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Literature summary for 1.1.1.85 extracted from

  • Watanabe, K.; Ohkuri, T.; Yokobori, S.; Yamagishi, A.
    Designing thermostable proteins: ancestral mutants of 3-isopropylmalate dehydrogenase designed by using a phylogenetic tree (2006), J. Mol. Biol., 355, 664-674.
    View publication on PubMed

Cloned(Commentary)

Cloned (Comment) Organism
gene leuB, DNA and amino acid sequence determination and analysis, phylogenetic analysis, expression of wild-type and mutants in Escherichia coli strain MA153 Thermus thermophilus

Protein Variants

Protein Variants Comment Organism
A335E site-directed mutagenesis, exchange of a cofactor binding residue to the residue of the ancestral mutant, the mutant shows increased thermal stability compared to the wild-type enzyme Thermus thermophilus
D184H site-directed mutagenesis, exchange of a cofactor binding residue to the residue of the ancestral mutant, the mutant shows increased thermal stability compared to the wild-type enzyme Thermus thermophilus
F53L site-directed mutagenesis, exchange of a cofactor binding residue to the residue of the ancestral mutant, the mutant shows decreased thermal stability compared to the wild-type enzyme Thermus thermophilus
H179K site-directed mutagenesis, exchange of a cofactor binding residue to the residue of the ancestral mutant, the mutant shows decreased thermal stability compared to the wild-type enzyme Thermus thermophilus
L134N site-directed mutagenesis, exchange of a cofactor binding residue to the residue of the ancestral mutant, the mutant shows increased thermal stability compared to the wild-type enzyme Thermus thermophilus
additional information designing thermostable 3-isopropylmalate dehydrogenase designed by using a phylogenetic tree and ancestral mutant sequences, overview Thermus thermophilus
P324T site-directed mutagenesis, exchange of a cofactor binding residue to the residue of the ancestral mutant, the mutant shows increased thermal stability compared to the wild-type enzyme Thermus thermophilus
P56E site-directed mutagenesis, exchange of a cofactor binding residue to the residue of the ancestral mutant, the mutant shows increased thermal stability compared to the wild-type enzyme Thermus thermophilus
R58L site-directed mutagenesis, exchange of a cofactor binding residue to the residue of the ancestral mutant, the mutant shows decreased thermal stability compared to the wild-type enzyme Thermus thermophilus
S261N site-directed mutagenesis, exchange of a cofactor binding residue to the residue of the ancestral mutant, the mutant shows decreased thermal stability compared to the wild-type enzyme Thermus thermophilus
T16V site-directed mutagenesis, exchange of a cofactor binding residue to the residue of the ancestral mutant, the mutant shows increased thermal stability compared to the wild-type enzyme Thermus thermophilus
V181T site-directed mutagenesis, exchange of a cofactor binding residue to the residue of the ancestral mutant, the mutant shows increased thermal stability compared to the wild-type enzyme Thermus thermophilus
V61I site-directed mutagenesis, exchange of a cofactor binding residue to the residue of the ancestral mutant, the mutant shows decreased thermal stability compared to the wild-type enzyme Thermus thermophilus

Metals/Ions

Metals/Ions Comment Organism Structure
Mg2+
-
Thermus thermophilus

Organism

Organism UniProt Comment Textmining
Thermus thermophilus Q5SIY4 gene leuB
-

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
threo-DL-isopropylmalate + NAD+
-
Thermus thermophilus 2-isopropyl-3-oxosuccinate + NADH
-
?

Subunits

Subunits Comment Organism
More three-dimensional structure analysis of wild-type and mutant enzymes Thermus thermophilus

Synonyms

Synonyms Comment Organism
IPMDH
-
Thermus thermophilus

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
50
-
assay at Thermus thermophilus

Temperature Stability [°C]

Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
85 95 thermal degeneration curves of wild-type and mutant enzymes at pH 7.6, overview Thermus thermophilus

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
7.6 8 assay at Thermus thermophilus

Cofactor

Cofactor Comment Organism Structure
NAD+
-
Thermus thermophilus