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Literature summary for 1.1.1.238 extracted from

  • Edenharder, R.; Pfutzner, A.
    Characterization of NADP-dependent 12beta-hydroxysteroid dehydrogenase from Clostridium paraputrificum (1988), Biochim. Biophys. Acta, 962, 362-370.
    View publication on PubMed

Application

Application Comment Organism
analysis potential application in quantification of 12beta-hydroxyl groups in di- and trisubstituted bile acids Clostridium paraputrificum

General Stability

General Stability Organism
most stable in presence of reducing agents e.g. dithiothreitol Clostridium paraputrificum

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.0837
-
7,12-Dioxo-5beta-cholanoate
-
Clostridium paraputrificum
0.0854
-
NADPH
-
Clostridium paraputrificum
0.0895
-
3,12-Dioxo-5beta-cholanoate
-
Clostridium paraputrificum
0.138
-
7alpha-Hydroxy-3,12-dioxo-5beta-cholanoate
-
Clostridium paraputrificum
0.141
-
3alpha-Hydroxy-12-oxo-5beta-cholanoate
-
Clostridium paraputrificum
0.142
-
3,7,12-trioxotaureate
-
Clostridium paraputrificum
0.144
-
3alpha-Hydroxy-7,12-dioxo-5beta-cholanoate
-
Clostridium paraputrificum
0.171
-
3,7,12-Trioxo-5beta-cholanoate
-
Clostridium paraputrificum
0.274
-
3alpha,7alpha-dihydroxy-12-oxo-5beta-cholanoate
-
Clostridium paraputrificum

Localization

Localization Comment Organism GeneOntology No. Textmining
membrane
-
Clostridium paraputrificum 16020
-

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
126000
-
gel filtration Clostridium paraputrificum

Organism

Organism UniProt Comment Textmining
Clostridium paraputrificum
-
-
-

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
2 4.4 depending on substrate Clostridium paraputrificum
2.2 6.4 depending on inducer Clostridium paraputrificum

Storage Stability

Storage Stability Organism
-20°C, lyophilized powder, at least 4 weeks Clostridium paraputrificum

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
3,12-dioxo-5beta-cholanoate + NADPH
-
Clostridium paraputrificum 12beta-hydroxy-3-oxo-5beta-cholanoate + NADP+
-
?
3,7,12-trioxo-5beta-cholanoate + NADPH
-
Clostridium paraputrificum 12beta-hydroxy-3,7-dioxo-5beta-cholanoate + NADP+
-
?
3,7,12-trioxotaureate + NADPH
-
Clostridium paraputrificum 12beta-hydroxy-3,7-dioxotaureate + NADP+
-
?
3alpha,7alpha-dihydroxy-12-oxo-5beta-cholanoate + NADPH
-
Clostridium paraputrificum 3alpha,7alpha,12beta-trihydroxy-5beta-cholanoate + NADP+
-
?
3alpha-hydroxy-12-oxo-5beta-cholanoate + NADPH reverse reaction 10% the rate of forward reaction Clostridium paraputrificum 3alpha,12beta-dihydroxy-5beta-cholanoate + NADP+
-
r
3alpha-hydroxy-7,12-dioxo-5beta-cholanoate + NADPH
-
Clostridium paraputrificum 3alpha,12beta-dihydroxy-7-oxo-5beta-cholanoate + NADP+
-
?
7,12-dioxo-5beta-cholanoate + NADPH
-
Clostridium paraputrificum 12beta-hydroxy-7-oxo-5beta-cholanoate + NADP+
-
?
7alpha-hydroxy-3,12-dioxo-5beta-cholanoate + NADPH
-
Clostridium paraputrificum 7alpha,12beta-dihydroxy-3-oxo-5beta-cholanoate + NADP+
-
?

Temperature Stability [°C]

Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
4
-
at least 6 h Clostridium paraputrificum
37
-
45 min, complete inactivation Clostridium paraputrificum
50
-
10 min, 95% inactivation, 15 min complete inactivation Clostridium paraputrificum

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
7.8
-
oxidation of 12beta-hydroxy bile acids Clostridium paraputrificum
10
-
reduction of 12-oxo bile acids Clostridium paraputrificum

pH Range

pH Minimum pH Maximum Comment Organism
7 11 about half maximum activity Clostridium paraputrificum

pH Stability

pH Stability pH Stability Maximum Comment Organism
8.5
-
most stable Clostridium paraputrificum

Cofactor

Cofactor Comment Organism Structure
NADP+
-
Clostridium paraputrificum
NADPH specific for, no activity with NADH Clostridium paraputrificum