Information on EC 4.2.1.53 - Oleate hydratase

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The enzyme appears in viruses and cellular organisms

EC NUMBER
COMMENTARY hide
4.2.1.53
-
RECOMMENDED NAME
GeneOntology No.
Oleate hydratase
REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
(R)-10-Hydroxystearate = oleate + H2O
show the reaction diagram
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
elimination
-
-
-
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SYSTEMATIC NAME
IUBMB Comments
(R)-10-hydroxystearate 10-hydro-lyase (oleate-forming)
Acts on a number of 10-hydroxy acids.
CAS REGISTRY NUMBER
COMMENTARY hide
9073-51-2
-
ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
(R)-10-Hydroxystearate
Oleate + H2O
show the reaction diagram
alpha-linolenate + H2O
(R)-10-hydroxy-12,15(Z,Z)-octadecadienoate
show the reaction diagram
alpha-linolenic acid
10-hydroxy-12(Z),15(Z)-octadecadienoic acid + 10,13-dihydroxy-15(Z)-octadecenoic acid
show the reaction diagram
alpha-linolenic acid
10-hydroxy-12,15(Z,Z)-octadecadienoic acid + H2O
show the reaction diagram
-
-
82% yield after 18 h
-
?
alpha-linolenic acid + H2O
10-hydroxy-12,15(Z,Z)-octadecadienoic acid
show the reaction diagram
gamma-linolenate + H2O
(R)-10-hydroxy-6,12-(Z,Z)-octadecadienoate
show the reaction diagram
gamma-linolenic acid
10-hydroxy-6(Z),12(Z)-octadecadienoic acid + 10,13-dihydroxy-6(Z)-octadecenoic acid
show the reaction diagram
gamma-linolenic acid + H2O
10-hydroxy-6,12(Z,Z)-octadecadienoic acid
show the reaction diagram
linoleate + H2O
(R)-10-hydroxy-12(Z)-octadecenoate
show the reaction diagram
linoleic acid
10-hydroxy-12(Z)-octadecenoic acid + 10,13-dihydroxyoctadecanoic acid
show the reaction diagram
linoleic acid + H2O
10-hydroxy-12(Z)-octadecenoic acid
show the reaction diagram
-
-
-
-
?
Linoleic acid + H2O
10D-Hydroxy-DELTA12-cis-octadecenoic acid
show the reaction diagram
-
-
-
-
myristoleate + H2O
(R)-10-hydroxymyristate
show the reaction diagram
myristoleic acid
10-hydroxytetradecanoic acid + H2O
show the reaction diagram
-
-
-
?
myristoleic acid + H2O
10-hydroxymyristic acid
show the reaction diagram
-
-
-
-
?
oleate + H2O
(R)-10-hydroxystearate
show the reaction diagram
Oleic acid
?
show the reaction diagram
-
-
-
-
-
Oleic acid
DELTA10-trans-octadecenoic acid
show the reaction diagram
oleic acid + H2O
10-hydroxyoctadecanoic acid
show the reaction diagram
oleic acid + H2O
10-hydroxystearic acid
show the reaction diagram
-
-
-
?
palmitoleate + H2O
(R)-10-hydroxypalmitate
show the reaction diagram
-
-
-
-
?
palmitoleic acid
10-hydroxyhexadecanoic acid + H2O
show the reaction diagram
-
-
-
?
Palmitoleic acid + H2O
10-Hydroxypalmitic acid
show the reaction diagram
ricinoleic acid
10,12-dihydroxystearic acid + H2O
show the reaction diagram
-
90% conversion
-
-
?
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
(R)-10-Hydroxystearate
Oleate + H2O
show the reaction diagram
oleate + H2O
(R)-10-hydroxystearate
show the reaction diagram
Oleic acid
?
show the reaction diagram
-
-
-
-
-
oleic acid + H2O
10-hydroxyoctadecanoic acid
show the reaction diagram
COFACTOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
FAD
required for activity, maximum activity in the presence of 0.2 M FAD
additional information
-
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
additional information
calcium has no role in the catalytic mechanism
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
ethanol
-
less than 60% residual activity at 6% (v/v) ethanol
methanol
-
45% residual activity at 8% (v/v) methanol
additional information
-
ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
EDTA
-
maximum activity at 2 M
ethanol
NaCl
-
maximum activity at 125 mM
Tween 40
additional information
addition of EDTA or EGTA (both at a 1000fold excess) has no effect on the activity
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KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.059
alpha-linolenic acid
at pH 6.5 and 25°C
0.77
gamma-linolenate
-
at pH 6.5 and 35°C
1.12
gamma-linolenic acid
at pH 6.5 and 25°C
0.58
linoleate
-
at pH 6.5 and 35°C
0.058
linoleic acid
at pH 6.5 and 25°C
0.3
myristoleic acid
at pH 6.5 and 25°C
0.46 - 0.54
oleate
0.078 - 0.56
oleic acid
0.76
palmitoleate
-
at pH 6.5 and 35°C
0.57
palmitoleic acid
at pH 6.5 and 25°C
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.13
alpha-linolenic acid
at pH 6.5 and 25°C
2.17
gamma-linolenate
-
at pH 6.5 and 35°C
2.67
gamma-linolenic acid
at pH 6.5 and 25°C
1.33
linoleate
-
at pH 6.5 and 35°C
0.13
linoleic acid
at pH 6.5 and 25°C
0.03
myristoleic acid
at pH 6.5 and 25°C
11.83 - 14.17
oleate
7.83
oleic acid
at pH 6.5 and 25°C
7.17
palmitoleate
-
at pH 6.5 and 35°C
3.83
palmitoleic acid
at pH 6.5 and 25°C
kcat/KM VALUE [1/mMs-1]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.42
alpha-linolenic acid
at pH 6.5 and 25°C
2.67
gamma-linolenate
-
at pH 6.5 and 35°C
4.5
gamma-linolenic acid
at pH 6.5 and 25°C
2.33
linoleate
-
at pH 6.5 and 35°C
0.33
linoleic acid
at pH 6.5 and 25°C
0.13
myristoleic acid
at pH 6.5 and 25°C
25.67 - 26.17
oleate
0.33
oleic acid
at pH 6.5 and 25°C
9.33
palmitoleate
-
at pH 6.5 and 35°C
22.67
palmitoleic acid
at pH 6.5 and 25°C
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
0.014
-
with myristoleic acid as substrate, at pH 6.5 and 35°C
0.033
-
with gamma-linolenic acid as substrate, at pH 6.5 and 35°C
0.14
at 30°C, in 20 mM Tris buffer, pH 8, 50 mM NaCl, with mixing speed of 1400 rpm
0.16
at 22°C, in 20 mM Tris buffer, pH 8, 50 mM NaCl, with mixing speed of 1400 rpm
0.17
at 30°C, in 20 mM Tris buffer, pH 8, 50 mM NaCl, with mixing speed of 1000 rpm
0.34
at 22°C, in 20 mM Tris buffer, pH 8, 50 mM NaCl, with mixing speed of 2200 rpm, in the presence of 2.5% isopropyl alcohol
0.39
at 22°C, in 20 mM Tris buffer, pH 8, 50 mM NaCl, with mixing speed of 2200 rpm
0.54
-
with alpha-linolenic acid as substrate, at pH 6.5 and 35°C
0.91
-
with linoleic acid as substrate, at pH 6.5 and 35°C
1.95
-
with palmitoleic acid as substrate, at pH 6.5 and 35°C
3.3
at pH 6.5 and 25°C
3.69
-
with oleate as substrate, at pH 6.5 and 35°C
20
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with alpha-linolenate as substrate, at pH 6.5 and 35°C
30
-
with myristoleate as substrate, at pH 6.5 and 35°C
110
-
with linoleate as substrate, at pH 6.5 and 35°C
160
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with gamma-linolenate as substrate, at pH 6.5 and 35°C
550
-
with palmitoleate as substrate, at pH 6.5 and 35°C
1580
-
with oleate as substrate, at pH 6.5 and 35°C
additional information
specific activity of the enzyme does not change significantly in the presence of 2.5% or 5% isopropyl alcohol, but it decreases at isopropyl alcohol concentrations higher than 10%
pH RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
5 - 7
pH 5: about 70% of maximal activity, pH 7: about about 75% of maximal activity
5.5 - 7.5
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
TEMPERATURE RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
15 - 35
40% activity at 15°C, 75% activity at 20°C, 100% activity at 25°C, 78% activity at 30°C, and 65% activity at 35°C
20 - 45
-
more than 60% activity between 20 and 45°C
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
68100
2 * 68100, calculated from amino acid sequence
69000
-
2 * 69000, SDS-PAGE
69200
-
2 * 69200, calculated from amino acid sequence
73487
1 * 73487, calculated from sequence
136000
138000
-
gel filtration
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
homodimer
monomer
1 * 73487, calculated from sequence
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
20 - 70
-
after 1 h incubation, the enzyme remains stable between 20 and 40°C (100% activity), at 45°C, 50°C, 55°C, 60°C, and 70°C it shows 80%, 70%, 45%, 20% and 0% residual activity, respectively
25 - 35
after 120 min, the enzyme activity is nearly unchanges at 25°C, but is reduced to only 25% at 35 °C
ORGANIC SOLVENT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
Methanol
-
100% activity in the presence of 1-4% (v/v) methanol
Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
by three steps of gel filtration. Recombinant oleate hydratase fused to an N-terminal His tag purified onh Ni-resin
His-Trap column chromatography
His-Trap column chromatography, and gel filtration
-
Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
expressed in Escherichia coli
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expressed in Escherichia coli BL21(DE3) cells
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expressed in Escherichia coli ER2566 cells
expressed in Escherichia coli ER2566, BL21(DE3), W3110, and MLK1067 cells
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into the pBAD/His A vector and expressed in Escherichia coli TOP10 cells
EXPRESSION
ORGANISM
UNIPROT
LITERATURE
expression is strongly upregulated in cells grown in the presence of oleic acid
ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
E50A
the mutant shows reduced activity and FAD binding compared to the wild type enzyme
S34A
the mutant shows reduced activity and FAD binding compared to the wild type enzyme
E50A
-
the mutant shows reduced activity and FAD binding compared to the wild type enzyme
-
S34A
-
the mutant shows reduced activity and FAD binding compared to the wild type enzyme
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Show AA Sequence (281 entries)
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