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Literature summary for 1.1.1.176 extracted from

  • Braun, M.; Lunsdorf, H.; Buckmann, A.F.
    12 alpha-hydroxysteroid dehydrogenase from Clostridium group P, strain C 48-50. Production, purification and characterization (1991), Eur. J. Biochem., 196, 439-450.
    View publication on PubMed

General Stability

General Stability Organism
glycerol, 50% v/v, stabilization Clostridium sp.

Inhibitors

Inhibitors Comment Organism Structure
Ag+
-
Clostridium sp.
Cu2+
-
Clostridium sp.
Hg2+
-
Clostridium sp.
p-chloromercuribenzoate
-
Clostridium sp.
pyridoxal 5'-phosphate protection by NADP+ or NADPH Clostridium sp.

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.0012
-
12-oxochenodeoxycholic acid
-
Clostridium sp.
0.0085
-
NADPH
-
Clostridium sp.
0.035
-
NADP+
-
Clostridium sp.
0.045
-
deoxycholic acid
-
Clostridium sp.
0.072
-
cholic acid
-
Clostridium sp.

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
26000
-
4 * 26000, SDS-PAGE Clostridium sp.
26000
-
4 * 26000, no disulfide bonds, SDS-PAGE, electron microscopy Clostridium sp.
98000 106000 native PAGE, analytical ultracentrifugation Clostridium sp.
102000 108000 native PAGE, analytical ultracentrifugation, FPLC gel filtration Clostridium sp.

Organism

Organism UniProt Comment Textmining
Clostridium sp.
-
group P, strain 48-50
-

Purification (Commentary)

Purification (Comment) Organism
-
Clostridium sp.

Storage Stability

Storage Stability Organism
-30°C, 6 months, 50% glycerol, no loss of activity Clostridium sp.
-30°C, concentrated form, at least 6 months Clostridium sp.

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
12-oxochenodeoxycholic acid + NADP+
-
Clostridium sp. (3alpha,5beta,7alpha,12alpha)-3,7,12-trihydroxycholan-24-oic acid + NADPH + H+
-
?
cholic acid + NAD(P)+ hardly accepts N6-modified NADP+ derivative Clostridium sp. 3alpha,7alpha-dihydroxy-12-oxo-5beta-cholan-24-oic acid + NAD(P)H
-
?
cholic acid + NAD(P)+ hardly accepts N6-modified NADP+ derivative Clostridium sp. 3alpha,7alpha-dihydroxy-12-oxo-5beta-cholan-24-oic acid + NAD(P)H
-
r
cholic acid + NAD(P)+ i.e. 3alpha,7alpha,12alpha-trihydroxy-5beta-cholan-24-oic acid Clostridium sp. 3alpha,7alpha-dihydroxy-12-oxo-5beta-cholan-24-oic acid + NAD(P)H
-
?
cholic acid + NAD(P)+ i.e. 3alpha,7alpha,12alpha-trihydroxy-5beta-cholan-24-oic acid Clostridium sp. 3alpha,7alpha-dihydroxy-12-oxo-5beta-cholan-24-oic acid + NAD(P)H
-
r
deoxycholic acid + NAD(P)+ i.e. 3alpha,12alpha-dihydroxy-5beta-cholan-24-oic acid Clostridium sp. 3alpha-hydroxy-12-oxo-5beta-cholan-24-oic acid + NAD(P)H + H+
-
?

Subunits

Subunits Comment Organism
tetramer 4 * 26000, SDS-PAGE Clostridium sp.
tetramer 4 * 26000, no disulfide bonds, SDS-PAGE, electron microscopy Clostridium sp.

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
additional information
-
optimum above 55°C, not measurable because enzyme is inactivated above 55°C Clostridium sp.

Temperature Stability [°C]

Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
25
-
half-life: 4 days, increases to 10 days by addition of NADP+ Clostridium sp.
30
-
half-life: 2 days, increases to 5 days by addition of NADP+ Clostridium sp.

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
additional information
-
reduction of 12-oxochenodeoxycholic acid optimal below pH 5.0, not measurable due to insolubility of substrate Clostridium sp.
8.5 9.5 oxidation of cholic acid Clostridium sp.

Cofactor

Cofactor Comment Organism Structure
NAD+
-
Clostridium sp.
NADP+
-
Clostridium sp.
NADPH
-
Clostridium sp.