Information on EC 1.1.1.159 - 7alpha-hydroxysteroid dehydrogenase

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The enzyme appears in viruses and cellular organisms

EC NUMBER
COMMENTARY hide
1.1.1.159
-
RECOMMENDED NAME
GeneOntology No.
7alpha-hydroxysteroid dehydrogenase
REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
cholate + NAD+ = 3alpha,12alpha-dihydroxy-7-oxo-5beta-cholan-24-oate + NADH + H+
show the reaction diagram
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
oxidation
-
-
-
-
redox reaction
-
-
-
-
reduction
-
-
-
-
PATHWAY
BRENDA Link
KEGG Link
MetaCyc Link
glycocholate metabolism (bacteria)
-
-
ursodeoxycholate biosynthesis (bacteria)
-
-
Secondary bile acid biosynthesis
-
-
SYSTEMATIC NAME
IUBMB Comments
7alpha-hydroxysteroid:NAD+ 7-oxidoreductase
Catalyses the oxidation of the 7alpha-hydroxy group of bile acids and alcohols both in their free and conjugated forms. The Bacteroides fragilis and Clostridium enzymes can also utilize NADP+.
CAS REGISTRY NUMBER
COMMENTARY hide
39361-64-3
-
ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
ATCC25285
-
-
Manually annotated by BRENDA team
AM-1
-
-
Manually annotated by BRENDA team
formerly Eubacterium sp. strain VP1
-
-
Manually annotated by BRENDA team
i.e. Pseudomonas testosteroni
UniProt
Manually annotated by BRENDA team
i.e. Pseudomonas testosteroni
UniProt
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
evolution
malfunction
physiological function
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
(3alpha,5beta)-3-hydroxy-7,12-dioxocholan-24-oic acid + NADH
(3alpha,5beta,7alpha)-3,7-dihydroxy-12-oxocholan-24-oic acid + NAD+
show the reaction diagram
-
-
-
-
r
12-ketochenodeoxycholic acid + NAD+
7,12-diketochenodeoxycholic acid + NADH
show the reaction diagram
3,7-diketocholic acid + NADH
3-ketocholic acid + NAD+
show the reaction diagram
-
high activity
-
-
r
6-fluoro-7-ketohyocholic acid + NADH
6-fluoro-3alpha,6alpha,7alpha-trihydroxy-5beta-cholan-24-oic acid + NAD+ + NAD+
show the reaction diagram
-
-
-
-
r
7,12-diketolithocholic acid + NADH
12-ketochenodeoxycholic acid + NAD+
show the reaction diagram
-
-
-
r
7-keto-lithocholic acid + NADH
cheno-deoxycholic acid + NAD+
show the reaction diagram
7-ketochenodeoxycholic acid + NADH
3alpha,7alpha-dihydroxy-5beta-cholanoic acid + NAD+
show the reaction diagram
-
-
-
-
r
7-ketohyocholic acid + NADH
3alpha,6alpha,7alpha-trihydroxy-5beta-cholan-24-oic acid + NAD+
show the reaction diagram
-
low activity
-
-
r
bile acids + NAD(P)+
?
show the reaction diagram
chenodeoxycholic acid + NAD(P)+
3alpha-hydroxy-7-oxo-5beta-cholan-24-oic acid + NAD(P)H
show the reaction diagram
chenodeoxycholic acid + NAD+
3alpha-hydroxy-7-oxo-5beta-cholan-24-oic acid + NADH
show the reaction diagram
chenodeoxycholic acid + NAD+
7-oxolithocholic acid + NADH + H+
show the reaction diagram
-
-
-
-
?
cholate + NAD+
3alpha,12alpha-dihydroxy-7-oxo-5beta-cholanate + NADH + H+
show the reaction diagram
cholic acid + NAD(P)+
3alpha,12alpha-dihydroxy-7-oxo-5beta-cholan-24-oic acid + NADH
show the reaction diagram
cholic acid + NAD+
3alpha,12alpha-dihydroxy-7-oxo-5beta-cholan-24-oic acid + NADH
show the reaction diagram
cholic acid + NADH
7-ketocholic acid + NAD+
show the reaction diagram
-
low activity in the reverse reaction
-
-
r
dehydrocholic acid + NADH
3,12-diketolithocholic acid + NAD+
show the reaction diagram
-
-
-
r
ethyl (R)-2-hydroxy-2-(3,5-difluorophenyl)acetate + NAD+
ethyl 2-oxo-2-(3,5-difluorophenyl)acetate + NADH
show the reaction diagram
-
-
-
-
?
ethyl (R)-2-hydroxy-3,3-dimethylbutyrate + NAD+
ethyl 2-oxo-3,3-dimethylbutyrate + NADH
show the reaction diagram
-
-
-
-
?
ethyl (R)-2-hydroxy-3-methylbutyrate + NAD+
ethyl 2-oxo-3-methylbutyrate + NADH
show the reaction diagram
-
-
-
-
?
ethyl 2-hydroxy-2-(3,4-dichlorophenyl)acetate + NAD+
ethyl 2-oxo-2-(3,4-dichlorophenyl)acetate + NADH
show the reaction diagram
-
-
-
-
?
ethyl 2-hydroxy-2-(4-bromophenyl)acetate + NAD+
ethyl 2-oxo-2-(4-bromophenyl)acetate + NADH
show the reaction diagram
-
-
-
-
?
ethyl 2-hydroxy-2-(4-chlorophenyl)acetate + NAD+
ethyl 2-oxo-2-(4-chlorophenyl)acetate + NADH
show the reaction diagram
-
-
-
-
?
ethyl 2-hydroxy-2-(4-cyanophenyl)acetate + NAD+
ethyl 2-oxo-2-(4-cyanophenyl)acetate + NADH
show the reaction diagram
-
high activity
-
-
?
ethyl 2-hydroxy-2-(4-fluorophenyl)acetate + NAD+
ethyl 2-oxo-2-(4-fluorophenyl)acetate + NADH
show the reaction diagram
-
-
-
-
?
ethyl 2-hydroxy-2-(4-methylphenyl)acetate + NAD+
ethyl 2-oxo-2-(4-methylphenyl)acetate + NADH
show the reaction diagram
-
-
-
-
?
ethyl 2-hydroxy-2-cyclohexylacetate + NAD+
ethyl 2-oxo-2-cyclohexylacetate + NADH
show the reaction diagram
-
best substrate
-
-
?
ethyl 2-hydroxy-2-phenylacetate + NAD+
ethyl 2-oxo-2-phenylacetate + NADH
show the reaction diagram
-
-
-
-
?
glycochenodeoxycholic acid + NAD(P)+
N-[3alpha-hydroxy-7,24-dioxocholan-24-yl]glycine + NAD(P)H
show the reaction diagram
glycocholic acid + NAD+
N-[3alpha,12alpha-dihydroxy-7,24-dioxo-5beta-cholan-24-yl]glycine + NADH
show the reaction diagram
lithocholic acid + NADPH + H+
ursodeoxycholic acid + NADP+
show the reaction diagram
methyl 2-hydroxy-2-phenylacetate + NAD+
methyl 2-oxo-2-phenylacetate + NADH
show the reaction diagram
-
-
-
-
?
methyl benzoylformate + NADH
?
show the reaction diagram
-
-
-
-
?
taurochenodeoxycholic acid + NAD+
2[[3alpha-hydroxy-7,24-dioxo-5beta-cholan-24-yl]amino]ethane sulfonic acid + NADH
show the reaction diagram
taurochenodeoxycholic acid + NADP+
? + NADPH + H+
show the reaction diagram
-
-
-
r
taurocholic acid + NAD+
2[[3alpha,12alpha-dihydroxy-7,24-dioxo-5beta-cholan-24-yl]amino]ethane sulfonic acid + NADH
show the reaction diagram
ursodeoxycholic acid + NAD+
7-oxolithocholic acid + NADH + H+
show the reaction diagram
-
-
-
-
?
additional information
?
-
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
7-keto-lithocholic acid + NADH
cheno-deoxycholic acid + NAD+
show the reaction diagram
-
in the gastrointestinal tract of host organisms
-
-
?
bile acids + NAD(P)+
?
show the reaction diagram
cholate + NAD+
3alpha,12alpha-dihydroxy-7-oxo-5beta-cholanate + NADH + H+
show the reaction diagram
additional information
?
-
COFACTOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
NADP+
thio-NAD+
-
utilized with the same activity as NAD+
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
CaCl2
-
activation
K+
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activates by 105.7% at 500 mM
KCl
-
activation
Na+
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activates by 64.7% at 200 mM
NaCl
-
activation
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
2-mercaptoethanol
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ascorbic acid
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chenodeoxycholate
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substrate inhibition at concentration above 1 mM
CoCl2
CuCl2
diethyl dicarbonate
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complete
FeCl3
glycochenodeoxycholate
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substrate inhibition at concentration above 1 mM
HgCl2
iodoacetic acid
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70% remaining activity at concentration 1 mM
N-bromosuccinimide
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complete
o-phenanthroline
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70% remaining acitvity at concentration 1 mM
p-chloromercuribenzoate
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80% remaining activity at concentration 0.3 mM
p-hydroxymercuribenzoate
complete inhibition at 0.0075 mM
potassium oxalate
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Sodium citrate
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sodium lauryl sulfate
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-
Sodium perchlorate
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Sodium periodate
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Sodium persulfate
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taurochenodeoxycholate
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substrate inhibition at concentration above 1 mM
Triton X-100
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-
ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
2-mercaptoethanol
-
activation
dithiothreitol
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activation
additional information
-
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.38
12-ketolithocholic acid
-
-
0.32 - 0.34
3alpha,7alpha,12alpha-trihydroxy-5beta-cholanoic acid
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-
0.32 - 0.34
3alpha,7alpha,12alpha-trihydroxy-5beta-cholanoyl glycine
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-
0.32 - 0.34
3alpha,7alpha,12alpha-trihydroxycholanoyl taurine
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-
0.1
3alpha,7alpha-dihydroxy-5beta-cholanoic acid
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-
0.1
3alpha,7alpha-Dihydroxy-5beta-cholanoyl glycine
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-
0.1
3alpha,7alpha-dihydroxy-5beta-cholanoyl taurine
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-
0.99
7,12-Diketolithocholic acid
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-
0.0065 - 0.43
chenodeoxycholic acid
0.09 - 1.2
cholic acid
0.7
Dehydrocholic acid
-
-
0.085
glycochenodeoxycholic acid
-
-
1 - 1.25
glycocholic acid
0.15 - 0.709
NAD+
0.25 - 1.3
NADH
0.34 - 1.77
NADP+
0.19 - 0.24
taurochenodeoxycholic acid
1 - 2
taurocholic acid
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
8.18
chenodeoxycholic acid
-
-
7.92 - 151
cholic acid
0.417
glycocholic acid
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-
13.76 - 58.55
NADP+
3.43
taurochenodeoxycholic acid
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-
3.13
taurocholic acid
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-
kcat/KM VALUE [1/mMs-1]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
33.18 - 40.1
NADP+
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
0.035
-
cell extract
0.11
-
purified enzyme
0.32
-
purified recombinant enzyme, substrate methyl benzoylformate
1.1
-
crude extract
45
-
deoxycholic acid or lithocholic acid
332
-
NADP+, recombinant enzyme from Escherichia coli
351
purified enzyme, 37C
374
-
NADP+
400
-
7,12-diketolithocholic acid, NADH
900
-
pH and temperature not specified in the publication
1600
-
cholic acid, NAD+
1700
-
12-ketolithocholic acid, NAD+
2348
-
taurocholic acid
2900
-
dehydrocholic acid, NADH
2916
-
glycocholic acid
3486
-
cholic acid
3743
-
taurochenodeoxycholic acid
3856
-
glycochenodeoxycholic acid
4164
-
chenodeoxycholic acid
additional information
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
5.3
-
dehydrocholic acid reduction
6.5
NADH oxidation
7 - 9
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NADP+-dependent activity
8.5 - 9
-
oxidation of 7alpha-hydroxy group
9 - 10
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oxidation of cholic acid
9.4 - 9.6
-
-
9.5 - 10
-
NAD+-dependent activity
10
-
oxidation of cholic acid
10.5
-
oxidation of cholic acid
additional information
-
value for reduction of dehydrocholic acid below 5.2, not measurable due to insolubility of substrate
pH RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
5.5 - 10.5
NAD reduction
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
22
-
assay at room temperature
28
-
substrate cholic acid
TEMPERATURE RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE
additional information
PDB
SCOP
CATH
ORGANISM
UNIPROT
Brucella melitensis biotype 1 (strain 16M / ATCC 23456 / NCTC 10094)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
25000
-
SDS-PAGE
28000
-
4 * 28000, SDS-PAGE
30000
-
? * 30000, SDS-PAGE
54000
-
gel filtration
80000
-
NAD+-specific form, gel filtration
104000
-
gel filtration
107100
-
sequence determination
108000
-
gel filtration
110000
120000
-
gel filtration
127000
-
NADP+-dependent form, gel filtration
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
tetramer
Crystallization/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
purified recombinant enzyme complexed with taurochenodeoxycholic acid and NADP+, sitting drop vapour diffusion method, mixing of 200 nl of 1.12 mM protein in 50 mM Tris-HCl, pH 8.0, 200 mM NaCl,and 5.6 mM of NADP+ and taurochenodeoxycholic acid, with 200 nl of reservoir solution containing 0.1 M HEPES, pH 7.5, and 25% PEG-3350, and equilibration against 0.03 ml reservoir solution, at 20C, X-ray diffraction structure determination and analysis at 2.0 A resolution
-
binary complex with NAD+, ternary complex with NADH and 7-oxoglycochenodeoxycholic acid as product, as well as possibly partially glycochenodeoxycholic acid as substrat
pH STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
5.7 - 10.5
-
5C overnight
286075
6 - 11
-
-
286075
8 - 9
-
-
286085
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
4 - 37
-
purified recombinant enzyme, no change in activity after incubation at 4C or 25C for 108 h, but dramatic decrease in activity at 37C
25
-
completely stable for 108 h
30
-
stable below
40
-
inactivation at
60
-
10 min stable
75
-
30 min, presence of substrates, NAD+-dependent form stable
80
-
10 min, complete inactivation
additional information
-
the enzyme is thermostable
GENERAL STABILITY
ORGANISM
UNIPROT
LITERATURE
inactivation by freezing
-
NAD+-dependent form lyophilized is stable to freezing/thawing, NADP+-dependent form lyophilized is unstable
-
stabilization by glutathione, EDTA
-
STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
4C, purified enzyme, 50 mM sodium phosphate, pH 7.5, stable for 2 days
-
Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
native enzyme 175fold by anion exchange chromatography, ammonium sulfate fractionation, hydrophobic interaction and affinity chromatography
-
partially purified by ion-exchange chromatography
-
recombinant enzyme from Escherichia coli by heat treatment at 55-60C, polyethyleneimine treatment, ammonium sulfate fractionation, and gel filtration, followed by lyophilization
-
recombinant from Escherichia coli, high yield
-
recombinant GST-tagged wild-type and mutant enzymes from Escherichia coli strain BL21(DE3) by glutathione affinity chromatography, anion exchange chromatography, and gel filtration
-
recombinant His-tagged enzyme from Escherichia coli strain BL21(DE3) pLysS by metal affinity chromatography
wild-type and recombinant from Escherichia coli
-
Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
CA 7alpha-HSDH, recombinant overexpression of GST-tagged wild-type and mutant enzymes in Escherichia coli strain BL21(DE3)
-
expressed in Escherichia coli
expression in Escherichia coli DH5alpha-MCR, DNA and amino acid sequence analysis
-
gene 7alpha-HSD, located 11.9 kb upstream of the hsdA gene, DNA and amino acid sequence determination and analysis, phylogenetic analysis, subcloning in Escherichia coli strain HB101, sequence comparisons, recombinant overexpression of His-tagged enzyme in Escherichia coli strain BL21(DE3) pLysS
gene from strain HB101 cloned in plasmid pSD1 and expressed in Escherichia coli strain DH1
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overexpression in Escherichia coli
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overexpression in Escherichia coli DH5alpha, amino acid sequence analysis and comparison with other dehydrogenases
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recombinant expression of wild-type and truncation mutant enzymes in Escherichia coli
-
site directed mutagenesis
-
ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
R16G
-
site-directed mutagenesis, kcat and Km of the R16G mutant increase by more than 4times and 5times compared with wild-type values, respectively, while the catalytic efficiency (kcat/Km) of R16G mutant decreases by 17.26% compared to the wild-type enzyme. The increase in Km indicates that affinity of R16G mutant toward NADP+ becomes weak, while the cofactor NADP(H) dissociates more easily from the binding site resulting in the increase in kcat
R194G
-
site-directed mutagenesis, the mutant shows slightly reduced catalytic efficiency compared with NADP+ compared to the wild-type enzyme
R38D
-
site-directed mutagenesis, inactive mutant
K163I
-
5.25% activity of wild-type activity
K163R
-
63.7% activity of wild-type activity
S146A
-
20.3% activity of wild-type activity
S146H
-
35.6% activity of wild-type activity
Y159F
-
no activity
Y159H
-
13.3% activity of wild-type activity
additional information
APPLICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
analysis
-
assay for stereospecific labeling of coenzymes NADP+ and NADPH
degradation
-
new integrated chemo-enzymatic synthesis of ursodeoxycholic acid starting from sodium cholate by 7alpha- and 12alpha-hydroxysteroid dehydrogenases
synthesis
Show AA Sequence (672 entries)
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