Information on EC 1.1.1.201 - 7beta-hydroxysteroid dehydrogenase (NADP+)

Please wait a moment until all data is loaded. This message will disappear when all data is loaded.
Specify your search results
Select one or more organisms in this record:


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

EC NUMBER
COMMENTARY hide
1.1.1.201
-
RECOMMENDED NAME
GeneOntology No.
7beta-hydroxysteroid dehydrogenase (NADP+)
REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
a 7beta-hydroxysteroid + NADP+ = a 7-oxosteroid + NADPH + H+
show the reaction diagram
; Catalyses the oxidation of the 7beta-hydroxyl group of bile salts such as ursodeoxycholate
-
-
-
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
oxidation
-
-
-
-
redox reaction
-
-
-
-
reduction
-
-
-
-
PATHWAY
BRENDA Link
KEGG Link
MetaCyc Link
Secondary bile acid biosynthesis
-
-
glycocholate metabolism (bacteria)
-
-
ursodeoxycholate biosynthesis (bacteria)
-
-
SYSTEMATIC NAME
IUBMB Comments
7beta-hydroxysteroid:NADP+ 7-oxidoreductase
Catalyses the oxidation of the 7beta-hydroxy group of bile acids such as ursodeoxycholate.
CAS REGISTRY NUMBER
COMMENTARY hide
79393-83-2
-
ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
formerly Eubacterium sp.
-
-
Manually annotated by BRENDA team
strain 25.11.c
-
-
Manually annotated by BRENDA team
strain 25.11.c
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
isolated from human feces
UniProt
Manually annotated by BRENDA team
isolated from human feces
UniProt
Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
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
3,7,12-triketo-5beta-cholanoic acid + NADPH
7beta-hydroxy-3,12-diketo-5beta-cholanoic acid + NADP+
show the reaction diagram
-
-
-
?
3,7,12-triketo-5beta-cholanoyl taurine + NADPH
7beta-hydroxy-3,12-diketo-5beta-cholanoyl taurine + NADP+
show the reaction diagram
-
-
-
?
3,7-diketo-5beta-cholanoic acid + NADPH
7beta-hydroxy-3-keto-5beta-cholanoic acid + NADP+
show the reaction diagram
3alpha,12alpha-dihydroxy-7-keto-5beta-cholanoic acid + NADPH
3alpha,7beta,12alpha-trihydroxy-5beta-cholanoic acid + NADP+
show the reaction diagram
3alpha,7beta,12alpha-trihydroxy-5beta-cholanoic acid + NADP+
3alpha,12alpha-dihydroxy-7-keto-5beta-cholanoic acid + NADPH
show the reaction diagram
3alpha,7beta-dihydroxy-5beta-cholan-24-oic acid + NADP+
7-oxo-lithocholic acid + NADPH + H+
show the reaction diagram
3alpha,7beta-dihydroxy-5beta-cholanoic acid + NADP+
3alpha,7keto-dihydroxy--5beta-cholanoic acid + NADPH
show the reaction diagram
3alpha,7beta-dihydroxy-5beta-cholanoic acid + NADP+
3alpha,7keto-dihydroxy-5beta-cholanoic acid + NADPH
show the reaction diagram
3alpha,7beta-dihydroxy-5beta-cholanoyl glycine + NADP+
3alpha-hydroxy-7-keto-5beta-cholanoyl glycine + NADPH
show the reaction diagram
3alpha,7beta-dihydroxy-5beta-cholanoyl taurine + NADP+
3alpha-hydroxy-7-keto-5beta-cholanoyl taurine + NADPH
show the reaction diagram
3alpha-hydroxy-7,12-diketo-5beta-cholanoic acid + NADPH
3alpha,7beta-dihydroxy-12-keto-5beta-cholanoic acid + NADP+
show the reaction diagram
3alpha-hydroxy-7-oxo-5beta-cholan-24-oic acid + NADPH + H+
3alpha,7beta-dihydroxy-5beta-cholan-24-oic acid + NADP+
show the reaction diagram
5-cholesten-3beta,7beta-diol + NADP+
5-cholesten-3beta-ol-7-one + NADPH
show the reaction diagram
-
i.e. 7beta-hydroxycholesterol
-
-
?
7,12-diketo-5beta-cholanoic acid + NADPH
7beta-hydroxy-12-keto-5beta-cholanoic acid + NADP+
show the reaction diagram
7,12-dioxo-ursodeoxycholic acid + NADPH + H+
12-oxo-ursodeoxycholic acid + NADP+
show the reaction diagram
7-ketolithocholic acid + NADPH + H+
ursodeoxycholic acid + NADP+
show the reaction diagram
7beta-hydroxycholesterol + NADP+
7-oxocholesterol + NADPH
show the reaction diagram
-
-
-
-
?
7beta-hydroxysteroid + NADP+
7-oxosteroid + NADPH + H+
show the reaction diagram
chenodeoxycholic acid + NADPH + H+
ursodeoxycholic acid + NADP+
show the reaction diagram
corticosterone + NADP+
21-hydroxy-pregn-4-ene-3,11,20-trione + NADPH
show the reaction diagram
-
oxidation at C-11
-
-
?
cortisone + NADP+
? + NADPH
show the reaction diagram
-
oxidation at C-11
-
-
?
dehydrocholic acid + NADPH + H+
12-oxo-ursodeoxycholic acid + NADP+
show the reaction diagram
dehydrocholic acid + NADPH + H+
? + NADP+ + H+
show the reaction diagram
lithocholic acid + NADPH + H+
ursodeoxycholic acid + NADP+
show the reaction diagram
lithocholic acid + NADPH + H+ |
ursodeoxycholic acid + NADP+
show the reaction diagram
-
-
-
-
?
ursodeoxycholic acid + NADP+
7-ketolithocholic acid + NADPH + H+
show the reaction diagram
ursodeoxycholic acid + NADP+
7-oxo-lithocholic acid + NADPH + 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
3,7,12-triketo-5beta-cholanoic acid + NADPH
7beta-hydroxy-3,12-diketo-5beta-cholanoic acid + NADP+
show the reaction diagram
-
-
-
?
3,7,12-triketo-5beta-cholanoyl taurine + NADPH
7beta-hydroxy-3,12-diketo-5beta-cholanoyl taurine + NADP+
show the reaction diagram
-
-
-
?
3,7-diketo-5beta-cholanoic acid + NADPH
7beta-hydroxy-3-keto-5beta-cholanoic acid + NADP+
show the reaction diagram
3alpha,12alpha-dihydroxy-7-keto-5beta-cholanoic acid + NADPH
3alpha,7beta,12alpha-trihydroxy-5beta-cholanoic acid + NADP+
show the reaction diagram
3alpha,7beta,12alpha-trihydroxy-5beta-cholanoic acid + NADP+
3alpha,12alpha-dihydroxy-7-keto-5beta-cholanoic acid + NADPH
show the reaction diagram
3alpha,7beta-dihydroxy-5beta-cholanoic acid + NADP+
3alpha,7keto-dihydroxy--5beta-cholanoic acid + NADPH
show the reaction diagram
3alpha,7beta-dihydroxy-5beta-cholanoyl glycine + NADP+
3alpha-hydroxy-7-keto-5beta-cholanoyl glycine + NADPH
show the reaction diagram
3alpha,7beta-dihydroxy-5beta-cholanoyl taurine + NADP+
3alpha-hydroxy-7-keto-5beta-cholanoyl taurine + NADPH
show the reaction diagram
3alpha-hydroxy-7,12-diketo-5beta-cholanoic acid + NADPH
3alpha,7beta-dihydroxy-12-keto-5beta-cholanoic acid + NADP+
show the reaction diagram
3alpha-hydroxy-7-oxo-5beta-cholan-24-oic acid + NADPH + H+
3alpha,7beta-dihydroxy-5beta-cholan-24-oic acid + NADP+
show the reaction diagram
7,12-diketo-5beta-cholanoic acid + NADPH
7beta-hydroxy-12-keto-5beta-cholanoic acid + NADP+
show the reaction diagram
7beta-hydroxycholesterol + NADP+
7-oxocholesterol + NADPH
show the reaction diagram
-
-
-
-
?
7beta-hydroxysteroid + NADP+
7-oxosteroid + NADPH + H+
show the reaction diagram
chenodeoxycholic acid + NADPH + H+
ursodeoxycholic acid + NADP+
show the reaction diagram
lithocholic acid + NADPH + H+
ursodeoxycholic acid + NADP+
show the reaction diagram
lithocholic acid + NADPH + H+ |
ursodeoxycholic acid + NADP+
show the reaction diagram
-
-
-
-
?
ursodeoxycholic acid + NADP+
7-oxo-lithocholic acid + NADPH + H+
show the reaction diagram
additional information
?
-
COFACTOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
NADPH
additional information
the enzyme shows specificity toward NADPH/NADP+ against NADH/NAD+
-
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
iodoacetate
-
complete inhibition at 0.05 mM
p-chloromercuribenzoic acid
-
complete inhibition at 0.001 mM
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.07
3,12-diketocholanoic acid
-
-
0.123
3,7,12-Triketo-5beta-cholanoic acid
-
-
0.145
3,7,12-triketo-5beta-cholanoyl taurine
-
-
0.087
3,7-Diketo-5beta-cholanoic acid
-
-
0.142
3alpha,12alpha-dihydroxy-7-keto-5beta-cholanoic acid
-
-
0.076
3alpha,7beta,12alpha-trihydroxy-5beta-cholanoic acid
-
-
0.974
3alpha,7beta-dihydroxy-5beta-cholan-24-oic acid
pH 10.0, 37°C, recombinant enzyme
-
0.009 - 0.108
3alpha,7beta-dihydroxy-5beta-cholanoic acid
0.238 - 0.909
3alpha,7beta-dihydroxy-5beta-cholanoyl glycine
0.286 - 1.639
3alpha,7beta-dihydroxy-5beta-cholanoyl taurine
0.124
3alpha-Hydroxy-7,12-diketo-5beta-cholanoic acid
-
-
0.055
3alpha-hydroxy-7-keto-5beta-cholanoic acid
-
-
0.0388
3alpha-hydroxy-7-oxo-5beta-cholan-24-oic acid
pH 6.0, 37°C, recombinant enzyme
0.0052
7-ketolithocholic acid
-
pH 8.0, 25°C
0.0092
Dehydrocholic acid
-
pH 8.0, 25°C
0.0053 - 0.4
NADP+
0.0045
NADPH
-
pH 8.0, 25°C
0.0062
Ursodeoxycholic acid
-
pH 8.0, 25°C
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
15.8
7-ketolithocholic acid
-
pH 8.0, 25°C
14.6
Dehydrocholic acid
-
pH 8.0, 25°C
15.7
NADP+
-
pH 8.0, 25°C
17.2
NADPH
-
pH 8.0, 25°C
19.6
Ursodeoxycholic acid
-
pH 8.0, 25°C
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
0.1
-
crude cell extract
0.276
-
purified enzyme
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
6
assay at, ursodeoxycholic acid formation
7.7
-
potassium buffer
9.5 - 10.5
-
-
9.8 - 10.5
-
-
11
-
glycine-NaOH buffer
pH RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
5 - 11.5
activity range
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE
additional information
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY hide
GeneOntology No.
LITERATURE
SOURCE
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
30000
-
2 * 30000, SDS-PAGE
34000
-
x * 34000, SDS-PAGE
45000
-
gel filtration
53000
-
gel filtration
56100
-
gel filtration
82000
-
gel filtration
115000
-
gel filtration
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
?
-
x * 34000, SDS-PAGE
homodimer
additional information
Crystallization/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
purified His-tagged enzyme complexed with NADP+, mixing of 10-15 mg/ml protein in 20% v/v glycerol, 20 mM potassium phosphate, pH 7.0, and 5 mM NADP+, with1 M sodium formate, 100 mM sodium acetate, pH 4.6, at room temperature, 3-4days, X-ray diffraction structure determination and analysis at 1.6 A resolution, modelling, molecular replacement using the the apoform structure of 7alpha-HSDH from Escherichia coli as search model, PDB ID 1AHH
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
20
-
50% loss of activity within 60 min
20 - 60
purified recombinant His-tagged enzyme, in 100 mM MES, pH 6.0, 30 min, reductive reaction, loss of 50% activity at 50°C, and inactivation at 60°C
GENERAL STABILITY
ORGANISM
UNIPROT
LITERATURE
heat-sensitive
-
presence of reducing agents protects against thermal denaturation
-
STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
-20°C, 50 mM potassium phosphate, pH 8.0, stable for at least 3 months with several times of freezing and thawing
-
Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
native enzyme from liver microsomes 300fold by ion exchange, hydroxylapatite, 2',5'-ADP affinity
-
recombinant His6-tagged enzyme from Escherichia coli strain JM109 by nickel affinity chromatography, dialysis, and ultrafiltration
recombinant N-terminally His6-tagged enzyme from Escherichia coli strain BL21(DE3) by nickel affinity chromatography, ultrafiltration, and gel filtration
to homogeneity, chromatography techniques
Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
expression in Escherichia coli
-
gene RUMGNA_02585, DNA and amino acid sequence determination and analysis, recombinant expression of N-terminally His6-tagged enzyme in Escherichia coli strain JM109
recombinant expression of N-terminally His6-tagged enzyme in Escherichia coli strain BL21(DE3)
ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
additional information
APPLICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
synthesis