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Literature summary for 2.3.3.13 extracted from

  • Wiegel, J.; Schlegel, H.G.
    alpha-Isopropylmalate synthase from Alcaligenes eutrophus H 16 I. Purification and general properties (1977), Arch. Microbiol., 112, 239-246.
    View publication on PubMed

Inhibitors

Inhibitors Comment Organism Structure
K2SO4 250 mM K2SO4, pH 7.5, 20% inhibition Cupriavidus necator
KCl 400 mM KCl, pH 8.2, 20% inhibition Cupriavidus necator

Metals/Ions

Metals/Ions Comment Organism Structure
Mn2+ 1 mM, 25% activation Cupriavidus necator

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
100000
-
in presence of Leu, gel filtration Cupriavidus necator
114500
-
in absence of Leu, gel filtration Cupriavidus necator

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
2-oxo-3-methylbutanoate + acetyl-CoA + H2O Cupriavidus necator first enzyme in biosynthesis of L-Leu ?
-
?
2-oxo-3-methylbutanoate + acetyl-CoA + H2O Cupriavidus necator H 16 first enzyme in biosynthesis of L-Leu ?
-
?

Organism

Organism UniProt Comment Textmining
Cupriavidus necator
-
-
-
Cupriavidus necator
-
Val-Ile double auxotrophic mutant of strain H 16
-
Cupriavidus necator H 16
-
-
-

Purification (Commentary)

Purification (Comment) Organism
-
Cupriavidus necator

Renatured (Commentary)

Renatured (Comment) Organism
irreversible denaturation after 5 min at 60°C Cupriavidus necator

Source Tissue

Source Tissue Comment Organism Textmining
additional information highest specific activity in cells harvested in the late exponential phase Cupriavidus necator
-

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
12.05
-
-
Cupriavidus necator

Storage Stability

Storage Stability Organism
30°C, pH 7.5, stable for more than 10 days Cupriavidus necator
freezing at -20°C and thawing, about 10% loss of activity. Storage at -20°C, 100 mM potassium phosphate, 0.2 mM 2-oxo-3-methylbutanoate, pH 7.5 or pH 8.0, no further loss of activity after 5 months Cupriavidus necator

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2-oxo-3-methylbutanoate + acetyl-CoA + H2O
-
Cupriavidus necator 3-hydroxy-4-methyl-3-carboxypentanoate + CoA
-
?
2-oxo-3-methylbutanoate + acetyl-CoA + H2O
-
Cupriavidus necator H 16 3-hydroxy-4-methyl-3-carboxypentanoate + CoA
-
?
2-oxo-3-methylbutanoate + acetyl-CoA + H2O first enzyme in biosynthesis of L-Leu Cupriavidus necator ?
-
?
2-oxo-3-methylbutanoate + acetyl-CoA + H2O first enzyme in biosynthesis of L-Leu Cupriavidus necator H 16 ?
-
?

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
44
-
-
Cupriavidus necator

Temperature Stability [°C]

Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
additional information
-
cold lability can be overcome by stabilizing the enzyme by buffer of increased ionic strength Cupriavidus necator
additional information
-
stabilization at low temperature with KCl Cupriavidus necator
60
-
5 min, in absence of substrates irreversible denaturation Cupriavidus necator
65
-
5 min, in presence of 2-oxo-3-methylbutanoate and acetyl-CoA, stable Cupriavidus necator

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
8 8.5 Tris-HCl buffer Cupriavidus necator
8.2
-
phosphate buffer Cupriavidus necator

pH Stability

pH Stability pH Stability Maximum Comment Organism
7
-
3 h, about 15% of maximal activity Cupriavidus necator
7.2 8.2 3 h, stable Cupriavidus necator
9
-
3 h, about 50% loss of activity Cupriavidus necator
9.5
-
3 h, about 30% of maximal activity Cupriavidus necator