Any feedback?
Please rate this page
(literature.php)
(0/150)

BRENDA support

Literature summary for 3.5.1.15 extracted from

  • Le Coq, J.; An, H.J.; Lebrilla, C.; Viola, R.E.
    Characterization of human aspartoacylase: the brain enzyme responsible for Canavan disease (2006), Biochemistry, 45, 5878-5884.
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

Cloned (Comment) Organism
stable expression of wild-type and mutant enzyme in Pichia pastoris, expression in Escherichia coli strain XL10 primarily in inclusion bodies, while the expression yields are lower in Pichia pastoris than in Escherichia coli, the purified enzyme is significantly more stable, the enzyme form has the same substrate specificity but is 150fold more active than the Escherichia coli-expressed enzyme Homo sapiens

Protein Variants

Protein Variants Comment Organism
N117Q site-directed mutagenesis, the mutant enzyme is less stable than the wild-type enzyme, while it shows similar catalytic properties and substrate specificity Homo sapiens

General Stability

General Stability Organism
the Escherichia coli-expressed enzyme form is quite unstable, losing a significant fraction of its catalytic activity within 24 h Homo sapiens

Inhibitors

Inhibitors Comment Organism Structure
N-acetylaspartic acid substrate inhibition at concentration above 0.2 mM Homo sapiens

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
additional information
-
additional information the enzyme shows sigmoidal behaviour and cooperative substrate bindng at low substrate concentration of 0.05-0.2 mM Homo sapiens
0.12
-
N-acetylaspartic acid pH 7.2, recombinant Pichia pastoris-expressed enzyme Homo sapiens
0.15
-
N-trifluoroacetylaspartic acid pH 7.2, recombinant Pichia pastoris-expressed enzyme Homo sapiens
0.21
-
N-trifluoroacetylaspartic acid pH 7.2, recombinant Escherichia coli-expressed enzyme Homo sapiens
0.36
-
N-acetylaspartic acid pH 7.2, recombinant Escherichia coli-expressed enzyme Homo sapiens

Metals/Ions

Metals/Ions Comment Organism Structure
Zn2+ required for activity, spectroscopical determination of metal content, the enzyme contains about 1 Zn2+ per enzyme subunit Homo sapiens

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
36000
-
2 * 36000, recombinant enzyme, SDS-PAGE Homo sapiens
73000
-
dynamic light scattering study Homo sapiens

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
N-acetylaspartic acid + H2O Homo sapiens enzyme mutations cause the Canavan disease L-asparatate + acetate
-
?

Organism

Organism UniProt Comment Textmining
Homo sapiens
-
-
-

Oxidation Stability

Oxidation Stability Organism
the Pichia pastoris-expressed enzyme shows sensitivity to oxidation over time and will precipitate if not kept under the proper reducing conditions, addition of 1 mM DTT reverses the precipitated state of the enzyme with no significant loss of activity Homo sapiens

Posttranslational Modification

Posttranslational Modification Comment Organism
glycoprotein carbohydrate determination by mass spectrometry, overview, the N-glycosylation site is N117 Homo sapiens

Purification (Commentary)

Purification (Comment) Organism
recombinant enzyme by metal affinity chromatography, dialysis, and anion exchange chromatography Homo sapiens

Renatured (Commentary)

Renatured (Comment) Organism
the Pichia pastoris-expressed wild-type enzyme shows sensitivity to oxidation over time and will precipitate if not kept under the proper reducing conditions, addition of 1 mM DTT reverses the precipitated state of the enzyme with no significant loss of activity Homo sapiens

Source Tissue

Source Tissue Comment Organism Textmining
brain
-
Homo sapiens
-

Storage Stability

Storage Stability Organism
4°C, purified wild-type enzyme, expressed Homo sapiens
the purified enzyme expressed in Pichia pastoris is significantly more stable than the Escherichia coli-expressed enzyme, losing less than 10% of its catalytic activity after 2 weeks when stored under conditions where the Escherichia coli-expressed enzyme becomes completely inactivated within 24 h, the Pichia pastoris-expressed enzyme shows sensitivity to oxidation over time and will precipitate if not kept under the proper reducing conditions Homo sapiens

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
N-acetylaspartic acid + H2O
-
Homo sapiens L-asparatate + acetate
-
?
N-acetylaspartic acid + H2O enzyme mutations cause the Canavan disease Homo sapiens L-asparatate + acetate
-
?
N-trifluoroacetyl-L-aspartate + H2O
-
Homo sapiens L-aspartate + trifluoroacetate
-
?

Subunits

Subunits Comment Organism
dimer 2 * 36000, recombinant enzyme, SDS-PAGE Homo sapiens
More quarternary structure, dynamic light scattering Homo sapiens

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
0.083
-
N-acetylaspartic acid pH 7.2, recombinant Escherichia coli-expressed enzyme Homo sapiens
1.2
-
N-trifluoroacetylaspartic acid pH 7.2, recombinant Escherichia coli-expressed enzyme Homo sapiens
12.7
-
N-acetylaspartic acid pH 7.2, recombinant Pichia pastoris-expressed enzyme Homo sapiens
116
-
N-trifluoroacetylaspartic acid pH 7.2, recombinant Pichia pastoris-expressed enzyme Homo sapiens

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
7.2
-
assay at Homo sapiens