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Literature summary for 1.1.1.50 extracted from

  • Yang, H.; Fang, Y.; Wang, Z.; Zhang, L.
    One step affinity recovery of 3alpha-hydroxysteroid dehydrogenase from cloned Escherichia coli (2015), J. Chromatogr. B, 991, 79-84.
    View publication on PubMed

Cloned(Commentary)

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 [Da] Molecular Weight Maximum [Da] Comment Organism
23000
-
-
Comamonas testosteroni
56000
-
about, gel filtration, recombinant enzyme Comamonas testosteroni

Natural Substrates/ Products (Substrates)

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

Organism UniProt Comment Textmining
Comamonas testosteroni P80702
-
-
Comamonas testosteroni ATCC 11996 P80702
-
-

Purification (Commentary)

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 [micromol/min/mg]

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 and Products (Substrate)

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

Subunits Comment Organism
homodimer non-covalent homodimer , 2 * 23000, about, recombinant enzyme, SDS-PAGE Comamonas testosteroni

Synonyms

Synonyms Comment Organism
3alpha-HSD
-
Comamonas testosteroni
3alpha-hydroxysteroid dehydrogenase
-
Comamonas testosteroni
hsdA
-
Comamonas testosteroni

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
25
-
assay at Comamonas testosteroni

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
11
-
assay at Comamonas testosteroni

Cofactor

Cofactor Comment Organism Structure
NAD+
-
Comamonas testosteroni
NADH
-
Comamonas testosteroni