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 |
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 |
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 |
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 | - |
? |
EC Number | Organism | UniProt | Comment | Textmining |
---|---|---|---|---|
1.14.14.17 | Rattus norvegicus | - |
- |
- |
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 |
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 |
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 | - |
? |
EC Number | Synonyms | Comment | Organism |
---|---|---|---|
1.14.14.17 | squalene epoxidase | - |
Rattus norvegicus |
EC Number | Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|---|
1.14.14.17 | 37 | - |
assay at | Rattus norvegicus |
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 |
EC Number | pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|---|
1.14.14.17 | 7.4 | - |
assay at | Rattus norvegicus |
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 |