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Literature summary for 4.1.3.32 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
Search for more literature for 4.1.3.32

Cloned(Commentary)

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 (Commentary)

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

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

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 [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

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 [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/ Products (Substrates)

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

Organism

Organism UniProt Comment Textmining
Aspergillus niger Q2L887
-
-

Purification (Commentary)

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 and Products (Substrate)

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

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

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

Temperature Range [°C]

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

Turnover Number [1/s]

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

pH Optimum Minimum pH Optimum Maximum Comment Organism
7.5
-
-
 Aspergillus niger
7.5
-
 activity assay Aspergillus niger

pH Range

pH Minimum pH Maximum Comment Organism
5 8
-
 Aspergillus niger

Ki Value [mM]

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

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

kcat/KM [mM/s]

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