Cloned (Comment) | Organism |
---|---|
gene ApalaDH, DNA and amino acid sequence determination and analysis, sequence comparisons, phylogenetic analysis and tree, recombinant expression of His-tagged enzyme in Escherichia coli strain BL21, subcloning in Escherichia coli strain DH5alpha | Aphanothece halophytica |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
glycine | GxRA activity is inhibited by 60% in presence of 20 mM glycine, but glycine does not inhibit the PvRA activity | Aphanothece halophytica | |
KCl | slightly enhances PvRA activity and it moderately promotes ALD activity. In contrast, increasing amounts of KCl gradually inhibits GxRA activity | Aphanothece halophytica | |
L-alanine | the PvRA activity of ApAlaDH is strongly inhibited by 2 mM L-alanine, 68% inhibition. The GxRA activity is completely inhibited in the presence of L-alanine (100% inhibition at 10 and 20 mM L-alanine) | Aphanothece halophytica | |
additional information | all enzyme reactions are inhibited to varying degrees by their products | Aphanothece halophytica | |
NaCl | NaCl does not inhibit PvRA activity up to 1 M, but inhibition is observed at 2.5 M. The activities of ALD and GxRA are significantly increased in the presence of NaCl at concentrations of 0.1-0.25 M | Aphanothece halophytica | |
pyruvate | the ALD activity of ApAlaDH is strongly inhibited by 2 mM L-alanine, 45% inhibition | Aphanothece halophytica |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.22 | - |
pyruvate | pH 9.0, temperature not specified in the publication | Aphanothece halophytica | |
0.72 | - |
L-alanine | pH 10.0, temperature not specified in the publication | Aphanothece halophytica | |
1.91 | - |
glyoxylate | pH 8.5, temperature not specified in the publication | Aphanothece halophytica |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
KCl | slightly enhances PvRA activity and it moderately promotes ALD activity. In contrast, increasing amounts of KCl gradually inhibits GxRA activity | Aphanothece halophytica |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
glyoxylate + NH3 + NADH + H+ | Aphanothece halophytica | - |
glycine + H2O + NAD+ | - |
ir | |
glyoxylate + NH3 + NADH + H+ | Aphanothece halophytica CM1 | - |
glycine + H2O + NAD+ | - |
ir | |
L-alanine + H2O + NAD+ | Aphanothece halophytica | - |
pyruvate + NH3 + NADH + H+ | - |
? | |
L-alanine + H2O + NAD+ | Aphanothece halophytica CM1 | - |
pyruvate + NH3 + NADH + H+ | - |
? | |
pyruvate + NH3 + NADH + H+ | Aphanothece halophytica | - |
L-alanine + H2O + NAD+ | - |
r | |
pyruvate + NH3 + NADH + H+ | Aphanothece halophytica CM1 | - |
L-alanine + H2O + NAD+ | - |
r |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Aphanothece halophytica | A0A3G4YK61 | - |
- |
Aphanothece halophytica CM1 | A0A3G4YK61 | - |
- |
Purification (Comment) | Organism |
---|---|
recombinant His-tagged enzyme from Escherichia coli strain BL21 by nickel affinity chromatography | Aphanothece halophytica |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
glyoxylate + NH3 + NADH + H+ | - |
Aphanothece halophytica | glycine + H2O + NAD+ | - |
ir | |
glyoxylate + NH3 + NADH + H+ | - |
Aphanothece halophytica CM1 | glycine + H2O + NAD+ | - |
ir | |
L-alanine + H2O + NAD+ | - |
Aphanothece halophytica | pyruvate + NH3 + NADH + H+ | - |
? | |
L-alanine + H2O + NAD+ | - |
Aphanothece halophytica CM1 | pyruvate + NH3 + NADH + H+ | - |
? | |
pyruvate + NH3 + NADH + H+ | - |
Aphanothece halophytica | L-alanine + H2O + NAD+ | - |
r | |
pyruvate + NH3 + NADH + H+ | - |
Aphanothece halophytica CM1 | L-alanine + H2O + NAD+ | - |
r |
Subunits | Comment | Organism |
---|---|---|
? | x * 42000, recombinant His-tagged enzyme, SDS-PAGE, x * 38532, sequence calculation | Aphanothece halophytica |
Synonyms | Comment | Organism |
---|---|---|
alanine oxidative deaminase | - |
Aphanothece halophytica |
ALD | - |
Aphanothece halophytica |
ApalaDH | - |
Aphanothece halophytica |
glyoxylate reductive aminase | - |
Aphanothece halophytica |
GxrA | - |
Aphanothece halophytica |
PvRA | - |
Aphanothece halophytica |
pyruvate reductive aminase | - |
Aphanothece halophytica |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
8.5 | - |
recombinant enzyme, reductive amination reaction with glyoxylate | Aphanothece halophytica |
8.8 | 9 | recombinant enzyme, reductive amination reaction with pyruvate | Aphanothece halophytica |
10 | 10.5 | recombinant enzyme, oxidative deamination reaction | Aphanothece halophytica |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
NAD+ | - |
Aphanothece halophytica | |
NADH | - |
Aphanothece halophytica |
Organism | Comment | Expression |
---|---|---|
Aphanothece halophytica | ApalaDH expression is upregulated by NaCl. PvRA and GxRA activities are detected in vivo and increase about 1.2- and 2.7fold upon salt stress, respectively | up |
General Information | Comment | Organism |
---|---|---|
physiological function | gene ApalaDH encodes a bifunctional protein catalyzing the reversible reaction of pyruvate to L-alanine via its pyruvate reductive aminase (PvRA) activity, the reaction of L-alanine to pyruvate via its alanine oxidative dehydrogenase activity, and the non-reversible reaction of glyoxylate to glycine via its glyoxylate reductive aminase (GxRA) activity. The assimilatory/dissimilatory roles of enzyme ApAlaDH from the halotolerant cyanobacterium Aphanothece halophytica are not only specific for L-alanine and pyruvate, but also, upon salt stress, include glyoxylate to generate glycine. ApAlaDH is a bifunctional enzyme | Aphanothece halophytica |