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Literature summary extracted from
- Human peroxisomal multifunctional enzyme type 2: site-directed mutagenesis studies show the importance of two protic residues for 2-enoyl-CoA hydratase 2 activity (2000), J. Biol. Chem., 275, 4965-4972.
Cloned(Commentary)
| EC Number | Cloned (Comment) | Organism |
|---|---|---|
| 1.1.1.36 | wild type (HsMFE-2) and its variants are expressed in Saccharomyces cerevisiae | Homo sapiens |
| 4.2.1.119 | wild type (HsMFE-2) and its variants are expressed in Saccharomyces cerevisiae, the recombinant HsMFE-2(dhdelta) and its variants are expressed in Escherichia coli BL21(DE3)pLysS | Homo sapiens |
Protein Variants
| EC Number | Protein Variants | Comment | Organism |
|---|---|---|---|
| 1.1.1.36 | D370A | site-directed mutagenesis | Homo sapiens |
| 1.1.1.36 | D490A | site-directed mutagenesis | Homo sapiens |
| 1.1.1.36 | D510A | site-directed mutagenesis, inactive | Homo sapiens |
| 1.1.1.36 | D517A | site-directed mutagenesis | Homo sapiens |
| 1.1.1.36 | E366A | site-directed mutagenesis, kcat/Km 100times lower than that of the wild type | Homo sapiens |
| 1.1.1.36 | E408A | site-directed mutagenesis | Homo sapiens |
| 1.1.1.36 | G16S | site-directed mutagenesis | Homo sapiens |
| 1.1.1.36 | H406A | site-directed mutagenesis | Homo sapiens |
| 1.1.1.36 | H515A | site-directed mutagenesis | Homo sapiens |
| 1.1.1.36 | H532A | site-directed mutagenesis | Homo sapiens |
| 1.1.1.36 | additional information | constructs are tested for complementation in Saccharomyces cerevisiae | Homo sapiens |
| 1.1.1.36 | Y347A | site-directed mutagenesis | Homo sapiens |
| 1.1.1.36 | Y410A | site-directed mutagenesis | Homo sapiens |
| 1.1.1.36 | Y505A | site-directed mutagenesis | Homo sapiens |
| 4.2.1.119 | D370A | reduced specific acitivity of 2-enoyl-CoA hydratase 2 when expressed in Saccharomyces cerevisiae | Homo sapiens |
| 4.2.1.119 | D490A | reduced specific acitivity of 2-enoyl-CoA hydratase 2 when expressed in Saccharomyces cerevisiae | Homo sapiens |
| 4.2.1.119 | D510A | inactive mutant enzyme | Homo sapiens |
| 4.2.1.119 | D517A | reduced specific acitivity of 2-enoyl-CoA hydratase 2 when expressed in Saccharomyces cerevisiae | Homo sapiens |
| 4.2.1.119 | E366A | kcat/Km 100times lower than that of the wild type | Homo sapiens |
| 4.2.1.119 | E408A | reduced specific acitivity of 2-enoyl-CoA hydratase 2 when expressed in Saccharomyces cerevisiae | Homo sapiens |
| 4.2.1.119 | G16S | reduced specific acitivity of 2-enoyl-CoA hydratase 2 when expressed in Saccharomyces cerevisiae | Homo sapiens |
| 4.2.1.119 | H406A | reduced specific acitivity of 2-enoyl-CoA hydratase 2 when expressed in Saccharomyces cerevisiae | Homo sapiens |
| 4.2.1.119 | H515A | inactive mutant enzyme | Homo sapiens |
| 4.2.1.119 | H532A | reduced specific acitivity of 2-enoyl-CoA hydratase 2 when expressed in Saccharomyces cerevisiae | Homo sapiens |
| 4.2.1.119 | additional information | mutant constructs are tested for complementation in Saccharomyces cerevisiae | Homo sapiens |
| 4.2.1.119 | Y347A | inactive mutant enzyme | Homo sapiens |
| 4.2.1.119 | Y410A | reduced specific acitivity of 2-enoyl-CoA hydratase 2 when expressed in Saccharomyces cerevisiae | Homo sapiens |
| 4.2.1.119 | Y505A | inactive mutant enzyme | Homo sapiens |
KM Value [mM]
| EC Number | KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|---|
| 4.2.1.119 | 0.0081 | - |
(2E)-2-decenoyl-CoA | wild type, pH 8 | Homo sapiens |  |
| 4.2.1.119 | 0.0089 | - |
(2E)-2-decenoyl-CoA | wild type, pH 5 | Homo sapiens | |
| 4.2.1.119 | 0.0092 | - |
(2E)-2-decenoyl-CoA | mutant enzyme E366A, pH 5 | Homo sapiens | |
| 4.2.1.119 | 0.0131 | - |
(2E)-2-decenoyl-CoA | mutant enzyme E366A, pH 8 | Homo sapiens | |
Localization
| EC Number | Localization | Comment | Organism | GeneOntology No. | Textmining |
|---|---|---|---|---|---|
| 1.1.1.36 | peroxisome | - |
Homo sapiens | 5777 | - |
| 4.2.1.119 | peroxisome | peroxisomal hydratase 2 together with (3R)-hydroxyacyl-CoA dehydrogenase, and hydratase 1 together with (3S)-hydroxyacyl-CoA dehydrogenase, are present as multifunctional enzymes. When present simultaneously in peroxisomes, beta-oxidation has two stereochemical possibilities | Homo sapiens | 5777 | - |
Molecular Weight [Da]
| EC Number | Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
|---|---|---|---|---|
| 4.2.1.119 | 45000 | - |
x * 45000, HsMFE-2(dhdelta), HsMFE-2(dhdelta, E366A), HsMFE-2(dhdelta, D510A), SDS-PAGE | Homo sapiens |
Natural Substrates/ Products (Substrates)
| EC Number | Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
|---|---|---|---|---|---|---|---|
| 4.2.1.119 | additional information | Homo sapiens | peroxisomal hydratase 2 together with (3R)-hydroxyacyl-CoA dehydrogenase, and peroxisomal hydratase 1 together with (3S)-hydroxyacyl-CoA dehydrogenase, are present as multifunctional enzymes. When present simultaneously in peroxisomes, beta-oxidation has two stereochemical possibilities | ? | - |
? | |
Organism
| EC Number | Organism | UniProt | Comment | Textmining |
|---|---|---|---|---|
| 1.1.1.36 | Homo sapiens | - |
alignment of the amino acid sequence of the hydratase 2 domain from human MFE-2 with those of other known hydratase 2 domains from eukaryotes shown | - |
| 4.2.1.119 | Homo sapiens | - |
- |
- |
Purification (Commentary)
| EC Number | Purification (Comment) | Organism |
|---|---|---|
| 4.2.1.119 | - |
Homo sapiens |
Specific Activity [micromol/min/mg]
| EC Number | Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
|---|---|---|---|---|
| 1.1.1.36 | 0.023 | - |
strain HsMFE-2(Y347A), (3R)-hydroxyacyl-CoA dehydrogenase | Homo sapiens |
| 1.1.1.36 | 0.023 | - |
strain HsMFE-2(Y410A), (3R)-hydroxyacyl-CoA dehydrogenase | Homo sapiens |
| 1.1.1.36 | 0.039 | - |
strain HsMFE-2(E408A), (3R)-hydroxyacyl-CoA dehydrogenase | Homo sapiens |
| 1.1.1.36 | 0.041 | - |
strain HsMFE-2(D490A), (3R)-hydroxyacyl-CoA dehydrogenase | Homo sapiens |
| 1.1.1.36 | 0.046 | - |
strain HsMFE-2(H406A), (3R)-hydroxyacyl-CoA dehydrogenase | Homo sapiens |
| 1.1.1.36 | 0.047 | - |
strain HsMFE-2(D370A), (3R)-hydroxyacyl-CoA dehydrogenase | Homo sapiens |
| 1.1.1.36 | 0.048 | - |
strain HsMFE-2(D517A), (3R)-hydroxyacyl-CoA dehydrogenase | Homo sapiens |
| 1.1.1.36 | 0.051 | - |
strain HsMFE-2(D510A), (3R)-hydroxyacyl-CoA dehydrogenase | Homo sapiens |
| 1.1.1.36 | 0.056 | - |
strain HsMFE-2(E366A), (3R)-hydroxyacyl-CoA dehydrogenase | Homo sapiens |
| 1.1.1.36 | 0.066 | - |
strain HsMFE-2(H532A), (3R)-hydroxyacyl-CoA dehydrogenase | Homo sapiens |
| 1.1.1.36 | 0.082 | - |
strain HsMFE-2, (3R)-hydroxyacyl-CoA dehydrogenase | Homo sapiens |
| 1.1.1.36 | 0.091 | - |
strain UTL-7A, (3R)-hydroxyacyl-CoA dehydrogenase | Homo sapiens |
| 4.2.1.119 | 0.12 | - |
strain HsMFE-2(D490A) | Homo sapiens |
| 4.2.1.119 | 0.16 | - |
strain HsMFE-2(G16S) | Homo sapiens |
| 4.2.1.119 | 0.2 | - |
strain HsMFE-2(H532A) | Homo sapiens |
| 4.2.1.119 | 0.21 | - |
strain HsMFE-2(D370A), strain HsMFE-2(H406A) | Homo sapiens |
| 4.2.1.119 | 0.24 | - |
strain HsMFE-2(Y410A) | Homo sapiens |
| 4.2.1.119 | 0.26 | - |
strain HsMFE-2(D517A), strain HsMFE-2(E408A) | Homo sapiens |
| 4.2.1.119 | 0.4 | - |
strain UTL-7A | Homo sapiens |
| 4.2.1.119 | 0.54 | - |
strain HsMFE-2 | Homo sapiens |
Substrates and Products (Substrate)
| EC Number | Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
|---|---|---|---|---|---|---|---|
| 4.2.1.119 | additional information | peroxisomal hydratase 2 together with (3R)-hydroxyacyl-CoA dehydrogenase, and peroxisomal hydratase 1 together with (3S)-hydroxyacyl-CoA dehydrogenase, are present as multifunctional enzymes. When present simultaneously in peroxisomes, beta-oxidation has two stereochemical possibilities | Homo sapiens | ? | - |
? | |
| 4.2.1.119 | trans-2-decenoyl-CoA + H2O | - |
Homo sapiens | (3R)-3-hydroxydecanoyl-CoA | - |
r | |
Subunits
| EC Number | Subunits | Comment | Organism |
|---|---|---|---|
| 4.2.1.119 | ? | x * 45000, HsMFE-2(dhdelta), HsMFE-2(dhdelta, E366A), HsMFE-2(dhdelta, D510A), SDS-PAGE | Homo sapiens |
Synonyms
| EC Number | Synonyms | Comment | Organism |
|---|---|---|---|
| 1.1.1.36 | (3R)-hydroxyacyl-CoA dehydrogenase | domain in human MFE-2 | Homo sapiens |
| 4.2.1.119 | 2-enoyl-CoA hydratase 2 | - |
Homo sapiens |
| 4.2.1.119 | 2-enoyl-CoA hydratase 2 | domain in human MFE-2 | Homo sapiens |
| 4.2.1.119 | MFE-2 | peroxisomal hydratase 2 together with (3R)-hydroxyacyl-CoA dehydrogenase is present as multifunctional enzyme | Homo sapiens |
Temperature Optimum [°C]
| EC Number | Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
|---|---|---|---|---|
| 4.2.1.119 | 22 | - |
assay at | Homo sapiens |
Turnover Number [1/s]
| EC Number | Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|---|
| 4.2.1.119 | 1.3 | - |
(2E)-2-decenoyl-CoA | mutant enzyme E366A, pH 5 | Homo sapiens | |
| 4.2.1.119 | 17.9 | - |
(2E)-2-decenoyl-CoA | mutant enzyme E366A, pH 8 | Homo sapiens | |
| 4.2.1.119 | 105 | - |
(2E)-2-decenoyl-CoA | wild type enzyme, pH 5 | Homo sapiens | |
| 4.2.1.119 | 196 | - |
(2E)-2-decenoyl-CoA | wild type enzyme, pH 8 | Homo sapiens | |
pH Range
| EC Number | pH Minimum | pH Maximum | Comment | Organism |
|---|---|---|---|---|
| 4.2.1.119 | 5 | 10 | pH dependence experiment is performed in 200 mM potassium phosphate buffer at pH values varying from 5 to 10 | Homo sapiens |
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