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Literature summary for 1.1.1.42 extracted from

  • Lv, P.; Tang, W.; Wang, P.; Cao, Z.; Zhu, G.
    Enzymatic characterization and functional implication of two structurally different isocitrate dehydrogenases from Xylella fastidiosa (2018), Biotechnol. Appl. Biochem., 65, 230-237 .
    View publication on PubMed
Search for more literature for 1.1.1.42

Cloned(Commentary)

Cloned (Comment) Organism
expressed in Escherichia coli Rosetta(DE3) cells Xylella fastidiosa

Protein Variants

Protein Variants Comment Organism
H590L/R601L the mutations greatly reduce the affinity for NADP+, but fail to improve the ability to use NAD+ so the mutant has similar affinities to NADP+ and NAD+ Xylella fastidiosa
K589T/H590L/R601L the mutations greatly reduce the affinity for NADP+, but fail to improve the ability to use NAD+ so the mutant has similar affinities to NADP+ and NAD+ Xylella fastidiosa

Inhibitors

Inhibitors Comment Organism Structure
Ca2+ 3.0% residual activity at 2 mM Xylella fastidiosa
Co2+ 8.9% residual activity at 2 mM Xylella fastidiosa
Cu2+ complete inhibition at 2 mM Xylella fastidiosa
K+ 3.9% residual activity at 2 mM Xylella fastidiosa
Na+ 5.2% residual activity at 2 mM Xylella fastidiosa
Ni2+ 3.8% residual activity at 2 mM Xylella fastidiosa
Zn2+ complete inhibition at 2 mM Xylella fastidiosa

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.001
-
 NAD+ wild type enzyme, at pH 8.0 and 25°C Xylella fastidiosa
0.959
-
 NAD+ mutant enzyme K589T/H590L/R601L, at pH 8.0 and 25°C Xylella fastidiosa
1.506
-
 NAD+ mutant enzyme H590L/R601L, at pH 8.0 and 25°C Xylella fastidiosa
2.455
-
 NAD+ wild type enzyme, at pH 8.0 and 25°C Xylella fastidiosa
2.695
-
 NAD+ mutant enzyme H590L/R601L, at pH 8.0 and 25°C Xylella fastidiosa
5.534
-
 NAD+ mutant enzyme K589T/H590L/R601L, at pH 8.0 and 25°C Xylella fastidiosa

Metals/Ions

Metals/Ions Comment Organism Structure
Mn2+ Mn2+ is the most favored divalent cation. Mn2+ can be partly replaced by Mg2+ (17.6% activity). 2 mM is used in assay conditions Xylella fastidiosa

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
82400
-
 MALDI-TOF mass spectrometry Xylella fastidiosa

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
isocitrate + NADP+ Xylella fastidiosa
-
 2-oxoglutarate + CO2 + NADPH + H+
-
 ?

Organism

Organism UniProt Comment Textmining
Xylella fastidiosa
-
-
-

Purification (Commentary)

Purification (Comment) Organism
metal affinity resin column chromatography Xylella fastidiosa

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
7.4
-
 with NAD+ as cosubstrate, at pH 8.0 and 25°C Xylella fastidiosa
73.2
-
 with NADP+ as cosubstrate, at pH 8.0 and 25°C Xylella fastidiosa

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
isocitrate + NAD+
-
 Xylella fastidiosa 2-oxoglutarate + CO2 + NADH + H+
-
 ?
isocitrate + NADP+
-
 Xylella fastidiosa 2-oxoglutarate + CO2 + NADPH + H+
-
 ?

Subunits

Subunits Comment Organism
monomer 1 * 80000, SDS-PAGE Xylella fastidiosa

Synonyms

Synonyms Comment Organism
NADP+-dependent IDH
-
 Xylella fastidiosa
NADPH-IDH
-
 Xylella fastidiosa

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
50
-
-
 Xylella fastidiosa

Temperature Stability [°C]

Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
35 42 the enzyme is stable below 35°C, but its activity decreases rapidly above 35°C. The enzyme is almost inactivated after incubation at 42.5°C for 20 min Xylella fastidiosa

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
8.9
-
 NAD+ mutant enzyme K589T/H590L/R601L, at pH 8.0 and 25°C Xylella fastidiosa
16.7
-
 NAD+ wild type enzyme, at pH 8.0 and 25°C Xylella fastidiosa
30.8
-
 NAD+ mutant enzyme H590L/R601L, at pH 8.0 and 25°C Xylella fastidiosa
56.1
-
 NAD+ mutant enzyme K589T/H590L/R601L, at pH 8.0 and 25°C Xylella fastidiosa
62.4
-
 NAD+ mutant enzyme H590L/R601L, at pH 8.0 and 25°C Xylella fastidiosa
96.5
-
 NAD+ wild type enzyme, at pH 8.0 and 25°C Xylella fastidiosa

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
7.8
-
 in the presence of Mn2+ Xylella fastidiosa

Cofactor

Cofactor Comment Organism Structure
NADP+ the enzyme shows 13800fold preferences for NADP+ over NAD+ Xylella fastidiosa

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
7
-
 NAD+ wild type enzyme, at pH 8.0 and 25°C Xylella fastidiosa
9
-
 NAD+ mutant enzyme K589T/H590L/R601L, at pH 8.0 and 25°C Xylella fastidiosa
10
-
 NAD+ mutant enzyme K589T/H590L/R601L, at pH 8.0 and 25°C Xylella fastidiosa
20
-
 NAD+ mutant enzyme H590L/R601L, at pH 8.0 and 25°C Xylella fastidiosa
23
-
 NAD+ mutant enzyme H590L/R601L, at pH 8.0 and 25°C Xylella fastidiosa
96500
-
 NAD+ wild type enzyme, at pH 8.0 and 25°C Xylella fastidiosa