |
0.13 | - |
(2S,3R)-2-methylisocitrate | wild type enzyme, at pH 7.5 and 25°C | Aspergillus niger | |
0.13 | - |
(2R,3S)-2-methylisocitrate | wild type enzyme, in 50 mM K+-HEPES (pH 7.5 and 25°C) | Aspergillus niger | |
0.13 | - |
(2S,3R)-2-methylisocitrate | wild type enzyme, in 50 mM K+-HEPES (pH 7.5 and 25°C) | Aspergillus niger | |
0.357 | - |
oxaloacetate | mutant enzyme P240S, at pH 7.5 and 25°C | Aspergillus niger | |
0.357 | - |
oxaloacetate | mutant P240S | Aspergillus niger | |
0.48 | - |
oxaloacetate | wild type | Aspergillus niger | |
0.48 | - |
oxaloacetate | direct optical method, K+-Hepes as buffer | Aspergillus niger | |
0.48 | - |
oxaloacetate | wild type enzyme, at pH 7.5 and 25°C | Aspergillus niger | |
0.505 | - |
oxaloacetate | direct optical method, imidazole as buffer | Aspergillus niger | |
0.79 | - |
(2R)-propyl-(3S)-methylmalate | - |
Aspergillus niger | |
0.79 | - |
(2R)-propyl-(3S)-methylmalate | wild type enzyme, at pH 7.5 and 25°C | Aspergillus niger | |
0.79 | - |
(2R)-propyl-(3S)-methylmalate | wild type enzyme, in 50 mM K+-HEPES (pH 7.5 and 25°C) | Aspergillus niger | |
2.64 | - |
(2R)-methylmalate | - |
Aspergillus niger | |
2.64 | - |
(2R)-methylmalate | wild type enzyme, at pH 7.5 and 25°C | Aspergillus niger | |
2.64 | - |
(2R)-methylmalate | wild type enzyme, in 50 mM K+-HEPES (pH 7.5 and 25°C) | Aspergillus niger | |
2.8 | - |
(2R,3S)-dimethylmalate | mutant enzyme P240T, at pH 7.5 and 25°C | Aspergillus niger | |
2.8 | - |
(2R,3S)-dimethylmalate | mutant P240T | Aspergillus niger | |
2.8 | - |
(2R,3S)-dimethylmalate | mutant enzyme P240T, in 50 mM K+-HEPES (pH 7.5 and 25°C) | Aspergillus niger | |
4.4 | - |
(2R)-ethylmalate | - |
Aspergillus niger | |
4.4 | - |
(2R)-ethylmalate | wild type enzyme, at pH 7.5 and 25°C | Aspergillus niger | |
4.4 | - |
(2R)-ethylmalate | wild type enzyme, in 50 mM K+-HEPES (pH 7.5 and 25°C) | Aspergillus niger | |
11.7 | - |
(2R,3S)-dimethylmalate | mutant enzyme P240S, at pH 7.5 and 25°C | Aspergillus niger | |
11.7 | - |
(2R,3S)-dimethylmalate | mutant P240S | Aspergillus niger | |
11.7 | - |
(2R,3S)-dimethylmalate | mutant enzyme P240S, in 50 mM K+-HEPES (pH 7.5 and 25°C) | Aspergillus niger | |
19.2 | - |
(2R,3S)-dimethylmalate | - |
Aspergillus niger | |
19.2 | - |
(2R,3S)-dimethylmalate | wild type | Aspergillus niger | |
19.2 | - |
(2R,3S)-dimethylmalate | wild type enzyme, at pH 7.5 and 25°C | Aspergillus niger | |
19.2 | - |
(2R,3S)-dimethylmalate | wild type enzyme, in 50 mM K+-HEPES (pH 7.5 and 25°C) | Aspergillus niger | |
63 | - |
(2R)-ethyl-(3S)-methylmalate | - |
Aspergillus niger | |
63 | - |
(2R)-ethyl-(3S)-methylmalate | wild type enzyme, at pH 7.5 and 25°C | Aspergillus niger | |
63 | - |
(2R)-ethyl-(3S)-methylmalate | wild type enzyme, in 50 mM K+-HEPES (pH 7.5 and 25°C) | Aspergillus niger |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7.5 | - |
- |
Aspergillus niger |
7.5 | - |
activity assay | 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 | - |
Aspergillus niger | |
0.0024 | - |
Phosphonopyruvate | in 50 mM K+-HEPES (pH 7.5 and 25°C) | Aspergillus niger | |
0.0025 | - |
3,3-difluoroxaloacetate | - |
Aspergillus niger | |
0.0025 | - |
3,3-difluoroxaloacetate | in 50 mM K+-HEPES (pH 7.5 and 25°C) | Aspergillus niger | |
1.6 | - |
oxalate | - |
Aspergillus niger | |
1.6 | - |
oxalate | in 50 mM K+-HEPES (pH 7.5 and 25°C) | Aspergillus niger | |
2.4 | - |
Phosphonopyruvate | - |
Aspergillus niger | |
2.5 | - |
3,3-difluoroxaloacetate | - |
Aspergillus niger | |
5 | - |
(S)-malate | Ki above 5 mM | Aspergillus niger | |
5 | - |
(R)-malate | Ki above 5 mM | Aspergillus niger | |
5 | - |
(2R,3S)-isocitrate | Ki above 5 mM | Aspergillus niger | |
5 | - |
(2E)-2-(phosphonooxy)but-2-enedioic acid | Ki above 5 mM | Aspergillus niger | |
5 | - |
(S)-malate | larger than 5 mM | Aspergillus niger | |
5 | - |
(R)-malate | larger than 5 mM | Aspergillus niger | |
5 | - |
(2R,3S)-isocitrate | larger than 5 mM | Aspergillus niger | |
5 | - |
carboxyphosphoenolpyruvate | larger than 5 mM | 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 |
---|---|---|---|---|---|
0.005 | - |
(2R,3S)-dimethylmalate | mutant enzyme C124A, in 50 mM K+-HEPES (pH 7.5 and 25°C) | Aspergillus niger | |
0.01 | - |
(2R,3S)-2-methylisocitrate | wild type enzyme, in 50 mM K+-HEPES (pH 7.5 and 25°C) | Aspergillus niger | |
0.01 | - |
(2S,3R)-2-methylisocitrate | wild type enzyme, in 50 mM K+-HEPES (pH 7.5 and 25°C) | Aspergillus niger | |
0.1 | - |
(2R)-ethylmalate | wild type enzyme, in 50 mM K+-HEPES (pH 7.5 and 25°C) | Aspergillus niger | |
0.3 | - |
(2R)-propyl-(3S)-methylmalate | wild type enzyme, in 50 mM K+-HEPES (pH 7.5 and 25°C) | Aspergillus niger | |
1 | - |
(2R)-methylmalate | wild type enzyme, in 50 mM K+-HEPES (pH 7.5 and 25°C) | Aspergillus niger | |
20 | - |
(2R,3S)-dimethylmalate | mutant enzyme D59S, in 50 mM K+-HEPES (pH 7.5 and 25°C) | Aspergillus niger | |
30 | - |
(2R,3S)-dimethylmalate | mutant enzyme C124S, in 50 mM K+-HEPES (pH 7.5 and 25°C) | Aspergillus niger | |
50 | - |
(2R)-ethyl-(3S)-methylmalate | wild type enzyme, in 50 mM K+-HEPES (pH 7.5 and 25°C) | Aspergillus niger | |
60 | - |
(2R,3S)-dimethylmalate | mutant enzyme D59A, in 50 mM K+-HEPES (pH 7.5 and 25°C) | Aspergillus niger | |
90 | - |
(2R,3S)-dimethylmalate | wild type enzyme, in 50 mM K+-HEPES (pH 7.5 and 25°C) | Aspergillus niger | |
700000 | - |
(2R,3S)-dimethylmalate | mutant enzyme P240T, in 50 mM K+-HEPES (pH 7.5 and 25°C) | Aspergillus niger | |
2000000 | - |
(2R,3S)-dimethylmalate | mutant enzyme P240S, in 50 mM K+-HEPES (pH 7.5 and 25°C) | Aspergillus niger |