BRENDA - Enzyme Database show
show all sequences of 1.1.1.176

12 alpha-hydroxysteroid dehydrogenase from Clostridium group P, strain C 48-50. Production, purification and characterization

Braun, M.; Lunsdorf, H.; Buckmann, A.F.; Eur. J. Biochem. 196, 439-450 (1991)

Data extracted from this reference:

General Stability
General Stability
Organism
glycerol, 50% v/v, stabilization
Clostridium sp.
Inhibitors
Inhibitors
Commentary
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
Commentary
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]
Commentary
Organism
26000
-
4 * 26000, no disulfide bonds, SDS-PAGE, electron microscopy; 4 * 26000, SDS-PAGE
Clostridium sp.
98000
106000
native PAGE, analytical ultracentrifugation
Clostridium sp.
102000
108000
native PAGE, analytical ultracentrifugation, FPLC gel filtration
Clostridium sp.
Organism
Organism
Primary Accession No. (UniProt)
Commentary
Textmining
Clostridium sp.
-
group P, strain 48-50
-
Purification (Commentary)
Commentary
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
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
12-oxochenodeoxycholic acid + NADP+
-
347737
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
347737
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
347737
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
347737
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
347737
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
347737
Clostridium sp.
3alpha-hydroxy-12-oxo-5beta-cholan-24-oic acid + NAD(P)H + H+
-
-
-
?
Subunits
Subunits
Commentary
Organism
tetramer
4 * 26000, no disulfide bonds, SDS-PAGE, electron microscopy; 4 * 26000, SDS-PAGE
Clostridium sp.
Temperature Optimum [°C]
Temperature Optimum [°C]
Temperature Optimum Maximum [°C]
Commentary
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]
Commentary
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
Commentary
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
Commentary
Organism
Structure
NAD+
-
Clostridium sp.
NADP+
-
Clostridium sp.
NADPH
-
Clostridium sp.
Cofactor (protein specific)
Cofactor
Commentary
Organism
Structure
NAD+
-
Clostridium sp.
NADP+
-
Clostridium sp.
NADPH
-
Clostridium sp.
General Stability (protein specific)
General Stability
Organism
glycerol, 50% v/v, stabilization
Clostridium sp.
Inhibitors (protein specific)
Inhibitors
Commentary
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] (protein specific)
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
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] (protein specific)
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
26000
-
4 * 26000, no disulfide bonds, SDS-PAGE, electron microscopy; 4 * 26000, SDS-PAGE
Clostridium sp.
98000
106000
native PAGE, analytical ultracentrifugation
Clostridium sp.
102000
108000
native PAGE, analytical ultracentrifugation, FPLC gel filtration
Clostridium sp.
Purification (Commentary) (protein specific)
Commentary
Organism
-
Clostridium sp.
Storage Stability (protein specific)
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) (protein specific)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
12-oxochenodeoxycholic acid + NADP+
-
347737
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
347737
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
347737
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
347737
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
347737
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
347737
Clostridium sp.
3alpha-hydroxy-12-oxo-5beta-cholan-24-oic acid + NAD(P)H + H+
-
-
-
?
Subunits (protein specific)
Subunits
Commentary
Organism
tetramer
4 * 26000, no disulfide bonds, SDS-PAGE, electron microscopy; 4 * 26000, SDS-PAGE
Clostridium sp.
Temperature Optimum [°C] (protein specific)
Temperature Optimum [°C]
Temperature Optimum Maximum [°C]
Commentary
Organism
additional information
-
optimum above 55°C, not measurable because enzyme is inactivated above 55°C
Clostridium sp.
Temperature Stability [°C] (protein specific)
Temperature Stability Minimum [°C]
Temperature Stability Maximum [°C]
Commentary
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 (protein specific)
pH Optimum Minimum
pH Optimum Maximum
Commentary
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.
Other publictions for EC 1.1.1.176
No.
1st author
Pub Med
title
organims
journal
volume
pages
year
Activating Compound
Application
Cloned(Commentary)
Crystallization (Commentary)
Engineering
General Stability
Inhibitors
KM Value [mM]
Localization
Metals/Ions
Molecular Weight [Da]
Natural Substrates/ Products (Substrates)
Organic Solvent Stability
Organism
Oxidation Stability
Posttranslational Modification
Purification (Commentary)
Reaction
Renatured (Commentary)
Source Tissue
Specific Activity [micromol/min/mg]
Storage Stability
Substrates and Products (Substrate)
Subunits
Temperature Optimum [°C]
Temperature Range [°C]
Temperature Stability [°C]
Turnover Number [1/s]
pH Optimum
pH Range
pH Stability
Cofactor
Ki Value [mM]
pI Value
IC50 Value
Activating Compound (protein specific)
Application (protein specific)
Cloned(Commentary) (protein specific)
Cofactor (protein specific)
Crystallization (Commentary) (protein specific)
Engineering (protein specific)
General Stability (protein specific)
IC50 Value (protein specific)
Inhibitors (protein specific)
Ki Value [mM] (protein specific)
KM Value [mM] (protein specific)
Localization (protein specific)
Metals/Ions (protein specific)
Molecular Weight [Da] (protein specific)
Natural Substrates/ Products (Substrates) (protein specific)
Organic Solvent Stability (protein specific)
Oxidation Stability (protein specific)
Posttranslational Modification (protein specific)
Purification (Commentary) (protein specific)
Renatured (Commentary) (protein specific)
Source Tissue (protein specific)
Specific Activity [micromol/min/mg] (protein specific)
Storage Stability (protein specific)
Substrates and Products (Substrate) (protein specific)
Subunits (protein specific)
Temperature Optimum [°C] (protein specific)
Temperature Range [°C] (protein specific)
Temperature Stability [°C] (protein specific)
Turnover Number [1/s] (protein specific)
pH Optimum (protein specific)
pH Range (protein specific)
pH Stability (protein specific)
pI Value (protein specific)
Expression
General Information
General Information (protein specific)
Expression (protein specific)
KCat/KM [mM/s]
KCat/KM [mM/s] (protein specific)
701240
Giovannini
7alpha- and 12alpha-Hydroxyste ...
Acinetobacter lwoffii
Steroids
73
1385-1390
2008
-
1
-
-
-
-
-
-
-
-
-
-
-
1
-
-
1
-
-
-
2
1
1
-
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-
-
1
-
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1
-
1
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1
-
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2
1
1
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
668040
Fossati
Exploitation of the alcohol de ...
Clostridium sp.
Biotechnol. Bioeng.
93
1216-1220
2006
-
-
-
-
-
-
2
-
-
-
-
-
-
1
-
-
-
-
-
1
1
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1
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1
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1
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1
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2
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1
1
-
1
-
-
-
-
-
1
-
-
-
-
-
-
-
-
-
347747
Panichajakul
-
Purification and characterizat ...
Brevibacterium fuscum
Microb. Util. Renewable Resour.
8
119-128
1993
-
-
-
-
-
-
-
-
-
-
2
-
-
1
-
-
1
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1
1
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2
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2
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1
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1
1
-
-
-
-
2
-
-
-
-
-
-
-
-
-
347748
Panichajakul
-
Bile acids induction of 12 alp ...
Brevibacterium fuscum
Microb. Util. Renewable Resour.
8
112-118
1993
-
-
-
-
-
-
-
-
-
-
-
1
-
1
-
-
-
-
-
-
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2
-
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1
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-
-
-
2
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
347737
Braun
12 alpha-hydroxysteroid dehydr ...
Clostridium sp.
Eur. J. Biochem.
196
439-450
1991
-
-
-
-
-
1
5
5
-
-
3
-
-
1
-
-
1
-
-
-
-
2
6
1
1
-
2
-
2
-
-
3
-
-
-
-
-
-
3
-
-
1
-
5
-
5
-
-
3
-
-
-
-
1
-
-
-
2
6
1
1
-
2
-
2
-
-
-
-
-
-
-
-
-
286074
Ottolina
Enzymatic synthesis of [4R-2H] ...
Clostridium sp.
Biochim. Biophys. Acta
998
173-178
1989
-
-
-
-
-
-
-
-
-
-
-
-
-
1
-
-
-
-
-
-
-
-
-
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2
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2
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-
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-
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-
286075
Kinoshita
-
Purification and properties of ...
Brevibacterium fuscum
J. Ferment. Technol.
66
145-152
1988
-
-
-
-
-
-
-
-
-
-
-
-
-
1
-
-
1
-
-
-
-
-
6
-
-
-
-
-
-
-
-
-
-
-
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-
-
-
-
-
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-
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-
-
-
-
-
-
-
-
1
-
-
-
-
6
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
347739
MacDonald
Affinity chromatography of bil ...
Clostridium sp.
J. Chromatogr.
259
154-158
1983
-
-
-
-
-
-
-
-
-
-
-
-
-
1
-
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1
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1
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-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
347740
MacDonald
Bile salt 3alpha- and 12alpha- ...
Eggerthella lenta
Appl. Environ. Microbiol.
37
992-1000
1979
-
-
-
-
-
-
-
5
-
-
1
-
-
1
-
-
-
-
-
-
-
1
7
-
-
-
-
-
1
1
-
1
-
-
-
-
-
-
1
-
-
-
-
-
-
5
-
-
1
-
-
-
-
-
-
-
-
1
7
-
-
-
-
-
1
1
-
-
-
-
-
-
-
-
347741
Harris
Partial purification and chara ...
Clostridium leptum
Biochim. Biophys. Acta
528
148-157
1978
-
-
-
-
-
-
-
6
-
-
1
-
-
2
-
-
1
-
-
-
-
1
6
-
-
-
1
-
1
-
-
1
-
-
-
-
-
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1
-
-
-
-
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-
6
-
-
1
-
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1
-
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1
6
-
-
-
1
-
1
-
-
-
-
-
-
-
-
-
347742
Mahony
Bile salt degradation by nonfe ...
Clostridium sp.
Appl. Environ. Microbiol.
34
419-423
1977
-
-
-
-
-
-
-
-
-
-
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-
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1
-
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1
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2
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2
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-
-
-
-
1
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
347743
MacDonald
NAD-dependent 3alpha- and 12al ...
Eggerthella lenta
Biochim. Biophys. Acta
489
466-476
1977
-
-
-
-
-
-
-
2
-
-
-
-
-
2
-
-
-
-
-
-
-
1
1
-
-
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1
-
1
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2
-
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2
-
-
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2
-
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-
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-
-
-
1
1
-
-
-
1
-
1
-
-
-
-
-
-
-
-
-
347744
MacDonald
3alpha-, 7alpha- and 12alpha-h ...
Clostridium perfringens
Biochim. Biophys. Acta
450
142-153
1976
-
-
-
-
-
-
-
1
-
-
-
-
-
3
-
-
-
-
-
1
-
-
4
-
-
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2
-
1
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1
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-
1
-
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1
-
-
-
-
-
-
-
-
-
1
-
-
4
-
-
-
2
-
1
-
-
-
-
-
-
-
-
-