Information on EC 1.14.13.101 - senecionine N-oxygenase

Please wait a moment until all data is loaded. This message will disappear when all data is loaded.
Specify your search results
Mark a special word or phrase in this record:
Select one or more organisms in this record:
Show additional data
Do not include text mining results
Include (text mining) results (more...)
Include results (AMENDA + additional results, but less precise; more...)


The expected taxonomic range for this enzyme is: Eukaryota

EC NUMBER
COMMENTARY
1.14.13.101
-
RECOMMENDED NAME
GeneOntology No.
senecionine N-oxygenase
REACTION
REACTION DIAGRAM
COMMENTARY
ORGANISM
UNIPROT
LITERATURE
senecionine + NADPH + H+ + O2 = senecionine N-oxide + NADP+ + H2O
show the reaction diagram
-
-
-
-
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
oxidation
-
-
-
-
redox reaction
-
-
-
-
reduction
-
-
-
-
PATHWAY
BRENDA Link
KEGG Link
MetaCyc Link
senecionine N-oxide biosynthesis
-
-
Tropane, piperidine and pyridine alkaloid biosynthesis
-
-
SYSTEMATIC NAME
IUBMB Comments
senecionine,NADPH:oxygen oxidoreductase (N-oxide-forming)
A flavoprotein. NADH cannot replace NADPH. While pyrrolizidine alkaloids of the senecionine and monocrotaline types are generally good substrates (e.g. senecionine, retrorsine and monocrotaline), the enzyme does not use ester alkaloids lacking an hydroxy group at C-7 (e.g. supinine and phalaenopsine), 1,2-dihydro-alkaloids (e.g. sarracine) or unesterified necine bases (e.g. senkirkine) as substrates [1]. Senecionine N-oxide is used by insects as a chemical defense: senecionine N-oxide is non-toxic, but it is bioactivated to a toxic form by the action of cytochrome P-450 oxidase when absorbed by insectivores.
SYNONYMS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
senecionine monooxygenase (N-oxide-forming)
-
-
-
-
CAS REGISTRY NUMBER
COMMENTARY
220581-68-0
-
ORGANISM
COMMENTARY
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
Amerila phaedra
-
-
-
Manually annotated by BRENDA team
Arctia intercalaris
-
-
-
Manually annotated by BRENDA team
no activity in Spodoptera littoralis
-
-
-
Manually annotated by BRENDA team
sinnabar moth
SwissProt
Manually annotated by BRENDA team
SUBSTRATE
PRODUCT                      
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate)
LITERATURE
(Substrate)
COMMENTARY
(Product)
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
atropine + NADPH + H+ + O2
atropine N-oxide + NADP+ + H2O
show the reaction diagram
-
-
-
-
?
atropine + NADPH + O2
atropine N-oxide + NADP+ + H2O
show the reaction diagram
D6CHF2
21% of the activity with monocrotaline
-
-
?
axillaridine + NADPH + H+ + O2
axillaridine N-oxide + NADP+ + H2O
show the reaction diagram
-
-
-
-
?
axillaridine + NADPH + O2
axillaridine N-oxide + NADP+ + H2O
show the reaction diagram
-
76% of the activity with senecionine
-
?
axillaridine + NADPH + O2
axillaridine N-oxide + NADP+ + H2O
show the reaction diagram
-
79% of the activity with senecionine
-
?
axillaridine + NADPH + O2
axillaridine N-oxide + NADP+ + H2O
show the reaction diagram
-
93% of the activity with senecionine
-
?
axillarine + NADPH + H+ + O2
axillarine N-oxide + NADP+ + H2O
show the reaction diagram
-
-
-
-
?
axillarine + NADPH + O2
axillarine N-oxide + NADP+ + H2O
show the reaction diagram
-
38% of the activity with senecionine
-
?
axillarine + NADPH + O2
axillarine N-oxide + NADP+ + H2O
show the reaction diagram
-
55% of the activity with senecionine
-
?
axillarine + NADPH + O2
axillarine N-oxide + NADP+ + H2O
show the reaction diagram
-
74% of the activity with senecionine
-
?
axillarine + NADPH + O2
axillarine N-oxide + NADP+ + H2O
show the reaction diagram
Q8MP06
74% of the activity with senecionine, native enzyme. 83% of the activity with senecionine, recombinant enzyme
-
?
axillarine + NADPH + O2
axillarine N-oxide + NADP+ + H2O
show the reaction diagram
D6CHF2
69% of the activity with monocrotaline
-
-
?
cysteamine + NADPH + H+ + O2
cysteamine N-oxide + NADP+ + H2O
show the reaction diagram
-
-
-
-
?
heliotrine + NADPH + H+ + O2
heliotrine N-oxide + NADP+ + H2O
show the reaction diagram
-
-
-
-
?
heliotrine + NADPH + O2
heliotrine N-oxide + NADP+ + H2O
show the reaction diagram
-
-
-
-
-
heliotrine + NADPH + O2
heliotrine N-oxide + NADP+ + H2O
show the reaction diagram
-
115% of the activity with senecionine
-
?
heliotrine + NADPH + O2
heliotrine N-oxide + NADP+ + H2O
show the reaction diagram
-
25% of the activity with senecionine
-
?
heliotrine + NADPH + O2
heliotrine N-oxide + NADP+ + H2O
show the reaction diagram
Q8MP06
25% of the activity with senecionine, native enzyme. 49% of the activity with senecionine, recombinant enzyme
-
?
heliotrine + NADPH + O2
heliotrine N-oxide + NADP+ + H2O
show the reaction diagram
-
86% of the activity with senecionine
-
?
heliotrine + NADPH + O2
heliotrine N-oxide + NADP+ + H2O
show the reaction diagram
D6CHF2
49% of the activity with monocrotaline
-
-
?
indicine + NADPH + H+ + O2
indicine N-oxide + NADP+ + H2O
show the reaction diagram
-
-
-
-
?
indicine + NADPH + O2
indicine N-oxide + NADP+ + H2O
show the reaction diagram
-
35% of the activity with senecionine
-
?
indicine + NADPH + O2
indicine N-oxide + NADP+ + H2O
show the reaction diagram
-
78% of the activity with senecionine
-
?
indicine + NADPH + O2
indicine N-oxide + NADP+ + H2O
show the reaction diagram
-
90% of the activity with senecionine
-
?
lycopsamine + NADPH + O2
lycopsamine N-oxide + NADP+ + H2O
show the reaction diagram
-
20% of the activity with senecionine
-
?
lycopsamine + NADPH + O2
lycopsamine N-oxide + NADP+ + H2O
show the reaction diagram
-
75% of the activity with senecionine
-
?
lycopsamine + NADPH + O2
lycopsamine N-oxide + NADP+ + H2O
show the reaction diagram
-
93% of the activity with senecionine
-
?
monocrotaline + NADPH + H+ + O2
monocrotaline N-oxide + NADP+ + H2O
show the reaction diagram
-
-
-
-
?
monocrotaline + NADPH + O2
monocrotaline N-oxide + NADP+ + H2O
show the reaction diagram
-
-
-
-
-
monocrotaline + NADPH + O2
monocrotaline N-oxide + NADP+ + H2O
show the reaction diagram
D6CHF2
-
-
-
?
monocrotaline + NADPH + O2
monocrotaline N-oxide + NADP+ + H2O
show the reaction diagram
-
103% of the activity with senecionine
-
?
monocrotaline + NADPH + O2
monocrotaline N-oxide + NADP+ + H2O
show the reaction diagram
-
110% of the activity with senecionine
-
?
monocrotaline + NADPH + O2
monocrotaline N-oxide + NADP+ + H2O
show the reaction diagram
-
92% of the activity with senecionine
-
?
nicotine + NADPH + H+ + O2
nicotine N-oxide + NADP+ + H2O
show the reaction diagram
-
-
-
-
?
phalaenopsine + NADPH + H+ + O2
phalaenopsine N-oxide + NADP+ + H2O
show the reaction diagram
-
-
-
-
?
phalaenopsine + NADPH + O2
phalaenopsine N-oxide + NADP+ + H2O
show the reaction diagram
D6CHF2
19% of the activity with monocrotaline
-
-
?
retronecine + NADPH + H+ + O2
retronecine N-oxide + NADP+ + H2O
show the reaction diagram
-
-
-
-
?
retrorsine + NADPH + H+ + O2
retrosine N-oxide + NADP+ + H2O
show the reaction diagram
-
-
-
-
?
retrorsine + NADPH + O2
retrorsine N-oxide + NADP+ + H2O
show the reaction diagram
-
93% of the activity with senecionine
-
?
retrorsine + NADPH + O2
retrorsine N-oxide + NADP+ + H2O
show the reaction diagram
-
97% of the activity with senecionine
-
?
retrorsine + NADPH + O2
retrorsine N-oxide + NADP+ + H2O
show the reaction diagram
-
as active as senecionine
-
?
rinderine + NADPH + H+ + O2
rinderine N-oxide + NADP+ + H2O
show the reaction diagram
-
-
-
-
?
rinderine + NADPH + O2
rinderine N-oxide + NADP+ + H2O
show the reaction diagram
-
23% of the activity with senecionine
-
?
rinderine + NADPH + O2
rinderine N-oxide + NADP+ + H2O
show the reaction diagram
-
79% of the activity with senecionine
-
?
rinderine + NADPH + O2
rinderine N-oxide + NADP+ + H2O
show the reaction diagram
-
81% of the activity with senecionine
-
?
rinderine + NADPH + O2
rinderine N-oxide + NADP+ + H2O
show the reaction diagram
D6CHF2
43% of the activity with monocrotaline
-
-
?
sarracine + NADPH + H+ + O2
sarracine N-oxide + NADP+ + H2O
show the reaction diagram
-
-
-
-
?
senecionine + NADPH + H+ + O2
senecionine N-oxide + NADP+ + H2O
show the reaction diagram
-
-
-
-
?
senecionine + NADPH + H+ + O2
senecionine N-oxide + NADP+ + H2O
show the reaction diagram
-
most efficient substrate
-
-
?
senecionine + NADPH + O2
senecionine N-oxide + NADP+ + H2O
show the reaction diagram
-
-
-
?
senecionine + NADPH + O2
senecionine N-oxide + NADP+ + H2O
show the reaction diagram
-
-
-
-
-
senecionine + NADPH + O2
senecionine N-oxide + NADP+ + H2O
show the reaction diagram
Q8MP06
-
-
?
senecionine + NADPH + O2
senecionine N-oxide + NADP+ + H2O
show the reaction diagram
Q8MP06
-
-
-
?
senecionine + NADPH + O2
senecionine N-oxide + NADP+ + H2O
show the reaction diagram
-
-
-
?
senecionine + NADPH + O2
senecionine N-oxide + NADP+ + H2O
show the reaction diagram
D6CHF2
61% of the activity with monocrotaline
-
-
?
seneciphylline + NADPH + H+ + O2
seneciphylline N-oxide + NADP+ + H2O
show the reaction diagram
-
second best substrate
-
-
?
seneciphylline + NADPH + O2
seneciphylline N-oxide + NADP+ + H2O
show the reaction diagram
-
117% of the activity with senecionine
-
?
seneciphylline + NADPH + O2
seneciphylline N-oxide + NADP+ + H2O
show the reaction diagram
-
119% of the activity with senecionine
-
?
seneciphylline + NADPH + O2
seneciphylline N-oxide + NADP+ + H2O
show the reaction diagram
-
95% of the activity with senecionine
-
?
seneciphylline + NADPH + O2
seneciphylline N-oxide + NADP+ + H2O
show the reaction diagram
Q8MP06
95% of the activity with senecionine, native enzyme. 93% of the activity with senecionine, recombinant enzyme
-
?
seneciphylline + NADPH + O2
seneciphylline N-oxide + NADP+ + H2O
show the reaction diagram
D6CHF2
59% of the activity with monocrotaline
-
-
?
senecivernine + NADPH + H+ + O2
senecivernine N-oxide + NADP+ + H2O
show the reaction diagram
-
-
-
-
?
senecivernine + NADPH + O2
senecivernine N-oxide + NADP+ + H2O
show the reaction diagram
-
107% of the activity with senecionine
-
?
senecivernine + NADPH + O2
senecivernine N-oxide + NADP+ + H2O
show the reaction diagram
-
113% of the activity with senecionine
-
?
senecivernine + NADPH + O2
senecivernine N-oxide + NADP+ + H2O
show the reaction diagram
-
81% of the activity with senecionine
-
?
triangularine + NADPH + O2
triangularine N-oxide + NADP+ + H2O
show the reaction diagram
-
60% of the activity with senecionine
-
?
monocrotaline + NADPH + O2
monocrotaline N-oxide + NADP+ + H2O
show the reaction diagram
Q8MP06
92% of the activity with senecionine, native enzyme. 94% of the activity with senecionine, recombinant enzyme
-
?
additional information
?
-
Q8MP06
the enzyme is highly specific for toxic pyrrolizidine alkaloids. No activity with: senkirkine, dimethylaniline, L-Pro, caffeine, atropine, supinidine, retronecine, phalaenopsine
-
?
additional information
?
-
-
the enzyme N-oxidizes only alkaloids with structural elements which are essential for hepatotoxic and genotoxic pyrrolizidine alkaloids, i.e. 1,2-double bond, esterification of the allylic hydroxyl group, presence of a second free or esterified hydroxyl group at carbon 7. No activity with: senkirkine, sarracine, supinine, phalaenopsine, retronecine, heliotridine, supinidine, isoretronecanol, nicotine, caffeine, tropane alkaloids, quinolizidine alkaloids, indole alkaloids, isoquinolines and synthetic heterocyclic tertiary amines
-
?
additional information
?
-
-
the enzyme N-oxidizes only alkaloids with structural elements which are essential for hepatotoxic and genotoxic pyrrolizidine alkaloids, i.e. 1,2-double bond, esterification of the allylic hydroxyl group, presence of a second free or esterified hydroxyl group at carbon 7. No activity with: triangularine, senkirkine, sarracine, supinine, phalaenopsine, retronecine, heliotridine, supinidine, isoretronecanol, nicotine, caffeine, tropane alkaloids, quinolizidine alkaloids, indole alkaloids, isoquinolines and synthetic heterocyclic tertiary amines
-
?
additional information
?
-
Q8MP06
the enzyme allows the larvae to feed on pyrrolizidine alkaloid-containing plants and to accumulate predation-deterrent pyrrolizidine alkaloids in the hemolymph
-
?
additional information
?
-
D6CHF2
the enzyme shows activity with glutathione (41% of the activity with monocrotaline)
-
-
-
additional information
?
-
-
no activity with senkirkine, heliotridine, supinidine, ephedrine, dimethylaniline, L-cysteine, hydroxylamine, and glutathione
-
-
-
NATURAL SUBSTRATES
NATURAL PRODUCTS
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate)
LITERATURE
(Substrate)
COMMENTARY
(Product)
LITERATURE
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
senecionine + NADPH + H+ + O2
senecionine N-oxide + NADP+ + H2O
show the reaction diagram
-
most efficient substrate
-
-
?
senecionine + NADPH + O2
senecionine N-oxide + NADP+ + H2O
show the reaction diagram
Q8MP06
-
-
-
?
seneciphylline + NADPH + H+ + O2
seneciphylline N-oxide + NADP+ + H2O
show the reaction diagram
-
second best substrate
-
-
?
additional information
?
-
Q8MP06
the enzyme allows the larvae to feed on pyrrolizidine alkaloid-containing plants and to accumulate predation-deterrent pyrrolizidine alkaloids in the hemolymph
-
?
COFACTOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
FAD
-
molar apoenzyme/FAD ratio of 0.4
FAD
Q8MP06
-
NADPH
-
strictly dependent on
NADPH
-
strictly dependent on, no reaction with NADH
NADPH
Q8MP06
dependent on
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
cytochrome c
-
-
ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
guanethidine
-
enhances activity
Metyrapone
-
enhances activity
n-octylamine
-
activity of solubilized enzyme is 1.6fold increased by 0.5 mM n-octylamine
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
0.00983
atropine
-
in 100 mM glycine/NaOH buffer pH 9.0, at 37C
0.0186
heliotrine
-
pH 7.0, 37C
0.0229
heliotrine
-
pH 7.0, 37C
0.2639
heliotrine
-
in 100 mM glycine/NaOH buffer pH 9.0, at 37C
0.284
heliotrine
-
pH 7.0, 37C
0.0029
monocrotaline
-
pH 7.0, 37C
0.0051
monocrotaline
-
pH 7.0, 37C
0.01193
monocrotaline
-
in 100 mM glycine/NaOH buffer pH 9.0, at 37C
0.0125
monocrotaline
-
pH 7.0, 37C
0.0013
NADPH
-
pH 7.0, pH 37C
0.8376
phalaenopsine
-
in 100 mM glycine/NaOH buffer pH 9.0, at 37C
0.00108
senecionine
-
in 100 mM glycine/NaOH buffer pH 9.0, at 37C
0.0014
senecionine
-
pH 7.0, 37C
0.0031
senecionine
-
pH 7.0, 37C
0.0048
senecionine
-
pH 7.0, 37C
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
0.37
atropine
-
in 100 mM glycine/NaOH buffer pH 9.0, at 37C
0.12
heliotrine
-
in 100 mM glycine/NaOH buffer pH 9.0, at 37C
0.96
monocrotaline
-
in 100 mM glycine/NaOH buffer pH 9.0, at 37C
0.13
phalaenopsine
-
in 100 mM glycine/NaOH buffer pH 9.0, at 37C
0.99
senecionine
-
in 100 mM glycine/NaOH buffer pH 9.0, at 37C
kcat/KM VALUE [1/mMs-1]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
37.64
atropine
-
in 100 mM glycine/NaOH buffer pH 9.0, at 37C
4789
0.454
heliotrine
-
in 100 mM glycine/NaOH buffer pH 9.0, at 37C
22428
80.5
monocrotaline
-
in 100 mM glycine/NaOH buffer pH 9.0, at 37C
13488
0.155
phalaenopsine
-
in 100 mM glycine/NaOH buffer pH 9.0, at 37C
117306
916.7
senecionine
-
in 100 mM glycine/NaOH buffer pH 9.0, at 37C
9685
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
3.3
D6CHF2
purified enzyme with senecionine as substrate
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
7.5
-
solubilized partially purified enzyme
8
D6CHF2
assay at
9
-
in potassium phosphate buffer (10 mM), Tris/HCl buffer (10 mM), and in glycine/NaOH buffer (100 mM)
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
37
D6CHF2
assay at
40 - 45
-
-
40 - 45
-
solubilized partially purified enzyme
pI VALUE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
4.9
-
determination on an Ampholine PAGplate pH 3.5-9.5 using a horizantal electrophoresis system
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
SOURCE
Amerila phaedra
-
-
Manually annotated by BRENDA team
-
from late instar larvae
Manually annotated by BRENDA team
Q8MP06
fat body and head
Manually annotated by BRENDA team
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY
GeneOntology No.
LITERATURE
SOURCE
additional information
-
not a microsomal enzyme
-
Manually annotated by BRENDA team
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
91200
-
gel filtration
728567
200000
-
gel filtration
651040
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
homodimer
-
2 * 47793, calculated from amino acid sequence
tetramer
-
4 * 51000, SDS-PAGE
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
37
-
20 min, solubilized enzyme is completely inactivated in absence of NADPH, preincubation the presence of NADPH has no effect
689412
Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
Ni2+-NTA agarose column chromatography
-
Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
expressing GgPNO without the signal peptide in at least a partially soluble and active form. Betaine is added to the medium and promotes its uptake by osmotic stress by sorbitol. Further improvements are achieved by coexpression with the Escherichia coli chaperones DnaK/DnaJ/GrpE by using the plasmid pRDKJG and by reducing the expression temperature to 4C
D6CHF2
expression in active form in Escherichia coli BL21
Q8MP06
the heterologous expression results in the formation of an insoluble and inactive protein. Despite the finding that protein solubilization and subsequent renaturation results in an active protein, the specific activity remains low. Active SNO is detected by using an an Sf9 insect cell expression system, but the yield of protein is too low for further biochemical characterization. Using an alignment of senecionine N-oxygenase of Tyria jacobaeae and of flavin-dependent monooxygenase-like sequences of the dipteran species Drosophila melanogaster and Anopheles gambiae, several new flavin-dependent monooxygenase sequences of arctiids and other Lepidoptera are idenified
Q8MP06
expressed in Escherichia coli BL21(DE3) cells
-