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Literature summary for 1.1.1.85 extracted from

  • Ohkuri, T.; Yamagishi, A.
    The effects of mutations at position 253 on the thermostability of the Bacillus subtilis 3-isopropylmalate dehydrogenase subunit interface (2007), J. Biochem., 141, 791-797.
    View publication on PubMed

Protein Variants

Protein Variants Comment Organism
E253A specific activity: 4.7 unit/mg, Km: 0.016 mM (3-isopropylmalate), 0.246 mM (NAD), kcat: 14.9/sec Bacillus subtilis
E253F specific activity: 7.6 unit/mg Bacillus subtilis
E253G specific activity: 5.9 unit/mg Bacillus subtilis
E253I specific activity: 7.4 unit/mg Bacillus subtilis
E253K when a lysine is the position 253 the protein dissociates Bacillus subtilis
E253L specific activity: 7.6 unit/mg, Km: 0.0122 mM (3-isopropylmalate), 0.474 mM (NAD), kcat: 13.7/sec Bacillus subtilis
E253Q specific activity: 7 unit/mg, Km: 0.0189 mM (3-isopropylmalate), 0.273 mM (NAD), kcat: 9.9/sec. this mutant is more thermostable than the other mutants Bacillus subtilis
E253V specific activity: 9 unit/mg, Km: 0.0244 mM (3-isopropylmalate), 0.171 mM (NAD), kcat: 12.4/sec Bacillus subtilis
E253W specific activity: 5.2 unit/mg, Km: 0.0254 mM (3-isopropylmalate), 0.244 mM (NAD), kcat: 8.3/sec Bacillus subtilis
additional information results show that, for residues with non-polar side chains, when positioned at residue 253, the thermostabilities of their corresponding mutants correlate positively with the relative hydrophobicities of the side chains Bacillus subtilis

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.009
-
3-isopropylmalate wild-type Bacillus subtilis
0.0122
-
3-isopropylmalate mutant Glu253Leu Bacillus subtilis
0.016
-
3-isopropylmalate mutant Glu253Ala Bacillus subtilis
0.0189
-
3-isopropylmalate mutant Glu253Gln Bacillus subtilis
0.0244
-
3-isopropylmalate mutant Glu253Val Bacillus subtilis
0.0254
-
3-isopropylmalate mutant Glu253Trp Bacillus subtilis
0.171
-
NAD+ mutant Glu253Val Bacillus subtilis
0.244
-
NAD+ mutant Glu253Trp Bacillus subtilis
0.246
-
NAD+ wild-type Bacillus subtilis
0.246
-
NAD+ mutant Glu253Ala Bacillus subtilis
0.273
-
NAD+ mutant Glu253Gln Bacillus subtilis
0.474
-
NAD+ mutant Glu253Leu Bacillus subtilis

Metals/Ions

Metals/Ions Comment Organism Structure
K+
-
Bacillus subtilis
Mn2+
-
Bacillus subtilis

Organism

Organism UniProt Comment Textmining
Bacillus subtilis
-
-
-

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
4.7
-
mutant E253A Bacillus subtilis
5.2
-
mutant E253W Bacillus subtilis
5.9
-
mutant E253G Bacillus subtilis
7
-
mutant E253Q Bacillus subtilis
7.4
-
mutant E253I Bacillus subtilis
7.6
-
mutant E253F Bacillus subtilis
7.6
-
mutant E253L Bacillus subtilis
9
-
mutant E253V Bacillus subtilis
13.9
-
wild-type Bacillus subtilis

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
3-isopropylmalate + NAD+
-
Bacillus subtilis 2-ketoisocaproate + NADH + CO2
-
?

Subunits

Subunits Comment Organism
dimer
-
Bacillus subtilis

Synonyms

Synonyms Comment Organism
3-isopropylmalate dehydrogenase
-
Bacillus subtilis
3-isopropylmalateDH
-
Bacillus subtilis
Bs3-isopropylmalateDH
-
Bacillus subtilis

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
40
-
assay at Bacillus subtilis

Temperature Stability [°C]

Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
additional information
-
all mutant Bs3-isopropylmalateDHs and the wild-type enzyme are irreversibly inactivated by heat treatments. A linear correlation between thermal stability and hydrophobicity exists for those 3-isopropylmalateDHs containing position 253 residues with hydrophobic side chains Bacillus subtilis

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
8.3
-
3-isopropylmalate mutant Glu253Trp Bacillus subtilis
9.5
-
3-isopropylmalate mutant Glu253Ala Bacillus subtilis
9.9
-
3-isopropylmalate mutant Glu253Gln Bacillus subtilis
12.4
-
3-isopropylmalate mutant Glu253Val Bacillus subtilis
13.7
-
3-isopropylmalate mutant Glu253Leu Bacillus subtilis
14.9
-
3-isopropylmalate wild-type Bacillus subtilis

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
7.6
-
assay at Bacillus subtilis