Activating Compound | Comment | Organism | Structure |
---|---|---|---|
2-mercaptoethanol | about 1575.34% activity at 1 mM | Pseudomonas denitrificans (nom. rej.) | |
dithiothreitol | about 182.11% activity at 1 mM | Pseudomonas denitrificans (nom. rej.) | |
EDTA | about 156.21% activity at 1 mM | Pseudomonas denitrificans (nom. rej.) |
Cloned (Comment) | Organism |
---|---|
expressed in Escherichia coli BL21(DE3) cells | Pseudomonas denitrificans (nom. rej.) |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
Ag+ | complete inhibition at 1 mM | Pseudomonas denitrificans (nom. rej.) | |
Co2+ | about 68.1% residual activity at 1 mM | Pseudomonas denitrificans (nom. rej.) | |
Cu2+ | about 15.67% residual activity at 1 mM | Pseudomonas denitrificans (nom. rej.) | |
Fe2+ | complete inhibition at 1 mM | Pseudomonas denitrificans (nom. rej.) | |
Hg2+ | complete inhibition at 1 mM | Pseudomonas denitrificans (nom. rej.) | |
K+ | about 92.11% residual activity at 1 mM | Pseudomonas denitrificans (nom. rej.) | |
Li+ | about 92.32% residual activity at 1 mM | Pseudomonas denitrificans (nom. rej.) | |
Mg2+ | about 87.34% residual activity at 1 mM | Pseudomonas denitrificans (nom. rej.) | |
NH4+ | about 90.17% residual activity at 1 mM | Pseudomonas denitrificans (nom. rej.) | |
Ni2+ | about 63.67% residual activity at 1 mM | Pseudomonas denitrificans (nom. rej.) | |
Zn2+ | about 16.13% residual activity at 1 mM | Pseudomonas denitrificans (nom. rej.) |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.12 | - |
(S)-3-hydroxyisobutyrate | at 37°C and pH 9.0 | Pseudomonas denitrificans (nom. rej.) | |
2.09 | - |
3-hydroxypropionic acid | at 37°C and pH 9.0 | Pseudomonas denitrificans (nom. rej.) | |
10.37 | - |
methyl DL-serine | at 37°C and pH 9.0 | Pseudomonas denitrificans (nom. rej.) | |
10.48 | - |
methyl (2S)-3-hydroxy-2-methylpropanoate | at 37°C and pH 9.0 | Pseudomonas denitrificans (nom. rej.) | |
31.84 | - |
L-serine | at 37°C and pH 9.0 | Pseudomonas denitrificans (nom. rej.) |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Mn2+ | about 127.2% activity at 1 mM | Pseudomonas denitrificans (nom. rej.) | |
additional information | not influenced by Na+ | Pseudomonas denitrificans (nom. rej.) |
Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|
25000 | - |
x * 25000, SDS-PAGE | Pseudomonas denitrificans (nom. rej.) |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Pseudomonas denitrificans (nom. rej.) | - |
- |
- |
Pseudomonas denitrificans (nom. rej.) ATCC 13867 | - |
- |
- |
Purification (Comment) | Organism |
---|---|
Ni-NTA column chromatography and Superdex 200 gel filtration | Pseudomonas denitrificans (nom. rej.) |
Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
---|---|---|---|
1.62 | - |
crude extract, at 37°C and pH 9.0 | Pseudomonas denitrificans (nom. rej.) |
58.67 | - |
after 36.22fold purification, at 37°C and pH 9.0 | Pseudomonas denitrificans (nom. rej.) |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
(S)-3-hydroxyisobutyrate + NAD+ | the enzyme is very specific to (S)-3-hydroxyisobutyrate | Pseudomonas denitrificans (nom. rej.) | 2-methyl-3-oxopropanoate + NADH + H+ | - |
? | |
(S)-3-hydroxyisobutyrate + NAD+ | the enzyme is very specific to (S)-3-hydroxyisobutyrate | Pseudomonas denitrificans (nom. rej.) ATCC 13867 | 2-methyl-3-oxopropanoate + NADH + H+ | - |
? | |
3-hydroxypropionic acid + NAD+ | - |
Pseudomonas denitrificans (nom. rej.) | malonate semialdehyde + NADH + H+ | - |
r | |
3-hydroxypropionic acid + NAD+ | - |
Pseudomonas denitrificans (nom. rej.) ATCC 13867 | malonate semialdehyde + NADH + H+ | - |
r | |
L-serine + NAD+ | - |
Pseudomonas denitrificans (nom. rej.) | 3-oxo-L-alanine + NADH + H+ | - |
r | |
L-serine + NAD+ | - |
Pseudomonas denitrificans (nom. rej.) ATCC 13867 | 3-oxo-L-alanine + NADH + H+ | - |
r | |
methyl (2S)-3-hydroxy-2-methylpropanoate + NAD+ | - |
Pseudomonas denitrificans (nom. rej.) | methyl (2S)-2-methyl-3-oxopropanoate + NADH + H+ | - |
r | |
methyl (2S)-3-hydroxy-2-methylpropanoate + NAD+ | - |
Pseudomonas denitrificans (nom. rej.) ATCC 13867 | methyl (2S)-2-methyl-3-oxopropanoate + NADH + H+ | - |
r | |
methyl DL-serine + NAD+ | - |
Pseudomonas denitrificans (nom. rej.) | methyl 3-oxo-DL-alanine + NADH + H+ | - |
r | |
additional information | no activity with (R)-3-hydroxyisobutyrate | Pseudomonas denitrificans (nom. rej.) | ? | - |
? | |
additional information | no activity with (R)-3-hydroxyisobutyrate | Pseudomonas denitrificans (nom. rej.) ATCC 13867 | ? | - |
? |
Subunits | Comment | Organism |
---|---|---|
? | x * 25000, SDS-PAGE | Pseudomonas denitrificans (nom. rej.) |
Synonyms | Comment | Organism |
---|---|---|
3-hydroxyisobutyrate dehydrogenase-I | - |
Pseudomonas denitrificans (nom. rej.) |
3HIBDHI | - |
Pseudomonas denitrificans (nom. rej.) |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
37 | - |
- |
Pseudomonas denitrificans (nom. rej.) |
Temperature Minimum [°C] | Temperature Maximum [°C] | Comment | Organism |
---|---|---|---|
30 | 45 | more than 50% activity between 30 and 40°C. Approximately 80% of the activity is lost at 45°C | Pseudomonas denitrificans (nom. rej.) |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
7.94 | - |
methyl DL-serine | at 37°C and pH 9.0 | Pseudomonas denitrificans (nom. rej.) | |
32.17 | - |
3-hydroxypropionic acid | at 37°C and pH 9.0 | Pseudomonas denitrificans (nom. rej.) | |
41.35 | - |
methyl (2S)-3-hydroxy-2-methylpropanoate | at 37°C and pH 9.0 | Pseudomonas denitrificans (nom. rej.) | |
47.16 | - |
L-serine | at 37°C and pH 9.0 | Pseudomonas denitrificans (nom. rej.) | |
70.81 | - |
(S)-3-hydroxyisobutyrate | at 37°C and pH 9.0 | Pseudomonas denitrificans (nom. rej.) |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
9 | - |
- |
Pseudomonas denitrificans (nom. rej.) |
pH Minimum | pH Maximum | Comment | Organism |
---|---|---|---|
8 | 10 | approximately 40% of the activity is retained at pH 11.0. At pH 7.0, the activity is reduced to 30% of the activity observed at pH 9.0 | Pseudomonas denitrificans (nom. rej.) |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
NAD+ | the enzyme prefers NAD+ over NADP+ as a cofactor | Pseudomonas denitrificans (nom. rej.) |
kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.75 | - |
methyl DL-serine | at 37°C and pH 9.0 | Pseudomonas denitrificans (nom. rej.) | |
1.46 | - |
L-serine | at 37°C and pH 9.0 | Pseudomonas denitrificans (nom. rej.) | |
4.04 | - |
methyl (2S)-3-hydroxy-2-methylpropanoate | at 37°C and pH 9.0 | Pseudomonas denitrificans (nom. rej.) | |
15.4 | - |
3-hydroxypropionic acid | at 37°C and pH 9.0 | Pseudomonas denitrificans (nom. rej.) | |
604.1 | - |
(S)-3-hydroxyisobutyrate | at 37°C and pH 9.0 | Pseudomonas denitrificans (nom. rej.) |