Information on EC 1.3.99.6 - 3-oxo-5beta-steroid 4-dehydrogenase

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

EC NUMBER
COMMENTARY
1.3.99.6
-
RECOMMENDED NAME
GeneOntology No.
3-oxo-5beta-steroid 4-dehydrogenase
REACTION
REACTION DIAGRAM
COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
a 3-oxo-5beta-steroid + acceptor = a 3-oxo-DELTA4-steroid + reduced acceptor
show the reaction diagram
-
-
-
-
REACTION TYPE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
oxidation
-
-
-
-
redox reaction
-
-
-
-
reduction
-
-
-
-
PATHWAY
KEGG Link
MetaCyc Link
Metabolic pathways
-
Steroid hormone biosynthesis
-
SYSTEMATIC NAME
IUBMB Comments
3-oxo-5beta-steroid:acceptor DELTA4-oxidoreductase
-
SYNONYMS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
3-keto-DELTA4-steroid-5beta-reductase
-
-
-
-
3-oxo-5beta-steroid DELTA4-dehydrogenase
-
-
-
-
3-oxo-DELTA4-steroid-5beta-reductase
-
-
-
-
5beta-POR
-
-
5beta-POR
Q6PQJ9
-
5beta-POR
Q078S6
-
5beta-reductase
-
-
-
-
AKR1D1
-
-
Aldo-keto reductase family 1 member D1
-
-
-
-
Delta(4)-3-ketosteroid 5-beta-reductase
-
-
-
-
DELTA-3-ketosteroid-5beta-reductase
-
-
-
-
DELTA4-3-ketosteroid-5beta-reductase
-
-
-
-
DELTA4-3-oxosteroid 5beta-reductase
-
-
-
-
DELTA4-3-oxosteroid 5beta-reductase
-
-
DELTA4-5beta-DH
-
-
-
-
EC 1.3.1.3
Q078S6
related
progesterone 5beta-reductase
-
-
progesterone 5beta-reductase
Q6PQJ9
-
progesterone 5beta-reductase
-
-
progesterone 5beta-reductase
Q078S6
-
steroid 5beta reductase
-
-
-
-
steroid 5beta-reductase
-
-
steroid DELTA-5beta-dehydrogenase
-
-
-
-
testosterone 5beta-reductase
-
-
-
-
CAS REGISTRY NUMBER
COMMENTARY
9067-97-4
-
ORGANISM
COMMENTARY
LITERATURE
SEQUENCE CODE
SEQUENCE DB
SOURCE
gene SRD5B1, enzyme AKR1D1, healthy persons and 3 different patients with neonatal onset cholestatic liver disease and normal gamma-glutamyl transpeptidase activity
-
-
Manually annotated by BRENDA team
SUBSTRATE
PRODUCT                      
REACTION DIAGRAM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
(Substrate)
LITERATURE
(Substrate)
COMMENTARY
(Product)
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
17beta-hydroxy-5beta-androstan-3-one + menaphthone
testosterone + reduced acceptor
show the reaction diagram
-
acceptor: phenazine methosulfate
-
-
?
17beta-hydroxy-5beta-androstan-3-one + menaphthone
testosterone + reduced acceptor
show the reaction diagram
-
acceptor: 1,2-naphthoquinone, 1,4-naphthoquinone, 1,4-benzoquinone, menadione or coenzyme Q10
-
-
?
21-hydroxy-5beta-pregnan-3,20-dione + acceptor
21-hydroxy-pregn-4-en-3,20-dione + reduced acceptor
show the reaction diagram
-
acceptor: 1,2-naphthoquinone, 1,4-naphthoquinone, 1,4-benzoquinone, menadione or coenzyme Q10
-
-
?
3,12-dioxo-5beta-cholan-24-oic acid + menaphthone
3,12-dioxo-4-cholen-24-oic acid + reduced menaphthone
show the reaction diagram
-
-
-
-
?
3,12-dioxo-5beta-cholan-24-oic acid + phenazine methosulfate
3,12-dioxo-4-cholen-24-oic acid + reduced phenazine methosulphate
show the reaction diagram
-
-
-
-
?
3,12-dioxo-5beta-cholan-24-oic acid methyl ester + menaphthone
3,12-dioxo-4-cholen-24 oic acid methyl ester + reduced menaphthone
show the reaction diagram
-
-
-
-
?
3,12-dioxo-5beta-cholan-24-oic acid methyl ester + phenazine methosulfate
3,12-dioxo-4-cholen-24 oic acid methyl ester + reduced phenazine methosulfate
show the reaction diagram
-
-
-
-
?
3-oxo-5beta-24-cholan-oic acid + phenazine methosulfate
3-oxo-4-cholen-24-oic acid + reduced phenazine methosulfate
show the reaction diagram
-
-
-
-
?
3-oxo-5beta-cholan-24-oic acid + menaphthone
3-oxo-4-cholen-24-oic acid + reduced menaphthone
show the reaction diagram
-
-
-
-
?
4-androstene-3,17-dione + NADPH
5beta-androstan-3,15-dione + NADP+
show the reaction diagram
Q078S6
-
-
-
?
4-androstene-3,17-dione + NADPH
5beta-androstan-3,15-dione + NADP+
show the reaction diagram
Q6PQJ9
-
-
-
?
4-pregnen-3,20-dione + NADH
5beta-pregnan-3,20-dione + NAD+
show the reaction diagram
-
-
-
-
?
4-pregnen-3,20-dione + NADH
5beta-pregnan-3,20-dione + NAD+
show the reaction diagram
-
i.e. progesterone
-
-
?
5beta-androst-3,17-dione + acceptor
androst-1,4-dien-3,17-dione + reduced acceptor
show the reaction diagram
-
acceptor: 1,2-naphthoquinone, 1,4-naphthoquinone, 1,4-benzoquinone, menadione or coenzyme Q10
-
-
?
5beta-androstan-3,17-dione + acceptor
4-androsten-3,17-dione + reduced acceptor
show the reaction diagram
-
acceptor: phenazine methosulfate
-
-
?
5beta-androstan-3,17-dione + acceptor
4-androsten-3,17-dione + reduced acceptor
show the reaction diagram
-
acceptor: nitroblue tetrazolium
-
-
?
5beta-androstan-3,17-dione + acceptor
4-androsten-3,17-dione + reduced acceptor
show the reaction diagram
-
acceptor: 1,2-naphthoquinone, 1,4-naphthoquinone, 1,4-benzoquinone, menadione or coenzyme Q10
-
-
?
5beta-androstane-3,17-dione + acceptor
androst-4-en-3,17-dione + reduced acceptor
show the reaction diagram
-
acceptor: 1,2-naphthoquinone, 1,4-naphthoquinone, 1,4-benzoquinone, menadione or coenzyme Q10
-
-
?
5beta-pregnan-3,20-dione + menaphthone
4-pregnen-3,20-dione + reduced menaphthone
show the reaction diagram
-
-
-
-
?
5beta-pregnan-3,20-dione + phenazine methosulfate
4-pregnen-3,20-dione + reduced menazine methosulfate
show the reaction diagram
-
-
-
-
?
cortexone + NADPH
21-trihydroxy-5beta-pregnane-3,20-dione + NADP+
show the reaction diagram
Q078S6
-
-
-
?
cortexone + NADPH
21-trihydroxy-5beta-pregnane-3,20-dione + NADP+
show the reaction diagram
Q6PQJ9
-
-
-
?
cortisol + NADPH
11beta,17,21-trihydroxy-5beta-pregnane-3,20-dione + NADP+
show the reaction diagram
Q078S6
-
-
-
?
progesterone + NADPH + H+
5beta-pregnane-3,20-dione + NADP+
show the reaction diagram
-
-
-
-
?
progesterone + NADPH + H+
5beta-pregnane-3,20-dione + NADP+
show the reaction diagram
Q078S6
-
-
-
ir
progesterone + NADPH + H+
5beta-pregnane-3,20-dione + NADP+
show the reaction diagram
-
-
-
-
?
progesterone + NADPH + H+
5beta-pregnane-3,20-dione + NADP+
show the reaction diagram
Q6PQJ9
5alpha-pregnane-3,20-dione, the 5alpha-isomer of the pregnane-3,20-dione is not formed
-
-
ir
progesterone + NADPH + H+
5beta-pregnane-3,20-dione + NADP+
show the reaction diagram
-
highest substrate specificity
-
-
ir
testosterone + NADH
17beta-hydroxy-5beta-androstan-3-one + NAD+
show the reaction diagram
-
-
-
-
-
testosterone + NADPH + H+
(5beta,17beta)-17-hydroxyandrostan-3-one + NADP+
show the reaction diagram
Q6PQJ9
-
-
-
?
cortisol + NADPH
11beta,17,21-trihydroxy-5beta-pregnane-3,20-dione + NADP+
show the reaction diagram
Q6PQJ9
-
-
-
?
additional information
?
-
-
no activity with: NAD+, NADP+, FMN and FAD, inducible by 3-oxo-5beta-cholan-24-oic acid
-
-
-
additional information
?
-
-
inducible by testosterone and progesterone
-
-
-
additional information
?
-
-
enzyme is involved in bile acid biosynthesis
-
-
-
additional information
?
-
-
enzyme is essential in bile acids formation, analysis of bile acid contents in urine and plasma from healthy persons and 3 different patients with neonatal onset cholestatic liver disease, physiolgical role of the enzyme, overview
-
-
-
additional information
?
-
Q6PQJ9
pregnenolone, 21-OH-pregenenolone and isoprogesterone are not accepted by recombinant 5beta-POR
-
-
-
additional information
?
-
-
the recombinant enzyme catalyzes the stereospecific reduction of the delta4-double bond of several steroids with a 3-oxo,delta4,5 structure
-
-
-
NATURAL SUBSTRATES
NATURAL PRODUCTS
REACTION DIAGRAM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
(Substrate)
LITERATURE
(Substrate)
COMMENTARY
(Product)
LITERATURE
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
additional information
?
-
-
inducible by 3-oxo-5beta-cholan-24-oic acid
-
-
-
additional information
?
-
-
inducible by testosterone and progesterone
-
-
-
additional information
?
-
-
enzyme is involved in bile acid biosynthesis
-
-
-
additional information
?
-
-
enzyme is essential in bile acids formation, analysis of bile acid contents in urine and plasma from healthy persons and 3 different patients with neonatal onset cholestatic liver disease, physiolgical role of the enzyme, overview
-
-
-
COFACTOR
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
NADPH
-
cannot be replaced by NADH
INHIBITORS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
iodoacetate
-
50 mM, partial
ACTIVATING COMPOUND
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
FMNH2
-
slightly stimulatory in combination with NADH
KM VALUE [mM]
KM VALUE [mM] Maximum
SUBSTRATE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
0.228
-
4-androstene-3,17-dione
-
-
2.193
-
4-androstene-3,17-dione
Q078S6
-
0.042
-
4-pregnen-3,20-dione
-
-
1.597
-
cortexone
-
-
2.373
-
cortexone
Q078S6
-
0.291
-
cortisol
-
-
2.079
-
cortisol
Q078S6
-
0.0079
-
NADPH
-
-
0.008
-
NADPH
-
-
0.015
-
NADPH
Q078S6
-
0.0391
-
progesterone
-
-
0.12
-
progesterone
-
-
0.12
-
progesterone
-
purified wild type enzyme
0.215
-
progesterone
Q078S6
-
SPECIFIC ACTIVITY [µmol/min/mg]
SPECIFIC ACTIVITY MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
1.89
-
-
purified wild type enzyme, using progesterone as substrate
additional information
-
-
-
pH OPTIMUM
pH MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
7.8
-
-
with progesterone as substrate
8.5
10.5
-
pH 8.5: about 40% of maximal activity, pH 10.5: about 20% of maximal activity
8.5
-
Q078S6
-
9
-
-
above pH 9.0, the rate of DELTA4-5beta-dehydrogenase reaction increases with pH to the limit where accurate measurement can be made, pH 9.0
pH RANGE
pH RANGE MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
TEMPERATURE OPTIMUM
TEMPERATURE OPTIMUM MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
40
-
-
with progesterone as substrate
TEMPERATURE RANGE
TEMPERATURE MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
31
43
-
31°C: about 70% of maximal activity, 43°C: about 60% of maximal activity
SOURCE TISSUE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
SOURCE
-
lowest expression
Manually annotated by BRENDA team
-
from patients late in pregnancy, before term labour, during term labour and within the first 24 h postpartum
Manually annotated by BRENDA team
-
maximally transcribed in mature flowers
Manually annotated by BRENDA team
-
maximally transcribed
Manually annotated by BRENDA team
-
of alcohol-fed animals
Manually annotated by BRENDA team
-
relative expression decreases significantly in association with labour by about 10fold
Manually annotated by BRENDA team
-
mRNA is expressed in hepatocyte-like cells of rats treated with copper depletion regimen but not in the normal pancreas of rat fed a normal diet
Manually annotated by BRENDA team
-
most highly expressed, relative expression decreases significantly in association with labour by about 2fold
Manually annotated by BRENDA team
-
at the base of the rosette
Manually annotated by BRENDA team
-
at the basal part of the leaf, lowest amount
Manually annotated by BRENDA team
LOCALIZATION
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
GeneOntology No.
LITERATURE
SOURCE
MOLECULAR WEIGHT
MOLECULAR WEIGHT MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
32000
-
-
gel filtration
80000
-
-
gel filtration
SUBUNITS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
?
-
x * 37000, SDS-PAGE
?
-
x * 43963, sequence analysis, x * 43000, purified enzyme, SDS–PAGE
?
-
x * 40000, SDS–PAGE
?
Q078S6
x * 45000, SDS–PAGE
Crystallization/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
5beta-POR, in the presence and absence of the cofactor NADP+, hanging drop vapour diffusion method, using 15% polyethylene glycol 4000, 0.1 M ammonium acetate, and 0.1 M sodium citrate, pH 5.6
-
by the hanging-drop method, at 2.7 A resolution, crystals belong to space group P43212, contain a single 5beta-POR molecule in the asymmetric unit and tend to diffract anisotropically, identification of six out of eight possible Se-atom positions
-
TEMPERATURE STABILITY
TEMPERATURE STABILITY MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
58
-
-
half-life: 10 min
GENERAL STABILITY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
glycerol stabilizes
-
phenazine methosulfate stabilizes
-
OXIDATION STABILITY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
STORAGE STABILITY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
-70°C, 20% glycerol, 1 week, less than 30% loss of activity
-
4°C, in neutral buffer, 20% glycerol, about 50% loss of activity after 1 month
-
Purification/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
by Ni-NTA affinity chromatography
-
nickel-Sepharose affinity column chromatography and Superdex 75 gel filtration
-
to homogeneity by Ni–NTA affinity chromatography
-
to apparent homogeneity by affinity chromatography
-
by Ni-NTA affinity chromatography
Q078S6
Cloned/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
expressed in Escherichia coli strain M15
-
overproduced by the pQE expression system and transformed into Escherichia coli strain B834(DE3)
-
Sph1/Sal1 5beta-POR fragment cloned into the pQE vector and transformed into Escherichia coli strain M15(pREP4)
-
full-length P5betaR cDNA overexpressed in Escherichia coli as a fusion protein with MBP or His-tag
-
gene SRD5B1, DNA sequence determination and analysis from healthy persons and 3 different patients with neonatal onset cholestatic liver disease and normal gamma-glutamyl transpeptidase activity, parents and siblings
-
SPHI /SALI IC5beta-POR fragment cloned into the pQE vector system and transformed into Escherichai coli strain M15(pREP4)
Q078S6
expression in Escherichia coli
-
ENGINEERING
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
Y179A
-
inactive
Y179F
-
inactive
P198L
-
naturally occurring mutation of C662T in patient MS, homozygous, patient shows giant cell hepatitis
L106F
-
naturally occurring mutation of C385T in patient RM, homozygous, patient shows giant cell hepatitis and prominent extramedullary haemopoiesis
additional information
-
in patient BH naturally occurs a frame shift due to a base deletion, 511delT, resulting in a premature stop codon in exon 5, patient shows giant cell hepatitis and prominent extramedullary haemopoiesis
APPLICATION
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
analysis
-
determination of the concentration of chenodeoxycholic acid by an enzyme kit consisting of 3alpha-hydroxysteroid dehydrogenase, 3-keto-5beta-steroid-DELTA4-dehydrogenase, NAD+, nitroblue tetrazolium, diaphorase and some other reagents
additional information
-
inspection of the MAD-phased electron-density map shows that 5beta-POR is a Rossmann-type reductase and the quality of the map is such that it is anticipated that a complete atomic model of 5beta-POR may readily be built
additional information
-
5beta-POR is highly conserved within the genus Digitalis and the respective genes and proteins share considerable homology to putative progesterone reductases from other plant species
additional information
-
P5betaR belongs to the short-chain dehydrogenase/reductase (SDR) superfamily, bearing no structural homology to its mammalian counterpart, which is a member of the aldo-keto reductase (AKR) superfamily
additional information
Q078S6
significant homology to the putative progesterone 5beta-reductases isolated from other plant species, such as Digitalis lanata (ca. 98% identical) and Arabidopsis thaliana (ca. 69% identical)