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Literature summary extracted from

  • Wu, M.; Tian, C.; Cheng, H.; Xu, L.; Wang, P.; Zhu, G.
    A novel type II NAD+-specific isocitrate dehydrogenase from the marine bacterium Congregibacter litoralis KT71 (2015), PLoS ONE, 10, e0125229.
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

EC Number Cloned (Comment) Organism
1.1.1.41 gene KT71_12905, DNA and amino acid sequence determination and analysis, sequence comparisons,, phylogenetic analysis and tree, recombinant expression of His6-tagged wild-type and mutant enzymes in Escherichia coli strain Rosetta (DE3) Congregibacter litoralis

Protein Variants

EC Number Protein Variants Comment Organism
1.1.1.41 D487R/L488H site-directed mutagenesis, the coenzyme specificity of a the ClIDH mutant is altered compared to the wild-type enzyme, and the preference of the mutant for NADP+ is approximately 24fold higher than that for NAD+, suggesting that ClIDH is an NAD+-specific ancestral enzyme in the type II IDH subgroup Congregibacter litoralis

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
1.1.1.41 0.157
-
 NADP+ mutant D487R/L488H, with Mn2+, pH 7.5, 35°C Congregibacter litoralis
1.1.1.41 0.263
-
 NAD+ wild-type enzyme, with Mg2+, pH 7.5, 35°C Congregibacter litoralis
1.1.1.41 0.309
-
 NAD+ wild-type enzyme, with Mn2+, pH 7.5, 35°C Congregibacter litoralis
1.1.1.41 0.664
-
 NAD+ mutant D487R/L488H, with Mn2+, pH 7.5, 35°C Congregibacter litoralis

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
1.1.1.41 Co2+ activates, 38.3% activity compared to Mn2+ Congregibacter litoralis
1.1.1.41 Mg2+ activates, 45.7% activity compared to Mn2+ Congregibacter litoralis
1.1.1.41 Mn2+ most effectivly activating cation Congregibacter litoralis
1.1.1.41 additional information enzyme ClIDH is dependent on divalent cations, the most effective being Mn2+. Poor activation by Ca2+, Zn2+, Ni2+, K+, Na+, Rb+, and Li+. No activation by Cu2+ Congregibacter litoralis

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
1.1.1.41 64000
-
-
 Congregibacter litoralis
1.1.1.41 390000
-
 recombinant His6-tagged enzyme, gel filtration and analytical ultracentrifugation Congregibacter litoralis

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
1.1.1.41 isocitrate + NAD+ Congregibacter litoralis
-
 2-oxoglutarate + CO2 + NADH + H+
-
 ?
1.1.1.41 isocitrate + NAD+ Congregibacter litoralis KT71
-
 2-oxoglutarate + CO2 + NADH + H+
-
 ?

Organism

EC Number Organism UniProt Comment Textmining
1.1.1.41 Congregibacter litoralis A4ADB8
-
-
1.1.1.41 Congregibacter litoralis KT71 A4ADB8
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
1.1.1.41 recombinant His6-tagged wild-type and mutant enzymes from Escherichia coli strain Rosetta (DE3) by metal affinity chromatography Congregibacter litoralis

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
1.1.1.41 isocitrate + NAD+
-
 Congregibacter litoralis 2-oxoglutarate + CO2 + NADH + H+
-
 ?
1.1.1.41 isocitrate + NAD+
-
 Congregibacter litoralis KT71 2-oxoglutarate + CO2 + NADH + H+
-
 ?
1.1.1.41 isocitrate + NADP+ activity with enzyme mutant D487R/L488H, not the wild-type enzyme Congregibacter litoralis 2-oxoglutarate + CO2 + NADPH + H+
-
 ?
1.1.1.41 isocitrate + NADP+ activity with enzyme mutant D487R/L488H, not the wild-type enzyme Congregibacter litoralis KT71 2-oxoglutarate + CO2 + NADPH + H+
-
 ?

Subunits

EC Number Subunits Comment Organism
1.1.1.41 homohexamer 6 * 64000, recombinant His6-tagged enzyme, SDS-PAGE Congregibacter litoralis

Synonyms

EC Number Synonyms Comment Organism
1.1.1.41 type II NAD+-specific isocitrate dehydrogenase
-
 Congregibacter litoralis

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
1.1.1.41 35
-
-
 Congregibacter litoralis

Temperature Range [°C]

EC Number Temperature Minimum [°C] Temperature Maximum [°C] Comment Organism
1.1.1.41 20 32 activity range, profile overview Congregibacter litoralis

Temperature Stability [°C]

EC Number Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
1.1.1.41 33
-
 purified recombinant enzyme, 20 min, loss of 50% activity Congregibacter litoralis
1.1.1.41 38
-
 purified recombinant enzyme, 20 min, loss of over 90% activity Congregibacter litoralis

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
1.1.1.41 9.7
-
 NAD+ mutant D487R/L488H, with Mn2+, pH 7.5, 35°C Congregibacter litoralis
1.1.1.41 36.7
-
 NAD+ wild-type enzyme, with Mg2+, pH 7.5, 35°C Congregibacter litoralis
1.1.1.41 55.1
-
 NADP+ mutant D487R/L488H, with Mn2+, pH 7.5, 35°C Congregibacter litoralis
1.1.1.41 84.7
-
 NAD+ wild-type enzyme, with Mn2+, pH 7.5, 35°C Congregibacter litoralis

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
1.1.1.41 7.5
-
-
 Congregibacter litoralis

Cofactor

EC Number Cofactor Comment Organism Structure
1.1.1.41 NAD+ the recombinant enzyme is NAD+-specific and shows no detectable activity with NADP+ Congregibacter litoralis

General Information

EC Number General Information Comment Organism
1.1.1.41 evolution phylogenetic analyses divide the IDH protein family into two subgroups: types I and II. Based on cofactor usage, IDHs are either NAD+-specific (NAD-IDH) or NADP+-specific (NADP-IDH). NADP-IDH evolved from NAD-IDH. Type I IDHs include NAD-IDHs and NADP-IDHs. Type II NAD-IDHs is identified from the marine bacterium Congregibacter litoralis KT71, i.e ClIDH. Evolutionary relationships between 151 IDHs from different organisms, overview Congregibacter litoralis

kcat/KM [mM/s]

EC Number kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
1.1.1.41 15
-
 NAD+ mutant D487R/L488H, with Mn2+, pH 7.5, 35°C Congregibacter litoralis
1.1.1.41 140
-
 NAD+ wild-type enzyme, with Mg2+, pH 7.5, 35°C Congregibacter litoralis
1.1.1.41 270
-
 NAD+ wild-type enzyme, with Mn2+, pH 7.5, 35°C Congregibacter litoralis
1.1.1.41 350
-
 NADP+ mutant D487R/L488H, with Mn2+, pH 7.5, 35°C Congregibacter litoralis