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Literature summary for 4.2.1.119 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
Search for more literature for 4.2.1.119

Application

Application Comment Organism
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)

Cloned (Comment) Organism
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

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
additional information
-
 additional information kinetic parameters of the enzyme are measured with concentrations of substrates from 5 to 200 microM Rattus norvegicus
4.6
-
 (2E)-decenoyl-CoA
-
 Rattus norvegicus
8.7
-
 (2E)-hexenoyl-CoA
-
 Rattus norvegicus
60
-
 crotonyl-CoA
-
 Rattus norvegicus

Localization

Localization Comment Organism GeneOntology No. Textmining
peroxisome
-
 Rattus norvegicus 5777
-

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
46000
-
 recombinant hydratase 2, SDS-PAGE Rattus norvegicus

Organism

Organism UniProt Comment Textmining
Rattus norvegicus
-
 Wistar rats
-

Purification (Commentary)

Purification (Comment) Organism
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

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
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
33.4
-
 pET-Hydr2 expressed in Escherichia coli, soluble extract of the cells Rattus norvegicus
48
-
 recombinant 46 kDa hydratase 2, last purification step: size exclusion Rattus norvegicus

Storage Stability

Storage Stability Organism
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)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
(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 ?
(2E)-decenoyl-CoA + H2O
-
 Rattus norvegicus (3R)-3-hydroxydecanoyl-CoA
-
 ?
(2E)-enoyl-CoA + H2O straight-chain Rattus norvegicus (3R)-hydroxyacyl-CoA
-
 ?
(2E)-hexenoyl-CoA + H2O
-
 Rattus norvegicus (3R)-3-hydroxyhexanoyl-CoA
-
 ?
Crotonyl-CoA + H2O
-
 Rattus norvegicus (3R)-3-Hydroxybutanoyl-CoA
-
 ?

Subunits

Subunits Comment Organism
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

Synonyms Comment Organism
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]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
additional information
-
 additional information kinetic parameters of the enzyme are measured with concentrations of substrates from 5 to 200 microM Rattus norvegicus
2.3
-
 crotonyl-CoA
-
 Rattus norvegicus
22.8
-
 (2E)-hexenoyl-CoA
-
 Rattus norvegicus
26
-
 (2E)-decenoyl-CoA
-
 Rattus norvegicus