Cloned (Comment) | Organism |
---|---|
expressed in Escherichia coli BL21 (DE3)-T1R cells | Metallosphaera sedula |
Crystallization (Comment) | Organism |
---|---|
enzyme in complex with NAD+, hanging drop vapor diffusion method, using 35% (v/v) 2-ethoxyethanol and 0.1M sodium cacodylate/hydrochloric acid pH 6.0 at 20°C.Enzyme as ternary complex with malate and NAD+, hanging drop vapor diffusion method, using 40% (w/v) polyethylene glycol 300, 0.1M CHES/sodium hydroxide pH 9.5, and 0.2 M sodium chloride | Metallosphaera sedula |
Protein Variants | Comment | Organism |
---|---|---|
E145A | the mutant shows severely reduced activity compared to the wild type enzyme | Metallosphaera sedula |
H172A | the mutant shows severely reduced activity compared to the wild type enzyme | Metallosphaera sedula |
N120A | the mutant shows severely reduced activity compared to the wild type enzyme | Metallosphaera sedula |
R148A | the mutant shows severely reduced activity compared to the wild type enzyme | Metallosphaera sedula |
R82A | the mutant shows severely reduced activity compared to the wild type enzyme | Metallosphaera sedula |
R88A | the mutant shows severely reduced activity compared to the wild type enzyme | Metallosphaera sedula |
T218A | the mutant shows severely reduced activity compared to the wild type enzyme | Metallosphaera sedula |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.123 | - |
NADH | at pH 7.0, temperature not specified in the publication | Metallosphaera sedula | |
0.165 | - |
NADPH | at pH 7.0, temperature not specified in the publication | Metallosphaera sedula |
Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|
133000 | - |
gel filtration | Metallosphaera sedula |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
(S)-malate + NAD+ | Metallosphaera sedula | - |
oxaloacetate + NADH + H+ | - |
r | |
(S)-malate + NADP+ | Metallosphaera sedula | - |
oxaloacetate + NADPH + H+ | - |
r | |
oxaloacetate + NADH + H+ | Metallosphaera sedula | - |
(S)-malate + NAD+ | - |
r | |
oxaloacetate + NADPH + H+ | Metallosphaera sedula | - |
(S)-malate + NADP+ | - |
r |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Metallosphaera sedula | A0A088E2H7 | - |
- |
Purification (Comment) | Organism |
---|---|
Ni-NTA agarose column chromatography and Sephacryl S-300 gel filtration | Metallosphaera sedula |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
(S)-malate + NAD+ | - |
Metallosphaera sedula | oxaloacetate + NADH + H+ | - |
r | |
(S)-malate + NADP+ | - |
Metallosphaera sedula | oxaloacetate + NADPH + H+ | - |
r | |
oxaloacetate + NADH + H+ | - |
Metallosphaera sedula | (S)-malate + NAD+ | - |
r | |
oxaloacetate + NADPH + H+ | - |
Metallosphaera sedula | (S)-malate + NADP+ | - |
r |
Subunits | Comment | Organism |
---|---|---|
homotetramer | - |
Metallosphaera sedula |
Synonyms | Comment | Organism |
---|---|---|
MDH | - |
Metallosphaera sedula |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.01 | - |
NADPH | at pH 7.0, temperature not specified in the publication | Metallosphaera sedula | |
0.02 | - |
NADH | at pH 7.0, temperature not specified in the publication | Metallosphaera sedula |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
NAD+ | the enzyme has a slightly higher affinity for NADH than for NADPH but can use both as cofactor | Metallosphaera sedula | |
NADP+ | the enzyme has a slightly higher affinity for NADH than for NADPH but can use both as cofactor | Metallosphaera sedula |