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Literature summary extracted from

  • Sasaki, M.; Uno, M.; Akanuma, S.; Yamagishi, A.
    Random mutagenesis improves the low-temperature activity of the tetrameric 3-isopropylmalate dehydrogenase from the hyperthermophile Sulfolobus tokodaii (2008), Protein Eng. Des. Sel., 21, 721-727.
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
1.1.1.85 gene leuB, expression of wild-type and mutant enzymes in Escherichia coli strain MA153 Sulfurisphaera tokodaii

Protein Variants

EC Number Protein Variants Comment Organism
1.1.1.85 F145S random mutagenesis, the mutation alters the Km values for NAD+ and/or the kcat value, the mutant shows reduced thermostability compared to the wild-type enzyme Sulfurisphaera tokodaii
1.1.1.85 F3S random mutagenesis, the mutation alters the Km values for NAD+ and/or the kcat value, the mutant shows reduced thermostability compared to the wild-type enzyme Sulfurisphaera tokodaii
1.1.1.85 F3S/L288P random mutagenesis, the mutation alters the Km values for NAD+ and/or the kcat value, the mutant shows reduced thermostability compared to the wild-type enzyme. The MA153 strain containing a doubly mutated leuB gene grows more rapidly than does Escherichia coli MA153 containing the wild-type leuB gene in leucine-free M9 minimal liquid medium Sulfurisphaera tokodaii
1.1.1.85 F3S/S232G random mutagenesis, the mutation alters the Km values for NAD+ and/or the kcat value, the mutant shows reduced thermostability compared to the wild-type enzyme. The MA153 strain containing a doubly mutated leuB gene grows more rapidly than does Escherichia coli MA153 containing the wild-type leuB gene in leucine-free M9 minimal liquid medium Sulfurisphaera tokodaii
1.1.1.85 I68L/I142T/K335E random mutagenesis, the mutation alters the Km values for NAD+ and/or the kcat value, the mutant shows reduced thermostability compared to the wild-type enzyme Sulfurisphaera tokodaii
1.1.1.85 L288P random mutagenesis, the mutation alters the Km values for NAD+ and/or the kcat value, the mutant shows reduced thermostability compared to the wild-type enzyme Sulfurisphaera tokodaii
1.1.1.85 L328S random mutagenesis, the mutation alters the Km values for NAD+ and/or the kcat value, the mutant shows reduced thermostability compared to the wild-type enzyme Sulfurisphaera tokodaii
1.1.1.85 L336P random mutagenesis, the mutation alters the Km values for NAD+ and/or the kcat value, the mutant shows reduced thermostability compared to the wild-type enzyme Sulfurisphaera tokodaii
1.1.1.85 R284G random mutagenesis, the mutation alters the Km values for NAD+ and/or the kcat value, the mutant shows reduced thermostability compared to the wild-type enzyme Sulfurisphaera tokodaii
1.1.1.85 Y309H random mutagenesis, the mutation alters the Km values for NAD+ and/or the kcat value, the mutant shows reduced thermostability compared to the wild-type enzyme Sulfurisphaera tokodaii

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
1.1.1.85 additional information
-
 additional information Michaelis-Menten kinetics of recombinant wild-type and mutant enzymes at different temperatures, overview Sulfurisphaera tokodaii
1.1.1.85 0.0039
-
 3-isopropylmalate pH 7.5, 70°C, recombinant mutant F3S/S232G Sulfurisphaera tokodaii
1.1.1.85 0.0044
-
 3-isopropylmalate pH 7.5, 70°C, recombinant mutant L288P Sulfurisphaera tokodaii
1.1.1.85 0.0055
-
 3-isopropylmalate pH 7.5, 70°C, recombinant wild-type enzyme Sulfurisphaera tokodaii
1.1.1.85 0.0067
-
 3-isopropylmalate pH 7.5, 70°C, recombinant mutant F3S/L288P Sulfurisphaera tokodaii
1.1.1.85 0.0076
-
 3-isopropylmalate pH 7.5, 70°C, recombinant mutant F3S Sulfurisphaera tokodaii
1.1.1.85 0.008
-
 3-isopropylmalate pH 7.5, 70°C, recombinant mutant R284G Sulfurisphaera tokodaii
1.1.1.85 0.22
-
 NAD+ pH 7.5, 70°C, recombinant mutant F3S/S232G Sulfurisphaera tokodaii
1.1.1.85 0.25
-
 NAD+ pH 7.5, 70°C, recombinant mutant R284G Sulfurisphaera tokodaii
1.1.1.85 0.37
-
 NAD+ pH 7.5, 70°C, recombinant mutant L288P Sulfurisphaera tokodaii
1.1.1.85 0.39
-
 NAD+ pH 7.5, 70°C, recombinant wild-type enzyme and mutant F3S Sulfurisphaera tokodaii
1.1.1.85 0.4
-
 NAD+ pH 7.5, 70°C, recombinant mutant F3S/L288P Sulfurisphaera tokodaii

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
1.1.1.85 Mg2+
-
 Sulfurisphaera tokodaii

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
1.1.1.85 (2R,3S)-3-isopropylmalate + NAD+ Sulfurisphaera tokodaii the enzyme is enzyme involved in leucine biosynthesis ?
-
 ?

Organism

EC Number Organism UniProt Comment Textmining
1.1.1.85 Sulfurisphaera tokodaii P50455 gene leuB
-

Purification (Commentary)

EC Number Purification (Comment) Organism
1.1.1.85 recombinant wild-type and mutant enzymes from Escherichia coli strain MA153 by anion exchange and hydrophobic interaction chromatography Sulfurisphaera tokodaii

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
1.1.1.85 (2R,3S)-3-isopropylmalate + NAD+
-
 Sulfurisphaera tokodaii ?
-
 ?
1.1.1.85 (2R,3S)-3-isopropylmalate + NAD+ the enzyme is enzyme involved in leucine biosynthesis Sulfurisphaera tokodaii ?
-
 ?

Subunits

EC Number Subunits Comment Organism
1.1.1.85 tetramer
-
 Sulfurisphaera tokodaii

Synonyms

EC Number Synonyms Comment Organism
1.1.1.85 IPMDH
-
 Sulfurisphaera tokodaii

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
1.1.1.85 95
-
-
 Sulfurisphaera tokodaii

Temperature Range [°C]

EC Number Temperature Minimum [°C] Temperature Maximum [°C] Comment Organism
1.1.1.85 25 95 2% of maximal activity at 25°C, maximal activity at 95°C Sulfurisphaera tokodaii

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
1.1.1.85 3.8
-
 3-isopropylmalate pH 7.5, 70°C, recombinant mutant F3S/S232G Sulfurisphaera tokodaii
1.1.1.85 6
-
 3-isopropylmalate pH 7.5, 70°C, recombinant mutant R284G Sulfurisphaera tokodaii
1.1.1.85 6.8
-
 3-isopropylmalate pH 7.5, 70°C, recombinant mutant F3S Sulfurisphaera tokodaii
1.1.1.85 6.9
-
 3-isopropylmalate pH 7.5, 70°C, recombinant wild-type enzyme Sulfurisphaera tokodaii
1.1.1.85 7.5
-
 3-isopropylmalate pH 7.5, 70°C, recombinant mutant L288P Sulfurisphaera tokodaii
1.1.1.85 7.8
-
 3-isopropylmalate pH 7.5, 70°C, recombinant mutant F3S/L288P Sulfurisphaera tokodaii

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
1.1.1.85 7.5
-
 assay at Sulfurisphaera tokodaii

Cofactor

EC Number Cofactor Comment Organism Structure
1.1.1.85 NAD+
-
 Sulfurisphaera tokodaii