Activating Compound | Comment | Organism | Structure |
---|---|---|---|
L-valine | VDH synthesis induced by L-valine, but severely repressed by ammonia | Streptomyces fradiae |
Cloned (Comment) | Organism |
---|---|
vdh gene encoding VDH2, only one gene responsible for VDH activity, VDH2 is the only active VDH | Streptomyces fradiae |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
Cu2+ | VDH2 : 1 mM, 35.5% inhibition | Streptomyces fradiae | |
iodoacetamide | VDH2: 1 mM, complete inhibition | Streptomyces fradiae | |
L-valine | VDH2: 20 mM instead of 10 mM in reaction mixture caused 22% loss of activity, 50 mM 40% loss of activity | Streptomyces fradiae | |
p-hydroxymercuribenzoate | VDH2: 0.01 mM, 70% loss of activity | Streptomyces fradiae |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.028 | - |
NADH | VDH2 | Streptomyces fradiae | |
0.04 | - |
NAD+ | VDH2 | Streptomyces fradiae | |
0.31 | - |
2-oxoisovalerate | VDH2 | Streptomyces fradiae | |
0.43 | - |
L-valine | VDH2 | Streptomyces fradiae | |
25.6 | - |
NH4+ | VDH2 | Streptomyces fradiae |
Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|
41000 | - |
2 * 41000, VDH2, SDS-PAGE | Streptomyces fradiae |
80000 | - |
VDH2, gel filtration | Streptomyces fradiae |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
L-valine + NAD(P)+ + H2O | Streptomyces fradiae | VDH2 rather than VDH1 plays a role in metabolism of branched chain amino acids and thus in tylosin biosynthesis, expression of VDH1 is unstable | 2-oxoisovalerate + NH3 + NAD(P)H | - |
? | |
L-valine + NAD(P)+ + H2O | Streptomyces fradiae | VDH is required for utilization of branched chain amino acids, the catabolism appears to be an alternative source of n-butyrate, 2-methylmalonate, and propionate needed for biosynthesis of macrolide and polyether antibiotics | 2-oxoisovalerate + NH3 + NAD(P)H | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Streptomyces fradiae | - |
- |
- |
Streptomyces fradiae | - |
two enzymes: VDH1 and VDH2 | - |
Streptomyces fradiae | - |
tylosin producer | - |
Streptomyces fradiae 30/3 | - |
- |
- |
Purification (Comment) | Organism |
---|---|
VDH2, 346fold purification | Streptomyces fradiae |
Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
---|---|---|---|
19.56 | - |
oxidative deamination | Streptomyces fradiae |
40.92 | - |
reductive amination | Streptomyces fradiae |
Storage Stability | Organism |
---|---|
-20°C, 50 mM Tris-HCl buffer, pH 7.4, 5 mM 2-mercaptoethanol, 1 mM EDTA, 30% v/v glycerol, 24 h, 90% loss of activity | Streptomyces fradiae |
4°C, 50 mM Tris-HCl buffer, pH 7.4, 5 mM 2-mercaptoethanol, 1 mM EDTA, 10% v/v glycerol, 48 h, stable | Streptomyces fradiae |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
L-2-aminobutyrate + H2O + NAD+ | VDH2: preferred substrate, 158% of the activity with L-valine | Streptomyces fradiae | 2-oxobutyrate + NADH + NH3 | VDH2: reductive amination, 100% of the activity with 2-oxoisovalerate | r | |
L-2-aminobutyrate + H2O + NAD+ | VDH2: preferred substrate, 158% 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 | VDH2: reductive amination, 100% 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+ | VDH2: 8.3% 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+ | VDH2: 8.3% of the activity with L-valine | Streptomyces fradiae | pyruvate + NH3 + NADH | VDH2: reductive amination, 23.3% of the activity with 2-oxoisovalerate | r | |
L-isoleucine + H2O + NAD+ | VDH2: 22% 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+ | VDH2: 22% of the activity with L-valine | Streptomyces fradiae | 3-methyl-2-oxopentanoate + NADH + NH3 | VDH2: reductive amination, 16.6% of the activity with 2-oxoisovalerate | r | |
L-leucine + H2O + NAD+ | VDH2: 24% 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-leucine + H2O + NAD+ | VDH2: 24% of the activity with L-valine | Streptomyces fradiae | 2-oxoisocaproate + NADH + NH3 | VDH2: reductive amination, 20% of the activity with 2-oxoisovalerate | r | |
L-norleucine + H2O + NAD+ | VDH2: 34.5% of the activity with L-valine | Streptomyces fradiae | 2-oxocaproate + NADH + NH3 | - |
r | |
L-norvaline + H2O + NAD+ | VDH2: 96% of the activity with L-valine | Streptomyces fradiae | 2-oxovalerate + NH3 + NADH | - |
r | |
L-valine + NAD(P)+ + H2O | preferred substrate | Streptomyces fradiae | 2-oxoisovalerate + NH3 + NAD(P)H | NADH | r | |
L-valine + NAD(P)+ + H2O | preferred substrate | 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 | preferred substrate | Streptomyces fradiae | 2-oxoisovalerate + NH3 + NAD(P)H | reductive amination: preferred substrate | 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 | reductive amination rate is twice the oxidative deamination rate | Streptomyces fradiae | 2-oxoisovalerate + NH3 + NAD(P)H | NADH | r | |
L-valine + NAD(P)+ + H2O | reductive amination rate is twice the oxidative deamination rate | 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 | reductive amination rate is twice the oxidative deamination rate | Streptomyces fradiae | 2-oxoisovalerate + NH3 + NAD(P)H | reductive amination: preferred substrate | r | |
L-valine + NAD(P)+ + H2O | VDH2 rather than VDH1 plays a role in metabolism of branched chain amino acids and thus in tylosin biosynthesis, expression of VDH1 is unstable | Streptomyces fradiae | 2-oxoisovalerate + NH3 + NAD(P)H | - |
? | |
L-valine + NAD(P)+ + H2O | VDH is required for utilization of branched chain amino acids, the catabolism appears to be an alternative source of n-butyrate, 2-methylmalonate, and propionate needed for biosynthesis of macrolide and polyether antibiotics | Streptomyces fradiae | 2-oxoisovalerate + NH3 + NAD(P)H | - |
? | |
L-valine + NAD(P)+ + H2O | preferred substrate | Streptomyces fradiae 30/3 | 2-oxoisovalerate + NH3 + NAD(P)H | NADH | r | |
L-valine + NAD(P)+ + H2O | preferred substrate | 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 | preferred substrate | Streptomyces fradiae 30/3 | 2-oxoisovalerate + NH3 + NAD(P)H | reductive amination: preferred substrate | r | |
L-valine + NAD(P)+ + H2O | NAD+ | Streptomyces fradiae 30/3 | 2-oxoisovalerate + NH3 + NAD(P)H | NADH | r | |
additional information | NH3 is the sole substrate as amino donor | Streptomyces fradiae | additional information | - |
? | |
additional information | NH3 is the sole substrate as amino donor | Streptomyces fradiae 30/3 | additional information | - |
? |
Subunits | Comment | Organism |
---|---|---|
dimer | 2 * 41000, VDH2, SDS-PAGE | Streptomyces fradiae |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
65 | - |
- |
Streptomyces fradiae |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
8.8 | - |
VDH2: reductive amination of 2-oxoisovalerate, 0.3 M Tris-HCl buffer | Streptomyces fradiae |
10.4 | - |
VDH2: oxidative deamination of L-valine, 0.3 M glycine-KCl-KOH buffer | Streptomyces fradiae |
pH Stability | pH Stability Maximum | Comment | Organism |
---|---|---|---|
6.3 | - |
VDH2: stable at pH 6.3 during purification step Reactive-Blue 2 Sepharose chromatography | Streptomyces fradiae |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
1,N6-etheno-NAD+ | VDH2: 70.5% of the activity with NAD+ | Streptomyces fradiae | |
3-pyridinealdehyde-NAD+ | VDH2: 47.2% of the activity with NAD+, VDH1: no activity | Streptomyces fradiae | |
Alpha-NAD+ | VDH2: 5.8% of the activity with NAD+ | Streptomyces fradiae | |
deamino-NAD+ | VDH2: 124% of the activity with NAD+ | Streptomyces fradiae | |
deamino-NAD+ | amino group in the adenine moiety of NAD+ is not essential | Streptomyces fradiae | |
additional information | VDH2: not with NADP+, compared to VDH1 with 10.8% of the activity with NAD+ | Streptomyces fradiae | |
NAD+ | NAD+ required, no use of NADP+ | Streptomyces fradiae | |
NAD+ | natural cofactor for oxidative deamination | Streptomyces fradiae | |
NADH | - |
Streptomyces fradiae |