Cloned (Comment) | Organism |
---|---|
gene mdh, DNA and amino acid sequence determination and analysis, phylogenetic analysis, expression of the His6-tagged enzyme in Escherichia coli strain BL21(DE3) | Pyrobaculum islandicum |
gene mdh, DNA and amino acid sequence determination and analysis, phylogenetic tree | Pyrobaculum islandicum |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
additional information | - |
additional information | activity kinetics of the native and recombinant proteins are identical | Pyrobaculum islandicum | |
0.015 | - |
oxaloacetate | pH 8.0, 80°C | Pyrobaculum islandicum | |
0.015 | - |
oxaloacetate | recombinant enzyme, pH 8.0, 80°C | Pyrobaculum islandicum | |
0.028 | - |
NAD+ | recombinant enzyme, pH 8.0, 80°C | Pyrobaculum islandicum | |
0.086 | - |
NADH | pH 8.0, 80°C | Pyrobaculum islandicum | |
0.086 | - |
NADH | recombinant enzyme, pH 8.0, 80°C | Pyrobaculum islandicum | |
0.5 | - |
L-malate | recombinant enzyme, pH 8.0, 80°C | Pyrobaculum islandicum |
Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|
33000 | - |
4 * 33000, SDS-PAGE | Pyrobaculum islandicum |
35000 | - |
4 * 35000, native enzyme, SDS-PAGE | Pyrobaculum islandicum |
150000 | - |
gel filtration | Pyrobaculum islandicum |
150000 | - |
native enzyme, gel filtration | Pyrobaculum islandicum |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
(S)-malate + NAD(P)+ | Pyrobaculum islandicum | - |
oxaloacetate + NAD(P)H | - |
r | |
(S)-malate + NAD+ | Pyrobaculum islandicum | - |
oxaloacetate + NADH + H+ | - |
r |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Pyrobaculum islandicum | - |
- |
- |
Pyrobaculum islandicum | - |
a hyperthermophilic archaeon, strain DSM 4184 | - |
Purification (Comment) | Organism |
---|---|
native enzyme 103fold by two steps of anion exchange chromatography, hydrophobic interaction chromatography, and gel filtration | Pyrobaculum islandicum |
native enzyme 103fold to homogeneity by two steps of anion exchange chromatography, hydrophobic interaction chromatography, and gel filtration, recombinant His6-tagged enzyme by nickel affinity chromatography from Escherichia coli strain BL21(DE3) | Pyrobaculum islandicum |
Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
---|---|---|---|
7 | - |
purified recombinant enzyme, oxidation reaction with oxaloacetate and NADPH | Pyrobaculum islandicum |
28 | - |
purified recombinant enzyme, reduction reaction with L-malate and NAD+ | Pyrobaculum islandicum |
310 | - |
purified native enzyme, reduction reaction with oxaloacetate and NADH | Pyrobaculum islandicum |
470 | - |
purified native enzyme | Pyrobaculum islandicum |
470 | - |
purified recombinant enzyme, reduction reaction with oxaloacetate and NADH | Pyrobaculum islandicum |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
(S)-malate + NAD(P)+ | - |
Pyrobaculum islandicum | oxaloacetate + NAD(P)H | - |
r | |
(S)-malate + NAD+ | - |
Pyrobaculum islandicum | oxaloacetate + NADH + H+ | - |
r |
Subunits | Comment | Organism |
---|---|---|
More | peptide mass fingerprinting, overview | Pyrobaculum islandicum |
tetramer | 4 * 33000, SDS-PAGE | Pyrobaculum islandicum |
tetramer | 4 * 35000, native enzyme, SDS-PAGE | Pyrobaculum islandicum |
Synonyms | Comment | Organism |
---|---|---|
malate dehydrogenase | - |
Pyrobaculum islandicum |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
80 | - |
assay at | Pyrobaculum islandicum |
92 | - |
probably above | Pyrobaculum islandicum |
Temperature Minimum [°C] | Temperature Maximum [°C] | Comment | Organism |
---|---|---|---|
40 | 92 | - |
Pyrobaculum islandicum |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
10 | - |
- |
Pyrobaculum islandicum |
pH Minimum | pH Maximum | Comment | Organism |
---|---|---|---|
5 | 11 | - |
Pyrobaculum islandicum |
5.5 | 11 | activity range | Pyrobaculum islandicum |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
additional information | no activity with malate and NADP+ | Pyrobaculum islandicum | |
NAD+ | dependent on | Pyrobaculum islandicum | |
NAD+ | preferred cofactor | Pyrobaculum islandicum | |
NADH | dependent on | Pyrobaculum islandicum | |
NADH | preferred cofactor | Pyrobaculum islandicum | |
NADP+ | 10% of the activity with NAD+ | Pyrobaculum islandicum | |
NADPH | 10% of the activity with NADH | Pyrobaculum islandicum | |
NADPH | less than 10% of maximal activity with NADH | Pyrobaculum islandicum |