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Literature summary extracted from

  • Narayanan, B.; Niu, W.; Joosten, H.J.; Li, Z.; Kuipers, R.K.; Schaap, P.J.; Dunaway-Mariano, D.; Herzberg, O.
    Structure and function of 2,3-dimethylmalate lyase, a PEP mutase/isocitrate lyase superfamily member (2009), J. Mol. Biol., 386, 486-503.
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
3.7.1.1 expressed in Escherichia coli BL21(DE3) cells Aspergillus niger
4.1.3.32 expressed in Escherichia coli Aspergillus niger
4.1.3.32 expressed in Escherichia coli BL21(DE3) cells Aspergillus niger
4.1.3.32 into the vector pGEM-T Easy and subsequently into pET3c for expression in Escherichia coli BL21DE3 cells Aspergillus niger

Crystallization (Commentary)

EC Number Crystallization (Comment) Organism
3.7.1.1 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
4.1.3.32 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
4.1.3.32 in complex with Mg2+, hanging drop vapour diffusion method Aspergillus niger
4.1.3.32 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

EC Number Protein Variants Comment Organism
3.7.1.1 C124A the mutant shows decreased kcat compared to the wild type enzyme Aspergillus niger
3.7.1.1 C124S the mutant shows decreased kcat compared to the wild type enzyme Aspergillus niger
3.7.1.1 D59A the mutant shows decreased kcat compared to the wild type enzyme Aspergillus niger
3.7.1.1 D59S the mutant shows decreased kcat compared to the wild type enzyme Aspergillus niger
3.7.1.1 P240S the mutant shows decreased kcat compared to the wild type enzyme Aspergillus niger
3.7.1.1 P240T the mutant shows decreased kcat compared to the wild type enzyme Aspergillus niger
4.1.3.32 C124A reduced activity Aspergillus niger
4.1.3.32 C124A site-directed mutant for testing the contributions made by key active-site residues Aspergillus niger
4.1.3.32 C124A the mutant shows decreased kcat compared to the wild type enzyme Aspergillus niger
4.1.3.32 C124S reduced activity Aspergillus niger
4.1.3.32 C124S site-directed mutant for testing the contributions made by key active-site residues Aspergillus niger
4.1.3.32 C124S the mutant shows decreased kcat compared to the wild type enzyme Aspergillus niger
4.1.3.32 D59A reduced activity Aspergillus niger
4.1.3.32 D59A site-directed mutant for testing the contributions made by key active-site residues Aspergillus niger
4.1.3.32 D59A the mutant shows decreased kcat compared to the wild type enzyme Aspergillus niger
4.1.3.32 D59S reduced activity Aspergillus niger
4.1.3.32 D59S site-directed mutant for testing the contributions made by key active-site residues Aspergillus niger
4.1.3.32 D59S the mutant shows decreased kcat compared to the wild type enzyme Aspergillus niger
4.1.3.32 P240S reduced activity Aspergillus niger
4.1.3.32 P240S site-directed mutant for testing the contributions made by key active-site residues Aspergillus niger
4.1.3.32 P240S the mutant shows decreased kcat compared to the wild type enzyme Aspergillus niger
4.1.3.32 P240T reduced activity Aspergillus niger
4.1.3.32 P240T site-directed mutant for testing the contributions made by key active-site residues Aspergillus niger
4.1.3.32 P240T the mutant shows decreased kcat compared to the wild type enzyme Aspergillus niger
4.1.3.32 Y44F inactive Aspergillus niger
4.1.3.32 Y44F site-directed mutant for testing the contributions made by key active-site residues Aspergillus niger

Inhibitors

EC Number Inhibitors Comment Organism Structure
3.7.1.1 3,3-difluoroxaloacetate tight binding competitive inhibitor Aspergillus niger
3.7.1.1 additional information (2R,3S)-isocitrate, (R)-malate, (S)-malate, and carboxyphosphoenolpyruvate are not inhibitors Aspergillus niger
3.7.1.1 oxalate weak binding competitive inhibitor Aspergillus niger
3.7.1.1 Phosphonopyruvate
-
Aspergillus niger
4.1.3.32 (2R,3S)-isocitrate
-
Aspergillus niger
4.1.3.32 (R)-malate
-
Aspergillus niger
4.1.3.32 (S)-malate
-
Aspergillus niger
4.1.3.32 3,3-difluoroxaloacetate tight binding competitive inhibitor Aspergillus niger
4.1.3.32 carboxyphosphoenolpyruvate
-
Aspergillus niger
4.1.3.32 additional information (2R,3S)-isocitrate, (R)-malate, (S)-malate, and carboxyphosphoenolpyruvate are not inhibitors Aspergillus niger
4.1.3.32 oxalate weak binding competitive inhibitor Aspergillus niger
4.1.3.32 Phosphonopyruvate
-
Aspergillus niger

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
3.7.1.1 0.027
-
oxaloacetate mutant enzyme C124S, in 50 mM K+-HEPES (pH 7.5 and 25°C) Aspergillus niger
3.7.1.1 0.22
-
oxaloacetate wild type enzyme, in 50 mM K+-HEPES (pH 7.5 and 25°C) Aspergillus niger
3.7.1.1 0.234
-
oxaloacetate wild type enzyme, in 0.1 M imidazole (pH 7.6 and 25°C) Aspergillus niger
3.7.1.1 1.11
-
oxaloacetate mutant enzyme P240S, in 50 mM K+-HEPES (pH 7.5 and 25°C) Aspergillus niger
3.7.1.1 1.22
-
oxaloacetate mutant enzyme P240T, in 50 mM K+-HEPES (pH 7.5 and 25°C) Aspergillus niger
3.7.1.1 20.14
-
oxaloacetate mutant enzyme C124A, in 50 mM K+-HEPES (pH 7.5 and 25°C) Aspergillus niger
4.1.3.32 0.027
-
oxaloacetate mutant C124S Aspergillus niger
4.1.3.32 0.027
-
oxaloacetate mutant enzyme C124S, at pH 7.5 and 25°C Aspergillus niger
4.1.3.32 0.036
-
(2R,3S)-dimethylmalate mutant C124A Aspergillus niger
4.1.3.32 0.036
-
(2R,3S)-dimethylmalate mutant enzyme C124A, at pH 7.5 and 25°C Aspergillus niger
4.1.3.32 0.036
-
(2R,3S)-dimethylmalate mutant enzyme C124A, in 50 mM K+-HEPES (pH 7.5 and 25°C) Aspergillus niger
4.1.3.32 0.142
-
oxaloacetate mutant C124A Aspergillus niger
4.1.3.32 0.142
-
oxaloacetate mutant enzyme C124A, at pH 7.5 and 25°C Aspergillus niger
4.1.3.32 0.22
-
(2R,3S)-dimethylmalate
-
Aspergillus niger
4.1.3.32 0.22
-
oxaloacetate wild type Aspergillus niger
4.1.3.32 0.22
-
(2R,3S)-dimethylmalate wild type Aspergillus niger
4.1.3.32 0.22
-
oxaloacetate direct optical method, K+-Hepes as buffer Aspergillus niger
4.1.3.32 0.22
-
oxaloacetate wild type enzyme, at pH 7.5 and 25°C Aspergillus niger
4.1.3.32 0.22
-
(2R,3S)-dimethylmalate wild type enzyme, at pH 7.5 and 25°C Aspergillus niger
4.1.3.32 0.22
-
(2R,3S)-dimethylmalate wild type enzyme, in 50 mM K+-HEPES (pH 7.5 and 25°C) Aspergillus niger
4.1.3.32 0.234
-
oxaloacetate direct optical method, imidazole as buffer Aspergillus niger
4.1.3.32 1.11
-
oxaloacetate mutant enzyme P240S, at pH 7.5 and 25°C Aspergillus niger
4.1.3.32 1.11
-
oxaloacetate mutant P240S Aspergillus niger
4.1.3.32 1.17
-
(2R)-ethyl-(3S)-methylmalate
-
Aspergillus niger
4.1.3.32 1.17
-
(2R)-ethyl-(3S)-methylmalate wild type enzyme, at pH 7.5 and 25°C Aspergillus niger
4.1.3.32 1.17
-
(2R)-ethyl-(3S)-methylmalate wild type enzyme, in 50 mM K+-HEPES (pH 7.5 and 25°C) Aspergillus niger
4.1.3.32 1.22
-
oxaloacetate mutant enzyme P240T, at pH 7.5 and 25°C Aspergillus niger
4.1.3.32 1.22
-
oxaloacetate mutant P240T Aspergillus niger
4.1.3.32 2.2
-
(2R)-methylmalate
-
Aspergillus niger
4.1.3.32 2.2
-
(2R)-methylmalate wild type enzyme, at pH 7.5 and 25°C Aspergillus niger
4.1.3.32 2.2
-
(2R)-methylmalate wild type enzyme, in 50 mM K+-HEPES (pH 7.5 and 25°C) Aspergillus niger
4.1.3.32 2.7
-
(2R)-propyl-(3S)-methylmalate
-
Aspergillus niger
4.1.3.32 2.7
-
(2R)-propyl-(3S)-methylmalate wild type enzyme, at pH 7.5 and 25°C Aspergillus niger
4.1.3.32 2.7
-
(2R)-propyl-(3S)-methylmalate wild type enzyme, in 50 mM K+-HEPES (pH 7.5 and 25°C) Aspergillus niger
4.1.3.32 3.6
-
(2R,3S)-dimethylmalate mutant C124S Aspergillus niger
4.1.3.32 3.6
-
(2R,3S)-dimethylmalate mutant enzyme C124S, at pH 7.5 and 25°C Aspergillus niger
4.1.3.32 3.6
-
(2R,3S)-dimethylmalate mutant enzyme C124S, in 50 mM K+-HEPES (pH 7.5 and 25°C) Aspergillus niger
4.1.3.32 4.1
-
(2R,3S)-dimethylmalate mutant enzyme P240T, at pH 7.5 and 25°C Aspergillus niger
4.1.3.32 4.1
-
(2R,3S)-dimethylmalate mutant P240T Aspergillus niger
4.1.3.32 4.1
-
(2R,3S)-dimethylmalate mutant enzyme P240T, in 50 mM K+-HEPES (pH 7.5 and 25°C) Aspergillus niger
4.1.3.32 5.9
-
(2R,3S)-dimethylmalate mutant enzyme P240S, at pH 7.5 and 25°C Aspergillus niger
4.1.3.32 5.9
-
(2R,3S)-dimethylmalate mutant P240S Aspergillus niger
4.1.3.32 5.9
-
(2R,3S)-dimethylmalate mutant enzyme P240S, in 50 mM K+-HEPES (pH 7.5 and 25°C) Aspergillus niger
4.1.3.32 6
-
(2R,3S)-dimethylmalate mutant D59S Aspergillus niger
4.1.3.32 6
-
(2R,3S)-dimethylmalate mutant enzyme D59S, at pH 7.5 and 25°C Aspergillus niger
4.1.3.32 6
-
(2R,3S)-dimethylmalate mutant enzyme D59S, in 50 mM K+-HEPES (pH 7.5 and 25°C) Aspergillus niger
4.1.3.32 7.4
-
(2R,3S)-dimethylmalate mutant D59A Aspergillus niger
4.1.3.32 7.4
-
(2R,3S)-dimethylmalate mutant enzyme D59A, at pH 7.5 and 25°C Aspergillus niger
4.1.3.32 7.4
-
(2R,3S)-dimethylmalate mutant enzyme D59A, in 50 mM K+-HEPES (pH 7.5 and 25°C) Aspergillus niger
4.1.3.32 12
-
(2R)-ethylmalate
-
Aspergillus niger
4.1.3.32 12
-
(2S,3R)-2-methylisocitrate
-
Aspergillus niger
4.1.3.32 12
-
(2R,3S)-methylisocitrate
-
Aspergillus niger
4.1.3.32 12
-
(2R,3S)-2-methylisocitrate wild type enzyme, at pH 7.5 and 25°C Aspergillus niger
4.1.3.32 12
-
(2R)-ethylmalate wild type enzyme, at pH 7.5 and 25°C Aspergillus niger
4.1.3.32 12
-
(2S,3R)-2-methylisocitrate wild type enzyme, at pH 7.5 and 25°C Aspergillus niger
4.1.3.32 12
-
(2R,3S)-2-methylisocitrate wild type enzyme, in 50 mM K+-HEPES (pH 7.5 and 25°C) Aspergillus niger
4.1.3.32 12
-
(2R)-ethylmalate wild type enzyme, in 50 mM K+-HEPES (pH 7.5 and 25°C) Aspergillus niger
4.1.3.32 12
-
(2S,3R)-2-methylisocitrate wild type enzyme, in 50 mM K+-HEPES (pH 7.5 and 25°C) Aspergillus niger

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
3.7.1.1 Mg2+ required for activity (5 mM) Aspergillus niger
4.1.3.32 Mg2+
-
Aspergillus niger
4.1.3.32 Mg2+ required for activity Aspergillus niger
4.1.3.32 Mg2+ required for activity (5 mM) Aspergillus niger
4.1.3.32 Mn2+
-
Aspergillus niger

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
3.7.1.1 31986
-
4 * 31986, mass spectrometry Aspergillus niger
3.7.1.1 32000
-
4 * 32000, SDS-PAGE Aspergillus niger
3.7.1.1 32117
-
4 * 32117, calculated from amino acid sequence Aspergillus niger
3.7.1.1 110000
-
gel filtration Aspergillus niger
4.1.3.32 31986
-
4 * 31986, mass spectrometry Aspergillus niger
4.1.3.32 31990
-
mass spectrometry Aspergillus niger
4.1.3.32 31990
-
determined by mass spectrometry Aspergillus niger
4.1.3.32 32000
-
SDS-PAGE Aspergillus niger
4.1.3.32 32000
-
determined by SDS-PAGE Aspergillus niger
4.1.3.32 32117
-
4 * 32117, calculated from amino acid sequence Aspergillus niger
4.1.3.32 32120
-
calculated from amino acid sequence Aspergillus niger
4.1.3.32 32120
-
theoretical Aspergillus niger
4.1.3.32 110000
-
gel filtration Aspergillus niger
4.1.3.32 110000
-
determined by gel-filtration chromatography Aspergillus niger

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
4.1.3.32 (2R,3S)-2,3-dimethylmalate Aspergillus niger
-
propanoate + pyruvate
-
?

Organism

EC Number Organism UniProt Comment Textmining
3.7.1.1 Aspergillus niger
-
-
-
4.1.3.32 Aspergillus niger Q2L887
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
3.7.1.1 Q-Sepharose column chromatography, ammonium sulfate precipitation, butyl Sepharose column chromatography, and phenyl Sepharose column chromatography Aspergillus niger
4.1.3.32 by using a column chromatography-based protocol Aspergillus niger
4.1.3.32 Q-Sepharose column chromatography, ammonium sulfate precipitation, butyl Sepharose column chromatography, and phenyl Sepharose column chromatography Aspergillus niger

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
3.7.1.1 oxaloacetate + H2O DMML also possesses significant oxaloacetate acetyl hydrolase activity Aspergillus niger oxalate + acetate
-
?
4.1.3.32 (2R)-ethyl-(3S)-methylmalate
-
Aspergillus niger ?
-
?
4.1.3.32 (2R)-ethylmalate
-
Aspergillus niger ?
-
?
4.1.3.32 (2R)-methylmalate
-
Aspergillus niger ?
-
?
4.1.3.32 (2R)-propyl-(3S)-methylmalate
-
Aspergillus niger ?
-
?
4.1.3.32 (2R,3S)-2,3-dimethylmalate
-
Aspergillus niger propanoate + pyruvate
-
?
4.1.3.32 (2R,3S)-2-methylisocitrate
-
Aspergillus niger ?
-
?
4.1.3.32 (2R,3S)-dimethylmalate
-
Aspergillus niger propanoate + pyruvate
-
?
4.1.3.32 (2R,3S)-isocitrate
-
Aspergillus niger ?
-
?
4.1.3.32 (2R,3S)-methylisocitrate
-
Aspergillus niger ?
-
?
4.1.3.32 (2S)-methylmalate
-
Aspergillus niger ?
-
?
4.1.3.32 (2S,3R)-2-methylisocitrate
-
Aspergillus niger ?
-
?
4.1.3.32 (3S)-isopropylmalate
-
Aspergillus niger ?
-
?
4.1.3.32 (R)-malate
-
Aspergillus niger ?
-
?
4.1.3.32 (S)-malate
-
Aspergillus niger ?
-
?
4.1.3.32 3,3-difluoroxaloacetate
-
Aspergillus niger ?
-
?
4.1.3.32 3-butylmalate
-
Aspergillus niger ?
-
?
4.1.3.32 carboxyphosphoenolpyruvate
-
Aspergillus niger ?
-
?
4.1.3.32 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 ?
-
?
4.1.3.32 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 ?
-
?
4.1.3.32 additional information DMML also possesses significant oxaloacetate acetyl hydrolase activity Aspergillus niger ?
-
?
4.1.3.32 oxaloacetate
-
Aspergillus niger ?
-
?
4.1.3.32 oxaloacetate
-
Aspergillus niger acetate + oxalate
-
?
4.1.3.32 phosphonopyruvate
-
Aspergillus niger ?
-
?

Subunits

EC Number Subunits Comment Organism
3.7.1.1 homotetramer 4 * 32000, SDS-PAGE Aspergillus niger
3.7.1.1 homotetramer 4 * 31986, mass spectrometry Aspergillus niger
3.7.1.1 homotetramer 4 * 32117, calculated from amino acid sequence Aspergillus niger
4.1.3.32 homotetramer
-
Aspergillus niger
4.1.3.32 homotetramer 4 * 32000, SDS-PAGE Aspergillus niger
4.1.3.32 homotetramer 4 * 31986, mass spectrometry Aspergillus niger
4.1.3.32 homotetramer 4 * 32117, calculated from amino acid sequence Aspergillus niger
4.1.3.32 tetramer x-ray crystallography Aspergillus niger

Synonyms

EC Number Synonyms Comment Organism
3.7.1.1 (2R,3S)-dimethylmalate lyase
-
Aspergillus niger
3.7.1.1 An07g08390
-
Aspergillus niger
3.7.1.1 DMML
-
Aspergillus niger
4.1.3.32 (2R,3S)-dimethylmalate lyase
-
Aspergillus niger
4.1.3.32 2,3-dimethylmalate lyase
-
Aspergillus niger
4.1.3.32 An07g08390
-
Aspergillus niger
4.1.3.32 DMML
-
Aspergillus niger

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
4.1.3.32 25
-
activity assay Aspergillus niger

Temperature Range [°C]

EC Number Temperature Minimum [°C] Temperature Maximum [°C] Comment Organism
4.1.3.32 4.5 7.5 activity drops at higher pH Aspergillus niger

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
3.7.1.1 0.00001
-
oxaloacetate mutant enzyme Y44F, in 50 mM K+-HEPES (pH 7.5 and 25°C) Aspergillus niger
3.7.1.1 0.081
-
oxaloacetate mutant enzyme C124S, in 50 mM K+-HEPES (pH 7.5 and 25°C) Aspergillus niger
3.7.1.1 0.082
-
oxaloacetate mutant enzyme P240T, in 50 mM K+-HEPES (pH 7.5 and 25°C) Aspergillus niger
3.7.1.1 0.125
-
oxaloacetate mutant enzyme C124A, in 50 mM K+-HEPES (pH 7.5 and 25°C) Aspergillus niger
3.7.1.1 0.357
-
oxaloacetate mutant enzyme P240S, in 50 mM K+-HEPES (pH 7.5 and 25°C) Aspergillus niger
3.7.1.1 0.48
-
oxaloacetate wild type enzyme, in 50 mM K+-HEPES (pH 7.5 and 25°C) Aspergillus niger
3.7.1.1 0.505
-
oxaloacetate wild type enzyme, in 0.1 M imidazole (pH 7.6 and 25°C) Aspergillus niger
4.1.3.32 0.00001
-
oxaloacetate mutant Y44F, smaller than 0.00001 s-1 Aspergillus niger
4.1.3.32 0.00001
-
(2R,3S)-dimethylmalate mutant Y44F, smaller than 0.00001 s-1 Aspergillus niger
4.1.3.32 0.00001
-
(S)-malate smaller than 0.00001 s-1 Aspergillus niger
4.1.3.32 0.00001
-
Phosphonopyruvate smaller than 0.00001 s-1 Aspergillus niger
4.1.3.32 0.00001
-
(R)-malate smaller than 0.00001 s-1 Aspergillus niger
4.1.3.32 0.00001
-
(2R,3S)-isocitrate smaller than 0.00001 s-1 Aspergillus niger
4.1.3.32 0.00001
-
carboxyphosphoenolpyruvate smaller than 0.00001 s-1 Aspergillus niger
4.1.3.32 0.00001
-
3,3-difluoroxaloacetate smaller than 0.00001 s-1 Aspergillus niger
4.1.3.32 0.00001
-
3-butylmalate smaller than 0.00001 s-1 Aspergillus niger
4.1.3.32 0.00001
-
(2S)-methylmalate smaller than 0.00001 s-1 Aspergillus niger
4.1.3.32 0.00001
-
(3S)-isopropylmalate smaller than 0.00001 s-1 Aspergillus niger
4.1.3.32 0.00001
-
(2R,3S)-dimethylmalate mutant enzyme Y44F, in 50 mM K+-HEPES (pH 7.5 and 25°C) Aspergillus niger
4.1.3.32 0.000121
-
(2R,3S)-dimethylmalate mutant C124S Aspergillus niger
4.1.3.32 0.000121
-
(2R,3S)-dimethylmalate mutant enzyme C124S, at pH 7.5 and 25°C Aspergillus niger
4.1.3.32 0.000121
-
(2R,3S)-dimethylmalate mutant enzyme C124S, in 50 mM K+-HEPES (pH 7.5 and 25°C) Aspergillus niger
4.1.3.32 0.00014
-
(2R,3S)-dimethylmalate mutant D59S Aspergillus niger
4.1.3.32 0.00014
-
(2R,3S)-dimethylmalate mutant enzyme D59S, at pH 7.5 and 25°C Aspergillus niger
4.1.3.32 0.00014
-
(2R,3S)-dimethylmalate mutant enzyme D59S, in 50 mM K+-HEPES (pH 7.5 and 25°C) Aspergillus niger
4.1.3.32 0.000178
-
(2R,3S)-dimethylmalate mutant C124A Aspergillus niger
4.1.3.32 0.000178
-
(2R,3S)-dimethylmalate mutant enzyme C124A, at pH 7.5 and 25°C Aspergillus niger
4.1.3.32 0.000178
-
(2R,3S)-dimethylmalate mutant enzyme C124A, in 50 mM K+-HEPES (pH 7.5 and 25°C) Aspergillus niger
4.1.3.32 0.000427
-
(2R,3S)-dimethylmalate mutant D59A Aspergillus niger
4.1.3.32 0.000427
-
(2R,3S)-dimethylmalate mutant enzyme D59A, at pH 7.5 and 25°C Aspergillus niger
4.1.3.32 0.000427
-
(2R,3S)-dimethylmalate mutant enzyme D59A, in 50 mM K+-HEPES (pH 7.5 and 25°C) Aspergillus niger
4.1.3.32 0.081
-
oxaloacetate mutant C124S Aspergillus niger
4.1.3.32 0.081
-
oxaloacetate mutant enzyme C124S, at pH 7.5 and 25°C Aspergillus niger
4.1.3.32 0.082
-
oxaloacetate mutant enzyme P240T, at pH 7.5 and 25°C Aspergillus niger
4.1.3.32 0.082
-
oxaloacetate mutant P240T Aspergillus niger
4.1.3.32 0.125
-
oxaloacetate mutant C124A Aspergillus niger
4.1.3.32 0.125
-
oxaloacetate mutant enzyme C124A, at pH 7.5 and 25°C Aspergillus niger
4.1.3.32 0.13
-
(2S,3R)-2-methylisocitrate
-
Aspergillus niger
4.1.3.32 0.13
-
(2R,3S)-methylisocitrate
-
Aspergillus niger
4.1.3.32 0.13
-
(2R,3S)-2-methylisocitrate wild type enzyme, at pH 7.5 and 25°C Aspergillus niger
4.1.3.32 0.13
-
(2S,3R)-2-methylisocitrate wild type enzyme, at pH 7.5 and 25°C Aspergillus niger
4.1.3.32 0.13
-
(2R,3S)-2-methylisocitrate wild type enzyme, in 50 mM K+-HEPES (pH 7.5 and 25°C) Aspergillus niger
4.1.3.32 0.13
-
(2S,3R)-2-methylisocitrate wild type enzyme, in 50 mM K+-HEPES (pH 7.5 and 25°C) Aspergillus niger
4.1.3.32 0.357
-
oxaloacetate mutant enzyme P240S, at pH 7.5 and 25°C Aspergillus niger
4.1.3.32 0.357
-
oxaloacetate mutant P240S Aspergillus niger
4.1.3.32 0.48
-
oxaloacetate wild type Aspergillus niger
4.1.3.32 0.48
-
oxaloacetate direct optical method, K+-Hepes as buffer Aspergillus niger
4.1.3.32 0.48
-
oxaloacetate wild type enzyme, at pH 7.5 and 25°C Aspergillus niger
4.1.3.32 0.505
-
oxaloacetate direct optical method, imidazole as buffer Aspergillus niger
4.1.3.32 0.79
-
(2R)-propyl-(3S)-methylmalate
-
Aspergillus niger
4.1.3.32 0.79
-
(2R)-propyl-(3S)-methylmalate wild type enzyme, at pH 7.5 and 25°C Aspergillus niger
4.1.3.32 0.79
-
(2R)-propyl-(3S)-methylmalate wild type enzyme, in 50 mM K+-HEPES (pH 7.5 and 25°C) Aspergillus niger
4.1.3.32 2.64
-
(2R)-methylmalate
-
Aspergillus niger
4.1.3.32 2.64
-
(2R)-methylmalate wild type enzyme, at pH 7.5 and 25°C Aspergillus niger
4.1.3.32 2.64
-
(2R)-methylmalate wild type enzyme, in 50 mM K+-HEPES (pH 7.5 and 25°C) Aspergillus niger
4.1.3.32 2.8
-
(2R,3S)-dimethylmalate mutant enzyme P240T, at pH 7.5 and 25°C Aspergillus niger
4.1.3.32 2.8
-
(2R,3S)-dimethylmalate mutant P240T Aspergillus niger
4.1.3.32 2.8
-
(2R,3S)-dimethylmalate mutant enzyme P240T, in 50 mM K+-HEPES (pH 7.5 and 25°C) Aspergillus niger
4.1.3.32 4.4
-
(2R)-ethylmalate
-
Aspergillus niger
4.1.3.32 4.4
-
(2R)-ethylmalate wild type enzyme, at pH 7.5 and 25°C Aspergillus niger
4.1.3.32 4.4
-
(2R)-ethylmalate wild type enzyme, in 50 mM K+-HEPES (pH 7.5 and 25°C) Aspergillus niger
4.1.3.32 11.7
-
(2R,3S)-dimethylmalate mutant enzyme P240S, at pH 7.5 and 25°C Aspergillus niger
4.1.3.32 11.7
-
(2R,3S)-dimethylmalate mutant P240S Aspergillus niger
4.1.3.32 11.7
-
(2R,3S)-dimethylmalate mutant enzyme P240S, in 50 mM K+-HEPES (pH 7.5 and 25°C) Aspergillus niger
4.1.3.32 19.2
-
(2R,3S)-dimethylmalate
-
Aspergillus niger
4.1.3.32 19.2
-
(2R,3S)-dimethylmalate wild type Aspergillus niger
4.1.3.32 19.2
-
(2R,3S)-dimethylmalate wild type enzyme, at pH 7.5 and 25°C Aspergillus niger
4.1.3.32 19.2
-
(2R,3S)-dimethylmalate wild type enzyme, in 50 mM K+-HEPES (pH 7.5 and 25°C) Aspergillus niger
4.1.3.32 63
-
(2R)-ethyl-(3S)-methylmalate
-
Aspergillus niger
4.1.3.32 63
-
(2R)-ethyl-(3S)-methylmalate wild type enzyme, at pH 7.5 and 25°C Aspergillus niger
4.1.3.32 63
-
(2R)-ethyl-(3S)-methylmalate wild type enzyme, in 50 mM K+-HEPES (pH 7.5 and 25°C) Aspergillus niger

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
3.7.1.1 7.5
-
-
Aspergillus niger
4.1.3.32 7.5
-
-
Aspergillus niger
4.1.3.32 7.5
-
activity assay Aspergillus niger

pH Range

EC Number pH Minimum pH Maximum Comment Organism
3.7.1.1 5 8
-
Aspergillus niger
4.1.3.32 5 8
-
Aspergillus niger

Ki Value [mM]

EC Number Ki Value [mM] Ki Value maximum [mM] Inhibitor Comment Organism Structure
3.7.1.1 0.0024
-
Phosphonopyruvate in 50 mM K+-HEPES (pH 7.5 and 25°C) Aspergillus niger
3.7.1.1 0.0025
-
3,3-difluoroxaloacetate in 50 mM K+-HEPES (pH 7.5 and 25°C) Aspergillus niger
3.7.1.1 1.6
-
oxalate in 50 mM K+-HEPES (pH 7.5 and 25°C) Aspergillus niger
4.1.3.32 0.0024
-
Phosphonopyruvate
-
Aspergillus niger
4.1.3.32 0.0024
-
Phosphonopyruvate in 50 mM K+-HEPES (pH 7.5 and 25°C) Aspergillus niger
4.1.3.32 0.0025
-
3,3-difluoroxaloacetate
-
Aspergillus niger
4.1.3.32 0.0025
-
3,3-difluoroxaloacetate in 50 mM K+-HEPES (pH 7.5 and 25°C) Aspergillus niger
4.1.3.32 1.6
-
oxalate
-
Aspergillus niger
4.1.3.32 1.6
-
oxalate in 50 mM K+-HEPES (pH 7.5 and 25°C) Aspergillus niger
4.1.3.32 2.4
-
Phosphonopyruvate
-
Aspergillus niger
4.1.3.32 2.5
-
3,3-difluoroxaloacetate
-
Aspergillus niger
4.1.3.32 5
-
(S)-malate Ki above 5 mM Aspergillus niger
4.1.3.32 5
-
(R)-malate Ki above 5 mM Aspergillus niger
4.1.3.32 5
-
(2R,3S)-isocitrate Ki above 5 mM Aspergillus niger
4.1.3.32 5
-
(2E)-2-(phosphonooxy)but-2-enedioic acid Ki above 5 mM Aspergillus niger
4.1.3.32 5
-
(S)-malate larger than 5 mM Aspergillus niger
4.1.3.32 5
-
(R)-malate larger than 5 mM Aspergillus niger
4.1.3.32 5
-
(2R,3S)-isocitrate larger than 5 mM Aspergillus niger
4.1.3.32 5
-
carboxyphosphoenolpyruvate larger than 5 mM Aspergillus niger

General Information

EC Number General Information Comment Organism
3.7.1.1 metabolism DMML is a key enzyme in bacterial nicotinate catabolism Aspergillus niger
4.1.3.32 metabolism DMML is a key enzyme in bacterial nicotinate catabolism Aspergillus niger

kcat/KM [mM/s]

EC Number kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
3.7.1.1 2
-
oxaloacetate wild type enzyme, in 0.1 M imidazole (pH 7.6 and 25°C) Aspergillus niger
3.7.1.1 2
-
oxaloacetate wild type enzyme, in 50 mM K+-HEPES (pH7.5 and 25°C) Aspergillus niger
3.7.1.1 70000
-
oxaloacetate mutant enzyme P240T, in 50 mM K+-HEPES (pH 7.5 and 25°C) Aspergillus niger
3.7.1.1 300000
-
oxaloacetate mutant enzyme P240S, in 50 mM K+-HEPES (pH 7.5 and 25°C) Aspergillus niger
3.7.1.1 900000
-
oxaloacetate mutant enzyme C124A, in 50 mM K+-HEPES (pH 7.5 and 25°C) Aspergillus niger
3.7.1.1 3000000
-
oxaloacetate mutant enzyme C124S, in 50 mM K+-HEPES (pH 7.5 and 25°C) Aspergillus niger
4.1.3.32 0.005
-
(2R,3S)-dimethylmalate mutant enzyme C124A, in 50 mM K+-HEPES (pH 7.5 and 25°C) Aspergillus niger
4.1.3.32 0.01
-
(2R,3S)-2-methylisocitrate wild type enzyme, in 50 mM K+-HEPES (pH 7.5 and 25°C) Aspergillus niger
4.1.3.32 0.01
-
(2S,3R)-2-methylisocitrate wild type enzyme, in 50 mM K+-HEPES (pH 7.5 and 25°C) Aspergillus niger
4.1.3.32 0.1
-
(2R)-ethylmalate wild type enzyme, in 50 mM K+-HEPES (pH 7.5 and 25°C) Aspergillus niger
4.1.3.32 0.3
-
(2R)-propyl-(3S)-methylmalate wild type enzyme, in 50 mM K+-HEPES (pH 7.5 and 25°C) Aspergillus niger
4.1.3.32 1
-
(2R)-methylmalate wild type enzyme, in 50 mM K+-HEPES (pH 7.5 and 25°C) Aspergillus niger
4.1.3.32 20
-
(2R,3S)-dimethylmalate mutant enzyme D59S, in 50 mM K+-HEPES (pH 7.5 and 25°C) Aspergillus niger
4.1.3.32 30
-
(2R,3S)-dimethylmalate mutant enzyme C124S, in 50 mM K+-HEPES (pH 7.5 and 25°C) Aspergillus niger
4.1.3.32 50
-
(2R)-ethyl-(3S)-methylmalate wild type enzyme, in 50 mM K+-HEPES (pH 7.5 and 25°C) Aspergillus niger
4.1.3.32 60
-
(2R,3S)-dimethylmalate mutant enzyme D59A, in 50 mM K+-HEPES (pH 7.5 and 25°C) Aspergillus niger
4.1.3.32 90
-
(2R,3S)-dimethylmalate wild type enzyme, in 50 mM K+-HEPES (pH 7.5 and 25°C) Aspergillus niger
4.1.3.32 700000
-
(2R,3S)-dimethylmalate mutant enzyme P240T, in 50 mM K+-HEPES (pH 7.5 and 25°C) Aspergillus niger
4.1.3.32 2000000
-
(2R,3S)-dimethylmalate mutant enzyme P240S, in 50 mM K+-HEPES (pH 7.5 and 25°C) Aspergillus niger