Any feedback?
Please rate this page
(literature.php)
(0/150)

BRENDA support

Literature summary for 1.1.1.50 extracted from

  • Chang, Y.H.; Huang, T.J.; Chuang, L.Y.; Hwang, C.C.
    Role of S114 in the NADH-induced conformational change and catalysis of 3alpha-hydroxysteroid dehydrogenase/carbonyl reductase from Comamonas testosteroni (2009), Biochim. Biophys. Acta, 1794, 1459-1466.
    View publication on PubMed
Search for more literature for 1.1.1.50

Cloned(Commentary)

Cloned (Comment) Organism
expressed in Escherichia coli BL21 (DE3) cells Comamonas testosteroni

Protein Variants

Protein Variants Comment Organism
S114A the mutation eliminates the hydrogen bonding interaction with P185, causing a conformational change in a nonproductive binding of NADH and a significant loss of activity, the mutant enzyme decreases 3100fold in V/Et value with no apparent change in Km value for substrates Comamonas testosteroni
S114A mutant enzyme exhibits a pronounced increase in the magnitude of ellipticity at 222 nm. S114A mutant enzyme decreases 3100fold in catalytic efficiency with no apparent change in Km for substrates. Addition of NADH to S114A mutant enzyme induces a secondary structural change Comamonas testosteroni

Inhibitors

Inhibitors Comment Organism Structure
NADH
-
 Comamonas testosteroni

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.0012
-
 5alpha-androstane-3,17-dione wild-type, pH 7.5, 25°C Comamonas testosteroni
0.0012
-
 5alpha-androstan-3,17-dione wild type enzyme, in 0.1 M HEPES at pH 7.5 Comamonas testosteroni
0.0031
-
 5alpha-androstan-3,17-dione mutant enzyme S114A, in 0.1 M HEPES at pH 7.5 Comamonas testosteroni
0.0031
-
 5alpha-androstane-3,17-dione mutant S114A, pH 7.5, 25°C Comamonas testosteroni
0.004
-
 NADH wild-type, pH 7.5, 25°C Comamonas testosteroni
0.004
-
 NADH wild type enzyme, in 0.1 M HEPES at pH 7.5 Comamonas testosteroni
0.0065
-
 NADH mutant enzyme S114A, in 0.1 M HEPES at pH 7.5 Comamonas testosteroni
0.0065
-
 NADH mutant S114A, pH 7.5, 25°C Comamonas testosteroni

Organism

Organism UniProt Comment Textmining
Comamonas testosteroni P80702
-
-

Purification (Commentary)

Purification (Comment) Organism
Ni-NTA column chromatography Comamonas testosteroni

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
5alpha-androstane-3,17-dione + NADH + H+
-
 Comamonas testosteroni androsterone + NAD+
-
 ?
androsterone + NAD+
-
 Comamonas testosteroni 5alpha-androstan-3,17-dione + NADH + H+
-
 ?

Synonyms

Synonyms Comment Organism
3alpha-hydroxysteroid dehydrogenase/carbonyl reductase
-
 Comamonas testosteroni

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
0.02
-
 NADH mutant S114A, pH 7.5, 25°C Comamonas testosteroni
63
-
 NADH wild-type, pH 7.5, 25°C Comamonas testosteroni

Cofactor

Cofactor Comment Organism Structure
NAD+
-
 Comamonas testosteroni
NADH NADH association to enzyme involves a bimolecular binding step and isomerization. The binding of NADH into a hydrophobic pocket in the active site restricts its motion and shields the fluorescence quenching from solvent Comamonas testosteroni

Ki Value [mM]

Ki Value [mM] Ki Value maximum [mM] Inhibitor Comment Organism Structure
0.0034
-
 NADH wild-type, pH 7.5, 25°C Comamonas testosteroni
0.006
-
 NADH mutant S114A, pH 7.5, 25°C Comamonas testosteroni

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
3.8
-
 NADH mutant S114A, pH 7.5, 25°C Comamonas testosteroni
6.5
-
 5alpha-androstane-3,17-dione mutant S114A, pH 7.5, 25°C Comamonas testosteroni
15000
-
 NADH wild-type, pH 7.5, 25°C Comamonas testosteroni
54000
-
 5alpha-androstane-3,17-dione wild-type, pH 7.5, 25°C Comamonas testosteroni