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

BRENDA support

Literature summary for 1.1.1.42 extracted from

  • Li, X.; Wang, P.; Ge, Y.; Wang, W.; Abbas, A.; Zhu, G.
    NADP+-specific isocitrate dehydrogenase from oleaginous yeast Yarrowia lipolytica CLIB122: biochemical characterization and coenzyme sites evaluation (2013), Appl. Biochem. Biotechnol., 171, 403-416.
    View publication on PubMed
Search for more literature for 1.1.1.42

Cloned(Commentary)

Cloned (Comment) Organism
idh2, DNA and amino acid sequence determination and anaysis, phylogenetic analysis, overexpression of His6-tagged wild-type and mutant enzymes in Escherichia coli strain Rosetta (DE3) Yarrowia lipolytica

Protein Variants

Protein Variants Comment Organism
R322D site-directed mutagenesis, the mutant shows an 41fold higher Km for NADP+ than the wild-type enzyme, and it shows NAD+-dependent activity in contrast to the wild-type enzyme Yarrowia lipolytica
R322D/H323I site-directed mutagenesis, inactive mutant Yarrowia lipolytica

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
additional information
-
 additional information kinetic analysis Yarrowia lipolytica
0.031
-
 NADP+ pH 7.5, 25°C, recombinant enzyme, with Mg2+ Yarrowia lipolytica
0.058
-
 NADP+ pH 7.5, 25°C, recombinant enzyme, with Mg2+ Yarrowia lipolytica
0.059
-
 isocitrate pH 7.5, 25°C, recombinant enzyme, with Mg2+ Yarrowia lipolytica
0.12
-
 isocitrate pH 7.5, 25°C, recombinant enzyme, with Mn2+ Yarrowia lipolytica
47
-
 NAD+ pH 7.5, 25°C, recombinant enzyme mutant R322D Yarrowia lipolytica

Metals/Ions

Metals/Ions Comment Organism Structure
Co2+ activates to 43% of the activity with Mg2+ Yarrowia lipolytica
Li+ activates to 24% of the activity with Mg2+ Yarrowia lipolytica
Mg2+ most favorable cofactor Yarrowia lipolytica
Mn2+ activates to 60% of the activity with Mg2+ Yarrowia lipolytica
additional information the enzyme is absolutely divalent cation dependent, no or poor activation by Ca2+, Zn2+, K+, Na+, and Rb+ Yarrowia lipolytica
Ni2+ activates to 10% of the activity with Mg2+ Yarrowia lipolytica

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
45000
-
 2 * 45000, recombinant enzyme, SDS-PAGE Yarrowia lipolytica
81300
-
 recombinant enzyme, gel filtration Yarrowia lipolytica

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
isocitrate + NADP+ Yarrowia lipolytica
-
 2-oxoglutarate + CO2 + NADPH + H+
-
 r
isocitrate + NADP+ Yarrowia lipolytica CLIB 122
-
 2-oxoglutarate + CO2 + NADPH + H+
-
 r

Organism

Organism UniProt Comment Textmining
Yarrowia lipolytica Q6C2Y4
-
-
Yarrowia lipolytica CLIB 122 Q6C2Y4
-
-

Purification (Commentary)

Purification (Comment) Organism
recombinant His6-tagged wild-type and mutant enzymes from Escherichia coli strain Rosetta (DE3) b metal affinity chromatography Yarrowia lipolytica

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
isocitrate + NAD+ low activity with mutant enzyme R322D, no activity with the wild-type enzyme Yarrowia lipolytica 2-oxoglutarate + CO2 + NADH + H+
-
 r
isocitrate + NAD+ low activity with mutant enzyme R322D, no activity with the wild-type enzyme Yarrowia lipolytica CLIB 122 2-oxoglutarate + CO2 + NADH + H+
-
 r
isocitrate + NADP+
-
 Yarrowia lipolytica 2-oxoglutarate + CO2 + NADPH + H+
-
 r
isocitrate + NADP+
-
 Yarrowia lipolytica CLIB 122 2-oxoglutarate + CO2 + NADPH + H+
-
 r

Subunits

Subunits Comment Organism
homodimer 2 * 45000, recombinant enzyme, SDS-PAGE Yarrowia lipolytica

Synonyms

Synonyms Comment Organism
NADP+-specific isocitrate dehydrogenase
-
 Yarrowia lipolytica
YlIDP
-
 Yarrowia lipolytica

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
55
-
 recombinant enzyme Yarrowia lipolytica

Temperature Stability [°C]

Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
45 55 purified recombinant enzyme, stable up to 45°C, enzyme activity drops quickly above this temperature, incubation at 50°C for 20 min causes a 40% loss of activity, while incubation at 55°C almost inactivates the enzyme Yarrowia lipolytica

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
0.38
-
 NAD+ pH 8.5, 55°C, recombinant mutant R322D Yarrowia lipolytica
4.24
-
 NADP+ pH 7.5, 25°C, recombinant mutant R322D, with Mg2+ Yarrowia lipolytica
72
-
 NADP+ pH 7.5, 25°C, recombinant wild-type enzyme, with Mg2+ Yarrowia lipolytica

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
8.5
-
 recombinant enzyme Yarrowia lipolytica

Cofactor

Cofactor Comment Organism Structure
additional information enzyme YlIDP is absolutely dependent on NADP+ and shows no NAD-dependent activity Yarrowia lipolytica
NADP+
-
 Yarrowia lipolytica
NADPH
-
 Yarrowia lipolytica

General Information

General Information Comment Organism
evolution phylogenetic analysis and evolutionary relationships between YlIDP and IDPs from other yeasts or yeast-related species Yarrowia lipolytica

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
1.75
-
 NADP+ pH 7.5, 25°C, recombinant mutant R322D, with Mg2+ Yarrowia lipolytica
1220
-
 NADP+ pH 7.5, 25°C, recombinant wild-type enzyme, with Mg2+ Yarrowia lipolytica