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Literature summary extracted from

  • Savino, S.; Ferrandi, E.E.; Forneris, F.; Rovida, S.; Riva, S.; Monti, D.; Mattevi, A.
    Structural and biochemical insights into 7beta-hydroxysteroid dehydrogenase stereoselectivity (2016), Proteins, 84, 859-865.
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
1.1.1.201 recombinant expression of N-terminally His6-tagged enzyme in Escherichia coli strain BL21(DE3) Collinsella aerofaciens

Crystallization (Commentary)

EC Number Crystallization (Comment) Organism
1.1.1.201 purified His-tagged enzyme complexed with NADP+, mixing of 10-15 mg/ml protein in 20% v/v glycerol, 20 mM potassium phosphate, pH 7.0, and 5 mM NADP+, with1 M sodium formate, 100 mM sodium acetate, pH 4.6, at room temperature, 3-4days, X-ray diffraction structure determination and analysis at 1.6 A resolution, modelling, molecular replacement using the the apoform structure of 7alpha-HSDH from Escherichia coli as search model, PDB ID 1AHH Collinsella aerofaciens

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
1.1.1.201 7beta-hydroxysteroid + NADP+ Collinsella aerofaciens
-
 7-oxosteroid + NADPH + H+
-
 r
1.1.1.201 7beta-hydroxysteroid + NADP+ Collinsella aerofaciens ATCC 25986
-
 7-oxosteroid + NADPH + H+
-
 r
1.1.1.201 ursodeoxycholic acid + NADP+ Collinsella aerofaciens
-
 7-oxo-lithocholic acid + NADPH + H+
-
 r
1.1.1.201 ursodeoxycholic acid + NADP+ Collinsella aerofaciens ATCC 25986
-
 7-oxo-lithocholic acid + NADPH + H+
-
 r

Organism

EC Number Organism UniProt Comment Textmining
1.1.1.201 Collinsella aerofaciens A4ECA9
-
-
1.1.1.201 Collinsella aerofaciens ATCC 25986 A4ECA9
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
1.1.1.201 recombinant N-terminally His6-tagged enzyme from Escherichia coli strain BL21(DE3) by nickel affinity chromatography, ultrafiltration, and gel filtration Collinsella aerofaciens

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
1.1.1.201 3alpha,7beta-dihydroxy-5beta-cholan-24-oic acid + NADP+ i.e. ursodeoxycholic acid, exclusive specificity for the hydroxyl group in beta configuration at the C-7 position Collinsella aerofaciens 7-oxo-lithocholic acid + NADPH + H+
-
 r
1.1.1.201 3alpha,7beta-dihydroxy-5beta-cholan-24-oic acid + NADP+ i.e. ursodeoxycholic acid, exclusive specificity for the hydroxyl group in beta configuration at the C-7 position Collinsella aerofaciens ATCC 25986 7-oxo-lithocholic acid + NADPH + H+
-
 r
1.1.1.201 7beta-hydroxysteroid + NADP+
-
 Collinsella aerofaciens 7-oxosteroid + NADPH + H+
-
 r
1.1.1.201 7beta-hydroxysteroid + NADP+ stereoselective with 7-beta-hydroxysteroids Collinsella aerofaciens 7-oxosteroid + NADPH + H+
-
 r
1.1.1.201 7beta-hydroxysteroid + NADP+
-
 Collinsella aerofaciens ATCC 25986 7-oxosteroid + NADPH + H+
-
 r
1.1.1.201 7beta-hydroxysteroid + NADP+ stereoselective with 7-beta-hydroxysteroids Collinsella aerofaciens ATCC 25986 7-oxosteroid + NADPH + H+
-
 r
1.1.1.201 additional information no activity with chenodeoxycholic acid, i.e. 3alpha,7alpha-dihydroxy-5beta-cholan-24-oic acid Collinsella aerofaciens ?
-
 ?
1.1.1.201 additional information no activity with chenodeoxycholic acid, i.e. 3alpha,7alpha-dihydroxy-5beta-cholan-24-oic acid Collinsella aerofaciens ATCC 25986 ?
-
 ?
1.1.1.201 ursodeoxycholic acid + NADP+
-
 Collinsella aerofaciens 7-oxo-lithocholic acid + NADPH + H+
-
 r
1.1.1.201 ursodeoxycholic acid + NADP+
-
 Collinsella aerofaciens ATCC 25986 7-oxo-lithocholic acid + NADPH + H+
-
 r

Subunits

EC Number Subunits Comment Organism
1.1.1.201 homodimer a dimer of identical subunits arranged with C2 symmetry Collinsella aerofaciens
1.1.1.201 More crystal structure analysis reveals a loop gating active site accessibility, the bases of the specificity for NADP+, and the general architecture of the steroid binding site Collinsella aerofaciens

Synonyms

EC Number Synonyms Comment Organism
1.1.1.201 7beta-HSDH
-
 Collinsella aerofaciens
1.1.1.201 7beta-hydroxysteroid dehydrogenase
-
 Collinsella aerofaciens

Cofactor

EC Number Cofactor Comment Organism Structure
1.1.1.201 NADP+ binding site structure analysis Collinsella aerofaciens
1.1.1.201 NADPH
-
 Collinsella aerofaciens

General Information

EC Number General Information Comment Organism
1.1.1.201 additional information crystal structure analysis reveals a loop gating active site accessibility, the bases of the specificity for NADP+, and the general architecture of the steroid binding site. Conserved residues Ser143 and Tyr156 of 7beta-HSDH are homologous with and structurally superimpose to Ser146 and Tyr159 of 7alpha-HSDH Collinsella aerofaciens