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Literature summary extracted from

  • Leese, C.; Fotheringham, I.; Escalettes, F.; Speight, R.; Grogan, G.
    Cloning, expression, characterisation and mutational analysis of L-aspartate oxidase from Pseudomonas putida (2013), J. Mol. Catal. B, 85-86, 17-22.
No PubMed abstract available

Cloned(Commentary)

EC Number Cloned (Comment) Organism
1.4.3.16 expressed in Escherichia coli BL21-Gold (DE3) cells Pseudomonas putida

Protein Variants

EC Number Protein Variants Comment Organism
1.4.3.16 H244A the mutant shows reduced catalytic efficiency compared to the wild type enzyme Pseudomonas putida
1.4.3.16 H351A inactive Pseudomonas putida
1.4.3.16 Q242A the mutant shows reduced catalytic efficiency compared to the wild type enzyme Pseudomonas putida
1.4.3.16 R290A the mutant shows reduced catalytic efficiency compared to the wild type enzyme Pseudomonas putida
1.4.3.16 R386A inactive Pseudomonas putida
1.4.3.16 S389A the mutant shows reduced catalytic efficiency compared to the wild type enzyme Pseudomonas putida
1.4.3.16 T259A inactive Pseudomonas putida

Inhibitors

EC Number Inhibitors Comment Organism Structure
1.4.3.16 L-asparagine
-
Pseudomonas putida

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
1.4.3.16 2.26
-
L-aspartate wild type enzyme, at pH 7.5, temperature not specified in the publication Pseudomonas putida
1.4.3.16 14.25
-
L-aspartate mutant enzyme S389A, at pH 7.5, temperature not specified in the publication Pseudomonas putida
1.4.3.16 26.11
-
L-aspartate mutant enzyme R290A, at pH 7.5, temperature not specified in the publication Pseudomonas putida
1.4.3.16 63.27
-
L-aspartate mutant enzyme Q242A, at pH 7.5, temperature not specified in the publication Pseudomonas putida
1.4.3.16 75.44
-
L-aspartate mutant enzyme H244A, at pH 7.5, temperature not specified in the publication Pseudomonas putida

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
1.4.3.16 59900
-
x * 59900, calculated from amino acid sequence Pseudomonas putida
1.4.3.16 60000
-
x * 60000, SDS-PAGE Pseudomonas putida

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
1.4.3.16 L-asparagine + H2O + O2 Pseudomonas putida low activity 4-amino-2,4-dioxobutanoate + NH3 + H2O2
-
?
1.4.3.16 L-aspartate + H2O + O2 Pseudomonas putida best substrate oxaloacetate + NH3 + H2O2
-
?
1.4.3.16 L-glutamate + H2O + O2 Pseudomonas putida low activity 2-oxopentanedioate + NH3 + H2O2
-
?

Organism

EC Number Organism UniProt Comment Textmining
1.4.3.16 Pseudomonas putida
-
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
1.4.3.16 HiTrap nickel chelating column chromatography and Superdex 75 gel filtration Pseudomonas putida

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
1.4.3.16 L-asparagine + H2O + O2 low activity Pseudomonas putida 4-amino-2,4-dioxobutanoate + NH3 + H2O2
-
?
1.4.3.16 L-aspartate + H2O + O2 best substrate Pseudomonas putida oxaloacetate + NH3 + H2O2
-
?
1.4.3.16 L-glutamate + H2O + O2 low activity Pseudomonas putida 2-oxopentanedioate + NH3 + H2O2
-
?

Subunits

EC Number Subunits Comment Organism
1.4.3.16 ? x * 60000, SDS-PAGE Pseudomonas putida
1.4.3.16 ? x * 59900, calculated from amino acid sequence Pseudomonas putida

Synonyms

EC Number Synonyms Comment Organism
1.4.3.16 LASPO
-
Pseudomonas putida

Temperature Stability [°C]

EC Number Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
1.4.3.16 40 50 the enzyme is stable for 60 min at up to 40°C, but rapid losses in activity are observed at 50°C after 5 min Pseudomonas putida

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
1.4.3.16 0.23
-
L-aspartate mutant enzyme R290A, at pH 7.5, temperature not specified in the publication Pseudomonas putida
1.4.3.16 2.28
-
L-aspartate mutant enzyme Q242A, at pH 7.5, temperature not specified in the publication Pseudomonas putida
1.4.3.16 7.83
-
L-aspartate mutant enzyme S389A, at pH 7.5, temperature not specified in the publication Pseudomonas putida
1.4.3.16 10.6
-
L-aspartate wild type enzyme, at pH 7.5, temperature not specified in the publication Pseudomonas putida
1.4.3.16 23.69
-
L-aspartate mutant enzyme H244A, at pH 7.5, temperature not specified in the publication Pseudomonas putida

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
1.4.3.16 7.4
-
-
Pseudomonas putida

pH Range

EC Number pH Minimum pH Maximum Comment Organism
1.4.3.16 6 8 about 40% activity at pH 6.0, about 50% activity at pH 8.0 Pseudomonas putida

Cofactor

EC Number Cofactor Comment Organism Structure
1.4.3.16 FAD
-
Pseudomonas putida

kcat/KM [mM/s]

EC Number kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
1.4.3.16 0.00884
-
L-aspartate mutant enzyme R290A, at pH 7.5, temperature not specified in the publication Pseudomonas putida
1.4.3.16 0.036
-
L-aspartate mutant enzyme Q242A, at pH 7.5, temperature not specified in the publication Pseudomonas putida
1.4.3.16 0.314
-
L-aspartate mutant enzyme H244A, at pH 7.5, temperature not specified in the publication Pseudomonas putida
1.4.3.16 0.549
-
L-aspartate mutant enzyme S389A, at pH 7.5, temperature not specified in the publication Pseudomonas putida
1.4.3.16 4.69
-
L-aspartate wild type enzyme, at pH 7.5, temperature not specified in the publication Pseudomonas putida