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Literature summary extracted from

  • Nagai, S.; Takada, Y.
    Analysis of amino acid residues involved in the thermal properties of isocitrate dehydrogenases from a psychrophilic bacterium, Colwellia maris, and a psychrotrophic bacterium, Pseudomonas psychrophila (2020), J. Biosci. Bioeng., 129, 284-290 .
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
1.1.1.42 expressed in enzyme-defective Escherichia coli strain DEK2004 Colwellia maris
1.1.1.42 expressed in enzyme-defective Escherichia coli strain DEK2004 Pseudomonas psychrophila

Protein Variants

EC Number Protein Variants Comment Organism
1.1.1.42 A741S the mutant exhibits higher specific activity and thermostability than the wild type enzyme Colwellia maris
1.1.1.42 E538L the mutant exhibits higher thermostability than wild type enzyme Colwellia maris
1.1.1.42 E596L the mutant exhibits higher specific activity and thermostability than the wild type enzyme Colwellia maris
1.1.1.42 L536E the mutant is more thermolabile than wild type enzyme Pseudomonas psychrophila
1.1.1.42 L594E the mutant is more thermolabile than wild type enzyme and has a lower specific activity at temperatures over 45°C Pseudomonas psychrophila
1.1.1.42 S739A the mutant is more thermolabile than wild type enzyme Pseudomonas psychrophila

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
1.1.1.42 0.01558
-
 isocitrate mutant enzyme L594E, at pH 8.0 and 20°C Pseudomonas psychrophila
1.1.1.42 0.02008
-
 isocitrate wild type enzyme, at pH 8.0 and 20°C Pseudomonas psychrophila
1.1.1.42 0.04666
-
 isocitrate mutant enzyme E596L, at pH 8.0 and 20°C Colwellia maris
1.1.1.42 0.06
-
 isocitrate wild type enzyme, at pH 8.0 and 20°C Colwellia maris
1.1.1.42 0.06742
-
 isocitrate mutant enzyme A741S, at pH 8.0 and 20°C Colwellia maris

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
1.1.1.42 Mg2+ 2 mM used in assay conditions Colwellia maris
1.1.1.42 Mg2+ 2 mM used in assay conditions Pseudomonas psychrophila
1.1.1.42 Mn2+ 0.67 mM used in assay conditions Colwellia maris
1.1.1.42 Mn2+ 0.67 mM used in assay conditions Pseudomonas psychrophila

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
1.1.1.42 isocitrate + NADP+ Colwellia maris
-
 2-oxoglutarate + CO2 + NADPH + H+
-
 ?
1.1.1.42 isocitrate + NADP+ Pseudomonas psychrophila
-
 2-oxoglutarate + CO2 + NADPH + H+
-
 ?

Organism

EC Number Organism UniProt Comment Textmining
1.1.1.42 Colwellia maris
-
-
-
1.1.1.42 Pseudomonas psychrophila
-
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
1.1.1.42 Ni-NTA agarose column chromatography Colwellia maris
1.1.1.42 Ni-NTA agarose column chromatography Pseudomonas psychrophila

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
1.1.1.42 isocitrate + NADP+
-
 Colwellia maris 2-oxoglutarate + CO2 + NADPH + H+
-
 ?
1.1.1.42 isocitrate + NADP+
-
 Pseudomonas psychrophila 2-oxoglutarate + CO2 + NADPH + H+
-
 ?

Subunits

EC Number Subunits Comment Organism
1.1.1.42 monomer 1 * 80000, His-tagged enzyme, SDS-PAGE Colwellia maris
1.1.1.42 monomer 1 * 80000, His-tagged enzyme, SDS-PAGE Pseudomonas psychrophila

Synonyms

EC Number Synonyms Comment Organism
1.1.1.42 IDH
-
 Colwellia maris
1.1.1.42 IDH
-
 Pseudomonas psychrophila
1.1.1.42 NADP+-dependent isocitrate dehydrogenase
-
 Colwellia maris
1.1.1.42 NADP+-dependent isocitrate dehydrogenase
-
 Pseudomonas psychrophila

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
1.1.1.42 30
-
-
 Colwellia maris
1.1.1.42 50
-
-
 Pseudomonas psychrophila

Temperature Stability [°C]

EC Number Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
1.1.1.42 30 50 after incubation at 25°C and 30°C, the enzyme activity loses 31% and 69% of ist activity, respectively, and no activity is detected after incubation at 50°C Colwellia maris
1.1.1.42 40 70 after incubation at temperatures up to 40°C, the wild type enzyme maintains its activity, but its residual activity is decreased by incubation at over 45°C and is lost after incubation at 70°C Pseudomonas psychrophila

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
1.1.1.42 69.88
-
 isocitrate wild type enzyme, at pH 8.0 and 20°C Colwellia maris
1.1.1.42 82.47
-
 isocitrate mutant enzyme A741S, at pH 8.0 and 20°C Colwellia maris
1.1.1.42 89.04
-
 isocitrate mutant enzyme E596L, at pH 8.0 and 20°C Colwellia maris
1.1.1.42 136.85
-
 isocitrate wild type enzyme, at pH 8.0 and 20°C Pseudomonas psychrophila
1.1.1.42 158.91
-
 isocitrate mutant enzyme L594E, at pH 8.0 and 20°C Pseudomonas psychrophila

Cofactor

EC Number Cofactor Comment Organism Structure
1.1.1.42 NADP+
-
 Colwellia maris
1.1.1.42 NADP+
-
 Pseudomonas psychrophila

kcat/KM [mM/s]

EC Number kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
1.1.1.42 1164
-
 isocitrate wild type enzyme, at pH 8.0 and 20°C Colwellia maris
1.1.1.42 1227
-
 isocitrate mutant enzyme A741S, at pH 8.0 and 20°C Colwellia maris
1.1.1.42 1914
-
 isocitrate mutant enzyme E596L, at pH 8.0 and 20°C Colwellia maris
1.1.1.42 6810
-
 isocitrate wild type enzyme, at pH 8.0 and 20°C Pseudomonas psychrophila
1.1.1.42 101970
-
 isocitrate mutant enzyme L594E, at pH 8.0 and 20°C Pseudomonas psychrophila