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Literature summary extracted from

  • Abe, I.; Abe, T.; Lou, W.; Masuoka, T.; Noguchi, H.
    Site-directed mutagenesis of conserved aromatic residues in rat squalene epoxidase (2007), Biochem. Biophys. Res. Commun., 352, 259-263.
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
1.14.14.17 DNA and amino acid sequence determination and analysis, expression of His-tagged wild-type and mutant enzymes in Escherichia coli strain BL21(DE3) Rattus norvegicus

Protein Variants

EC Number Protein Variants Comment Organism
1.14.14.17 F203A site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme Rattus norvegicus
1.14.14.17 F223A site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme, the F223A mutant no longer accepts (3S)2,3-oxidosqualene as a substrate Rattus norvegicus
1.14.14.17 F228A site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme Rattus norvegicus
1.14.14.17 F287A site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme Rattus norvegicus
1.14.14.17 F305A site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme Rattus norvegicus
1.14.14.17 F375A site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme Rattus norvegicus
1.14.14.17 F476A site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme Rattus norvegicus
1.14.14.17 F491A site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme Rattus norvegicus
1.14.14.17 F522A site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme Rattus norvegicus
1.14.14.17 F523A site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme Rattus norvegicus
1.14.14.17 Y194A site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme Rattus norvegicus
1.14.14.17 Y209A site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme Rattus norvegicus
1.14.14.17 Y334A site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme Rattus norvegicus
1.14.14.17 Y473A site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme, the mutant converts (3S)2,3-oxidosqualene to (3S,22S)2,3-22,23-dioxidosqualene twice more efficiently than wild-type enzyme Rattus norvegicus
1.14.14.17 Y493A site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme Rattus norvegicus
1.14.14.17 Y528A site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme Rattus norvegicus

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
1.14.14.17 0.00425
-
 (3S)-squalene-2,3-epoxide pH 7.4, 37°C, recombinant His-tagged wild-type enzyme Rattus norvegicus
1.14.14.17 0.00987
-
 squalene pH 7.4, 37°C, recombinant His-tagged wild-type enzyme Rattus norvegicus

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
1.14.14.17 squalene + AH2 + O2 Rattus norvegicus rate-limiting step in chloesterol biosynthesis (3S)-squalene-2,3-epoxide + A + H2O
-
 ?

Organism

EC Number Organism UniProt Comment Textmining
1.14.14.17 Rattus norvegicus
-
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
1.14.14.17 recombinant His-tagged wild-type and mutant enzymes from Escherichia coli strain BL21(DE3) by nickel affinity chromatography Rattus norvegicus

Reaction

EC Number Reaction Comment Organism Reaction ID
1.14.14.17 squalene + [reduced NADPH-hemoprotein reductase] + O2 = (3S)-2,3-epoxy-2,3-dihydrosqualene + [oxidized NADPH-hemoprotein reductase] + H2O aromatic amino acid residues located at the substrate-binding domain of the active-site, e.g. Ph223 and Tyr473, control the stereochemical course of the enzyme reaction, mechanism of regio- and stereo-specific epoxidation of squalene to (3S)2,3-oxidosqualene, overview Rattus norvegicus
a

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
1.14.14.17 (3S)-squalene-2,3-epoxide + AH2 + O2 the wild-type enzyme also catalyzes conversion of (3S)2,3-oxidosqualene to (3S,22S)-2,3-22,23-dioxidosqualene Rattus norvegicus (3S,22S)-2,3-22,23-dioxidosqualene + A + H2O
-
 ?
1.14.14.17 squalene + AH2 + O2 rate-limiting step in chloesterol biosynthesis Rattus norvegicus (3S)-squalene-2,3-epoxide + A + H2O
-
 ?
1.14.14.17 squalene + AH2 + O2 aromatic amino acid residues located at the substrate-binding domain of the active-site, e.g. Ph223 and Tyr473, control the stereochemical course of the enzyme reaction, mechanism of regio- and stereo-specific epoxidation of squalene to (3S)2,3-oxidosqualene, overview Rattus norvegicus (3S)-squalene-2,3-epoxide + A + H2O
-
 ?

Synonyms

EC Number Synonyms Comment Organism
1.14.14.17 squalene epoxidase
-
 Rattus norvegicus

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
1.14.14.17 37
-
 assay at Rattus norvegicus

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
1.14.14.17 0.0075
-
 (3S)-squalene-2,3-epoxide pH 7.4, 37°C, recombinant His-tagged wild-type enzyme Rattus norvegicus
1.14.14.17 0.076
-
 squalene pH 7.4, 37°C, recombinant His-tagged wild-type enzyme Rattus norvegicus

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
1.14.14.17 7.4
-
 assay at Rattus norvegicus

Cofactor

EC Number Cofactor Comment Organism Structure
1.14.14.17 FAD sequence determination of the FAD-binding site with the dinucleotide-binding GXGXXG motif, and DG and GD motif Rattus norvegicus