Information on EC 4.2.1.92 - hydroperoxide dehydratase

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

EC NUMBER
COMMENTARY hide
4.2.1.92
-
RECOMMENDED NAME
GeneOntology No.
hydroperoxide dehydratase
REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
(9Z,11E,15Z)-(13S)-hydroperoxyoctadeca-9,11,15-trienoate = (9Z,15Z)-(13S)-12,13-epoxyoctadeca-9,11,15-trienoate + H2O
show the reaction diagram
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
C-O bond cleavage by elimination of water
-
-
-
-
PATHWAY
BRENDA Link
KEGG Link
MetaCyc Link
alpha-Linolenic acid metabolism
-
-
Biosynthesis of secondary metabolites
-
-
jasmonic acid biosynthesis
-
-
Metabolic pathways
-
-
SYSTEMATIC NAME
IUBMB Comments
(9Z,11E,15Z)-(13S)-hydroperoxyoctadeca-9,11,15-trienoate 12,13-hydro-lyase [(9Z,15Z)-(13S)-12,13-epoxyoctadeca-9,11,15-trienoate-forming]
Acts on a number of unsaturated fatty-acid hydroperoxides, forming the corresponding allene oxides. The product of the above reaction is unstable and is acted upon by EC 5.3.99.6, allene-oxide cyclase, to form the cyclopentenone derivative (15Z)-12-oxophyto-10,15-dienoate (OPDA), which is the first cyclic and biologically active metabolite in the jasmonate biosynthesis pathway [3]. The enzyme from many plants belongs to the CYP-74 family of P-450 monooxygenases [4].
CAS REGISTRY NUMBER
COMMENTARY hide
118390-60-6
-
37318-50-6
-
ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
garlic
-
-
Manually annotated by BRENDA team
gene ATEG_02036; gene ATEG_02036
UniProt
Manually annotated by BRENDA team
gene ATEG_02036; gene ATEG_02036
UniProt
Manually annotated by BRENDA team
oat
-
-
Manually annotated by BRENDA team
green bell pepper variety
-
-
Manually annotated by BRENDA team
susceptible and resistant pearl millet cultivars to Sclerospora graminicola infection, IP18292, ICMP451-P6, ICMR-01004, IP18293, H77/833-2, 700651, P310-17, 7042S, 843B and 852B, gene PgAOS1
-
-
Manually annotated by BRENDA team
var. lanatus, gene HPLCl
-
-
Manually annotated by BRENDA team
gene CmHPL
-
-
Manually annotated by BRENDA team
gene FOXB_01332
UniProt
Manually annotated by BRENDA team
gene FOXB_01332
UniProt
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
i.e. Botryodiplodia theobromae, no activity in strain CBS 122127
-
-
Manually annotated by BRENDA team
i.e. Botryodiplodia theobromae, no activity in strain CBS 122127
-
-
Manually annotated by BRENDA team
Japanese Honeysuckle
UniProt
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
gene Npun_R5468
UniProt
Manually annotated by BRENDA team
Picual and Arbequina cultivars
-
-
Manually annotated by BRENDA team
Passiflora f. edulis flavicarpa
UniProt
Manually annotated by BRENDA team
french bean
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
gene 13-hpl
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
eggplant
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
evolution
malfunction
-
loss-of-function mutant hpl3-1 produces disease-resembling lesions spreading through the whole leaves. Mutant hpl3-1 plants exhibit enhanced induction of jasmonic acid, trypsin proteinase inhibitors and other volatiles, but decreased levels of green leaf volatiles including (Z)-3-hexen-1-ol. Mutant hpl3-1 plants are more attractive to a BPH egg parasitoid, Anagrus nilaparvatae, than the wild-type, most likely as a result of increased release of BPH-induced volatiles. Mutant hpl3-1 plants also show increased resistance to bacterial blight (Xanthomonas oryzae pv. oryzae)
metabolism
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
(12R)-hydroperoxy-(5Z,8Z,10E,14Z)-eicosatetraenoic acid
(8R)-hydroxy-(11R,12R)-epoxyeicosa-(5Z,9E,14Z)-trienoic acid + H2O
show the reaction diagram
(12S)-hydroperoxy-(5Z,8Z,10E,14Z)-eicosatetraenoic acid
(10R)-hydroxy-(11S,12S)-epoxyeicosa-(5Z,8Z,14Z)-trienoic acid + (8R)-hydroxy-(11S,12S)-epoxyeicosa-(5Z,9E,14Z)-trienoic acid + H2O
show the reaction diagram
-
-
the former being a isomer of hepoxilin A3, ration of products is 2:1
?
(13R)-hydroperoxy-(10E,12Z)-octadecadienoic
11-hydroxy-12(13)-epoxy-octadecadienoic acid + 9-hydroxy-12(13)-epoxy-octadecadienoic acid + H2O
show the reaction diagram
(13R)-hydroperoxylinolenic acid
13-ketolinolenic acid + threo-11-hydroxy-(12R,13R)-epoxy-(9Z,15Z)-octadecadienoic acid + erythro-11-hydroxy-(12R,13R)-epoxy-(9Z,15Z)-octadecadienoic acid
show the reaction diagram
the G316A mutant converts (13R)-hydroperoxylinolenic acid to 13-ketolinolenic acid (with an apparent Km of 0.01 mM) and to epoxyalcohols viz. erythro- and threo-11-hydroxy-(12R,13R)-epoxy-(9Z,15Z)-octadecadienoic acids and one of the corresponding cis-epoxides as major products
-
-
?
(13S)-hydroperoxy-(10E,12Z)-octadecadienoic
11-hydroxy-12(13)-epoxy-octadecadienoic acid + 9-hydroxy-12(13)-epoxy-octadecadienoic acid + H2O
show the reaction diagram
(13S)-hydroperoxy-(9Z,11E)-octadecadienoic acid
(12,13S)-oxido-(9Z,11)-octadecadienoic acid + H2O
show the reaction diagram
(13S)-hydroperoxy-(9Z,11E)-octadecadienoic acid
(12S,13S)-oxido-(9Z,11)-octadecadienoic acid + H2O
show the reaction diagram
(13S)-hydroperoxy-(9Z,11E)-octadecatrienoic acid
(12,13S)-epoxy-(9Z,11,15Z)-octadecatrienoic acid + H2O
show the reaction diagram
-
-
12-oxo-13-hydroxy-(9Z,15Z)-octadecadienoic acid + 12-oxo-(10,15Z)-phytodienoic acid, formed by spontaneous chemical or enzyme-catalyzed cyclization
?
(13S)-hydroperoxy-(9Z,11Z,15Z)-octadecatrienoic acid
(12,13S)-epoxy-(9Z,11Z,15Z)-octadecatrienoic acid + H2O
show the reaction diagram
by AOS1
-
-
?
(13S)-hydroperoxyoctadecadienoic acid
13-hydroxy-12-oxo-9(Z)-octadecenoic acid + 12-oxo-10-phytoenoic acid
show the reaction diagram
-
-
13-hydroxy-12-oxo-(9Z)-octadecenoic acid is the major product
-
?
(13S)-hydroperoxyoctadecatrienoic acid
13-hydroxy-12-oxo-(9Z,15Z)-octadecadienoic acid + 12-oxo-(10Z,15Z)-phytodienoic acid
show the reaction diagram
-
-
OsAOS1 produces a 1:1 racemic mixture of cis-(+)/cis-(-)-12-oxo-(10Z,15Z)-phytodienoic acid via non-enzymatic cyclization of 12,13-epoxy-9,11,15-octadecatrienoic acid, an unstable allene oxide
-
?
(13S,9Z,11E,15Z)-13-hydroperoxy-9,11,15-octadecatrienoic acid
?
show the reaction diagram
-
is converted to an allene oxide
-
-
?
(15S)-hydroperoxyeicosatetraenoic acid
(15S)-hydroxy-11,12-epoxyeicosatrienoic acid + H2O
show the reaction diagram
-
-
-
-
?
(5Z,8Z,11E,13Z,15S)-15-hydroperoxyicosa-5,8,11,13-tetraenoic acid
14,15-hepoxilin A3 + 14,15-hepoxilin B3 + H2O
show the reaction diagram
(8R)-hydroperoxy-(9Z,12Z)-octadecadienoic acid
(7S,8S)-dihydroxy-(9Z,12Z)-octadecadienoic acid + H2O
show the reaction diagram
(8R)-hydroperoxyeicosatetraenoic acid
?
show the reaction diagram
(8R)-hydroperoxylinoleic acid + H2O
(5S,8R)-dihydroxylinoleic acid
show the reaction diagram
-
-
-
-
?
(8R)-hydroperoxylinoleic acid + H2O
(8R,11S)-dihydroxylinoleic acid
show the reaction diagram
-
-
-
-
?
(8R)-hydroxyeicosatetraenoic acid
(8R)-hydroxy-(9R,10R)-trans-epoxy-eicosa-(5Z,11Z,14Z)-trienoic acid
show the reaction diagram
-
the native, unmodified enzyme shows no activity with (8R)-hydroxyeicosatetraenoic acid, but the oxidized enzyme epoxidizes the substrate analogue stereospecifically on the 9,10-double bond to form (8R)-hydroxy-(9R,10R)-trans-epoxy-eicosa-(5Z,11Z,14Z)-trienoic acid as the predominant product, via Compound I (FeV=O) intermediate
erythro and threo diastereomers of (8R)-hydroxy-9,10-trans-epoxy-eicosa-5Z,11Z,14Z-trienoate
-
?
(9R)-hydroperoxy-(10E,12Z,15Z)-octadecatrienoic acid
9-hydroxy-10-oxo-(12Z,15Z)-octadecadienoic acid
show the reaction diagram
(9S)-hydroperoxy-(10E,12Z)-octadecadienoic acid
(9S,10)-10-epoxy-(10Z,12Z)-octadecadienoic acid + H2O
show the reaction diagram
recombinant FOXB_01332 oxidizes 18:2n-6 to (9S)-hydroperoxy-10(E),12(Z)-octadecadienoic acid by hydrogen abstraction and antarafacial insertion of molecular oxygen and sequentially to an allene oxide, (90S)-(10)-epoxy-10,12(Z)-octadecadienoic acid, which gives nonenzymatic hydrolysis products alpha- and gamma-ketols
-
-
?
(9S)-hydroperoxy-(10E,12Z)-octadecadienoic acid
(9S,10)-epoxy-(10Z,12Z)-octadecadienoic acid + H2O
show the reaction diagram
(9S)-hydroperoxy-(10E,12Z)-octadecadienoic acid
10-oxo-(9R)-hydroxy-cis-12-octadecadienoic acid + H2O
show the reaction diagram
(9S)-hydroperoxy-(10E,12Z)-octadecadienoic acid
9-oxononanoic acid + (3Z)-nonenal
show the reaction diagram
-
-
-
-
?
(9S)-hydroperoxy-(10E,12Z,15Z)-octadecatrienoic acid
(9S,10)-epoxy-(10Z,12Z,15Z)-octadecatrienoic acid + H2O
show the reaction diagram
-
-
-
?
(9S)-hydroperoxyoctadecadienoic acid
?
show the reaction diagram
-
-
-
-
?
(9S)-hydroperoxyoctadecatrienoic acid
?
show the reaction diagram
-
-
-
-
?
(9S,10E,12Z)-9-hydroperoxy-10,12-octadecadienoic acid
?
show the reaction diagram
(9Z,11E)-(9R)-hydroperoxyoctadeca-9,11-dienoate
(9R,10)-10-epoxy-(10Z,12Z)-octadecadienoic acid
show the reaction diagram
(9Z,11E,13S,15Z)-13-hydroperoxy-9,11,15-octadecatrienoic acid
?
show the reaction diagram
13-hydroperoxy-9,11,15-octadecatrienoic acid
?
show the reaction diagram
-
-
-
-
?
13-hydroperoxy-9,11-octadecadienoic acid
(10E,12Z)-12-[(3S)-3-methyl-3-propyloxiran-2-ylidene]dodec-10-enoic acid + H2O
show the reaction diagram
-
wild-type AOS activity
-
-
?
13-hydroperoxy-9,11-octadecadienoic acid
(10E,12Z)-13-[(2-hydroxypentan-2-yl)oxy]trideca-10,12-dienoic acid + H2O
show the reaction diagram
-
activity of AOS mutant F137L
-
-
?
13-hydroperoxy-9,11-octadecadienoic acid
?
show the reaction diagram
-
-
-
-
?
13-hydroperoxylinoleic acid
(Z)-3-hexanal + 12-oxododecenoic acid
show the reaction diagram
-
-
-
-
?
13-hydroperoxylinoleic acid
?
show the reaction diagram
-
-
-
-
?
13-hydroperoxylinolenic acid
12-oxo-cis-10,15-phytodienoic acid + 9-hydroxy-12-oxo-cis-15-trans-11-octadecadienoic acid + H2O
show the reaction diagram
-
-
the latter is a minor product of approximately 5%
?
9-hydroperoxylinolenic acid
13-hydroxy-10-oxo-cis-15-trans-11-octadecadienoic acid + H2O
show the reaction diagram
-
-
-
?
alpha-linolenic acid
(E,Z)-3,6-nonadienal + 9-oxononanoic acid
show the reaction diagram
-
cleavage of 9-hydroperoxy fatty acid
-
-
?
heptadecatrienoic acid
12-hydroperoxyheptadecatrienoic acid + (8Z,10S,11Z,14Z)-10-hydroperoxyheptadeca-8,11,14-trienoic acid
show the reaction diagram
the G316A mutant transforms 17:3n-3 to both 12-hydroperoxyheptadecatrienoic acid (about 93%) and 10-hydroperoxyheptadecatrienoic acid (7%)
-
-
?
linoleate (10S)-10-hydroperoxide
(8E)-10-oxodecenoic acid + 1-octen-3-ol + (2Z)-octen-1-ol
show the reaction diagram
the catalase-related hemoprotein reacts rapidly and specifically with linoleate 10S-hydroperoxide with a hydroperoxide lyase activity specific for the 10S-hydroperoxy enantiomer
in a 3:1 ratio, strict enzymatic control in formation of the 3R alcohol configuration with 99% enantiomeric excess
-
?
linoleic acid
(Z)-3-nonenal + 9-oxononanoic acid
show the reaction diagram
-
cleavage of 9-hydroperoxy fatty acid
-
-
?
linoleic acid
?
show the reaction diagram
Mn-LO G316A metabolizeS 18:2n-6 to (11S)-hydroperoxyoctadecadienoic acid and (13R)-hydroperoxyoctadecadienoic acid in approximately the same relative amounts as the native enzyme, and (13R)-hydroperoxyoctadecadienoic acid accumulates as the end product
-
-
?
linolenic acid
(13R)-hydroperoxyoctadecatrienoic acid + (11S)-hydroperoxyoctadecatrienoic acid
show the reaction diagram
Mn-LO G316A metabolizes 18:3n-3 to (13R)-hydroperoxyoctadecatrienoic acid and (11S)-hydroperoxyoctadecatrienoic acid
-
-
?
linolic acid hydroperoxide + H2O
9,12,13-trihydroxy-trans-10-octadecenoic acid + 9,10,13-trihydroxy-trans-11-octadecenoic acid
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
(13S)-hydroperoxy-(9Z,11E)-octadecadienoic acid
(12,13S)-oxido-(9Z,11)-octadecadienoic acid + H2O
show the reaction diagram
(8R)-hydroperoxyeicosatetraenoic acid
?
show the reaction diagram
-
the substrate is converted to an unstable allene oxide product via the FeIV-OH intermediate, Compound II, the native enzyme shows a strong preference for its natural substrate
-
-
?
(8R)-hydroperoxylinoleic acid + H2O
(5S,8R)-dihydroxylinoleic acid
show the reaction diagram
-
-
-
-
?
(8R)-hydroperoxylinoleic acid + H2O
(8R,11S)-dihydroxylinoleic acid
show the reaction diagram
-
-
-
-
?
(9R)-hydroperoxy-(10E,12Z,15Z)-octadecatrienoic acid
9-hydroxy-10-oxo-(12Z,15Z)-octadecadienoic acid
show the reaction diagram
(9S)-hydroperoxy-(10E,12Z)-octadecadienoic acid
(9S,10)-epoxy-(10Z,12Z)-octadecadienoic acid + H2O
show the reaction diagram
(9Z,11E)-(9R)-hydroperoxyoctadeca-9,11-dienoate
(9R,10)-10-epoxy-(10Z,12Z)-octadecadienoic acid
show the reaction diagram
13-hydroperoxy-9,11-octadecadienoic acid
(10E,12Z)-12-[(3S)-3-methyl-3-propyloxiran-2-ylidene]dodec-10-enoic acid + H2O
show the reaction diagram
-
wild-type AOS activity
-
-
?
13-hydroperoxy-9,11-octadecadienoic acid
(10E,12Z)-13-[(2-hydroxypentan-2-yl)oxy]trideca-10,12-dienoic acid + H2O
show the reaction diagram
-
activity of AOS mutant F137L
-
-
?
13-hydroperoxylinoleic acid
(Z)-3-hexanal + 12-oxododecenoic acid
show the reaction diagram
-
-
-
-
?
alpha-linolenic acid
(E,Z)-3,6-nonadienal + 9-oxononanoic acid
show the reaction diagram
-
cleavage of 9-hydroperoxy fatty acid
-
-
?
linoleic acid
(Z)-3-nonenal + 9-oxononanoic acid
show the reaction diagram
-
cleavage of 9-hydroperoxy fatty acid
-
-
?
additional information
?
-
COFACTOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
cytochrome
P450 6003B
-
cytochrome P450
heme
-
ferric heme iron
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
heme
-
CYP74 heme is strongly hold in place than usual via ionic interactions with its proprionate groups
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
CH3HgI
-
-
imidazole
-
imidazole may coordinate to ferric heme iron, triggering a heme-iron transition from high spin state to low spin state
iodoacetic acid
-
-
N-bromosuccinimide
-
-
nordihydroguaiaretic acid
p-chloromercuribenzoate
-
-
additional information
-
mechanism based suicide inactivation is a common feature of hydroperoxide lyases and many other enzymes of the P450 family
-
ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
cytochrome P450
-
AOS is a non-classical cytochrome P450, which, like other CYP74 enzymes, does not require molecular oxygen and NADPH-dependent P450 reductase
-
additional information
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.0593
(13S)-hydroperoxy-(9Z,11Z,15Z)-octadecatrienoic acid
with 0.0001 mg purified AOS1, in 1 mL of buffer containing 50 mM sodium phosphate, pH 6.0
0.0123
(13S)-hydroperoxyoctadecadienoic acid
-
pH 7.5, 22C
0.0125
(13S)-hydroperoxyoctadecatrienoic acid
-
pH 7.5, 22C
-
0.0069
(9S)-hydroperoxyoctadecadienoic acid
-
pH 7.5, 22C
-
0.0086
(9S)-hydroperoxyoctadecatrienoic acid
-
pH 7.5, 22C
-
0.024
linoleate hydroperoxide
-
-
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
494.5
(13S)-hydroperoxyoctadecadienoic acid
Oryza sativa
-
pH 7.5, 22C
537.8
(13S)-hydroperoxyoctadecatrienoic acid
Oryza sativa
-
pH 7.5, 22C
-
44.9
(9S)-hydroperoxyoctadecadienoic acid
Oryza sativa
-
pH 7.5, 22C
-
17.5
(9S)-hydroperoxyoctadecatrienoic acid
Oryza sativa
-
pH 7.5, 22C
-
kcat/KM VALUE [1/mMs-1]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
39600
(13S)-hydroperoxyoctadecadienoic acid
Oryza sativa
-
pH 7.5, 22C
197758
43700
(13S)-hydroperoxyoctadecatrienoic acid
Oryza sativa
-
pH 7.5, 22C
197759
6500
(9S)-hydroperoxyoctadecadienoic acid
Oryza sativa
-
pH 7.5, 22C
197760
2000
(9S)-hydroperoxyoctadecatrienoic acid
Oryza sativa
-
pH 7.5, 22C
197761
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
2.32
-
purified recombinant His-tagged enzyme, solubilized with Triton X-100, pH 7.0, 22C
5.44
-
purified recombinant His-tagged enzyme, solubilized with dodecyl maltoside, pH 7.0, 22C
pH RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
5 - 7.8
-
about 20% of maximum activity at pH 5 and pH 7.8
5.5 - 9
-
about 50% of maximum activity at pH 5.5 and pH 9.0
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
20
-
assay at
pI VALUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
8.4
calculated from amino acid sequence
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE
-
quiescent barley
Manually annotated by BRENDA team
-
maximum enzyme abundance in vascular as well as in the epidermal regions
Manually annotated by BRENDA team
-
maximum enzyme abundance in vascular as well as in the epidermal regions
Manually annotated by BRENDA team
-
of germinating barley
Manually annotated by BRENDA team
additional information
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY hide
GeneOntology No.
LITERATURE
SOURCE
-
rhomboid proteases AtRBL8 and AtRBL9 in the chloroplast envelope affect the level of allene oxide synthase in Arabidopsis thaliana
Manually annotated by BRENDA team
-
rhomboid proteases AtRBL8 and AtRBL9 in the chloroplast envelope affect the level of allene oxide synthase in Arabidopsis thaliana
Manually annotated by BRENDA team
additional information
-
equally distributed between the high speed supernatant and particulate fractions
-
Manually annotated by BRENDA team
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
40000 - 45000
-
gel filtration, 2 bands of enzyme activity, MW 220000 and MW 40000-45000
53500
sequence analysis; sequence analysis
59200
calculated from amino acid sequence
133000
recombinant C-terminally His6-tagged, V5-tagged enzyme
220000
-
gel filtration, 2 bands of enzyme activity, MW 220000 and MW 40000-45000
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
additional information
-
secondary structure analysis by circular dichroism, the enzyme consists of 13.53% alpha-helix, 32.73% beta-sheet, 21.76% turn and 31.13% unordered, overview
POSTTRANSLATIONAL MODIFICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
additional information
-
rhomboid proteases AtRBL8 and AtRBL9 in the chloroplast envelope affect the level of allene oxide synthase in Arabidopsis thaliana
Crystallization/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
AOS free or in complex with 12R,13S-vernolic acid, sitting drop vapour diffusion method, using 100 mM Tris-HCl, pH 7.5, 15-20% polyethylene glycol 3350, and 39 mM nonanoyl-N-hydroxyethylglucamide
-
is a typical P450, the loop forms a compact beta-turn and incorporates the most typical P450 consensus sequence, FXXG, followed by three residues (XRX) followed by CXG encompassing the cysteine. CYP74 heme is strongly hold in place than usual via ionic interactions with its proprionate groups
-
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
10 - 80
-
purified enzyme, thermal denaturation at 80C, unfolding starts at about 40C
20
-
pH 7: 10% loss of activity after 20 h, pH 5: 60% loss of activity after 20 h
45
-
pH 6, 60 min, 35% loss of activity
60
-
pH 6, 60 min, complete loss of activity
Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
AOS1 purified on Ni-NTA resin
Ni-NTA column chromatography
-
recombinant His-tagged enzyme from Escherichia coli membranes by solubilization with Triton X-100 or dodecyl maltoside, the latter results in higher enzyme activity, followed by nickel affinity chromatography
-
recombinant N- or C-terminal His6-tagged enzyme from Escherichia coli strain BL21(DE3) by nickel affinity chromatography and dialysis
Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
AOS2 cloned and overexpressed in Escherichia coli; whole ORF of AOS1 amplified and ligated into cloning vector pBluescript SK II(+), sequenced and ligated int vector pET23a and overexpressed in Escherichia coli BL21
expressed in Escherichia coli BL21(DE3) cells
-
expression in Pichia pastoris
gene 13-hpl, sequence comparisons, functional expression in Escherichia coli partly in inculsion bodies, fusion of the maltose binding protein malE to the 5'-truncated 13-hpl gene improves functional expression to some extent
-
gene ATEG_02036, sequence comparisons with other enzymes, cloning and functional expression of the enzyme with V5 epitope and His6-tag from plasmid pIZ/V5-His in Spodoptera frugiperda Sf9 cells and in Escherichia coli
gene CmHPL, expression in Nicotiana benthamiana leaves via infiltration method with a aviral vector, real-time PCR expression analysis over 10 days after infiltration with increase of C-9 aldehyde production, overview
-
gene FOXB_01332, phylogenetic tree, expression of wild-type and mutant enzymes
gene HPLCl, overexpression in Saccharomyces cerevisiae strain BY4743 isogenic
-
gene Npun_R5468, expression of the enzyme with N- or C-terminal His6 tags in Escherichia coli strain BL21(DE3)
gene OsHPL3, DNA and amino acid sequence determination, analysis, and comparison
-
gene PgAOS1, phylogenetic analysis, quantitative enzyme expression analysis in different cultivars with and without infection by Sclerospora graminicola
-
overexpression of His-tagged enzyme in Escherichia coli membranes
-
EXPRESSION
ORGANISM
UNIPROT
LITERATURE
expression of AOS1 is higher than that of AOS2; expression of AOS1 is higher than that of AOS2
high temperature of 35C decreases HPL gene expression in fruit mesocarp
-
induction of allene oxide synthase activity in both susceptible and resistant pearl millet cultivars during Sclerospora graminicola infection with higher induction observed in the resistant cultivar
-
low temperature and wounding upregulates the enzyme and increases HPL enzyme activity, at 15C, a strong and transient increase in OeHPL gene expression level occurs in fruit mesocarp, mechanical damage increases the enzyme activity level
-
temperature, light, wounding and water regime regulate olive the 13-HPL gene at transcriptional level
-
ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
F137L/S155A
C1073S
site-directed mutagenesis, the substitution in the P450 domain abolishes allene oxide synthase activity
N964D
site-directed mutagenesis, the mutation markedly reduces allene oxide synthase activity
N964V
site-directed mutagenesis, the mutation markedly reduces allene oxide synthase activity
C1073S
-
site-directed mutagenesis, the substitution in the P450 domain abolishes allene oxide synthase activity
-
N964D
-
site-directed mutagenesis, the mutation markedly reduces allene oxide synthase activity
-
N964V
-
site-directed mutagenesis, the mutation markedly reduces allene oxide synthase activity
-
E946V
site-directed mutagenesis, mutant Glu946Val oxidized 18:2n-6 as the native enzyme with formation of the alpha-ketol as the main product after 30 min
F416W
site-directed mutagenesis, the catalytic residue mutant shows an unaltered product profile compared to the wild-type enzyme
N921V
site-directed mutagenesis, the mutant shows activity similar to the wild-type enzyme
S949A
site-directed mutagenesis, the mutant also forms alpha-ketol like the native enzyme
E946V
-
site-directed mutagenesis, mutant Glu946Val oxidized 18:2n-6 as the native enzyme with formation of the alpha-ketol as the main product after 30 min
-
F416W
-
site-directed mutagenesis, the catalytic residue mutant shows an unaltered product profile compared to the wild-type enzyme
-
N921V
-
site-directed mutagenesis, the mutant shows activity similar to the wild-type enzyme
-
S949A
-
site-directed mutagenesis, the mutant also forms alpha-ketol like the native enzyme
-
G316A
the G316A mutant increases the oxygenation at the n-8 carbon of 17:3n-3 and at the n-10 carbon of the C17 and C18 fatty acids (from 12% to 711%). The most striking effect of the G316A mutant is a 2-, 7-, and 15-fold increase in transformation of the n-6 hydroperoxides of 19:3n-3, 18:3n-3, and 17:3n-3, respectively, to keto fatty acids and epoxyalcohols. G316A mutant augments the hydroperoxide isomerase activity by positioning the hydroperoxy group at the n-6 carbon of n-3 fatty acids closer to the reduced catalytic metal
G316S
inactive mutant enzyme
G316T
inactive mutant enzyme
G316V
inactivfe mutant enzyme
E292G
-
site-directed mutagenesis, the divinyl ether synthase enzyme mutant E292G shows allene oxide synthase activity, producing the typical allene oxide synthase product alpha-ketol and 12-oxo-10,15-phytodienoic acid, and lacks divinyl ether synthase activity in contrast to the wild-type divinyl ether synthase CYP74B16
V379F
-
the divinyl ether synthase enzyme mutant V379F shows allene oxide synthase activity, producing the typical allene oxide synthase product alpha-ketol, and lacks divinyl ether synthase activity in contrast to the wild-type divinyl ether synthase CYP74D3
additional information
APPLICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
food industry
-
the enzyme produces aldehydes that are used as flavours in foods and beverages
medicine
-
enzyme is involved in skin differentiation and etiology of ichthyosiform diseases