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Literature summary for 1.1.1.41 extracted from

  • Romkina, A.; Kiriukhin, M.
    Biochemical and molecular characterization of the isocitrate dehydrogenase with dual coenzyme specificity from the obligate methylotroph Methylobacillus flagellatus (2017), PLoS ONE, 12, e0176056 .
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

Cloned (Comment) Organism
expressed in Escherichia coli BL21(DE3) cells Methylobacillus flagellatus

Inhibitors

Inhibitors Comment Organism Structure
Ca2+ complete inhibition at 2 mM (pH 6.0) Methylobacillus flagellatus
Cu2+ complete inhibition at 2 mM (pH 6.0) Methylobacillus flagellatus
additional information the enzyme is unaffected by the addition of 5 mM L-glutamate, L-glutamine, 2-oxoglutarate, oxaloacetate, cis-aconitate, citrate, pyruvate, malate, fumarate, succinate, CoA, AcCoA, NADH, and NADPH or 2 mM ADP and AMP Methylobacillus flagellatus
Zn2+ NAD+-linked activity at pH 8.5 is strongly inhibited by Zn2+ but partially restored at pH 6.0 Methylobacillus flagellatus

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.009
-
isocitrate at pH 8.5 and 42°C Methylobacillus flagellatus
0.113
-
NAD+ at pH 8.5 and 42°C Methylobacillus flagellatus

Metals/Ions

Metals/Ions Comment Organism Structure
Mg2+ Mg2+ can act as a significant substitute by providing up to 58-75% of the enzyme's maximal activity Methylobacillus flagellatus
Mn2+ Mn2+ enhances the activity the most effectively Methylobacillus flagellatus

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
83600
-
gel filtration Methylobacillus flagellatus

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
isocitrate + NAD+ Methylobacillus flagellatus
-
2-oxoglutarate + CO2 + NADH + H+
-
r
isocitrate + NAD+ Methylobacillus flagellatus ATCC51484
-
2-oxoglutarate + CO2 + NADH + H+
-
r

Organism

Organism UniProt Comment Textmining
Methylobacillus flagellatus
-
-
-
Methylobacillus flagellatus ATCC51484
-
-
-

Purification (Commentary)

Purification (Comment) Organism
Ni-NTA affinity resin column chromatography Methylobacillus flagellatus

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
isocitrate + APAD+ 55% activity compared to NAD+ Methylobacillus flagellatus 2-oxoglutarate + CO2 + APADH
-
r
isocitrate + APAD+ 55% activity compared to NAD+ Methylobacillus flagellatus ATCC51484 2-oxoglutarate + CO2 + APADH
-
r
isocitrate + NAD+
-
Methylobacillus flagellatus 2-oxoglutarate + CO2 + NADH + H+
-
r
isocitrate + NAD+ 100% activity Methylobacillus flagellatus 2-oxoglutarate + CO2 + NADH + H+
-
r
isocitrate + NAD+
-
Methylobacillus flagellatus ATCC51484 2-oxoglutarate + CO2 + NADH + H+
-
r
isocitrate + NAD+ 100% activity Methylobacillus flagellatus ATCC51484 2-oxoglutarate + CO2 + NADH + H+
-
r
isocitrate + NHD+ 4.0% activity compared to NAD+ Methylobacillus flagellatus 2-oxoglutarate + CO2 + NHDH
-
r
isocitrate + NHD+ 4.0% activity compared to NAD+ Methylobacillus flagellatus ATCC51484 2-oxoglutarate + CO2 + NHDH
-
r

Subunits

Subunits Comment Organism
homodimer 2 * 45000, SDS-PAGE Methylobacillus flagellatus
homodimer 2 * 44600, calculated from amino acid sequence Methylobacillus flagellatus

Synonyms

Synonyms Comment Organism
IDH
-
Methylobacillus flagellatus
NAD+-specific IDH
-
Methylobacillus flagellatus

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
60
-
-
Methylobacillus flagellatus

Temperature Stability [°C]

Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
50 75 the enzyme retains full activity at 50°C and loses ca. 50% of its activity after 1 h of incubation at 75°C Methylobacillus flagellatus

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
166
-
NAD+ at pH 8.5 and 42°C Methylobacillus flagellatus

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
8.5
-
-
Methylobacillus flagellatus

Cofactor

Cofactor Comment Organism Structure
NAD+ the catalytic efficiency for NAD+ is 5times higher for NAD+ than for NADP+ Methylobacillus flagellatus

pI Value

Organism Comment pI Value Maximum pI Value
Methylobacillus flagellatus isoelectric focusing
-
5.5
Methylobacillus flagellatus calculated from amino acid sequence
-
5.9

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
1500
-
NAD+ at pH 8.5 and 42°C Methylobacillus flagellatus