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Literature summary extracted from

  • Yamada, T.; Akutsu, N.; Miyazaki, K.; Kakinuma, K.; Yoshida, M.; Oshima, T.
    Purification, catalytic properties, and thermal stability of threo-Ds-3-isopropylmalate dehydrogenase coded by leuB gene from an extreme thermophile, Thermus thermophilus strain HB8 (1990), J. Biochem., 108, 449-456.
    View publication on PubMed

Crystallization (Commentary)

EC Number Crystallization (Comment) Organism
1.1.1.85 recombinant enzyme expressed in Escherichia coli, ammonium sulfate precipitation Thermus thermophilus

General Stability

EC Number General Stability Organism
1.1.1.85 complete loss of activity in presence of 4 M urea Thermus thermophilus

Inhibitors

EC Number Inhibitors Comment Organism Structure
1.1.1.85 1,2-Cyclohexanediamine-N,N,N',N'-tetraacetate
-
 Thermus thermophilus

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
1.1.1.85 0.08
-
 threo-Ds-3-Isopropylmalate 60°C Thermus thermophilus
1.1.1.85 0.63
-
 NAD+ 60°C Thermus thermophilus

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
1.1.1.85 K+ in 1.1 M potassium phosphate buffer, pH 7.6, the activity is over 20times that in 0.1 M sodium phosphate. Optimal KCl concentration is above 2 M, enhances activity 20times Thermus thermophilus
1.1.1.85 Mg2+ divalent cation required, most active in presence of 0.1 mM Mn2+ or 1 mM Mg2+ Thermus thermophilus
1.1.1.85 Mn2+ divalent cation required, most active in presence of 0.1 mM Mn2+ or 1 mM Mg2+ Thermus thermophilus
1.1.1.85 NH4+ NH4Cl, optimal concentration is about 0.4 M, enhances activity 18times Thermus thermophilus
1.1.1.85 Rb+ RbCl enhances activity Thermus thermophilus

Organism

EC Number Organism UniProt Comment Textmining
1.1.1.85 Thermus thermophilus
-
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
1.1.1.85 gene cloned in Escherichia coli Thermus thermophilus

Renatured (Commentary)

EC Number Renatured (Comment) Organism
1.1.1.85 enzyme solution containing 3.2 M urea is diluted 10times with a urea-free buffer. In samples containing 0.2 mM MnCl2 the activity is restored to 55-60%, full activity is recovered in absence of MnCl2 Thermus thermophilus

Storage Stability

EC Number Storage Stability Organism
1.1.1.85 -20°C, stable for at least 6 months when frozen quickly, 40% inactivation when frozen slowly Thermus thermophilus
1.1.1.85 4°C , 50 mM potassium phosphate buffer pH 7.6, 0.5 mM EDTA, 6 months, stable Thermus thermophilus

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
1.1.1.85 (2R,3S)-3-isopropylmalate + NAD+
-
 Thermus thermophilus 2-oxoisocaproate + NADH + H+ + CO2
-
 ?

Subunits

EC Number Subunits Comment Organism
1.1.1.85 dimer 2 * 35000-40000, SDS-PAGE Thermus thermophilus

Temperature Stability [°C]

EC Number Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
1.1.1.85 additional information
-
 high concentrations of KCl considerably protect the enzyme from irreversible thermal denaturation Thermus thermophilus
1.1.1.85 87
-
 denaturation temperature is higher than 87°C Thermus thermophilus

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
1.1.1.85 7.2
-
 at 75°C, in presence of K+ Thermus thermophilus
1.1.1.85 9
-
 75°C, without K+ Thermus thermophilus

Cofactor

EC Number Cofactor Comment Organism Structure
1.1.1.85 NAD+
-
 Thermus thermophilus