Activating Compound | Comment | Organism | Structure |
---|---|---|---|
2-mercaptoethanol | presence of 2-mercaptoethanol or dithiothreitol at 10 mM required for maximal activity | Streptomyces fradiae | |
dithiothreitol | presence of 2-mercaptoethanol or dithiothreitol at 10 mM required for maximal activity | Streptomyces fradiae |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
Hg2+ | HgCl2: strong inhibition, partially abolished in presence of 2-mercaptoethanol | Streptomyces fradiae | |
p-chloromercuribenzoate | strong inhibition, partially abolished in presence of 2-mercaptoethanol | Streptomyces fradiae |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.029 | - |
NAD+ | - |
Streptomyces fradiae | |
0.05 | - |
NADH | - |
Streptomyces fradiae | |
0.67 | - |
L-leucine | - |
Streptomyces fradiae | |
0.8 | - |
2-oxoisovalerate | - |
Streptomyces fradiae | |
0.82 | - |
2-oxoisocaproate | - |
Streptomyces fradiae | |
1 | - |
L-valine | - |
Streptomyces fradiae | |
1 | - |
2-oxobutyrate | - |
Streptomyces fradiae | |
1.18 | - |
L-isoleucine | - |
Streptomyces fradiae | |
1.33 | - |
L-norvaline | - |
Streptomyces fradiae | |
1.94 | - |
2-oxovalerate | - |
Streptomyces fradiae | |
2.23 | - |
2-oxocaproate | - |
Streptomyces fradiae | |
3.07 | - |
L-2-aminobutyrate | - |
Streptomyces fradiae | |
3.33 | - |
2-oxo-3-methylvalerate | - |
Streptomyces fradiae | |
6.68 | - |
L-norleucine | - |
Streptomyces fradiae | |
22 | - |
NH4+ | - |
Streptomyces fradiae | |
25 | - |
pyruvate | - |
Streptomyces fradiae | |
28.2 | - |
L-alanine | - |
Streptomyces fradiae |
Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|
18000 | - |
12 * 18000, SDS-PAGE | Streptomyces fradiae |
215000 | - |
equilibrium ultracentrifugation | Streptomyces fradiae |
218000 | - |
gel filtration | Streptomyces fradiae |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
L-valine + NAD(P)+ + H2O | Streptomyces fradiae | regulatory enzyme involved in biosynthesis of n-butyrate, a building unit of the oligoketide antibiotic tylosin, NAD+ as natural cofactor for oxidative deamination | 2-oxoisovalerate + NH3 + NAD(P)H | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Streptomyces fradiae | - |
- |
- |
Streptomyces fradiae 30/3 | - |
- |
- |
Purification (Comment) | Organism |
---|---|
1508fold purification | Streptomyces fradiae |
Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
---|---|---|---|
additional information | - |
- |
Streptomyces fradiae |
Storage Stability | Organism |
---|---|
-20°C, 0.2 M Tris-HCl buffer, pH 7.4, 3 months, no loss of activity | Streptomyces fradiae |
-20°C, 70% (NH4)2SO4 suspension in 0.2 M Tris-HCl buffer, 5 months, stable | Streptomyces fradiae |
4°C, 0.2 M Tris-HCl buffer, pH 7.4, 24 h, no loss of activity | Streptomyces fradiae |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
L-2-aminobutyrate + H2O + NAD+ | 68.9% of the activity with L-valine | Streptomyces fradiae | 2-oxobutyrate + NADH + NH3 | reductive amination: 78% of the activity with 2-oxoisovalerate | r | |
L-2-aminobutyrate + H2O + NAD+ | 68.9% of the activity with L-valine | Streptomyces fradiae | 2-oxobutyrate + NADH + NH3 | 2-oxobutyrate is identical with alpha-ketobutyrate | r | |
L-2-aminobutyrate + H2O + NAD+ | L-2-aminobutyrate is identical with L-alpha-aminobutyrate | Streptomyces fradiae | 2-oxobutyrate + NADH + NH3 | reductive amination: 78% of the activity with 2-oxoisovalerate | r | |
L-2-aminobutyrate + H2O + NAD+ | L-2-aminobutyrate is identical with L-alpha-aminobutyrate | Streptomyces fradiae | 2-oxobutyrate + NADH + NH3 | 2-oxobutyrate is identical with alpha-ketobutyrate | r | |
L-alanine + H2O + NAD+ | 14.7% of the activity with L-valine | Streptomyces fradiae | pyruvate + NH3 + NADH | pyruvate is identical with alpha-ketopropanoate and 2-oxopropanoate | r | |
L-alanine + H2O + NAD+ | 14.7% of the activity with L-valine | Streptomyces fradiae | pyruvate + NH3 + NADH | reductive amination: 31.6% of the activity with 2-oxoisovalerate | r | |
L-isoleucine + H2O + NAD+ | 29.4% of the activity with L-valine | Streptomyces fradiae | 3-methyl-2-oxopentanoate + NADH + NH3 | 2-oxo-3-methylvalerate is identical with alpha-keto-beta-methylpentanoate | r | |
L-isoleucine + H2O + NAD+ | 29.4% of the activity with L-valine | Streptomyces fradiae | 3-methyl-2-oxopentanoate + NADH + NH3 | reductive amination: 23.2% of the activity with 2-oxoisovalerate | r | |
L-leucine + H2O + NAD+ | 24.5% of the activity with L-valine | Streptomyces fradiae | 2-oxoisocaproate + NADH + NH3 | reductive amination: 20% of the activity with 2-oxoisovalerate | r | |
L-leucine + H2O + NAD+ | 24.5% of the activity with L-valine | Streptomyces fradiae | 2-oxoisocaproate + NADH + NH3 | 2-oxoisocaproate is identical with 2-oxo-4-methylpentanoate, 2-oxoisohexanoate and alpha-ketoisocaproate | r | |
L-norleucine + H2O + NAD+ | 51.5% of the activity with L-valine | Streptomyces fradiae | 2-oxocaproate + NADH + NH3 | 2-oxocaproate is identical with 2-oxohexanoate and alpha-keto-n-hexanoate | r | |
L-norleucine + H2O + NAD+ | 51.5% of the activity with L-valine | Streptomyces fradiae | 2-oxocaproate + NADH + NH3 | reductive amination: 43.8% of the activity with 2-oxoisovalerate | r | |
L-norvaline + H2O + NAD+ | 98% of the activity with L-valine | Streptomyces fradiae | 2-oxovalerate + NH3 + NADH | reductive amination: 79.5% of the activity with 2-oxoisovalerate | r | |
L-norvaline + H2O + NAD+ | 98% of the activity with L-valine | Streptomyces fradiae | 2-oxovalerate + NH3 + NADH | 2-oxovalerate is identical with 2-oxopentanoate and alpha-ketovalerate | r | |
L-valine + NAD(P)+ + H2O | NAD+ | Streptomyces fradiae | 2-oxoisovalerate + NH3 + NAD(P)H | NADH | r | |
L-valine + NAD(P)+ + H2O | NAD+ | Streptomyces fradiae | 2-oxoisovalerate + NH3 + NAD(P)H | 2-oxoisovalerate is identical with 3-methyl-2-oxobutanoate and alpha-ketoisovalerate | r | |
L-valine + NAD(P)+ + H2O | NAD+ | Streptomyces fradiae | 2-oxoisovalerate + NH3 + NAD(P)H | reductive amination: preferred substrate | r | |
L-valine + NAD(P)+ + H2O | NADP+ | Streptomyces fradiae | 2-oxoisovalerate + NH3 + NAD(P)H | NADH | r | |
L-valine + NAD(P)+ + H2O | NADP+ | Streptomyces fradiae | 2-oxoisovalerate + NH3 + NAD(P)H | 2-oxoisovalerate is identical with 3-methyl-2-oxobutanoate and alpha-ketoisovalerate | r | |
L-valine + NAD(P)+ + H2O | NADP+ | Streptomyces fradiae | 2-oxoisovalerate + NH3 + NAD(P)H | reductive amination: preferred substrate | r | |
L-valine + NAD(P)+ + H2O | regulatory enzyme involved in biosynthesis of n-butyrate, a building unit of the oligoketide antibiotic tylosin, NAD+ as natural cofactor for oxidative deamination | Streptomyces fradiae | 2-oxoisovalerate + NH3 + NAD(P)H | - |
? | |
L-valine + NAD(P)+ + H2O | NAD+ | Streptomyces fradiae 30/3 | 2-oxoisovalerate + NH3 + NAD(P)H | NADH | r | |
L-valine + NAD(P)+ + H2O | NAD+ | Streptomyces fradiae 30/3 | 2-oxoisovalerate + NH3 + NAD(P)H | 2-oxoisovalerate is identical with 3-methyl-2-oxobutanoate and alpha-ketoisovalerate | r | |
L-valine + NAD(P)+ + H2O | NAD+ | Streptomyces fradiae 30/3 | 2-oxoisovalerate + NH3 + NAD(P)H | reductive amination: preferred substrate | r | |
L-valine + NAD(P)+ + H2O | NADP+ | Streptomyces fradiae 30/3 | 2-oxoisovalerate + NH3 + NAD(P)H | NADH | r | |
L-valine + NAD(P)+ + H2O | NADP+ | Streptomyces fradiae 30/3 | 2-oxoisovalerate + NH3 + NAD(P)H | 2-oxoisovalerate is identical with 3-methyl-2-oxobutanoate and alpha-ketoisovalerate | r |
Subunits | Comment | Organism |
---|---|---|
dodecamer | 12 * 18000, SDS-PAGE | Streptomyces fradiae |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
30 | - |
assay at | Streptomyces fradiae |
65 | - |
- |
Streptomyces fradiae |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
8.7 | 8.9 | reductive amination of 2-oxoisovalerate, in presence of 0.3 M Tris-HCl buffer | Streptomyces fradiae |
10.5 | 10.7 | oxidative deamination of L-valine, in presence of 0.3 M glycine-KCl-KOH buffer | Streptomyces fradiae |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
1,N6-etheno-NAD+ | 64.4% of the activity with NAD+ | Streptomyces fradiae | |
3-acetylpyridine-NAD+ | 64.5% of the activity with NAD+ | Streptomyces fradiae | |
Alpha-NAD+ | 3.6% of the activity with NAD+ | Streptomyces fradiae | |
deamino-NAD+ | VDH2: 124% of the activity with NAD+ | Streptomyces fradiae | |
deamino-NAD+ | 93.9% of the activity with NAD+ | Streptomyces fradiae | |
NAD+ | natural cofactor for oxidative deamination | Streptomyces fradiae | |
NADH | NADH could not be replaced with NADPH for the reductive amination | Streptomyces fradiae | |
NADP+ | 10.8% of the activity with NAD+ | Streptomyces fradiae | |
nicotinamide guanine dinucleotide | 92.9% of the activity with NAD+ | Streptomyces fradiae |