EC Number | Application | Comment | Organism |
---|---|---|---|
4.2.1.119 | biotechnology | recombinant 46 kDa hydratase 2 survives in a purified form under storage, thus being the first protein of this type amenable to application as a tool in metabolic studies | Rattus norvegicus |
EC Number | Cloned (Comment) | Organism |
---|---|---|
4.2.1.107 | expressed in Escherichia coli BL21(DE3) plysS cells, for analysis the role of the hydratase in the beta-oxidation | Rattus norvegicus |
4.2.1.119 | a truncated version (amino acid residues 318-735) of perMFE-2 is expressed in Escherichia coli BL21(DE3) plysS cells as a recombinant protein | Rattus norvegicus |
EC Number | General Stability | Organism |
---|---|---|
4.2.1.107 | still active after 3 h of incubation in the enzyme assay | Rattus norvegicus |
EC Number | KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|---|
4.2.1.119 | additional information | - |
additional information | kinetic parameters of the enzyme are measured with concentrations of substrates from 5 to 200 microM | Rattus norvegicus | |
4.2.1.119 | 4.6 | - |
(2E)-decenoyl-CoA | - |
Rattus norvegicus | |
4.2.1.119 | 8.7 | - |
(2E)-hexenoyl-CoA | - |
Rattus norvegicus | |
4.2.1.119 | 60 | - |
crotonyl-CoA | - |
Rattus norvegicus |
EC Number | Localization | Comment | Organism | GeneOntology No. | Textmining |
---|---|---|---|---|---|
4.2.1.17 | peroxisome | - |
Rattus norvegicus | 5777 | - |
4.2.1.107 | peroxisome | - |
Rattus norvegicus | 5777 | - |
4.2.1.119 | peroxisome | - |
Rattus norvegicus | 5777 | - |
EC Number | Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|---|
4.2.1.107 | 46000 | - |
SDS-PAGE | Rattus norvegicus |
4.2.1.107 | 46580 | - |
amino acid sequence analysis | Rattus norvegicus |
4.2.1.119 | 46000 | - |
recombinant hydratase 2, SDS-PAGE | Rattus norvegicus |
EC Number | Organism | UniProt | Comment | Textmining |
---|---|---|---|---|
4.2.1.17 | Rattus norvegicus | - |
- |
- |
4.2.1.107 | Rattus norvegicus | - |
- |
- |
4.2.1.119 | Rattus norvegicus | - |
Wistar rats | - |
EC Number | Purification (Comment) | Organism |
---|---|---|
4.2.1.107 | to homogeneity, 13% yield, anion- and cation exchange chromatography, and gel filtration on Superdex 200 HR | Rattus norvegicus |
4.2.1.119 | recombinant protein purified from the cell extract to apparent homogeneity by three chromatographic steps on anion-exchange, cation-exchange and size-exclusion columns | Rattus norvegicus |
EC Number | Source Tissue | Comment | Organism | Textmining |
---|---|---|---|---|
4.2.1.17 | liver | - |
Rattus norvegicus | - |
4.2.1.107 | liver | - |
Rattus norvegicus | - |
EC Number | Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
---|---|---|---|---|
4.2.1.107 | 48 | - |
recombinant enzyme, pH 7.0 | Rattus norvegicus |
4.2.1.119 | additional information | - |
activity is below the detection limit of the assay system when using extracts from non-transformed cells or cells transformed with the vector only | Rattus norvegicus |
4.2.1.119 | 33.4 | - |
pET-Hydr2 expressed in Escherichia coli, soluble extract of the cells | Rattus norvegicus |
4.2.1.119 | 48 | - |
recombinant 46 kDa hydratase 2, last purification step: size exclusion | Rattus norvegicus |
EC Number | Storage Stability | Organism |
---|---|---|
4.2.1.119 | The purified enzyme can be stored as an active enzyme for at least half a year at 4°C or frozen at -20°C. | Rattus norvegicus |
EC Number | Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|---|
4.2.1.17 | (2E)-enoyl-CoA + H2O | - |
Rattus norvegicus | (3S)-hydroxyacyl-CoA | - |
? | |
4.2.1.107 | (24E)-3alpha,7alpha,12alpha-trihydroxy-5beta-cholest-24-enoyl-CoA + H2O | the equilibrium lies in the direction of the dehydration | Rattus norvegicus | (24R,25R)-3alpha,7alpha,12alpha,24-tetrahydroxy-5beta-cholestanoyl-CoA | only the (24R,25R) diastomer as product, analyzed using reverse-phase HPLC | ? | |
4.2.1.119 | (24E)-3alpha,7alpha,12alpha-trihydroxy-5beta-cholest-24-enoyl-CoA + H2O | reaction of the recombinant enzyme, protein converted rapidly | Rattus norvegicus | (24R,25R)-3alpha,7alpha,12alpha,24-tetrahydroxy-5beta-cholestanoyl-CoA | a physiological intermediate in bile acid synthesis | ? | |
4.2.1.119 | (2E)-decenoyl-CoA + H2O | - |
Rattus norvegicus | (3R)-3-hydroxydecanoyl-CoA | - |
? | |
4.2.1.119 | (2E)-enoyl-CoA + H2O | straight-chain | Rattus norvegicus | (3R)-hydroxyacyl-CoA | - |
? | |
4.2.1.119 | (2E)-hexenoyl-CoA + H2O | - |
Rattus norvegicus | (3R)-3-hydroxyhexanoyl-CoA | - |
? | |
4.2.1.119 | Crotonyl-CoA + H2O | - |
Rattus norvegicus | (3R)-3-Hydroxybutanoyl-CoA | - |
? |
EC Number | Subunits | Comment | Organism |
---|---|---|---|
4.2.1.119 | monomer | size-exclusion chromatography on a Superdex 200 HR column gives a native molecular mass of 59 kDa, suggesting that the recombinant protein is monomeric | Rattus norvegicus |
EC Number | Synonyms | Comment | Organism |
---|---|---|---|
4.2.1.17 | 2-enoyl-CoA hydratase 1 | - |
Rattus norvegicus |
4.2.1.17 | perMFE-1 | multifunctional enzyme, cf. 5.3.3.8 and EC 1.1.1.35, second multifunctional enzyme in rat liver peroxisome perMFE-2, cf. EC 4.2.1.107 and EC 4.2.1.119 | Rattus norvegicus |
4.2.1.107 | 46 kDa hydratase 2 | - |
Rattus norvegicus |
4.2.1.107 | perMFE-2 | multifunctional enzyme, cf. EC 4.2.1.119, second multifunctional enzyme in rat liver peroxisome perMFE-1, cf. EC 1.1.1.35 and EC 5.3.3.8 | Rattus norvegicus |
4.2.1.119 | 2-enoyl-CoA hydratase | monofunctional, has not been observed as a wild-type protein. Part of perMFE-2 (2-enoyl-CoA hydratase 2/(R)-3-hydroxyacyl-CoA dehydrogenase) | Rattus norvegicus |
EC Number | Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|---|
4.2.1.119 | additional information | - |
additional information | kinetic parameters of the enzyme are measured with concentrations of substrates from 5 to 200 microM | Rattus norvegicus | |
4.2.1.119 | 2.3 | - |
crotonyl-CoA | - |
Rattus norvegicus | |
4.2.1.119 | 22.8 | - |
(2E)-hexenoyl-CoA | - |
Rattus norvegicus | |
4.2.1.119 | 26 | - |
(2E)-decenoyl-CoA | - |
Rattus norvegicus |
EC Number | pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|---|
4.2.1.107 | 7 | - |
assay at | Rattus norvegicus |
EC Number | General Information | Comment | Organism |
---|---|---|---|
4.2.1.107 | physiological function | product of the reaction (24R,25R)-3alpha,7alpha,12alpha,24-tetrahydroxy-5beta-cholestanoyl-CoA is characterized as a physiological intermediate in bile acid synthesis, in the beta-oxidative side-chain modifications | Rattus norvegicus |