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Literature summary for 1.4.1.1 extracted from

  • Phogosee, S.; Hibino, T.; Kageyama, H.; Waditee-Sirisattha, R.
    Bifunctional alanine dehydrogenase from the halotolerant cyanobacterium Aphanothece halophytica characterization and molecular properties (2018), Arch. Microbiol., 200, 719-727 .
    View publication on PubMed
Search for more literature for 1.4.1.1

Cloned(Commentary)

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

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 [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

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/ Products (Substrates)

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

Organism UniProt Comment Textmining
Aphanothece halophytica A0A3G4YK61
-
-
Aphanothece halophytica CM1 A0A3G4YK61
-
-

Purification (Commentary)

Purification (Comment) Organism
recombinant His-tagged enzyme from Escherichia coli strain BL21 by nickel affinity chromatography Aphanothece halophytica

Substrates and Products (Substrate)

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

Subunits Comment Organism
? x * 42000, recombinant His-tagged enzyme, SDS-PAGE, x * 38532, sequence calculation Aphanothece halophytica

Synonyms

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

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

Cofactor Comment Organism Structure
NAD+
-
 Aphanothece halophytica
NADH
-
 Aphanothece halophytica

Expression

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

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