Cloned (Comment) | Organism |
---|---|
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 | Comment | Organism |
---|---|---|
F203A | site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme | Rattus norvegicus |
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 |
F228A | site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme | Rattus norvegicus |
F287A | site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme | Rattus norvegicus |
F305A | site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme | Rattus norvegicus |
F375A | site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme | Rattus norvegicus |
F476A | site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme | Rattus norvegicus |
F491A | site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme | Rattus norvegicus |
F522A | site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme | Rattus norvegicus |
F523A | site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme | Rattus norvegicus |
Y194A | site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme | Rattus norvegicus |
Y209A | site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme | Rattus norvegicus |
Y334A | site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme | Rattus norvegicus |
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 |
Y493A | site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme | Rattus norvegicus |
Y528A | site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme | Rattus norvegicus |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.00425 | - |
(3S)-squalene-2,3-epoxide | pH 7.4, 37°C, recombinant His-tagged wild-type enzyme | Rattus norvegicus | |
0.00987 | - |
squalene | pH 7.4, 37°C, recombinant His-tagged wild-type enzyme | Rattus norvegicus |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
squalene + AH2 + O2 | Rattus norvegicus | rate-limiting step in chloesterol biosynthesis | (3S)-squalene-2,3-epoxide + A + H2O | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Rattus norvegicus | - |
- |
- |
Purification (Comment) | Organism |
---|---|
recombinant His-tagged wild-type and mutant enzymes from Escherichia coli strain BL21(DE3) by nickel affinity chromatography | Rattus norvegicus |
Reaction | Comment | Organism | Reaction ID |
---|---|---|---|
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 |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
(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 | - |
? | |
squalene + AH2 + O2 | rate-limiting step in chloesterol biosynthesis | Rattus norvegicus | (3S)-squalene-2,3-epoxide + A + H2O | - |
? | |
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 | Comment | Organism |
---|---|---|
squalene epoxidase | - |
Rattus norvegicus |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
37 | - |
assay at | Rattus norvegicus |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.0075 | - |
(3S)-squalene-2,3-epoxide | pH 7.4, 37°C, recombinant His-tagged wild-type enzyme | Rattus norvegicus | |
0.076 | - |
squalene | pH 7.4, 37°C, recombinant His-tagged wild-type enzyme | Rattus norvegicus |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7.4 | - |
assay at | Rattus norvegicus |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
FAD | sequence determination of the FAD-binding site with the dinucleotide-binding GXGXXG motif, and DG and GD motif | Rattus norvegicus |