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Literature summary for 1.1.1.50 extracted from

  • Shiota, A.; Inaba, S.; Oda, M.
    Effects of active site residues of 3alpha-hydroxysteroid dehydrogenase from Pseudomonas sp. b-0831 on its catalysis and cofactor binding (2018), Biosci. Biotechnol. Biochem., 82, 1702-1707 .
    View publication on PubMed
Search for more literature for 1.1.1.50

Cloned(Commentary)

Cloned (Comment) Organism
overoverexpression with an N-terminal His-tag in Escherichia coli Pseudomonas sp. B-0831

Protein Variants

Protein Variants Comment Organism
K157A the NADH binding affinity of K157A mutant is much lower than that of the wild-type, mainly due to loss of a hydrogen bond Pseudomonas sp. B-0831
S114A mutant shows higher Km and lower kcat values in both oxidation and reduction reactions Pseudomonas sp. B-0831
S114A/Y153F double mutation results in a significant decrease in kcat relative to the single mutant Y153F Pseudomonas sp. B-0831
Y153F mutant shows higher Km and lower kcat values in both oxidation and reduction reactions. Loss of hydrogen bonding with NADH upon the Y153F mutation results in increased enthalpy change Pseudomonas sp. B-0831

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
51
-
 dehydrocholate 37°C, pH 8.0, wild-type enzyme Pseudomonas sp. B-0831
56
-
 cholate 37°C, pH 8.0, wild-type enzyme Pseudomonas sp. B-0831
78
-
 cholate 37°C, pH 8.0, mutant enzyme S114A/Y153F Pseudomonas sp. B-0831
96
-
 cholate 37°C, pH 8.0, mutant enzyme K157A Pseudomonas sp. B-0831
239
-
 dehydrocholate 37°C, pH 8.0, mutant enzyme S114A Pseudomonas sp. B-0831
468
-
 cholate 37°C, pH 8.0, mutant enzyme Y153F Pseudomonas sp. B-0831
586
-
 cholate 37°C, pH 8.0, mutant enzyme S114A Pseudomonas sp. B-0831
2489
-
 dehydrocholate 37°C, pH 8.0, mutant enzyme Y153F Pseudomonas sp. B-0831

Organism

Organism UniProt Comment Textmining
Pseudomonas sp. B-0831 Q59718
-
-

Purification (Commentary)

Purification (Comment) Organism
-
 Pseudomonas sp. B-0831

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
cholate + NAD+
-
 Pseudomonas sp. B-0831 7alpha,12alpha-dihydroxy-5-beta-cholan-3-one-24-oic acid + NADH + H+
-
 ?
dehydrocholate + NADH + H+
-
 Pseudomonas sp. B-0831 ?
-
 ?

Synonyms

Synonyms Comment Organism
3alpha-hydroxysteroid dehydrogenase
-
 Pseudomonas sp. B-0831
Ps3alphaHSD
-
 Pseudomonas sp. B-0831

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
0.045
-
 cholate 37°C, pH 8.0, mutant enzyme K157A Pseudomonas sp. B-0831
0.12
-
 cholate 37°C, pH 8.0, mutant enzyme S114A/Y153F Pseudomonas sp. B-0831
0.62
-
 cholate 37°C, pH 8.0, mutant enzyme Y153F Pseudomonas sp. B-0831
0.8
-
 cholate 37°C, pH 8.0, mutant enzyme S114A Pseudomonas sp. B-0831
0.89
-
 dehydrocholate 37°C, pH 8.0, mutant enzyme S114A Pseudomonas sp. B-0831
1.1
-
 dehydrocholate 37°C, pH 8.0, mutant enzyme Y153F Pseudomonas sp. B-0831
23
-
 cholate 37°C, pH 8.0, wild-type enzyme Pseudomonas sp. B-0831
48
-
 dehydrocholate 37°C, pH 8.0, wild-type enzyme Pseudomonas sp. B-0831