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Literature summary for 4.2.1.17 extracted from

  • Feng, Y.; Hofstein, H.A.; Zwahlen, J.; Tonge, P.J.
    Effect of mutagenesis on the stereochemistry of enoyl-CoA hydratase (2002), Biochemistry, 41, 12883-12890.
    View publication on PubMed
Search for more literature for 4.2.1.17

Protein Variants

Protein Variants Comment Organism
A98P kcat is decreased 3400fold compared to wild type and KM is increased 13fold, mutant enzyme has a severely compromised ability for catalyzing the formation of (3R)-3-hydroxybutanoyl-CoA Rattus norvegicus
E144D 60fold decreases in kcat with little change in KM Rattus norvegicus
E144Q 3000fold decreases in kcat with little change in KM. The mutant is unable to catalyze the formation of (3R)-3-hydroxybutanoyl-CoA even when the incubation is extended to 4 days Rattus norvegicus
E164D 1200fold decreases in kcat with little change in KM. First-order rate constant for the formation of (3R)-3-hydroxybutanoyl-CoA is similar to wild-type value Rattus norvegicus
E164Q 340000fold decreases in kcat with little change in KM. While wild-type enoyl-CoA hydratase catalyzes the rapid interconversion of substrate and the (3S)-3-hydroxybutanoyl-CoA product relative to the rate of (3R)-3-hydroxybutanoyl-CoA formation, E164Q catalyzes the formation of both product enantiomers at similar rates Rattus norvegicus
G141P 1600000fold decrease in kcat with no change in KM, mutant enzyme has a severely compromised ability for catalyzing the formation of (3R)-3-hydroxybutanoyl-CoA Rattus norvegicus

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.003
-
 crotonyl-CoA pH 7.4, 25°C, mutant enzyme G141P Rattus norvegicus
0.015
-
 crotonyl-CoA pH 7.4, 25°C, wild-type enzyme Rattus norvegicus
0.025
-
 crotonyl-CoA pH 7.4, 25°C, mutant enzyme E144Q Rattus norvegicus
0.032
-
 crotonyl-CoA pH 7.4, 25°C, mutant enzyme E164D Rattus norvegicus
0.041
-
 crotonyl-CoA pH 7.4, 25°C, mutant enzyme E164Q Rattus norvegicus
0.118
-
 3'-dephosphocrotonyl-CoA pH 7.4, 25°C, mutant enzyme E144D Rattus norvegicus
0.195
-
 crotonyl-CoA pH 7.4, 25°C, mutant enzyme A98P Rattus norvegicus

Localization

Localization Comment Organism GeneOntology No. Textmining
mitochondrion
-
 Rattus norvegicus 5739
-

Organism

Organism UniProt Comment Textmining
Rattus norvegicus
-
 recombinant
-

Purification (Commentary)

Purification (Comment) Organism
recombinant, wild-type and mutant enzymes Rattus norvegicus

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
3'-dephosphocrotonyl-CoA + H2O
-
 Rattus norvegicus ?
-
 ?
crotonyl-CoA + H2O i.e. (E)-2-butenoyl-CoA. Reaction is catalyzed with a stereospecificity of 1 in 400000. The enzyme catalyzes the rapid interconversion of substrate and the (3S)-3-hydroxybutanoyl-CoA product relative to the rate of (3R)-3-hydroxybutanoyl-CoA formation. Formation of the correct product enantiomer requires an intact oxyanion hole and optimal positioning of the substrate with respect to two catalytic glutamates (E144 and E164) in the active site Rattus norvegicus (3S)-3-hydroxybutanoyl-CoA
-
 r

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
25
-
 assay at Rattus norvegicus

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
0.0011
-
 crotonyl-CoA pH 7.4, 25°C, mutant enzyme G141P Rattus norvegicus
0.0053
-
 crotonyl-CoA pH 7.4, 25°C, mutant enzyme E164Q Rattus norvegicus
0.53
-
 crotonyl-CoA pH 7.4, 25°C, mutant enzyme A98P Rattus norvegicus
0.6
-
 crotonyl-CoA pH 7.4, 25°C, mutant enzyme E144Q Rattus norvegicus
1.5
-
 crotonyl-CoA pH 7.4, 25°C, mutant enzyme E164D Rattus norvegicus
26
-
 3'-dephosphocrotonyl-CoA pH 7.4, 25°C, mutant enzyme E144D Rattus norvegicus
1790
-
 crotonyl-CoA pH 7.4, 25°C, wild-type enzyme Rattus norvegicus

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
7.4
-
 assay at Rattus norvegicus