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Literature summary 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

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

Inhibitors

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

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
1.1.1.176 0.0012
-
 12-oxochenodeoxycholic acid
-
 Clostridium sp.
1.1.1.176 0.0085
-
 NADPH
-
 Clostridium sp.
1.1.1.176 0.035
-
 NADP+
-
 Clostridium sp.
1.1.1.176 0.045
-
 deoxycholic acid
-
 Clostridium sp.
1.1.1.176 0.072
-
 cholic acid
-
 Clostridium sp.

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
1.1.1.176 26000
-
 4 * 26000, SDS-PAGE Clostridium sp.
1.1.1.176 26000
-
 4 * 26000, no disulfide bonds, SDS-PAGE, electron microscopy Clostridium sp.
1.1.1.176 98000 106000 native PAGE, analytical ultracentrifugation Clostridium sp.
1.1.1.176 102000 108000 native PAGE, analytical ultracentrifugation, FPLC gel filtration Clostridium sp.

Organism

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

Purification (Commentary)

EC Number Purification (Comment) Organism
1.1.1.176
-
 Clostridium sp.

Storage Stability

EC Number Storage Stability Organism
1.1.1.176 -30°C, 6 months, 50% glycerol, no loss of activity Clostridium sp.
1.1.1.176 -30°C, concentrated form, at least 6 months Clostridium sp.

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
1.1.1.176 12-oxochenodeoxycholic acid + NADP+
-
 Clostridium sp. (3alpha,5beta,7alpha,12alpha)-3,7,12-trihydroxycholan-24-oic acid + NADPH + H+
-
 ?
1.1.1.176 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
-
 ?
1.1.1.176 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
1.1.1.176 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
-
 ?
1.1.1.176 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
1.1.1.176 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

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

Temperature Optimum [°C]

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

Temperature Stability [°C]

EC Number Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
1.1.1.176 25
-
 half-life: 4 days, increases to 10 days by addition of NADP+ Clostridium sp.
1.1.1.176 30
-
 half-life: 2 days, increases to 5 days by addition of NADP+ Clostridium sp.

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
1.1.1.176 additional information
-
 reduction of 12-oxochenodeoxycholic acid optimal below pH 5.0, not measurable due to insolubility of substrate Clostridium sp.
1.1.1.176 8.5 9.5 oxidation of cholic acid Clostridium sp.

Cofactor

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