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Literature summary 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)

EC Number Cloned (Comment) Organism
3.5.1.15 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

EC Number Protein Variants Comment Organism
3.5.1.15 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

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

Inhibitors

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

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
3.5.1.15 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
3.5.1.15 0.12
-
 N-acetylaspartic acid pH 7.2, recombinant Pichia pastoris-expressed enzyme Homo sapiens
3.5.1.15 0.15
-
 N-trifluoroacetylaspartic acid pH 7.2, recombinant Pichia pastoris-expressed enzyme Homo sapiens
3.5.1.15 0.21
-
 N-trifluoroacetylaspartic acid pH 7.2, recombinant Escherichia coli-expressed enzyme Homo sapiens
3.5.1.15 0.36
-
 N-acetylaspartic acid pH 7.2, recombinant Escherichia coli-expressed enzyme Homo sapiens

Metals/Ions

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

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
3.5.1.15 36000
-
 2 * 36000, recombinant enzyme, SDS-PAGE Homo sapiens
3.5.1.15 73000
-
 dynamic light scattering study Homo sapiens

Natural Substrates/ Products (Substrates)

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

Organism

EC Number Organism UniProt Comment Textmining
3.5.1.15 Homo sapiens
-
-
-

Oxidation Stability

EC Number Oxidation Stability Organism
3.5.1.15 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

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

Purification (Commentary)

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

Renatured (Commentary)

EC Number Renatured (Comment) Organism
3.5.1.15 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

EC Number Source Tissue Comment Organism Textmining
3.5.1.15 brain
-
 Homo sapiens
-

Storage Stability

EC Number Storage Stability Organism
3.5.1.15 4°C, purified wild-type enzyme, expressed Homo sapiens
3.5.1.15 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)

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

Subunits

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

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
3.5.1.15 0.083
-
 N-acetylaspartic acid pH 7.2, recombinant Escherichia coli-expressed enzyme Homo sapiens
3.5.1.15 1.2
-
 N-trifluoroacetylaspartic acid pH 7.2, recombinant Escherichia coli-expressed enzyme Homo sapiens
3.5.1.15 12.7
-
 N-acetylaspartic acid pH 7.2, recombinant Pichia pastoris-expressed enzyme Homo sapiens
3.5.1.15 116
-
 N-trifluoroacetylaspartic acid pH 7.2, recombinant Pichia pastoris-expressed enzyme Homo sapiens

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
3.5.1.15 7.2
-
 assay at Homo sapiens