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Literature summary for 1.4.3.16 extracted from

  • Tedeschi, G.; Nonnis, S.; Strumbo, B.; Cruciani, G.; Carosati, E.; Negri, A.
    On the catalytic role of the active site residue E121 of E. coli L-aspartate oxidase (2010), Biochimie, 92, 1335-1342.
    View publication on PubMed

Cloned(Commentary)

Cloned (Comment) Organism
expression of wild-type and mutant enzymes in Escherichia coli strain BL21(DE3) Escherichia coli

Protein Variants

Protein Variants Comment Organism
E121A site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme, catalytically inactive against either 3-OH-erythro- or 3-OH-threo-L-aspartate, but is active with N-acetyl- and N-formyl-L-aspartate Escherichia coli
E121D site-directed mutagenesis, the mutant shows highly reduced activity compared to the wild-type enzyme, catalytically inactive against either 3-OH-erythro- or 3-OH-threo-L-aspartate Escherichia coli
E121K site-directed mutagenesis, the mutant shows highly reduced activity compared to the wild-type enzyme, catalytically inactive against either 3-OH-erythro- or 3-OH-threo-L-aspartate Escherichia coli
E121Q site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme, catalytically inactive against either 3-OH-erythro- or 3-OH-threo-L-aspartate Escherichia coli

Inhibitors

Inhibitors Comment Organism Structure
D-Aspartate
-
Escherichia coli

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.038
-
L-aspartate pH 8.0, 25°C, recombinant mutant E121A Escherichia coli
0.048
-
L-aspartate pH 8.0, 25°C, recombinant wild-type enzyme Escherichia coli
0.61
-
L-aspartate pH 8.0, 25°C, recombinant mutant E121Q Escherichia coli
2.9
-
L-aspartate pH 8.0, 25°C, recombinant mutant E121K Escherichia coli
3.2
-
N-acetyl-L-aspartate pH 8.0, 25°C, recombinant mutant E121A Escherichia coli
3.7
-
N-formyl-L-aspartate pH 8.0, 25°C, recombinant wild-type enzyme Escherichia coli
4.7
-
3-hydroxy-erythro-L-aspartate pH 8.0, 25°C, recombinant wild-type enzyme Escherichia coli
17
-
N-acetyl-L-aspartate pH 8.0, 25°C, recombinant wild-type enzyme Escherichia coli
17.5
-
N-formyl-L-aspartate pH 8.0, 25°C, recombinant mutant E121A Escherichia coli
26.8
-
L-aspartate pH 8.0, 25°C, recombinant mutant E121D Escherichia coli

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
L-aspartate + fumarate Escherichia coli
-
iminosuccinate + succinate
-
?
L-aspartate + O2 Escherichia coli
-
iminosuccinate + H2O2
-
?
additional information Escherichia coli the enzyme oxidizes L-aspartate both under aerobic and anaerobic conditions using oxygen as well as fumarate as electron acceptor. No activity with 3-hydroxy-threo-L-aspartate ?
-
?

Organism

Organism UniProt Comment Textmining
Escherichia coli
-
-
-

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
3-hydroxy-erythro-L-aspartate + O2
-
Escherichia coli 2-amino-3-hydroxy-2-butenedioic acid + H2O2
-
?
L-aspartate + fumarate
-
Escherichia coli iminosuccinate + succinate
-
?
L-aspartate + O2
-
Escherichia coli iminosuccinate + H2O2
-
?
additional information the enzyme oxidizes L-aspartate both under aerobic and anaerobic conditions using oxygen as well as fumarate as electron acceptor. No activity with 3-hydroxy-threo-L-aspartate Escherichia coli ?
-
?
additional information the enzyme oxidizes L-aspartate both under aerobic and anaerobic conditions using oxygen as well as fumarate as electron acceptor. Catalytic role of the active site residue E121, substrate specificity of wild-type and mutant enzymes, molecular docking studies, role of R290, overview. E121 interacts favourably with the charged amino group of the substrate and different ligands might assume different orientations in the active site of the enzyme, binding modes for L-aspartate, overview Escherichia coli ?
-
?
N-acetyl-L-aspartate + O2
-
Escherichia coli ? + H2O2
-
?
N-formyl-L-aspartate + O2
-
Escherichia coli ? + H2O2
-
?

Subunits

Subunits Comment Organism
More three dimensional structure of holo-wild-type-LASPO, modelling, overview Escherichia coli

Synonyms

Synonyms Comment Organism
LASPO
-
Escherichia coli
More LASPO belongs to the succinate dehydrogenase/fumarate reductase family Escherichia coli

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
25
-
assay at Escherichia coli

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
0.0013
-
N-formyl-L-aspartate pH 8.0, 25°C, recombinant mutant E121A Escherichia coli
0.0035
-
N-acetyl-L-aspartate pH 8.0, 25°C, recombinant wild-type enzyme Escherichia coli
0.0055
-
N-acetyl-L-aspartate pH 8.0, 25°C, recombinant mutant E121A Escherichia coli
0.0077
-
N-formyl-L-aspartate pH 8.0, 25°C, recombinant wild-type enzyme Escherichia coli
0.173
-
3-hydroxy-erythro-L-aspartate pH 8.0, 25°C, recombinant wild-type enzyme Escherichia coli
0.173
-
L-aspartate pH 8.0, 25°C, recombinant mutant E121A Escherichia coli
0.202
-
L-aspartate pH 8.0, 25°C, recombinant mutant E121Q Escherichia coli
0.215
-
L-aspartate pH 8.0, 25°C, recombinant mutant E121D Escherichia coli
0.223
-
L-aspartate pH 8.0, 25°C, recombinant mutant E121K Escherichia coli
0.267
-
L-aspartate pH 8.0, 25°C, recombinant wild-type enzyme Escherichia coli

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
8
-
assay at Escherichia coli

Cofactor

Cofactor Comment Organism Structure
FAD E121 belongs to a polypeptide stretch, residues 119-127, involved in the interdomain contact between the FAD and the capping domains and is hydrogen bound to G51, itself belonging to a loop of residues 49-57 playing a key role in coenzyme binding Escherichia coli

General Information

General Information Comment Organism
additional information LASPO displays strong primary and tertiary structure similarity with the flavin containing subunit of the proteins belonging to the succinate dehydrogenase/fumarate reductase family. The similarity extends to the active site residues, with LASPO differing from the other enzymes of the family only for the presence of a conserved glutamate 121, which is substituted by apolar amino acids in the other enzymes Escherichia coli
physiological function LASPO is a flavoenzyme catalyzing the first step in the de novo biosynthesis of NAD+. The enzyme oxidizes L-aspartate both under aerobic and anaerobic conditions using oxygen as well as fumarate as electron acceptor Escherichia coli

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
0.00007
-
N-formyl-L-aspartate pH 8.0, 25°C, recombinant mutant E121A Escherichia coli
0.00021
-
N-acetyl-L-aspartate pH 8.0, 25°C, recombinant wild-type enzyme Escherichia coli
0.0017
-
N-acetyl-L-aspartate pH 8.0, 25°C, recombinant mutant E121A Escherichia coli
0.00208
-
N-formyl-L-aspartate pH 8.0, 25°C, recombinant wild-type enzyme Escherichia coli
0.008
-
L-aspartate pH 8.0, 25°C, recombinant mutant E121D Escherichia coli
0.0368
-
3-hydroxy-erythro-L-aspartate pH 8.0, 25°C, recombinant wild-type enzyme Escherichia coli
0.077
-
L-aspartate pH 8.0, 25°C, recombinant mutant E121K Escherichia coli
0.331
-
L-aspartate pH 8.0, 25°C, recombinant mutant E121Q Escherichia coli
4.55
-
L-aspartate pH 8.0, 25°C, recombinant mutant E121A Escherichia coli
5.56
-
L-aspartate pH 8.0, 25°C, recombinant wild-type enzyme Escherichia coli