Information on EC 1.14.99.22 - ecdysone 20-monooxygenase

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The enzyme appears in viruses and cellular organisms

EC NUMBER
COMMENTARY hide
1.14.99.22
-
RECOMMENDED NAME
GeneOntology No.
ecdysone 20-monooxygenase
REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
ecdysone + AH2 + O2 = 20-hydroxyecdysone + A + H2O
show the reaction diagram
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
oxidation
redox reaction
-
-
-
-
reduction
PATHWAY
BRENDA Link
KEGG Link
MetaCyc Link
ecdysone and 20-hydroxyecdysone biosynthesis
-
-
Insect hormone biosynthesis
-
-
SYSTEMATIC NAME
IUBMB Comments
ecdysone,hydrogen-donor:oxygen oxidoreductase (20-hydroxylating)
An enzyme from insect fat body or malpighian tubules involving a heme-thiolate protein (P-450). NADPH can act as ultimate hydrogen donor.
CAS REGISTRY NUMBER
COMMENTARY hide
55071-97-1
-
ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
females
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
canton S strain
-
-
Manually annotated by BRENDA team
flesh-fly
-
-
Manually annotated by BRENDA team
crayfish
-
-
Manually annotated by BRENDA team
Parasarcophaga argyrostoma
flesh-fly
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
cotton leafworm
-
-
Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
malfunction
RNAi-mediated knockdown of Shade negatively affects ecdysone-20-hydroxylation in the desert locust
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
alpha-ecdysone + NADPH + O2
20-hydroxyecdysone + NADP+ + H2O
show the reaction diagram
ecdysone + AH2 + O2
20-hydroxyecdysone + A + H2O
show the reaction diagram
ecdysone + NADPH + H+ + O2
20-hydroxyecdysone + NADP+ + H2O
show the reaction diagram
ecdysone + NADPH + O2
20-hydroxyecdysone + NADP+ + H2O
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
ecdysone + AH2 + O2
20-hydroxyecdysone + A + H2O
show the reaction diagram
ecdysone + NADPH + H+ + O2
20-hydroxyecdysone + NADP+ + H2O
show the reaction diagram
-
-
-
-
?
ecdysone + NADPH + O2
20-hydroxyecdysone + NADP+ + H2O
show the reaction diagram
additional information
?
-
-
E20-M preferentially uses NADPH in catalyzing ecdysone conversion to the physiologically active molting hormone, 20-hydroxyecdysone
-
-
-
COFACTOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
cytochrome P450
-
Ferredoxin
-
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
1,9-Dideoxyforskolin
1H-[1,2,4]oxadizolo[4,3-a]-quinoxalin-1-one
-
a guanylate cyclase inhibitor, applied with ecdysone agonist RH-5849 results in 63% reduced E20M activity
2-isopropoxy-1,4-naphthoquinone
2-methoxy-1,4-naphthoquinone
2-propoxy-1,4-naphthoquinone
20-hydroxyecdysone
6-anilinoquinoline-5,6-quinone
-
LY-38583, a guanylate cyclase inhibitor, applied with ecdysone agonist RH-5849 results in 80% reduced E20M activity, addition of guanylate cyclase activator S levels the activity to 78% of control activity
7-O-Hemisuccinyl-7-deacetyl-forskolin
diquat dibromide
ecdysterone
-
competitive inhibition
forskolin
KCN
-
25% inhibition
KK-42
-
i.e. 1-benzyl-5[(E)-2,6-dimethyl-1,5-heptadienyl]imidazole
methylene blue
-
a guanylate cyclase inhibitor, applied with ecdysone agonist RH-5849 results in 86% reduced E20M activity, reversed by addition of dibutyryl-cGMP/theophylline to 87% of control activity
Metyrapone
NADP+
-
competitive inhibition
Non-ionic detergents
-
Tween 20, Tween 80 and Triton X-100
-
Oxidized cytochrome c
-
reversed by KCN
-
Tris/HCl-buffer
-
-
Tween 80
additional information
-
ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
20-hydroxyecdysone
-
injection of 20-hydroxyecdysone stimulates ecdysone 20-hydroxylase activity in the midgut
ATP
-
increase of activity
dibutyryl-cGMP
-
activates E20M activity by 50fold, diminished by injections of guanylate cyclase inhibitors alone or combined with ecdysone agonist RH-5849
Juvenile hormone
-
-
L-dihydroxyphenylalanine
-
-
octopamine
additional information
-
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.00125
alpha-ecdysone
-
-
0.000000367 - 0.025
ecdysone
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
0.00000000617
Parasarcophaga argyrostoma
-
fat body
0.0000000103
-
fat body
0.00000041
-
mid- and hindgut
0.00000075
-
microsomes, fat body
0.00000115
-
malphigian tubules
0.000003
-
microsomes, malphigian tubules
0.00000562
-
malphigian tubules, 10000 x g pellet
0.0000197
-
fat body
0.0000466
-
mitochondrial enzyme system
0.0001721
-
midgut
0.0021
-
microsomes
27
-
recombinant GST-fusion protein, purified
additional information
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
6.8 - 8
-
potassium phosphate buffer
pH RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
6 - 8.5
-
-
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE
nondiapause egg
Manually annotated by BRENDA team
-
in the pupa of Bombyx mori, the rectal sac is an essential organ that operates as a repository for degraded ecdysteroids, and it can be distended by administration of 20-hydroxyecdysone. rectal sac distention appears 4 days after 20-hydroxyecdysone administration
Manually annotated by BRENDA team
additional information
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY hide
GeneOntology No.
LITERATURE
SOURCE
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
60000
-
? * 60000, SDS-PAGE
77700
-
? * 77700, sequence determination
103000
-
? * 103000, SDS-PAGE, recombinant GST-fusion protein
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
GENERAL STABILITY
ORGANISM
UNIPROT
LITERATURE
bovine serum albumin stabilizes
-
instable, linear activity under standard assay conditions for 10 min
-
STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
-30C, microsomal preparations with 30% v/v glycerol
-
-80C, quick frozen membranes, 15% glycerol v/v, at least 1 month
-
4C to -20C storage for prolonged periods not possible due to unstability of the enzyme, appreciable loss of activity
-
Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
not: purification of the enzyme from microsomes failed due to unstability of the enzyme
-
recombinant GST-fusion protein
-
solubilization of the enzyme from microsomes with detergent Synperonic NP10
-
Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
cDNA isolated from non-diapause eggs, expression in Escherichia coli BL21 as GST-fusion protein
-
cloned CYP6H1 cDNA is probably identical with purified enzyme
-
expression in S2 cells
gene SgShd, DNA and amino acid sequence determination and analysis
into a pSTBlue-1 Acceptor vector for sequencing, recombinant baculovirus harboring the BmE20OHase cDNA is constructed using the BAC-TO-BAC baculovirus expression system
EXPRESSION
ORGANISM
UNIPROT
LITERATURE
ecdysone 20-hydroxylase expression is not induced by 20-hydroxyecdysone
-
L-dihydroxyphenylalanine and octopamine feeding increases ecdysone 20-monooxygenase activity by a factor of 1.6 and 1.7, respectively
-
ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
additional information