Any feedback?
Please rate this page
(literature.php)
(0/150)

BRENDA support

Literature summary for 1.1.1.42 extracted from

  • Huang, S.; Cheng, H.; Wang, P.; Zhu, G.
    Biochemical characterization and complete conversion of coenzyme specificity of isocitrate dehydrogenase from bifidobacterium longum (2016), Int. J. Mol. Sci., 17, 296.
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

Cloned (Comment) Organism
gene icd, phylogenetic analysis, recombinant expression of His6-tagged enzyme in Escherichia coli strain Rosetta (DE3) Bifidobacterium longum subsp. infantis

Protein Variants

Protein Variants Comment Organism
D253A/S257K/K260Q/R314D/H315I/T327A site-directed mutagenesis, the mutant shows a witch in cofactor specificity from NADP+ to NAD+ Bifidobacterium longum subsp. infantis
additional information the coenzyme specificity of BlIDH can be completely reversed from NADP+ to NAD+ by a factor of 2387 by replacing six residues. The loss in NADP+-dependent activity might result from the removal of favorable interactions between Arg314 or His315 and the 2'-phosphate group Bifidobacterium longum subsp. infantis
R314D site-directed mutagenesis Bifidobacterium longum subsp. infantis
R314D/H315I/T327A site-directed mutagenesis Bifidobacterium longum subsp. infantis

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
additional information
-
additional information kinetics analysis, overview Bifidobacterium longum subsp. infantis
0.01945
-
NADP+ pH 8.0, 25°C, recombinant wild-type enzyme Bifidobacterium longum subsp. infantis
0.13
-
NAD+ pH 8.0, 25°C, recombinant mutant D253A/S257K/K260Q/R314D/H315I/T327A Bifidobacterium longum subsp. infantis
0.324
-
NADP+ pH 8.0, 25°C, recombinant mutant D253A/S257K/K260Q/R314D/H315I/T327A Bifidobacterium longum subsp. infantis
3.58
-
NAD+ pH 8.0, 25°C, recombinant wild-type enzyme Bifidobacterium longum subsp. infantis

Metals/Ions

Metals/Ions Comment Organism Structure
Co2+ 14.3% activity compared to Mn2+ Bifidobacterium longum subsp. infantis
Mg2+ 69.2% activity compared to Mn2+ Bifidobacterium longum subsp. infantis
Mn2+ best activating cation Bifidobacterium longum subsp. infantis
additional information the enzyme is dependent on divalent cations, overview. No activity with Zn2+, Cu2+, Ca2+, and Li+,very low activity with K+, Na+, Ni2+, and Rb+ Bifidobacterium longum subsp. infantis

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
45000
-
-
Bifidobacterium longum subsp. infantis
83000
-
recombinant His6-tagged enzyme, gel filtration Bifidobacterium longum subsp. infantis

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
isocitrate + NADP+ Bifidobacterium longum subsp. infantis
-
2-oxoglutarate + CO2 + NADPH + H+
-
r
isocitrate + NADP+ Bifidobacterium longum subsp. infantis ATCC 15697
-
2-oxoglutarate + CO2 + NADPH + H+
-
r

Organism

Organism UniProt Comment Textmining
Bifidobacterium longum subsp. infantis B7GQR3
-
-
Bifidobacterium longum subsp. infantis ATCC 15697 B7GQR3
-
-

Purification (Commentary)

Purification (Comment) Organism
recombinant His6-tagged enzyme from Escherichia coli strain Rosetta (DE3) by Co2+ affinity chromatography Bifidobacterium longum subsp. infantis

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
isocitrate + NAD+ engineered enzyme mutant D253A/S257K/K260Q/R314D/H315I/T327A, cf. EC 1.1.1.41 Bifidobacterium longum subsp. infantis 2-oxoglutarate + CO2 + NADH + H+
-
r
isocitrate + NAD+ engineered enzyme mutant D253A/S257K/K260Q/R314D/H315I/T327A, cf. EC 1.1.1.41 Bifidobacterium longum subsp. infantis ATCC 15697 2-oxoglutarate + CO2 + NADH + H+
-
r
isocitrate + NADP+
-
Bifidobacterium longum subsp. infantis 2-oxoglutarate + CO2 + NADPH + H+
-
r
isocitrate + NADP+
-
Bifidobacterium longum subsp. infantis ATCC 15697 2-oxoglutarate + CO2 + NADPH + H+
-
r

Subunits

Subunits Comment Organism
homodimer 2 * 45000, recombinant His6-tagged enzyme, SDS-PAGE Bifidobacterium longum subsp. infantis

Synonyms

Synonyms Comment Organism
ICD
-
Bifidobacterium longum subsp. infantis
NADP+-dependent IDH
-
Bifidobacterium longum subsp. infantis
NADP-IDH
-
Bifidobacterium longum subsp. infantis

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
60
-
with Mn2+ Bifidobacterium longum subsp. infantis
65
-
with Mg2+ Bifidobacterium longum subsp. infantis

Temperature Stability [°C]

Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
45
-
enzyme BlIDH retains 50% of maximal activity after incubation at 45°C for 20 min with either Mn2+ or Mg2+ Bifidobacterium longum subsp. infantis

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
0.307
-
NADP+ pH 8.0, 25°C, recombinant mutant D253A/S257K/K260Q/R314D/H315I/T327A Bifidobacterium longum subsp. infantis
0.518
-
NAD+ pH 8.0, 25°C, recombinant mutant D253A/S257K/K260Q/R314D/H315I/T327A Bifidobacterium longum subsp. infantis
11.7
-
NAD+ pH 8.0, 25°C, recombinant wild-type enzyme Bifidobacterium longum subsp. infantis
36.4
-
NADP+ pH 8.0, 25°C, recombinant wild-type enzyme Bifidobacterium longum subsp. infantis

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
7.5
-
with Mn2+ Bifidobacterium longum subsp. infantis
8
-
with Mg2+ Bifidobacterium longum subsp. infantis

Cofactor

Cofactor Comment Organism Structure
additional information the enzyme shows a 567 and 193fold preference for NADP+ over NAD+ in the presence of Mg2+ and Mn2+, respectively. The coenzyme specificity of BlIDH can be completely reversed from NADP+ to NAD+ by a factor of 2387 by replacing six residues by rational mutagenesis Bifidobacterium longum subsp. infantis
NADP+
-
Bifidobacterium longum subsp. infantis
NADPH
-
Bifidobacterium longum subsp. infantis

General Information

General Information Comment Organism
evolution based on the phylogenetic analysis, IDHs can be divided into three subfamilies: Type I IDHs, Type II IDHs and monomeric IDHs. The enzyme BlIDH from Bifidobacterium longum belongs to the type II subfamily. NAD+ use is an ancestral trait and NADP+ use by bacterial IDHs arose on or about the time that eukaryotic mitochondria first appeared, some 3.5 billion years ago Bifidobacterium longum subsp. infantis
physiological function NADP+-dependent IDH generates NADPH, which provides the reducing power for biosynthesis, maintains the redox state of the cell, and takes part in CO2 assimilation Bifidobacterium longum subsp. infantis

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
0.95
-
NAD+ pH 8.0, 25°C, recombinant mutant D253A/S257K/K260Q/R314D/H315I/T327A Bifidobacterium longum subsp. infantis
3
-
NAD+ pH 8.0, 25°C, recombinant wild-type enzyme Bifidobacterium longum subsp. infantis
4
-
NAD+ pH 8.0, 25°C, recombinant mutant D253A/S257K/K260Q/R314D/H315I/T327A Bifidobacterium longum subsp. infantis
1870
-
NAD+ pH 8.0, 25°C, recombinant wild-type enzyme Bifidobacterium longum subsp. infantis