Information on EC 1.1.1.176 - 12alpha-hydroxysteroid dehydrogenase

New: Word Map on EC 1.1.1.176
Please wait a moment until all data is loaded. This message will disappear when all data is loaded.
Specify your search results
Mark a special word or phrase in this record:
Select one or more organisms in this record:
Show additional data
Do not include text mining results
Include (text mining) results (more...)
Include results (AMENDA + additional results, but less precise; more...)


The expected taxonomic range for this enzyme is: Bacteria

EC NUMBER
COMMENTARY
1.1.1.176
-
RECOMMENDED NAME
GeneOntology No.
12alpha-hydroxysteroid dehydrogenase
REACTION
REACTION DIAGRAM
COMMENTARY
ORGANISM
UNIPROT
LITERATURE
cholate + NADP+ = 3alpha,7alpha-dihydroxy-12-oxo-5beta-cholan-24-oate + NADPH + H+
show the reaction diagram
-
-
-
-
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
redox reaction
-
-
-
-
PATHWAY
BRENDA Link
KEGG Link
MetaCyc Link
glycocholate metabolism (bacteria)
-
-
SYSTEMATIC NAME
IUBMB Comments
12alpha-hydroxysteroid:NADP+ 12-oxidoreductase
Catalyses the oxidation of the 12alpha-hydroxy group of bile acids, both in their free and conjugated form. Also acts on bile alcohols.
SYNONYMS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
12alpha-HSDH
-
-
12alpha-HSDH
-
-
12alpha-hydroxy steroid dehydrogenase
-
-
-
-
dehydrogenase, 12alpha-hydroxy steroid
-
-
-
-
NAD-dependent 12alpha-hydroxysteroid dehydrogenase
-
-
-
-
NADP-12alpha-hydroxysteroid dehydrogenase
-
-
-
-
NADP-dependent 12alpha-hydroxysteroid dehydrogenase
-
-
CAS REGISTRY NUMBER
COMMENTARY
61642-40-8
-
SUBSTRATE
PRODUCT                      
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate)
LITERATURE
(Substrate)
COMMENTARY
(Product)
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
12-ketochenodeoxycholic acid + NADH
cholic acid + NAD+
show the reaction diagram
-
-
-
r
12-oxochenodeoxycholic acid + NADP+
(3alpha,5beta,7alpha,12alpha)-3,7,12-trihydroxycholan-24-oic acid + NADPH + H+
show the reaction diagram
-
-
-
-
?
7alpha,12alpha-dihydroxy-5beta-cholanoate + NAD+
7alpha-hydroxy-12-oxo-5beta-cholanoate + NADH
show the reaction diagram
-
-
-
-
?
allocholate + NAD+
3alpha,7alpha-dihydroxy-12-oxo-5alpha-cholan-24-oic acid + NADH
show the reaction diagram
-
-
-
-
?
allodeoxycholate + NAD+
3alpha-hydroxy-12-oxo-5alpha-cholan-24-oic acid + NADH
show the reaction diagram
-
-
-
-
?
cholic acid + NAD(P)+
3alpha,7alpha-dihydroxy-12-oxo-5beta-cholan-24-oic acid + NAD(P)H
show the reaction diagram
-
-
-
-
?
cholic acid + NAD(P)+
3alpha,7alpha-dihydroxy-12-oxo-5beta-cholan-24-oic acid + NAD(P)H
show the reaction diagram
-
-
-
?
cholic acid + NAD(P)+
3alpha,7alpha-dihydroxy-12-oxo-5beta-cholan-24-oic acid + NAD(P)H
show the reaction diagram
-
-
-
-
r
cholic acid + NAD(P)+
3alpha,7alpha-dihydroxy-12-oxo-5beta-cholan-24-oic acid + NAD(P)H
show the reaction diagram
-
-
-
-
?
cholic acid + NAD(P)+
3alpha,7alpha-dihydroxy-12-oxo-5beta-cholan-24-oic acid + NAD(P)H
show the reaction diagram
-
-
-
-
?
cholic acid + NAD(P)+
3alpha,7alpha-dihydroxy-12-oxo-5beta-cholan-24-oic acid + NAD(P)H
show the reaction diagram
-
hardly accepts N6-modified NADP+ derivative
-
-
?, r
cholic acid + NAD(P)+
3alpha,7alpha-dihydroxy-12-oxo-5beta-cholan-24-oic acid + NAD(P)H
show the reaction diagram
-
i.e. 3alpha,7alpha,12alpha-trihydroxy-5beta-cholan-24-oic acid
-
-
?, r
cholic acid + NADP+
12-ketochenodeoxycholic acid + NADPH
show the reaction diagram
-
the assay is performed using an NADP+-regenerating system with acetone, NADPH, 2-propanol and alcohol dehydrogenase from Lactobacillus brevis or Thermoanaerobium brockii, overview
-
-
r
deoxycholic acid + NAD(P)+
3alpha-hydroxy-12-oxo-5beta-cholan-24-oic acid + NAD(P)H + H+
show the reaction diagram
-
i.e. 3alpha,12alpha-dihydroxy-5beta-cholan-24-oic acid
-
-
?
deoxycholic acid + NAD(P)+
3alpha-hydroxy-12-oxo-5beta-cholan-24-oic acid + NAD(P)H + H+
show the reaction diagram
-
i.e. 3alpha,12alpha-dihydroxy-5beta-cholan-24-oic acid
-
-
?
deoxycholic acid + NAD(P)+
3alpha-hydroxy-12-oxo-5beta-cholan-24-oic acid + NAD(P)H + H+
show the reaction diagram
-
i.e. 3alpha,12alpha-dihydroxy-5beta-cholan-24-oic acid
-
-
?
deoxycholic acid + NAD(P)+
3alpha-hydroxy-12-oxo-5beta-cholan-24-oic acid + NAD(P)H + H+
show the reaction diagram
-
i.e. 3alpha,12alpha-dihydroxy-5beta-cholan-24-oic acid
-
-
r
deoxycholic acid + NAD(P)+
3alpha-hydroxy-12-oxo-5beta-cholan-24-oic acid + NAD(P)H + H+
show the reaction diagram
-
i.e. 3alpha,12alpha-dihydroxy-5beta-cholan-24-oic acid
-
-
?
deoxycholic acid + NAD(P)+
3alpha-hydroxy-12-oxo-5beta-cholan-24-oic acid + NAD(P)H + H+
show the reaction diagram
-
i.e. 3alpha,12alpha-dihydroxy-5beta-cholan-24-oic acid
-
-
?
deoxycholic acid + NAD+
3alpha-hydroxy-12-oxo-5beta-cholan-24-oic acid + NADH + H+
show the reaction diagram
-
-
-
-
?
glycocholic acid + NAD+
3alpha,7alpha-dihydroxy-12-oxo-5beta-cholanoyl glycine + NADH
show the reaction diagram
-
i.e. 3alpha,7alpha,12alpha-trihydroxy-5beta-cholanoyl glycine
-
-
?
glycocholic acid + NAD+
3alpha,7alpha-dihydroxy-12-oxo-5beta-cholanoyl glycine + NADH
show the reaction diagram
-
i.e. 3alpha,7alpha,12alpha-trihydroxy-5beta-cholanoyl glycine
-
-
?
glycocholic acid + NAD+
3alpha,7alpha-dihydroxy-12-oxo-5beta-cholanoyl glycine + NADH
show the reaction diagram
-
i.e. 3alpha,7alpha,12alpha-trihydroxy-5beta-cholanoyl glycine
-
-
?
glycocholic acid + NAD+
3alpha,7alpha-dihydroxy-12-oxo-5beta-cholanoyl glycine + NADH
show the reaction diagram
-
i.e. 3alpha,7alpha,12alpha-trihydroxy-5beta-cholanoyl glycine
-
-
?
glycodeoxycholic acid + NAD+
3alpha-hydroxy-12-oxo-5beta-cholanoyl glycine + NADH
show the reaction diagram
-
i.e. 3alpha,12alpha-dihydroxy-5beta-cholanoyl glycine
-
-
?
glycodeoxycholic acid + NAD+
3alpha-hydroxy-12-oxo-5beta-cholanoyl glycine + NADH
show the reaction diagram
-
i.e. 3alpha,12alpha-dihydroxy-5beta-cholanoyl glycine
-
-
?
glycodeoxycholic acid + NAD+
3alpha-hydroxy-12-oxo-5beta-cholanoyl glycine + NADH
show the reaction diagram
-
i.e. 3alpha,12alpha-dihydroxy-5beta-cholanoyl glycine
-
-
?
glycodeoxycholic acid + NAD+
3alpha-hydroxy-12-oxo-5beta-cholanoyl glycine + NADH
show the reaction diagram
-
i.e. 3alpha,12alpha-dihydroxy-5beta-cholanoyl glycine
-
-
?
taurocholic acid + NAD(P)+
3alpha,7alpha-dihydroxy-12-oxo-5beta-cholanoyl taurine + NAD(P)H
show the reaction diagram
-
-
-
-
-
taurocholic acid + NAD(P)+
3alpha,7alpha-dihydroxy-12-oxo-5beta-cholanoyl taurine + NAD(P)H
show the reaction diagram
-
i.e. 3alpha,7alpha,12alpha-trihydroxy-5beta-cholanoyl taurine
-
-
?
taurocholic acid + NAD(P)+
3alpha,7alpha-dihydroxy-12-oxo-5beta-cholanoyl taurine + NAD(P)H
show the reaction diagram
-
i.e. 3alpha,7alpha,12alpha-trihydroxy-5beta-cholanoyl taurine
-
-
?
taurocholic acid + NAD(P)+
3alpha,7alpha-dihydroxy-12-oxo-5beta-cholanoyl taurine + NAD(P)H
show the reaction diagram
-
i.e. 3alpha,7alpha,12alpha-trihydroxy-5beta-cholanoyl taurine
-
-
?
taurodeoxycholic acid + NAD+
3alpha-hydroxy-12-oxo-5beta-cholanoyl taurine + NADH
show the reaction diagram
-
-
-
-
?
taurodeoxycholic acid + NAD+
3alpha-hydroxy-12-oxo-5beta-cholanoyl taurine + NADH
show the reaction diagram
-
-
-
-
?
NATURAL SUBSTRATES
NATURAL PRODUCTS
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate)
LITERATURE
(Substrate)
COMMENTARY
(Product)
LITERATURE
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
12-ketochenodeoxycholic acid + NADH
cholic acid + NAD+
show the reaction diagram
-
-
-
r
COFACTOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
NAD+
-
10% of NADP+-activity
NADP+
-
low activity
NADPH
-
B-stereospecific
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
acetone
-
at 25% v/v and above
p-chloromercuribenzoate
-
-
pyridoxal 5'-phosphate
-
protection by NADP+ or NADPH
additional information
-
no substrate inhibition at high concentration of cholic acid
-
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
0.0012
12-oxochenodeoxycholic acid
-
-
0.028
3alpha,12alpha-dihydroxy-5beta-cholanoate
-
-
0.8
3alpha,12alpha-dihydroxy-5beta-cholanoate
-
-
1.6
3alpha,12alpha-dihydroxy-5beta-cholanoate
-
-
0.17
3alpha,12alpha-dihydroxy-5beta-cholanoyl glycine
-
-
0.57
3alpha,12alpha-dihydroxy-5beta-cholanoyl glycine
-
-
0.44
3alpha,12alpha-dihydroxy-5beta-cholanoyl taurine
-
-
0.059
3alpha,7alpha,12alpha-trihydroxy-5beta-cholanoate
-
-
7
3alpha,7alpha,12alpha-trihydroxy-5beta-cholanoate
-
-
0.25
3alpha,7alpha,12alpha-trihydroxy-5beta-cholanoyl glycine
-
-
2.7
3alpha,7alpha,12alpha-trihydroxy-5beta-cholanoyl glycine
-
-
0.03
3alpha,7alpha,12alpha-trihydroxy-5beta-cholanoyl methyl ester
-
3alpha,12alpha-dihydroxy-5beta-cholanoate
4
3alpha,7alpha,12alpha-trihydroxy-5beta-cholanoyl taurine
-
-
0.1
7alpha,12alpha-dihydroxy-5beta-cholanoate
-
-
0.072
cholic acid
-
-
0.045
deoxycholic acid
-
-
0.024
NAD+
-
-
0.035
NADP+
-
-
0.0085
NADPH
-
-
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
0.027
-
cell extract
0.3
-
purified enzyme
additional information
-
assay method development using an NADP+-regenerating system with acetone, NADPH, 2-propanol and alcohol dehydrogenase from Lactobacillus brevis or Thermoanaerobium brockii, optimization, overview
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
5 - 6
-
reduction of dehydrocholic acid
8 - 10.5
-
oxidation of cholic acid
8.5 - 10.5
-
oxidation of bile acids
8.5 - 9
-
oxidation of cholic acid
8.5 - 9.5
-
oxidation of cholic acid
10.5
-
oxidation of bile acids
10.5
-
oxidation of deoxycholic acid
additional information
-
reduction of 12-oxochenodeoxycholic acid optimal below pH 5.0, not measurable due to insolubility of substrate
pH RANGE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
8 - 10.5
-
-
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
additional information
-
optimum above 55C, not measurable because enzyme is inactivated above 55C
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
SOURCE
-
enzyme in 50% glycerol-containing solution
Manually annotated by BRENDA team
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
98000 - 106000
-
native PAGE, analytical ultracentrifugation
347737
102000 - 108000
-
native PAGE, analytical ultracentrifugation, FPLC gel filtration
347737
110000
-
SDS-PAGE, gel filtration
347747
125000
-
gel filtration
347740
225000
-
gel filtration
347741
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
dimer
-
1 * 58000 + 1* 64000, SDS-PAGE
tetramer
-
4 * 26000, SDS-PAGE, 4 * 26000, no disulfide bonds, SDS-PAGE, electron microscopy
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
0 - 4
-
half-life: 11 days
347741
25
-
half-life: 4 days, increases to 10 days by addition of NADP+
347737
30
-
half-life: 2 days, increases to 5 days by addition of NADP+
347737
43
-
slow inactivation
347744
50
-
30 min, inactivation
347743
56
-
rapid inactivation
347744
GENERAL STABILITY
ORGANISM
UNIPROT
LITERATURE
glycerol, 50% v/v, stabilization
-
STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
4C, 0.5 ml TEA-buffer, 3 days
-
-20C, 20 days, 50% loss of activity
-
-30C, 6 months, 50% glycerol, no loss of activity
-
-30C, concentrated form, at least 6 months
-
-20C, 3 months, 5% loss of activity
-
-20C, lyophilized, 4 months
-
Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
partially purified by ion-exchange chromatography
-
APPLICATION
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
degradation
-
new integrated chemo-enzymatic synthesis of ursodeoxycholic acid starting from sodium cholate by 7alpha- and 12alpha-hydroxysteroid dehydrogenases