Cloned (Comment) | Organism |
---|---|
expression of wild-type and mutants in Escherichia coli BL21(DE3) | Zea mays |
Protein Variants | Comment | Organism |
---|---|---|
K225I | site-directed mutagenesis, mutation of a conserved residue involved in catalysis and substrate binding, mutant shows highly reduced activity and a 10fold higher partitioning ratio of oxaloacetate and malate compared to the wild-type enzyme, preference for reduction of oxaloacetate instead of decarboxylation | Zea mays |
K435L/K436L | site-directed mutagenesis, mutation of residues which are important in cofactor binding, over 6fold increased Ki for 2'-AMP, and 1.7fold decreased Ki for 5'-AMP, and increased activity with NAD+ compared to the wild-type enzyme | Zea mays |
R237L | site-directed mutagenesis, mutation of a conserved residue involved in catalysis and substrate binding, mutant shows and an over 100fold higher partitioning ratio of oxaloacetate and malate compared to the wild-type enzyme, preference for reduction of oxaloacetate instead of decarboxylation | Zea mays |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
2'-AMP | competitive | Zea mays | |
5'-AMP | competitive | Zea mays |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.008 | - |
NADP+ | wild-type enzyme, pH 8.0, 30°C | Zea mays | |
0.073 | - |
NADP+ | mutant K435L/K436L, pH 8.0, 30°C | Zea mays | |
0.123 | - |
NADP+ | mutant K225I, pH 8.0, 30°C | Zea mays | |
0.23 | - |
L-malate | wild-type enzyme, pH 8.0, 30°C | Zea mays | |
0.29 | - |
NADP+ | mutant R237L, pH 8.0, 30°C | Zea mays | |
0.31 | - |
L-malate | mutant K435L/K436L, pH 8.0, 30°C | Zea mays | |
1.9 | - |
NAD+ | mutant K435L/K436L, pH 8.0, 30°C | Zea mays | |
2.6 | - |
L-malate | mutant K225I, pH 8.0, 30°C | Zea mays | |
2.9 | - |
L-malate | mutant R237L, pH 8.0, 30°C | Zea mays | |
8.1 | - |
NAD+ | wild-type enzyme, pH 8.0, 30°C | Zea mays |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Mg2+ | - |
Zea mays |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Zea mays | - |
- |
- |
Purification (Comment) | Organism |
---|---|
recombinant wild-type and mutants from Escherichia coli to homogeneity | Zea mays |
Reaction | Comment | Organism | Reaction ID |
---|---|---|---|
(S)-malate + NADP+ = pyruvate + CO2 + NADPH + H+ | the conserved residues Arg237 and Lys225 are involved in catalysis and substrate binding as proton acceptors | Zea mays |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
(S)-malate + NAD+ | - |
Zea mays | pyruvate + CO2 + NADH | - |
? | |
(S)-malate + NADP+ | - |
Zea mays | pyruvate + CO2 + NADPH | - |
? | |
Oxaloacetate | - |
Zea mays | Pyruvate + CO2 | - |
? |
Synonyms | Comment | Organism |
---|---|---|
NADP+-dependent malic enzyme | - |
Zea mays |
NADP+-ME | - |
Zea mays |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
30 | - |
assay at | Zea mays |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.38 | - |
NADP+ | mutant R237L, pH 8.0, 30°C | Zea mays | |
1.1 | - |
NADP+ | mutant K225I, pH 8.0, 30°C | Zea mays | |
13.5 | - |
NAD+ | wild-type enzyme, pH 8.0, 30°C | Zea mays | |
18.4 | - |
NAD+ | mutant K435L/K436L, pH 8.0, 30°C | Zea mays | |
181.1 | - |
NADP+ | mutant K435L/K436L, pH 8.0, 30°C | Zea mays | |
201.3 | - |
NADP+ | wild-type enzyme, pH 8.0, 30°C | Zea mays |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
8 | - |
assay at | Zea mays |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
NAD+ | very low activity with | Zea mays | |
NADP+ | cofactor specificity is determined by Lys435/Lys436 interacting with the 2'-phosphate group of the ribose ring | Zea mays |
Ki Value [mM] | Ki Value maximum [mM] | Inhibitor | Comment | Organism | Structure |
---|---|---|---|---|---|
0.105 | - |
2'-AMP | wild-type enzyme, pH 8.0, 30°C | Zea mays | |
0.256 | - |
5'-AMP | mutant K435L/K436L, pH 8.0, 30°C | Zea mays | |
0.439 | - |
5'-AMP | wild-type enzyme, pH 8.0, 30°C | Zea mays | |
0.688 | - |
2'-AMP | mutant K435L/K436L, pH 8.0, 30°C | Zea mays |