Cloned (Comment) | Organism |
---|---|
gene hsdA, recombinant expression of His-tagged and untagged enzyme in Escherchia coli strain BL21(DE3) | Comamonas testosteroni |
Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|
23000 | - |
- |
Comamonas testosteroni |
56000 | - |
about, gel filtration, recombinant enzyme | Comamonas testosteroni |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
a 3alpha-hydroxysteroid + NAD+ | Comamonas testosteroni | - |
a 3-oxosteroid + NADH + H+ | - |
r | |
a 3alpha-hydroxysteroid + NAD+ | Comamonas testosteroni ATCC 11996 | - |
a 3-oxosteroid + NADH + H+ | - |
r | |
additional information | Comamonas testosteroni | the enzyme catalyzes reversibly the oxidoreduction of 3alpha-hydroxyl groups of steroid hormones | ? | - |
? | |
additional information | Comamonas testosteroni ATCC 11996 | the enzyme catalyzes reversibly the oxidoreduction of 3alpha-hydroxyl groups of steroid hormones | ? | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Comamonas testosteroni | P80702 | - |
- |
Comamonas testosteroni ATCC 11996 | P80702 | - |
- |
Purification (Comment) | Organism |
---|---|
recombinant His-tagged and untagged enzyme from Escherchia coli strain BL21(DE3) nickel affinity chromatography, and by deoxycholic acid affinity chromatography, respectively, the latter to 94-98% purity. Deoxycholic acid is linked to the resin Sepharose 4B with the aid of the spacers as cyanuric chloride and ethanediamine. Adsorption analysis by adapting the adsorption isotherm to a specific binding model, the Scatchard analysis model, overview. Comparison of purification methods between synthetic affinity medium and Ni2+-Sepharose column | Comamonas testosteroni |
Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
---|---|---|---|
3.94 | - |
recombinant His-tagged enzyme purified via Ni2+ affinity chromatography, pH 11.0, 25°C | Comamonas testosteroni |
15.4 | - |
recombinant untagged enzyme purified via deoxycholic acid affinity chromatography, pH 11.0, 25°C | Comamonas testosteroni |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
a 3alpha-hydroxysteroid + NAD+ | - |
Comamonas testosteroni | a 3-oxosteroid + NADH + H+ | - |
r | |
a 3alpha-hydroxysteroid + NAD+ | - |
Comamonas testosteroni ATCC 11996 | a 3-oxosteroid + NADH + H+ | - |
r | |
deoxycholic acid + NADH | - |
Comamonas testosteroni | 12alpha-hydroxy-3-oxo-5beta-cholan-24-oic acid + NAD+ | - |
r | |
deoxycholic acid + NADH | - |
Comamonas testosteroni ATCC 11996 | 12alpha-hydroxy-3-oxo-5beta-cholan-24-oic acid + NAD+ | - |
r | |
additional information | the enzyme catalyzes reversibly the oxidoreduction of 3alpha-hydroxyl groups of steroid hormones | Comamonas testosteroni | ? | - |
? | |
additional information | the enzyme catalyzes reversibly the oxidoreduction of 3alpha-hydroxyl groups of steroid hormones | Comamonas testosteroni ATCC 11996 | ? | - |
? |
Subunits | Comment | Organism |
---|---|---|
homodimer | non-covalent homodimer , 2 * 23000, about, recombinant enzyme, SDS-PAGE | Comamonas testosteroni |
Synonyms | Comment | Organism |
---|---|---|
3alpha-HSD | - |
Comamonas testosteroni |
3alpha-hydroxysteroid dehydrogenase | - |
Comamonas testosteroni |
hsdA | - |
Comamonas testosteroni |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
25 | - |
assay at | Comamonas testosteroni |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
11 | - |
assay at | Comamonas testosteroni |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
NAD+ | - |
Comamonas testosteroni | |
NADH | - |
Comamonas testosteroni |