Information on EC 1.1.1.176 - 12alpha-hydroxysteroid dehydrogenase

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The expected taxonomic range for this enzyme is: Bacteria, Eukaryota

EC NUMBER
COMMENTARY hide
1.1.1.176
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RECOMMENDED NAME
GeneOntology No.
12alpha-hydroxysteroid dehydrogenase
REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
cholate + NADP+ = 3alpha,7alpha-dihydroxy-12-oxo-5beta-cholan-24-oate + NADPH + H+
show the reaction diagram
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REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
redox reaction
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PATHWAY
BRENDA Link
KEGG Link
MetaCyc Link
Secondary bile acid biosynthesis
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glycocholate metabolism (bacteria)
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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.
CAS REGISTRY NUMBER
COMMENTARY hide
61642-40-8
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ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
group P C 48-50
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-
Manually annotated by BRENDA team
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Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
additional information
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
12-ketochenodeoxycholic acid + NADH
cholic acid + NAD+
show the reaction diagram
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r
12-oxochenodeoxycholic acid + NADP+
(3alpha,5beta,7alpha,12alpha)-3,7,12-trihydroxycholan-24-oic acid + NADPH + H+
show the reaction diagram
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-
-
-
?
7alpha,12alpha-dihydroxy-5beta-cholanoate + NAD+
7alpha-hydroxy-12-oxo-5beta-cholanoate + NADH
show the reaction diagram
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-
-
-
?
allocholate + NAD+
3alpha,7alpha-dihydroxy-12-oxo-5alpha-cholan-24-oic acid + NADH
show the reaction diagram
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-
-
-
?
allodeoxycholate + NAD+
3alpha-hydroxy-12-oxo-5alpha-cholan-24-oic acid + NADH
show the reaction diagram
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-
-
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?
cholate + NAD+
3alpha,7alpha-dihydroxy-12-oxo-5beta-cholanate + NADH + H+
show the reaction diagram
cholate + NADP+
3alpha,7alpha-dihydroxy-12-oxo-5beta-cholanate + NADH + H+
show the reaction diagram
cholate + NADP+
3alpha,7alpha-dihydroxy-12-oxo-5beta-cholanate + NADPH + 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 + NADP+
12-ketochenodeoxycholic acid + NADPH
show the reaction diagram
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the assay is performed using an NADP+-regenerating system with acetone, NADPH, 2-propanol and alcohol dehydrogenase from Lactobacillus brevis or Thermoanaerobium brockii, overview
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r
deoxycholate + NADP+
12-oxo-lithocholic acid + NADPH + H+
show the reaction diagram
deoxycholic acid + NAD(P)+
3alpha-hydroxy-12-oxo-5beta-cholan-24-oic acid + NAD(P)H + H+
show the reaction diagram
deoxycholic acid + NAD+
3alpha-hydroxy-12-oxo-5beta-cholan-24-oic acid + NADH + H+
show the reaction diagram
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-
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?
glycocholic acid + NAD+
3alpha,7alpha-dihydroxy-12-oxo-5beta-cholanoyl glycine + NADH
show the reaction diagram
glycodeoxycholic acid + NAD+
3alpha-hydroxy-12-oxo-5beta-cholanoyl glycine + NADH
show the reaction diagram
taurocholic acid + NAD(P)+
3alpha,7alpha-dihydroxy-12-oxo-5beta-cholanoyl taurine + NAD(P)H
show the reaction diagram
taurodeoxycholic acid + NAD+
3alpha-hydroxy-12-oxo-5beta-cholanoyl taurine + NADH
show the reaction diagram
additional information
?
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NATURAL SUBSTRATES
NATURAL PRODUCTS
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
12-ketochenodeoxycholic acid + NADH
cholic acid + NAD+
show the reaction diagram
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r
cholate + NAD+
3alpha,7alpha-dihydroxy-12-oxo-5beta-cholanate + NADH + H+
show the reaction diagram
cholate + NADP+
3alpha,7alpha-dihydroxy-12-oxo-5beta-cholanate + NADPH + H+
show the reaction diagram
additional information
?
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COFACTOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
NADP+
NADPH
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
acetone
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at 25% v/v and above
p-chloromercuribenzoate
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pyridoxal 5'-phosphate
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protection by NADP+ or NADPH
additional information
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no substrate inhibition at high concentration of cholic acid
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KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.0012
12-oxochenodeoxycholic acid
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0.028 - 1.6
3alpha,12alpha-dihydroxy-5beta-cholanoate
0.17 - 0.57
3alpha,12alpha-dihydroxy-5beta-cholanoyl glycine
0.44
3alpha,12alpha-dihydroxy-5beta-cholanoyl taurine
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0.059 - 7
3alpha,7alpha,12alpha-trihydroxy-5beta-cholanoate
0.25 - 2.7
3alpha,7alpha,12alpha-trihydroxy-5beta-cholanoyl glycine
0.03
3alpha,7alpha,12alpha-trihydroxy-5beta-cholanoyl methyl ester
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3alpha,12alpha-dihydroxy-5beta-cholanoate
4
3alpha,7alpha,12alpha-trihydroxy-5beta-cholanoyl taurine
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0.1
7alpha,12alpha-dihydroxy-5beta-cholanoate
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0.072
cholic acid
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0.045
deoxycholic acid
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0.024
NAD+
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0.035
NADP+
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0.0085
NADPH
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SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
0.027
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cell extract
0.3
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purified enzyme
additional information
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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 hide
LITERATURE
5 - 6
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reduction of dehydrocholic acid
8 - 10.5
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oxidation of cholic acid
8.5 - 9
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oxidation of cholic acid
8.5 - 9.5
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oxidation of cholic acid
8.5 - 10.5
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oxidation of bile acids
additional information
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reduction of 12-oxochenodeoxycholic acid optimal below pH 5.0, not measurable due to insolubility of substrate
pH RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
8 - 10.5
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TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
additional information
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optimum above 55°C, not measurable because enzyme is inactivated above 55°C
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE
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enzyme in 50% glycerol-containing solution
Manually annotated by BRENDA team
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
26000
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4 * 26000, no disulfide bonds, SDS-PAGE, electron microscopy; 4 * 26000, SDS-PAGE
58000
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1 * 58000 + 1* 64000, SDS-PAGE
98000 - 106000
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native PAGE, analytical ultracentrifugation
102000 - 108000
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native PAGE, analytical ultracentrifugation, FPLC gel filtration
110000
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SDS-PAGE, gel filtration
125000
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gel filtration
225000
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gel filtration
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
dimer
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1 * 58000 + 1* 64000, SDS-PAGE
tetramer
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4 * 26000, no disulfide bonds, SDS-PAGE, electron microscopy; 4 * 26000, SDS-PAGE
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
0 - 4
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half-life: 11 days
25
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half-life: 4 days, increases to 10 days by addition of NADP+
30
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half-life: 2 days, increases to 5 days by addition of NADP+
43
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slow inactivation
50
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30 min, inactivation
56
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rapid inactivation
GENERAL STABILITY
ORGANISM
UNIPROT
LITERATURE
glycerol, 50% v/v, stabilization
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STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
-20°C, 20 days, 50% loss of activity
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-20°C, 3 months, 5% loss of activity
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-20°C, lyophilized, 4 months
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-30°C, 6 months, 50% glycerol, no loss of activity
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-30°C, concentrated form, at least 6 months
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4°C, 0.5 ml TEA-buffer, 3 days
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Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
partially purified by ion-exchange chromatography
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ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
additional information
APPLICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
degradation
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new integrated chemo-enzymatic synthesis of ursodeoxycholic acid starting from sodium cholate by 7alpha- and 12alpha-hydroxysteroid dehydrogenases
synthesis