Activating Compound | Comment | Organism | Structure |
---|---|---|---|
2-mercaptoethanol | 14% activation at 10 mM | Cupriavidus necator | |
DTT | 25% activation at 10 mM | Cupriavidus necator |
Cloned (Comment) | Organism |
---|---|
DNA and amino acid seuence determination and analysis, sequence comparisons, phylogenetic tree, expression of His-tagged enzyme in Escherichia coli strains JM109 and BL21(DE3) | Cupriavidus necator |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
4-chloromercuribenzonic acid | 94% inhibition at 10 mM | Cupriavidus necator | |
calcium pantothenate | slight inhibition | Cupriavidus necator | |
FeCl2 | 32% inhibition at 1 mM after 60 min | Cupriavidus necator | |
FeCl3 | 73% inhibition at 1 mM after 60 min | Cupriavidus necator | |
HgCl2 | 77% inhibition at 1 mM after 60 min | Cupriavidus necator | |
iodoacetamide | 99% inhibition at 10 mM | Cupriavidus necator | |
iodoacetic acid | 52% inhibition at 10 mM | Cupriavidus necator | |
K3[Fe(CN)6] | 25% inhibition at 10 mM | Cupriavidus necator | |
additional information | poor inhibition by K[Fe(CN)6], no inhibition by ethylene diamine tetraacetic acid and ethylene glycol tetraacetic acid, and by trypsin inhibitor T-9378 | Cupriavidus necator | |
N-ethylmaleimide | complete inhibition | Cupriavidus necator | |
NaN3 | 88% inhibition at 10 mM | Cupriavidus necator | |
phenazinemethosulfate | complete inhibition | Cupriavidus necator | |
phenylmethanesulfonyl fluoride | 63% inhibition at 10 mM | Cupriavidus necator | |
SnCl2 | 16% inhibition at 1 mM after 60 min | Cupriavidus necator |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.1 | - |
NAD+ | pH 10.0, 30°C, native enzyme | Cupriavidus necator | |
11.6 | - |
L-threonine | pH 10.0, 30°C, native enzyme | Cupriavidus necator |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
AlCl3 | 18% activation at 1 mM | Cupriavidus necator | |
BaCl2 | 13% activation at 1 mM | Cupriavidus necator | |
CoCl2 | 15% activation at 1 mM | Cupriavidus necator | |
CrCl3 | 10% activation at 1 mM | Cupriavidus necator | |
CsCl | 17% activation at 1 mM | Cupriavidus necator | |
CuSO4 | 15% activation at 1 mM | Cupriavidus necator | |
MgSO4 | 12% activation at 1 mM | Cupriavidus necator | |
MnSO4 | 12% activation at 1 mM | Cupriavidus necator | |
additional information | no effect at 1 mM by AgNO3, MgCl2, and 10 mM EDTA and ethylene glycol tetraacetic acid | Cupriavidus necator | |
Na2MoO4 | 13% activation at 1 mM | Cupriavidus necator | |
NiCl2 | 19% activation at 1 mM | Cupriavidus necator | |
PbCl2 | 18% activation at 1 mM | Cupriavidus necator | |
ZnSO4 | 9% activation at 1 mM | Cupriavidus necator |
Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|
34628 | - |
2 x 37200, native enzyme, SDS-PAGE, x * 34628, sequence calculation | Cupriavidus necator |
79400 | - |
native enzyme, gel filtration | Cupriavidus necator |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
L-threonine + NAD+ | Cupriavidus necator | - |
(2S)-2-amino-3-oxobutanoate + NADH + H+ | - |
? | |
L-threonine + NAD+ | Cupriavidus necator NBRC 102504 | - |
(2S)-2-amino-3-oxobutanoate + NADH + H+ | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Cupriavidus necator | E5RQ20 | - |
- |
Cupriavidus necator NBRC 102504 | E5RQ20 | - |
- |
Purification (Comment) | Organism |
---|---|
recombinant His-tagged enzyme 4.2fold from Escherichia coli strain BL21(DE3) by nickel affinity chromatography, native enzyme 75.4fold by anion exchange and hydrophobic interaction chromatography, dialysis, hydroxyapatite chromatography, followed by gel filtration | Cupriavidus necator |
Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
---|---|---|---|
42.2 | - |
purified native enzyme, pH 10.0, 30°C | Cupriavidus necator |
65 | - |
purified recombinant enzyme, pH 10.0, 30°C | Cupriavidus necator |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
DL-2-amino-3-hydroxyvalerate + NAD+ | - |
Cupriavidus necator | DL-2-amino-3-oxopentanoate + NADH + H+ | - |
? | |
DL-2-amino-3-hydroxyvalerate + NAD+ | - |
Cupriavidus necator NBRC 102504 | DL-2-amino-3-oxopentanoate + NADH + H+ | - |
? | |
L-threonine + NAD+ | - |
Cupriavidus necator | (2S)-2-amino-3-oxobutanoate + NADH + H+ | - |
? | |
L-threonine + NAD+ | - |
Cupriavidus necator NBRC 102504 | (2S)-2-amino-3-oxobutanoate + NADH + H+ | - |
? | |
additional information | L-threonine and DL-2-amino-3-hydroxyvalerate are the only substrates for ThrDH among other L-amino acids, alcohols, and amino alcohols, substrate specificity, overview | Cupriavidus necator | ? | - |
? | |
additional information | L-threonine and DL-2-amino-3-hydroxyvalerate are the only substrates for ThrDH among other L-amino acids, alcohols, and amino alcohols, substrate specificity, overview | Cupriavidus necator NBRC 102504 | ? | - |
? |
Subunits | Comment | Organism |
---|---|---|
dimer | 2 x 37200, native enzyme, SDS-PAGE, x * 34628, sequence calculation | Cupriavidus necator |
Synonyms | Comment | Organism |
---|---|---|
ThrDH | - |
Cupriavidus necator |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
75 | - |
- |
Cupriavidus necator |
Temperature Minimum [°C] | Temperature Maximum [°C] | Comment | Organism |
---|---|---|---|
25 | 80 | activity range, profile overview | Cupriavidus necator |
Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
---|---|---|---|
25 | 75 | quite stable at, rapid loss of activity above 80°C | Cupriavidus necator |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
38 | - |
NAD+ | pH 10.0, 30°C, native enzyme | Cupriavidus necator | |
43.8 | - |
L-threonine | pH 10.0, 30°C, native enzyme | Cupriavidus necator |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
10 | - |
glycine-KOH buffer | Cupriavidus necator |
pH Minimum | pH Maximum | Comment | Organism |
---|---|---|---|
6 | 12 | activity range, pH-dependent activity varies in different buffer systems, profile overview | Cupriavidus necator |
pH Stability | pH Stability Maximum | Comment | Organism |
---|---|---|---|
4 | 11 | the purified enzyme is very stable from pH 6.0 to pH 11.0 in the glycine-KOH system, the enzyme is unstable in sodium acetate buffer, pH 4.0-5.0, and Na2CO3-NaHCO3 buffer, pH 10.0-11.0 | Cupriavidus necator |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
NAD+ | dependent on | Cupriavidus necator |
General Information | Comment | Organism |
---|---|---|
evolution | the enzyme belongs to the extended short-chain alcohol dehydrogenase superfamily | Cupriavidus necator |
additional information | the enzyme possesses a glycine-rich NAD+-binding domain at the N terminus and conserved catalytic triad of YxxxK residues | Cupriavidus necator |