Cloned (Comment) | Organism |
---|---|
gene ddh, DNA and amino acid sequence determination and analysis, sequence comparison, subcloning and expression in Escherichia coli strains XL1 Blue and BL21(DE3) in inclusion bodies | Haloferax mediterranei |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.031 | - |
NADPH | pH 8.5, 40°C, recombinant enzyme, substrate 2-oxoisocaproate | Haloferax mediterranei | |
0.046 | - |
NADPH | pH 8.5, 40°C, recombinant enzyme, substrate 2-oxoisoleucine | Haloferax mediterranei | |
0.054 | - |
NADPH | pH 8.5, 40°C, recombinant enzyme, substrate 2-oxobutyrate | Haloferax mediterranei | |
0.233 | - |
NADH | pH 8.5, 40°C, recombinant enzyme, substrate 2-oxoisocaproate | Haloferax mediterranei | |
0.33 | - |
NADH | pH 8.5, 40°C, recombinant enzyme, substrate 2-oxoisoleucine | Haloferax mediterranei | |
0.5 | - |
NADH | pH 8.5, 40°C, recombinant enzyme, substrate 2-oxobutyrate | Haloferax mediterranei | |
1.31 | - |
2-oxoisoleucine | pH 8.5, 40°C, recombinant enzyme, cofactor NADPH | Haloferax mediterranei | |
3.77 | - |
2-oxoisocaproate | pH 8.5, 40°C, recombinant enzyme, cofactor NADPH | Haloferax mediterranei | |
11.9 | - |
2-oxoisoleucine | pH 8.5, 40°C, recombinant enzyme, cofactor NADH | Haloferax mediterranei | |
13.45 | - |
2-oxobutyrate | pH 8.5, 40°C, recombinant enzyme, cofactor NADPH | Haloferax mediterranei | |
13.9 | - |
2-oxoisocaproate | pH 8.5, 40°C, recombinant enzyme, cofactor NADH | Haloferax mediterranei | |
21.96 | - |
pyruvate | pH 5.0, 40°C, recombinant enzyme, cofactor NADPH | Haloferax mediterranei | |
106 | - |
2-oxobutyrate | pH 8.5, 40°C, recombinant enzyme, cofactor NADH | Haloferax mediterranei |
Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|
29500 | - |
4 * 33336, sequence calculation, 4 * 29500, recombinant enzyme, CTAB-PAGE, 4 * 47000, SDS-PAGE | Haloferax mediterranei |
33336 | - |
4 * 33336, sequence calculation, 4 * 29500, recombinant enzyme, CTAB-PAGE, 4 * 47000, SDS-PAGE | Haloferax mediterranei |
47000 | - |
4 * 33336, sequence calculation, 4 * 29500, recombinant enzyme, CTAB-PAGE, 4 * 47000, SDS-PAGE | Haloferax mediterranei |
111000 | - |
recombinant enzyme, gel filtration | Haloferax mediterranei |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Haloferax mediterranei | Q2VEQ7 | gene ddh; strain R4, gene ddh | - |
Haloferax mediterranei R-4 / ATCC 33500 | Q2VEQ7 | gene ddh; strain R4, gene ddh | - |
Purification (Comment) | Organism |
---|---|
recombinant enzyme partially from Escherichia coli strain BL21(DE3) inclusion bodies by solubilization with 8 M urea in DTT-containing buffer and refolding by rapid 10fold dilution | Haloferax mediterranei |
Reaction | Comment | Organism | Reaction ID |
---|---|---|---|
an (R)-2-hydroxycarboxylate + NADP+ = a 2-oxocarboxylate + NADPH + H+ | the catalytic triad is probably formed by Arg226, Glu255, and His274 | Haloferax mediterranei |
Renatured (Comment) | Organism |
---|---|
refolding of recombinant enzyme from Escherichia coli strain BL21(DE3) inclusion bodies, solubilized by 8 M urea in DTT-containing buffer, by rapid 10fold dilution, optimally in presence of 4 M NaCl at pH 8.0 and 37°C | Haloferax mediterranei |
Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
---|---|---|---|
additional information | - |
substrate and cofactor specificity, overview | Haloferax mediterranei |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
2-oxo-4-methylpentanoate + NAD(P)H | - |
Haloferax mediterranei | (2R)-2-hydroxy-4-methylpentanoate + NAD(P)+ | - |
ir | |
2-oxo-4-methylpentanoate + NAD(P)H | - |
Haloferax mediterranei R-4 / ATCC 33500 | (2R)-2-hydroxy-4-methylpentanoate + NAD(P)+ | - |
ir | |
2-oxobutyrate + NAD(P)H | - |
Haloferax mediterranei | (2R)-2-hydroxybutyrate + NAD(P)+ | - |
ir | |
2-oxobutyrate + NAD(P)H | - |
Haloferax mediterranei R-4 / ATCC 33500 | (2R)-2-hydroxybutyrate + NAD(P)+ | - |
ir | |
2-oxobutyrate + NADH + H+ | - |
Haloferax mediterranei | (2R)-2-hydroxybutyrate + NAD+ | - |
r | |
2-oxoisocaproate + NADH + H+ | - |
Haloferax mediterranei | 2-D-hydroxyisocaproate + NAD+ | - |
r | |
2-oxoisoleucine + NAD(P)H | - |
Haloferax mediterranei | 2-D-hydroxyisoleucine + NAD(P)+ | - |
ir | |
2-oxoisoleucine + NAD(P)H | - |
Haloferax mediterranei R-4 / ATCC 33500 | 2-D-hydroxyisoleucine + NAD(P)+ | - |
ir | |
2-oxoisoleucine + NADH + H+ | - |
Haloferax mediterranei | 2-D-hydroxyisoleucine + NAD+ | - |
r | |
additional information | no activity with 2-D-hydroxyacids and L-2-hydroxyisocaproic acid in oxidation reaction | Haloferax mediterranei | ? | - |
? | |
additional information | no activity with 2-D-hydroxyacids and L-2-hydroxyisocaproic acid in oxidation reaction | Haloferax mediterranei R-4 / ATCC 33500 | ? | - |
? | |
pyruvate + NADPH + H+ | - |
Haloferax mediterranei | D-lactate + NADP+ | - |
ir | |
pyruvate + NADPH + H+ | - |
Haloferax mediterranei R-4 / ATCC 33500 | D-lactate + NADP+ | - |
ir |
Subunits | Comment | Organism |
---|---|---|
tetramer | 4 * 33336, sequence calculation, 4 * 29500, recombinant enzyme, CTAB-PAGE, 4 * 47000, SDS-PAGE | Haloferax mediterranei |
Synonyms | Comment | Organism |
---|---|---|
2-D-hydroxyacid dehydrogenase | - |
Haloferax mediterranei |
D-2-hydroxyacid dehydrogenase | - |
Haloferax mediterranei |
D2-HDH | - |
Haloferax mediterranei |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
40 | - |
assay at, recombinant refolded enzyme | Haloferax mediterranei |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
5 | - |
assay at, with substrate pyruvate | Haloferax mediterranei |
8.5 | - |
assay at, all substrates, except for pyruvate | Haloferax mediterranei |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
NADH | - |
Haloferax mediterranei | |
NADPH | preferred cofactor | Haloferax mediterranei |