Cloned (Comment) | Organism |
---|---|
expression in Escherichia coli JM109 | Klebsiella aerogenes |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
CuCl2 | 1 mM, 52% inhibition | Klebsiella aerogenes | |
HgCl2 | 1 mM, 38% inhibition | Klebsiella aerogenes | |
L-Ala | uncompetitive inhibition with respect to NADH | Klebsiella aerogenes | |
NAD+ | product inhibition | Klebsiella aerogenes |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.16 | - |
NAD+ | - |
Klebsiella aerogenes | |
0.22 | - |
pyruvate | - |
Klebsiella aerogenes | |
0.47 | - |
L-Ala | - |
Klebsiella aerogenes | |
66.7 | - |
NH4+ | - |
Klebsiella aerogenes |
Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|
39807 | - |
6 * 39807, calculation from nucleotide sequence | Klebsiella aerogenes |
40000 | - |
6 * 40000, SDS-PAGE | Klebsiella aerogenes |
245000 | - |
gel filtration | Klebsiella aerogenes |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Klebsiella aerogenes | - |
- |
- |
Purification (Comment) | Organism |
---|---|
- |
Klebsiella aerogenes |
Reaction | Comment | Organism | Reaction ID |
---|---|---|---|
L-alanine + H2O + NAD+ = pyruvate + NH3 + NADH + H+ | sequential ordered binary-ternary mechanism. NAD+ binds first to the enzyme, followed by L-Ala. The products are released in the order: NH4+, pyruvate and NADH | Klebsiella aerogenes |
Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
---|---|---|---|
92 | - |
- |
Klebsiella aerogenes |
Storage Stability | Organism |
---|---|
4°C, 60% saturation with ammonium sulfate, pH 7.2, stable for over 6 months | Klebsiella aerogenes |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
L-2-aminobutanoate + H2O + NAD+ | 8.7% of the activity with L-Ala | Klebsiella aerogenes | 2-oxobutanoate + NH3 + NADH + H+ | - |
r | |
L-2-aminobutanoate + H2O + NAD+ | in the reverse reaction 2-oxobutanoate reacts with 2.0% of the activity with pyruvate | Klebsiella aerogenes | 2-oxobutanoate + NH3 + NADH + H+ | - |
r | |
L-Ala + H2O + 3-acetylpyridine-NAD+ | 43% of the activity with NAD+ | Klebsiella aerogenes | pyruvate + H2O + 3-acetylpyridine-NADH | - |
? | |
L-Ala + H2O + deamino-NAD+ | 4.4% of the activity with NAD+ | Klebsiella aerogenes | pyruvate + NH3 + deamino-NADH | - |
? | |
L-Ala + H2O + NAD+ | - |
Klebsiella aerogenes | pyruvate + NH3 + NADH | - |
r | |
L-Ala + H2O + NADP+ | 7% of the activity with NAD+ | Klebsiella aerogenes | pyruvate + NH3 + NADPH | - |
? | |
L-Ala + H2O + nicotinamide guanine dinucleotide | 69.7% of the activity with NAD+ | Klebsiella aerogenes | ? | - |
? | |
L-norvaline + H2O + NAD+ | - |
Klebsiella aerogenes | 2-oxopentanoate + NH3 + NADH | - |
r | |
L-Ser + H2O + NAD+ | in the reverse reaction 3-hydroxypyruvate reacts with 14.2% of the activity with pyruvate | Klebsiella aerogenes | 3-hydroxypyruvate + NH3 + NADH | - |
r | |
L-Ser + H2O + NAD+ | 4.1% of the activity with L-Ala | Klebsiella aerogenes | 3-hydroxypyruvate + NH3 + NADH | - |
r | |
L-Val + H2O + NAD+ | 1.5% of the activity with L-Ala | Klebsiella aerogenes | 3-methyl-2-oxobutanoate + NH3 + NADH | - |
? |
Subunits | Comment | Organism |
---|---|---|
hexamer | 6 * 40000, SDS-PAGE | Klebsiella aerogenes |
hexamer | 6 * 39807, calculation from nucleotide sequence | Klebsiella aerogenes |
Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
---|---|---|---|
30 | - |
pH 6.0-10.5, 10 min, stable | Klebsiella aerogenes |
50 | - |
pH 7.4, 10 min, stable up to | Klebsiella aerogenes |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
8.7 | - |
amination of pyruvate | Klebsiella aerogenes |
10.9 | - |
deamination of L-Ala | Klebsiella aerogenes |
pH Stability | pH Stability Maximum | Comment | Organism |
---|---|---|---|
6 | 10.5 | 30°C, 10 min, most stable in the pH-range | Klebsiella aerogenes |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
3-acetylpyridine-NAD+ | 43% of the activity with NAD+ | Klebsiella aerogenes | |
deamino-NAD+ | 81% of the activity with NAD+ | Klebsiella aerogenes | |
NAD+ | - |
Klebsiella aerogenes | |
NADH | - |
Klebsiella aerogenes | |
NADP+ | 7% of the activity with NAD+ | Klebsiella aerogenes | |
nicotinamide guanine dinucleotide | - |
Klebsiella aerogenes |