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Literature summary for 1.4.1.1 extracted from

  • Chowdhury, E.K.; Saitoh, T.; Nagata, S.; Ashiuchi, M.; Misono, H.
    Alanine dehydrogenase from Enterobacter aerogenes: purification, characterization, and primary structure (1998), Biosci. Biotechnol. Biochem., 62, 2357-2363.
    View publication on PubMed
Search for more literature for 1.4.1.1

Cloned(Commentary)

Cloned (Comment) Organism
expression in Escherichia coli JM109 Klebsiella aerogenes

Inhibitors

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 [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 [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

Organism UniProt Comment Textmining
Klebsiella aerogenes
-
-
-

Purification (Commentary)

Purification (Comment) Organism
-
 Klebsiella aerogenes

Reaction

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
a

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
92
-
-
 Klebsiella aerogenes

Storage Stability

Storage Stability Organism
4°C, 60% saturation with ammonium sulfate, pH 7.2, stable for over 6 months Klebsiella aerogenes

Substrates and Products (Substrate)

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

Subunits Comment Organism
hexamer 6 * 40000, SDS-PAGE Klebsiella aerogenes
hexamer 6 * 39807, calculation from nucleotide sequence Klebsiella aerogenes

Temperature Stability [°C]

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

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 pH Stability Maximum Comment Organism
6 10.5 30°C, 10 min, most stable in the pH-range Klebsiella aerogenes

Cofactor

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