Cloned (Comment) | Organism |
---|---|
recombinant expression of N-terminally His6-tagged enzyme in Escherichia coli strain BL21(DE3) | Collinsella aerofaciens |
Crystallization (Comment) | Organism |
---|---|
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 | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
7beta-hydroxysteroid + NADP+ | Collinsella aerofaciens | - |
7-oxosteroid + NADPH + H+ | - |
r | |
7beta-hydroxysteroid + NADP+ | Collinsella aerofaciens ATCC 25986 | - |
7-oxosteroid + NADPH + H+ | - |
r | |
ursodeoxycholic acid + NADP+ | Collinsella aerofaciens | - |
7-oxo-lithocholic acid + NADPH + H+ | - |
r | |
ursodeoxycholic acid + NADP+ | Collinsella aerofaciens ATCC 25986 | - |
7-oxo-lithocholic acid + NADPH + H+ | - |
r |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Collinsella aerofaciens | A4ECA9 | - |
- |
Collinsella aerofaciens ATCC 25986 | A4ECA9 | - |
- |
Purification (Comment) | Organism |
---|---|
recombinant N-terminally His6-tagged enzyme from Escherichia coli strain BL21(DE3) by nickel affinity chromatography, ultrafiltration, and gel filtration | Collinsella aerofaciens |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
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 | |
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 | |
7beta-hydroxysteroid + NADP+ | - |
Collinsella aerofaciens | 7-oxosteroid + NADPH + H+ | - |
r | |
7beta-hydroxysteroid + NADP+ | stereoselective with 7-beta-hydroxysteroids | Collinsella aerofaciens | 7-oxosteroid + NADPH + H+ | - |
r | |
7beta-hydroxysteroid + NADP+ | - |
Collinsella aerofaciens ATCC 25986 | 7-oxosteroid + NADPH + H+ | - |
r | |
7beta-hydroxysteroid + NADP+ | stereoselective with 7-beta-hydroxysteroids | Collinsella aerofaciens ATCC 25986 | 7-oxosteroid + NADPH + H+ | - |
r | |
additional information | no activity with chenodeoxycholic acid, i.e. 3alpha,7alpha-dihydroxy-5beta-cholan-24-oic acid | Collinsella aerofaciens | ? | - |
? | |
additional information | no activity with chenodeoxycholic acid, i.e. 3alpha,7alpha-dihydroxy-5beta-cholan-24-oic acid | Collinsella aerofaciens ATCC 25986 | ? | - |
? | |
ursodeoxycholic acid + NADP+ | - |
Collinsella aerofaciens | 7-oxo-lithocholic acid + NADPH + H+ | - |
r | |
ursodeoxycholic acid + NADP+ | - |
Collinsella aerofaciens ATCC 25986 | 7-oxo-lithocholic acid + NADPH + H+ | - |
r |
Subunits | Comment | Organism |
---|---|---|
homodimer | a dimer of identical subunits arranged with C2 symmetry | Collinsella aerofaciens |
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 | Comment | Organism |
---|---|---|
7beta-HSDH | - |
Collinsella aerofaciens |
7beta-hydroxysteroid dehydrogenase | - |
Collinsella aerofaciens |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
NADP+ | binding site structure analysis | Collinsella aerofaciens | |
NADPH | - |
Collinsella aerofaciens |
General Information | Comment | Organism |
---|---|---|
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 |