BRENDA - Enzyme Database

Overexpression, purification, crystallization and data collection of 3-methylaspartase from Clostridium tetanomorphum

Asuncion, M.; Barlow, J.N.; Pollard, J.; Staines, A.G.; McMahon, S.A.; Blankenfeldt, W.; Gani, D.; Naismith, J.H.; Acta Crystallogr. Sect. D 57, 731-733 (2001)

Data extracted from this reference:

Cloned(Commentary)
EC Number
Commentary
Organism
4.3.1.2
expression in Escherichia coli
Clostridium tetanomorphum
Crystallization (Commentary)
EC Number
Crystallization
Organism
4.3.1.2
sitting-drop vapour-diffusion, 0.0045 ml protein sample and 0.0045 ml precipitant, crystals are obtained using 20-25% polyethylene glycol 6000, 100 mM sodium acetate, pH 7.0, 25 mM Tris-HCl as precipitant solution and 16-22% ethylene glycol as additive, crystals diffract to 2.0 A
Clostridium tetanomorphum
KM Value [mM]
EC Number
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
4.3.1.2
0.58
-
L-threo-3-methylaspartate
recombinant 3-methylaspartase
Clostridium tetanomorphum
Molecular Weight [Da]
EC Number
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
4.3.1.2
45538
-
2 * 45538, mass spectrometry
Clostridium tetanomorphum
Natural Substrates/ Products (Substrates)
EC Number
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
4.3.1.2
L-erythro-3-methylaspartate
Clostridium tetanomorphum
i.e. (2S,3R)-3-methylaspartate, slow syn elimination
mesaconate + NH3
-
Clostridium tetanomorphum
r
4.3.1.2
L-threo-3-methylaspartate
Clostridium tetanomorphum
-
mesaconate + NH3
-
Clostridium tetanomorphum
r
Organism
EC Number
Organism
Primary Accession No. (UniProt)
Commentary
Textmining
4.3.1.2
Clostridium tetanomorphum
-
-
-
Purification (Commentary)
EC Number
Commentary
Organism
4.3.1.2
recombinant 3-methylaspartase
Clostridium tetanomorphum
Substrates and Products (Substrate)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
4.3.1.2
L-erythro-3-methylaspartate
i.e. (2S,3R)-3-methylaspartate, slow syn elimination
649166
Clostridium tetanomorphum
mesaconate + NH3
-
649166
Clostridium tetanomorphum
?
4.3.1.2
L-erythro-3-methylaspartate
i.e. (2S,3R)-3-methylaspartate, slow syn elimination
649166
Clostridium tetanomorphum
mesaconate + NH3
-
649166
Clostridium tetanomorphum
r
4.3.1.2
L-threo-3-methylaspartate
-
649166
Clostridium tetanomorphum
mesaconate + NH3
-
649166
Clostridium tetanomorphum
r
4.3.1.2
L-threo-3-methylaspartate
i.e. (2S,3S)-3-methylaspartate
649166
Clostridium tetanomorphum
mesaconate + NH3
-
649166
Clostridium tetanomorphum
r
Subunits
EC Number
Subunits
Commentary
Organism
4.3.1.2
dimer
2 * 45538, mass spectrometry
Clostridium tetanomorphum
Turnover Number [1/s]
EC Number
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
Organism
Structure
4.3.1.2
240
-
L-threo-3-methylaspartate
recombinant 3-methylaspartase
Clostridium tetanomorphum
Cloned(Commentary) (protein specific)
EC Number
Commentary
Organism
4.3.1.2
expression in Escherichia coli
Clostridium tetanomorphum
Crystallization (Commentary) (protein specific)
EC Number
Crystallization
Organism
4.3.1.2
sitting-drop vapour-diffusion, 0.0045 ml protein sample and 0.0045 ml precipitant, crystals are obtained using 20-25% polyethylene glycol 6000, 100 mM sodium acetate, pH 7.0, 25 mM Tris-HCl as precipitant solution and 16-22% ethylene glycol as additive, crystals diffract to 2.0 A
Clostridium tetanomorphum
KM Value [mM] (protein specific)
EC Number
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
4.3.1.2
0.58
-
L-threo-3-methylaspartate
recombinant 3-methylaspartase
Clostridium tetanomorphum
Molecular Weight [Da] (protein specific)
EC Number
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
4.3.1.2
45538
-
2 * 45538, mass spectrometry
Clostridium tetanomorphum
Natural Substrates/ Products (Substrates) (protein specific)
EC Number
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
4.3.1.2
L-erythro-3-methylaspartate
Clostridium tetanomorphum
i.e. (2S,3R)-3-methylaspartate, slow syn elimination
mesaconate + NH3
-
Clostridium tetanomorphum
r
4.3.1.2
L-threo-3-methylaspartate
Clostridium tetanomorphum
-
mesaconate + NH3
-
Clostridium tetanomorphum
r
Purification (Commentary) (protein specific)
EC Number
Commentary
Organism
4.3.1.2
recombinant 3-methylaspartase
Clostridium tetanomorphum
Substrates and Products (Substrate) (protein specific)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
4.3.1.2
L-erythro-3-methylaspartate
i.e. (2S,3R)-3-methylaspartate, slow syn elimination
649166
Clostridium tetanomorphum
mesaconate + NH3
-
649166
Clostridium tetanomorphum
?
4.3.1.2
L-erythro-3-methylaspartate
i.e. (2S,3R)-3-methylaspartate, slow syn elimination
649166
Clostridium tetanomorphum
mesaconate + NH3
-
649166
Clostridium tetanomorphum
r
4.3.1.2
L-threo-3-methylaspartate
-
649166
Clostridium tetanomorphum
mesaconate + NH3
-
649166
Clostridium tetanomorphum
r
4.3.1.2
L-threo-3-methylaspartate
i.e. (2S,3S)-3-methylaspartate
649166
Clostridium tetanomorphum
mesaconate + NH3
-
649166
Clostridium tetanomorphum
r
Subunits (protein specific)
EC Number
Subunits
Commentary
Organism
4.3.1.2
dimer
2 * 45538, mass spectrometry
Clostridium tetanomorphum
Turnover Number [1/s] (protein specific)
EC Number
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
Organism
Structure
4.3.1.2
240
-
L-threo-3-methylaspartate
recombinant 3-methylaspartase
Clostridium tetanomorphum