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Literature summary extracted from

  • Aoshima, M.; Oshima, T.
    Stabilization of Escherichia coli isopropylmalate dehydrogenase by single amino acid substitution (1997), Protein Eng., 10, 249-254.
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
1.1.1.85 the sequence motif is introduced into a mesophilic Escherichia coli isopropylmalate dehydrogenase, one by one. The introduction of the whole motif leads the mesophilic enzyme to be more unstable whereas substitution of only one amino acid residue in the motif thermostabilizes the enzyme Thermus thermophilus

Protein Variants

EC Number Protein Variants Comment Organism
1.1.1.85 L204F higher thermostability compared to wild-type enzyme, denaturation rates at 68°C and 70°C are slower Thermus thermophilus

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+ Thermus thermophilus key enzyme in leucine biosynthesis 2-oxoisocaproate + NADH + H+ + CO2
-
 ?

Organism

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

Purification (Commentary)

EC Number Purification (Comment) Organism
1.1.1.85 gene cloned in Escherichia coli 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
-
 ?
1.1.1.85 (2R,3S)-3-isopropylmalate + NAD+
-
 Escherichia coli 2-oxoisocaproate + NADH + H+ + CO2
-
 ?
1.1.1.85 (2R,3S)-3-isopropylmalate + NAD+ key enzyme in leucine biosynthesis Thermus thermophilus 2-oxoisocaproate + NADH + H+ + CO2
-
 ?

Temperature Stability [°C]

EC Number Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
1.1.1.85 additional information
-
 mutant enzyme L204F shows higher thermostability compared to wild-type enzyme, denaturation rates at 68°C and 70°C are slower for the mutant enzyme than for wild-type enzyme Thermus thermophilus
1.1.1.85 87
-
 the thermophilic IMPDH was succesfully stabilized by replacing Ala172 with more hydrophobic residues like Glu, Asn and Gln without significant change in the enzymatic activity Thermus thermophilus

Cofactor

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