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Literature summary extracted from

  • Qin, Y.; Haapalainen, A.M.; Conry, D.; Cuebas, D.A.; Hiltunen, J.K.; Novikov, D.K.
    Recombinant 2-enoyl-CoA hydratase derived from rat peroxisomal multifunctional enzyme 2: role of the hydratase reaction in bile acid synthesis (1997), Biochem. J., 328, 377-382.
No PubMed abstract available

Application

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

Cloned(Commentary)

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

General Stability

EC Number General Stability Organism
4.2.1.107 still active after 3 h of incubation in the enzyme assay Rattus norvegicus

KM Value [mM]

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

Localization

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
-

Molecular Weight [Da]

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

Organism

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
-

Purification (Commentary)

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

Source Tissue

EC Number Source Tissue Comment Organism Textmining
4.2.1.17 liver
-
 Rattus norvegicus
-
4.2.1.107 liver
-
 Rattus norvegicus
-

Specific Activity [micromol/min/mg]

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

Storage Stability

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

Substrates and Products (Substrate)

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
-
 ?

Subunits

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

Synonyms

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

Turnover Number [1/s]

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

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
4.2.1.107 7
-
 assay at Rattus norvegicus

General Information

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