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Information on EC 3.1.8.1 - aryldialkylphosphatase and Organism(s) Saccharolobus solfataricus and UniProt Accession Q97VT7

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EC Tree
     3 Hydrolases
         3.1 Acting on ester bonds
             3.1.8 Phosphoric-triester hydrolases
                3.1.8.1 aryldialkylphosphatase
IUBMB Comments
Acts on organophosphorus compounds (such as paraoxon) including esters of phosphonic and phosphinic acids. Inhibited by chelating agents; requires divalent cations for activity. Previously regarded as identical with EC 3.1.1.2 arylesterase.
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Saccharolobus solfataricus
UNIPROT: Q97VT7
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Word Map
The taxonomic range for the selected organisms is: Saccharolobus solfataricus
The enzyme appears in selected viruses and cellular organisms
Synonyms
pon-1, serum paraoxonase, phosphotriesterase, organophosphorus hydrolase, dfpase, serum paraoxonase 1, pon 1, methyl parathion hydrolase, organophosphate hydrolase, hupon1, more
SYNONYM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
aryldialkylphosphatase
-
lactonase/phosphotriesterase
-
organophosphate hydrolase
-
phosphotriesterase-like lactonase
-
SsoPox
A-esterase
-
-
-
-
aryltriphosphatase
-
-
-
-
esterase B1
-
-
-
-
esterase E4
-
-
-
-
esterase, organophosphate
-
-
-
-
esterase, paraoxon
-
-
-
-
esterase, pirimiphos-methyloxon
-
-
-
-
HuPON1
-
-
-
-
OPA anhydrase
-
-
-
-
OPH
-
-
-
-
organophosphate hydrolase
-
-
-
-
organophosphorus acid anhydrase
-
-
-
-
organophosphorus hydrolase
-
-
-
-
paraoxon hydrolase
-
-
-
-
paraoxonase
-
-
-
-
phosphotriesterase
pirimiphos-methyloxon esterase
-
-
-
-
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
hydrolysis of phosphoric triester
-
-
-
-
SYSTEMATIC NAME
IUBMB Comments
aryltriphosphate dialkylphosphohydrolase
Acts on organophosphorus compounds (such as paraoxon) including esters of phosphonic and phosphinic acids. Inhibited by chelating agents; requires divalent cations for activity. Previously regarded as identical with EC 3.1.1.2 arylesterase.
CAS REGISTRY NUMBER
COMMENTARY hide
117698-12-1
-
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
(+)-cyclosarin + H2O
methyl-phosphonic acid monofluoride + cyclohexanol
show the reaction diagram
wild type enzyme and mutant enzyme W263F hydrolyze the (+)-enantiomer approximately 3 and 4.5 times faster than the (-)-enantiomer
-
-
?
chlorpyrifos + H2O
O,O-diethylphosphorothioate + 3,5,6-trichloropyridin-2-ol
show the reaction diagram
low activity, cf. EC 3.1.8.2
-
-
?
coumaphos + H2O
O,O-diethylphosphorothioate + 3-chloro-7-hydroxy-4-methyl-2H-chromen-2-one
show the reaction diagram
cf. EC 3.1.8.2
-
-
?
cyclohexylmethylphosphonofluoridate + H2O
?
show the reaction diagram
i.e. cyclosarin
-
-
?
diethyl-paraoxon + H2O
diethyl phosphate + 4-nitrophenol
show the reaction diagram
diethyl-parathion + H2O
diethyl thiophosphate + 4-nitrophenol
show the reaction diagram
dimethyl-paraoxon + H2O
dimethyl phosphate + 4-nitrophenol
show the reaction diagram
dimethyl-parathion + H2O
dimethyl thiophosphate + 4-nitrophenol
show the reaction diagram
ethyl dimethylphosphoramidocyanidate + H2O
?
show the reaction diagram
i.e. tabun
-
-
?
fenitrothion + H2O
O,O-diethylphosphorothioate + 3-methyl-4-nitrophenol
show the reaction diagram
-
-
-
?
fensulfothion + H2O
O,O-diethylphosphorothioate + 4-(methylsulfinyl)phenol
show the reaction diagram
some enzyme mutants are also capable of degrading fensulfothion, which is reported to be an inhibitor for the wild-type enzyme, as well as others that are not substrates of the starting template or previously reported W263 mutants
-
-
?
malathion + H2O
?
show the reaction diagram
-
-
-
?
malathion + H2O
O,O-diethylphosphorothioate + diethyl 2-mercaptosuccinate
show the reaction diagram
low activity
-
-
?
methyl paraoxon + H2O
4-nitrophenol + dimethyl phosphate
show the reaction diagram
-
-
-
?
methyl parathion + H2O
4-nitrophenol + dimethyl thiophosphate
show the reaction diagram
-
-
-
?
O-ethyl-S-(2-diisopropylaminoethyl)methylphosphonothiolate + H2O
?
show the reaction diagram
i.e. VX
-
-
?
O-isopropyl methylphosphonofluoridate + H2O
?
show the reaction diagram
i.e. sarin
-
-
?
paraoxon + H2O
4-nitrophenol + diethyl phosphate
show the reaction diagram
paraoxon + H2O
diethyl phosphate + 4-nitrophenol
show the reaction diagram
high activity with enzyme mutant C258L/I261F/W263A
-
-
?
ethyl paraoxon + H2O
?
show the reaction diagram
-
-
-
-
?
methyl paraoxon + H2O
4-nitrophenol + dimethyl phosphate
show the reaction diagram
-
-
-
?
methyl paraoxon + H2O
4-nitrophenol + dimethylphosphate
show the reaction diagram
-
the enzyme from Sulfolobus solfataricus has a low paraoxonase activity
-
-
?
paraoxon + H2O
4-nitrophenol + diethyl phosphate
show the reaction diagram
-
the enzyme from Sulfolobus solfataricus has a low paraoxonase activity
-
-
?
paraoxon + H2O
diethylphosphate + 4-nitrophenol
show the reaction diagram
paraoxon is the best substrate for the purified arylesterase
-
-
?
additional information
?
-
NATURAL SUBSTRATE
NATURAL PRODUCT
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
diethyl-paraoxon + H2O
diethyl phosphate + 4-nitrophenol
show the reaction diagram
-
-
-
?
diethyl-parathion + H2O
diethyl thiophosphate + 4-nitrophenol
show the reaction diagram
-
-
-
?
dimethyl-paraoxon + H2O
dimethyl phosphate + 4-nitrophenol
show the reaction diagram
-
-
-
?
dimethyl-parathion + H2O
dimethyl thiophosphate + 4-nitrophenol
show the reaction diagram
-
-
-
?
additional information
?
-
the phosphotriesterase-like lactonase enzyme is bifunctional showing lactonase (EC 3.1.1.81) and phosphotriesterase (EC 3.1.8.1 and 3.1.8.2) activities
-
-
?
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
Fe2+
bound at the alpha-site
Co2+
-
the enzyme depends on divalent metal ions with Co2+, Mg2+, and Ni2+ being the most effective
Mg2+
-
the enzyme depends on divalent metal ions with Co2+, Mg2+, and Ni2+ being the most effective
Ni2+
-
the enzyme depends on divalent metal ions with Co2+, Mg2+, and Ni2+ being the most effective
additional information
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
fensulfothion
2-propanol
55% residual activity in the presence of 90% (v/v) 2-propanol, after 60 min at 70°C
acetonitrile
43% residual activity in the presence of 90% (v/v) acetonitrile, after 60 min at 70°C
diethyl dicarbonate
6% residual activity in the presence of 5 mM diethyl dicarbonate, after 30 min at 30°C
diisopropylfluorophosphate
complete inactivation at 5 mM diisopropylfluorophosphate, after 30 min at 30°C
dimethyl sulfoxide
71% residual activity in the presence of 90% (v/v) dimethyl sulfoxide, after 60 min at 70°C
eserine
92% residual activity in the presence of 5 mM eserine, after 30 min at 30°C
ethanol
63% residual activity in the presence of 90% (v/v) ethanol, after 60 min at 70°C
Hg2+
complete inactivation at 5 mM Hg2+, after 30 min at 30°C
methanol
48% residual activity in the presence of 90% (v/v) methanol, after 60 min at 70°C
NaCl
76% residual activity in the presence of 2 M NaCl, after 60 min at 70°C
o-phenanthroline
-
inactivation
p-chloromercuribenzoate
23% residual activity in the presence of 5 mM p-chloromercuribenzoate, after 30 min at 30°C
petroleum ether
-
petroleum ether slightly reduces the activity down to 78%
-
Phenylglyoxal
97% residual activity in the presence of 5 mM phenylglyoxal, after 30 min at 30°C
phenylmethylsulfonyl fluoride
23% residual activity in the presence of 5 mM phenylmethylsulfonyl fluoride, after 30 min at 30°C
SDS
about half of the activity is retained at 1% SDS after incubation for 60 min at 70°C, the addition of 5% SDS completely eliminates the enzyme activity at 70°C
Toluene
-
toluene slightly reduces the activity down to 74%
Triton X-100
11% residual activity in the presence of 5% (v/v) Tween 80, after 60 min at 70°C
Tween 80
21% residual activity in the presence of 5% (v/v) Tween 80, after 60 min at 70°C
Urea
88% residual activity in the presence of 8 M urea, after 60 min at 70°C
xylene
-
xylene decreases the enzyme activity by more than 75%
ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
commercial soap LM1
wild-type SsoPox, at 70°C, 13fold enhancement of hydrolytic activity in the presence of 0.05% LM1, and for mutant C258L/I261F/W263A at 65°C 1.3fold enhancement
-
sodium deoxycholate
0.05%, approximately 33times catalytic efficiency enhancement compared to that without detergent
3-[(3-cholamidopropyl)-dimethylammonio]-1-propanesulfonate
the enzyme is activated by 87% by incubation with 1% (w/v) 3-[(3-cholamidopropyl)-dimethylammonio]-1-propanesulfonate, at 30°C
dimethyl sulfoxide
21% increase of activity at 50% (v/v) dimethyl sulfoxide, at 30°C
ethanol
15% increase of activity at 50% (v/v) ethanol, at 30°C
Lubrol
the enzyme is activated by 14% by incubation with 1% Lubrol, at 30°C
-
Methylcyclohexane
-
about 250% activity at 100% (v/v)
paraoxon
115 increase of activita in the presence of 0.5 mM paraoxon, after 30 min at 30°C
pyridoxal 5'-phosphate
110% activity in the presence of 5 mM pyridoxal 5'-phosphate, after 30 min at 30°C
Tween 20
the enzyme is activated by 67% by incubation for 60 min with 1% (v/v) Tween 20 at 30°C
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.107 - 1.586
diethyl-paraoxon
0.16
malathion
25°C, pH 9.0
2.14
methyl paraoxon
25°C, pH 9.0
0.12
methyl parathion
25°C, pH 9.0
0.064 - 24.25
paraoxon
0.205 - 0.246
methyl paraoxon
0.005 - 0.06
paraoxon
additional information
additional information
-
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
6.91 - 71.05
diethyl-paraoxon
0.00089
malathion
25°C, pH 9.0
2.71
methyl paraoxon
25°C, pH 9.0
0.0011
methyl parathion
25°C, pH 9.0
0.24 - 12.59
paraoxon
1.3 - 342
methyl paraoxon
0.24 - 597
paraoxon
kcat/KM VALUE [1/mMs-1]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
18.19 - 97.5
diethyl-paraoxon
0.0056
malathion
25°C, pH 9.0
1.27
methyl paraoxon
25°C, pH 9.0
0.0091
methyl parathion
25°C, pH 9.0
0.52 - 9.01
paraoxon
1.2 - 25
ethyl paraoxon
Ki VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
1.23
fensulfothion
70°C, pH 9.0
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
0.42
purified recombinant enzyme mutant W263F, pH 8.5, 65°C, substrate diethyl-paraoxon
12.8
purified recombinant enzyme mutant W263F, pH 8.5, 65°C, substrate diethyl-paraoxon
122.6
substrate paraoxon, enzyme mutant C258L/I261F/W263A, pH and temperature not specified in the publication
123.6
purified recombinant enzyme mutant C258L/I261F/W263A, pH 8.5, 65°C, substrate diethyl-paraoxon
0.00042
-
purified recombinant enzyme, substrate paraoxon
0.0022
-
purified recombinant enzyme, substrate methyl paraoxon
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
8 - 8.5
8.3
lactonase activity assay at
8.5
enzyme mutant C258L/I261F/W263A
7 - 9
-
broad optimum
pH RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
5 - 9
activity range, enzyme mutant SsoPox 3 M
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
65
enzyme mutant C258L/I261F/W263A
65 - 75
enzyme mutant SsoPox 3 M
85
enzyme mutant W263F
90
above, wild-type enzyme
TEMPERATURE RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
10 - 100
activity range, enzyme mutant SsoPox 3 M
ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
evolution
enzyme SsoPox belongs to the phosphotriesterase-like lactonase (PLL) family of enzymes. SsoPox shares only about 30% sequence identity with phosphotriesterases (PTEs) but all amino acids coordinating the binuclear metal-centre are conserved. The coexistence of lactonase and phosphotriesterase activities has been already reported for many members of PLL family
physiological function
SsoPox is a thermostable phosphotriesterase-like lactonase (PLL) that hydrolyses lactones (primary activity) and, at a lower rate, neurotoxic organophosphorus compounds (promiscuous activity)
metabolism
-
the enzyme exhibits promiscuous phosphotriesterase activity for the degradation of organophosphorous chemicals including insecticides and chemical warfare agents
additional information
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
34000
x * 34000, SDS-PAGE
34000
-
x * 34000, SDS-PAGE, x * 35565, sequence calculation, the enzyme is in a monomer-dimer equilibrium
35175
1 * 35175, MALDI-TOF mass spectrometry
35180
MALDI-TOF mass spectrometry
35565
-
x * 34000, SDS-PAGE, x * 35565, sequence calculation, the enzyme is in a monomer-dimer equilibrium
48000 - 50000
-
recombinant enzyme, gel filtration
SUBUNIT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
?
x * 34000, SDS-PAGE
dimer
monomer
1 * 35175, MALDI-TOF mass spectrometry
monomer or dimer
-
x * 34000, SDS-PAGE, x * 35565, sequence calculation, the enzyme is in a monomer-dimer equilibrium
additional information
CRYSTALLIZATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
crystal structure analysis of SsoPox in the apo form and in complex with a quorum-sensing lactone mimic at 2.6 and 2.0A resolution, respectively, overview
hanging drop vapour diffusion method
purified recombinant wild-type enzyme and mutant asA6 in open conformation, asA6 in closed conformation, asB5, asC6, asD6, and asA1, hanging drop vapor diffusion method, mixing of 500 nl protein solution, containing protein in 50 mM HEPES, pH 8.0, 150 mM NaCl, and 0.2 mM CoCl2, and 500 nl reservoir solution, containing 20-30% w/v PEG 8000, and 50 mM Tris-HCl, pH 8.0, a few days, 4°C, X-ray diffraction structure determination and analysis at 1.4-2.95 A resolution
purified recombinant enzyme, hanging-drop vapour-diffusion technique, 5.8 mg/ml protein in 20 mM HEPES, pH 8.5, 0.2 mM CoCl2 and 0.2 M NaCl, 0.001-0.002 ml of protein and reservoir solutions are mixed and equilibrated against 0.8 ml reservoir solution containing 15-18% w/v PEG 8000 and 50 mM Tris-HCl buffer pH 8.0, 1 week at 4°C, X-ray diffraction structure determination and analysis at 2.54 A resolution
-
PROTEIN VARIANTS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
C258A
site-directed mutagenesis, active site mutant, the mutant shows altered substrate specificity compared to the wild-type enzyme
C258L
site-directed mutagenesis, active site mutant, the mutant shows altered substrate specificity compared to the wild-type enzyme
C258L/I261F/W263A
D141T
site-directed mutagenesis, the mutant enzyme shows increased phosphotriesterase activity compared to the wild-type
F46L/C258A/W263M/I280T
site-directed mutagenesis, mutant alphasA6, the mutant shows altered substrate specificity compared to the wild-type enzyme
I767T
site-directed mutagenesis, active site mutant, the mutant shows altered substrate specificity compared to the wild-type enzyme
L130P
site-directed mutagenesis, active site mutant, the mutant shows altered substrate specificity compared to the wild-type enzyme
L228M
site-directed mutagenesis
L72I/Y99F/I122L/L228M/F229S/W263L
site-directed mutagenesis, mutant alphasC6, the mutant shows altered substrate specificity compared to the wild-type enzyme
V27A/D141T
site-directed mutagenesis, the mutant enzyme shows increased phosphotriesterase activity compared to the wild-type
V27A/I76T/Y97W/Y99F/L130P/L226V
site-directed mutagenesis, mutant alphasB5, the mutant shows altered substrate specificity compared to the wild-type enzyme
V27A/I76T/Y97W/Y99F/L130P/L226V/W263I
site-directed mutagenesis, mutant alphasB5 W263I, the mutant shows altered substrate specificity compared to the wild-type enzyme
V27A/I76T/Y97W/Y99F/L130P/L226V/W263L
site-directed mutagenesis, mutant alphasB5 W263L, the mutant shows altered substrate specificity compared to the wild-type enzyme
V27A/I76T/Y97W/Y99F/L130P/L226V/W263M
site-directed mutagenesis, mutant alphasB5 W263M, the mutant shows altered substrate specificity compared to the wild-type enzyme
V27A/Y97W/L228M/W263M
site-directed mutagenesis, the mutant alphasD6 enzyme demonstrates a large increase in catalytic efficiencies compared to the wild-type enzyme, with increases of 2210fold, 163fold, 58fold, and 16fold against methyl-parathion, malathion, ethyl-paraoxon, and methyl-paraoxon, respectively
V27A/Y97W/Y99F
site-directed mutagenesis, the mutant enzyme shows increased phosphotriesterase activity compared to the wild-type
W263F
W263I
site-directed mutagenesis, active site mutant, the mutant shows altered substrate specificity compared to the wild-type enzyme
W263L
site-directed mutagenesis, active site mutant, the mutant shows altered substrate specificity compared to the wild-type enzyme
W263N
site-directed mutagenesis, active site mutant, the mutant shows altered substrate specificity compared to the wild-type enzyme
W263Q
kcat/Km for paraoxon is 3.5fold lower than wild-type value
Y97W/I261F
kcat/Km for paraoxon is 1.4fold higher than wild-type value
Y97W/I98F/I261F
kcat/Km for paraoxon is 1.2fold higher than wild-type value
Y97W/W263F
kcat/Km for paraoxon is 1.6fold higher than wild-type value
Y97W/W263Q
kcat/Km for paraoxon is 1.7fold lower than wild-type value
Y99F
site-directed mutagenesis, active site mutant, the mutant shows altered substrate specificity compared to the wild-type enzyme
C105S
mutant shows 24.0% activity compared to the wild type enzyme
C107S
mutant shows 30.8% activity compared to the wild type enzyme
C129S
mutant shows 3.6% activity compared to the wild type enzyme
D251N
mutant shows 0.6% activity compared to the wild type enzyme
H281N
mutant shows 19.4% activity compared to the wild type enzyme
S156A
mutant shows 0.012% activity compared to the wild type enzyme
W263I
-
the mutant has increased lactonase and phosphotriesterase activities compared to the wild type
additional information
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
106
estimated denaturation temperature
65
purified recombinant enzyme mutant C258L/I261F/W263A, pH 8.5, 6.6 h, completely stable
75
purified recombinant enzyme mutant C258L/I261F/W263A, pH 8.5, loss of 10% activity after 6.6 h
85
purified recombinant enzyme mutant C258L/I261F/W263A, pH 8.5, loss of 30% activity after 6.6 h, t1/2 is 5.8 h
90
purified recombinant enzyme mutant C258L/I261F/W263A, pH 8.5, loss of 40% activity after 5.0 h, t1/2 is 2.2 h
95
purified recombinant enzyme mutant C258L/I261F/W263A, pH 8.5, loss of 40% activity after 1.5 h, t1/2 is 25 min
100
-
purified recombinant enzyme, half-life is 90 min
50 - 94
the enzyme retains 52% of its activity after 50 h of incubation at 90°C, the enzyme is rapidly inactivated above 94°C, most of the enzyme activity is maintained for 5 days at 50°C, the enzyme activity gradually decreases with time at higher temperatures, approximately 70% of the enzyme activity remains after 5 days at 70°C, and 52% of it is still preserved after 50 h at 90°C, however, at 90°C after 5 days, the enzyme is completely inactivated
70 - 85
-
purified recombinant enzyme, 4 h, completely stable
95
-
purified recombinant enzyme, half-life is 4 h
additional information
GENERAL STABILITY
ORGANISM
UNIPROT
LITERATURE
stable for up to 40 h in 0.2% SDS at 25°C
with the liquid enzyme, the remaining activity decreases down to 40% at the highest dose of beta-radiation (100 kGy)
-
ORGANIC SOLVENT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
guanidine-HCl
the conformational stability of SsoPox against the denaturing action of guanidine-HCl has been investigated at 25°C, pH 8.0, 20 mM Tris–HCl buffer, by performing CD and fluorescence measurements. The transition curves obtained by recording the molar ellipticity at 222 nm (detecting the secondary structure stability) present two inflection points, at 2.6 M guanidine-HCl and about 4.8 M guanidine-HCl, respectively, with a plateau at about 3 M guanidine-HCl. SsoPox is markedly more resistant to the denaturing action of guanidine-HCl with respect to the mesophilic counterpart. Overall guanidine-HCl-induced denaturation of SsoPox is not a reversible process in all the investigated experimental conditions: upon suitable dilution of fully denatured samples, there is not a complete recovery of the far-UV CD spectrum or fluorescence emission spectrum of the native enzyme
Triton X-100
after incubation for 60 min with Triton X-100 the enzyme activity is remarkably decreased with increasing temperature and concentration
urea
the purified enzyme shows high stability against urea
additional information
-
the enzyme activity is not affected by acetone, acetonitrile, butanone, butyl acetate, chloroform, dichloromethane, diethyl ether, ethanol, ethyl acetate, isopropanol, methanol, and methoxypropanol
OXIDATION STABILITY
ORGANISM
UNIPROT
LITERATURE
original color intensity after 23 days of storage in water or 20 mM sodium HEPES buffer (pH 8.0)
724003
storage under dry conditions is less favorable for the stability of the enzyme, as the amount of 4-nitrophenolate produced from 1 mM paraoxon is reduced to 60 to 75% of the initial amount after 23 days
724003
when the enzyme is immobilized on nanoalumina membranes 25% of the activity is retained upon immobilization. When tested with 1 mM paraoxon, the membranes produced at least 90% of their
724003
PURIFICATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
recombinant enzyme mutant SsoPox 3 M from Escherichia coli strain BL21(DE3) 13.10fold by heat treatment at 66-72°C, by response surface methodology, and ultrafiltration using an automatic tangential flow filtration system, followed by anion exchange chromatography. The ultrafiltration step is preferred compared to a gel filtration step
recombinant enzyme from Escherichia coli
-
recombinant enzyme from Escherichia coli by ultracentrifugation, anion exchange chromatography, and hydrophobic interaction chromatography
-
recombinant enzyme is purified by DEAE-cellulose column chromatography, butyl-Sepharose column chromatography, Q-Sepharose column chromatography, and hydroxyapatite column chromatography
CLONED (Commentary)
ORGANISM
UNIPROT
LITERATURE
expression in Escherichia coli
gene ssopox and gene ssopox-pte, sequence comparisons, recombinant expression of wild-type, point mutation, and chimeric mutant enzymes in Escherichia coli strains TOP10 and BL21(DE3)
overexpressed in Pseudomonas putida KT2440
plasmid library screening, recombinant wild-type and mutant enzymes from Escherichia coli BL21(DE3)-pGro7/GroEL (TaKaRa) chaperone expressing strain by ammonium sulfate fractionation, desalting gel filtration, and gel filtration
recombinant expression of mutant enzyme SsoPox 3 M in Escherichia coli strain BL21(DE3), induction by galactose, method optimization
DNA and amino acid sequence determination and analysis, overexpression in Escherichia coli strain HB101 and BL21(DE3)
-
expressed in Escherichia coli BL21(DE3) cells
expression in Escherichia coli
-
RENATURED/Commentary
ORGANISM
UNIPROT
LITERATURE
the conformational stability of SsoPox against the denaturing action of guanidine-HCl has been investigated at 25°C, pH 8.0, 20 mM Tris–HCl buffer, by performing CD and fluorescence measurements. The transition curves obtained by recording the molar ellipticity at 222 nm (detecting the secondary structure stability) present two inflection points, at 2.6 M guanidine-HCl and about 4.8 M guanidine-HCl, respectively, with a plateau at about 3 M guanidine-HCl. SsoPox is markedly more resistant to the denaturing action of guanidine-HCl with respect to the mesophilic counterpart. Overall guanidine-HCl-induced denaturation of SsoPox is not a reversible process in all the investigated experimental conditions: upon suitable dilution of fully denatured samples, there is not a complete recovery of the far-UV CD spectrum or fluorescence emission spectrum of the native enzyme
APPLICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
environmental protection
thermostable phosphotriesterase-like lactonases (PLLs) are able to degrade organophosphates and can be potentially employed as bioremediation tools and bioscavengers. The enzyme is employable in cleaning organophosphates from different surfaces like glass, tissues, and fruits, also in presence of surfactants and even when dissolved in tap water
synthesis
efficient production of enzyme in Escherichia coli, by designing high cell density cultivations and a membrane-based downstream process. In fed batches, enzyme production is increased by 69fold up to 4660 U/l, using galactose as inducer. the process is scalable from 2.5 up to 150 l. An enzyme recovery of 77% with a purity grade of 80% can be reached
environmental protection
-
enzymes showing phosphotriesterase activity are capable of hydrolysing the organophosphate phosphotriesters, a class of synthetic compounds employed worldwide both as insecticides and chemical warfare agents. Thermostable enzymes able to hydrolyse organophosphate phosphotriesters are considered good candidates for the set-up of efficient detoxification tools
REF.
AUTHORS
TITLE
JOURNAL
VOL.
PAGES
YEAR
ORGANISM (UNIPROT)
PUBMED ID
SOURCE
Elias, M.; Dupuy, J.; Merone, L.; Lecomte, C.; Rossi, M.; Masson, P.; Manco, G.; Chabriere, E.
Crystallization and preliminary X-ray diffraction analysis of the hyperthermophilic Sulfolobus solfataricus phosphotriesterase
Acta Crystallogr. Sect. F
63
553-555
2007
Saccharolobus solfataricus, Saccharolobus solfataricus MT-4 / DSM 5833
Manually annotated by BRENDA team
Merone, L.; Mandrich, L.; Rossi, M.; Manco, G.
A thermostable phosphotriesterase from the archaeon Sulfolobus solfataricus: cloning, overexpression and properties
Extremophiles
9
297-305
2005
Saccharolobus solfataricus, Saccharolobus solfataricus MT-4 / DSM 5833
Manually annotated by BRENDA team
Park, Y.J.; Yoon, S.J.; Lee, H.B.
A novel thermostable arylesterase from the archaeon Sulfolobus solfataricus P1: purification, characterization, and expression
J. Bacteriol.
190
8086-8095
2008
Saccharolobus solfataricus (B5BLW5), Saccharolobus solfataricus P1 (B5BLW5)
Manually annotated by BRENDA team
Elias, M.; Dupuy, J.; Merone, L.; Mandrich, L.; Porzio, E.; Moniot, S.; Rochu, D.; Lecomte, C.; Rossi, M.; Masson, P.; Manco, G.; Chabriere, E.
Structural basis for natural lactonase and promiscuous phosphotriesterase activities
J. Mol. Biol.
379
1017-1028
2008
Saccharolobus solfataricus (Q97VT7), Saccharolobus solfataricus
Manually annotated by BRENDA team
Del Vecchio, P.; Elias, M.; Merone, L.; Graziano, G.; Dupuy, J.; Mandrich, L.; Carullo, P.; Fournier, B.; Rochu, D.; Rossi, M.; Masson, P.; Chabriere, E.; Manco, G.
Structural determinants of the high thermal stability of SsoPox from the hyperthermophilic archaeon Sulfolobus solfataricus
Extremophiles
13
461-470
2009
Saccharolobus solfataricus (Q97VT7), Saccharolobus solfataricus, Saccharolobus solfataricus P2 (Q97VT7)
Manually annotated by BRENDA team
Ng, F.S.; Wright, D.M.; Seah, S.Y.
Characterization of a phosphotriesterase-like lactonase from Sulfolobus solfataricus and its immobilization for disruption of quorum sensing
Appl. Environ. Microbiol.
77
1181-1186
2010
Saccharolobus solfataricus (Q97VT7), Saccharolobus solfataricus, Saccharolobus solfataricus P2 (Q97VT7)
Manually annotated by BRENDA team
Merone, L.; Mandrich, L.; Porzio, E.; Rossi, M.; Mller, S.; Reiter, G.; Worek, F.; Manco, G.
Improving the promiscuous nerve agent hydrolase activity of a thermostable archaeal lactonase
Bioresour. Technol.
101
9204-9212
2010
Saccharolobus solfataricus (Q97VT7), Saccharolobus solfataricus, Saccharolobus solfataricus P2 (Q97VT7)
Manually annotated by BRENDA team
Mandrich, L.; Merone, L.; Manco, G.
Hyperthermophilic phosphotriesterases/lactonases for the environment and human health
Environ. Technol.
31
1115-1127
2010
Sulfolobus acidocaldarius, Saccharolobus solfataricus
Manually annotated by BRENDA team
Hiblot, J.; Gotthard, G.; Chabriere, E.; Elias, M.
Characterisation of the organophosphate hydrolase catalytic activity of SsoPox
Sci. Rep.
2
779
2012
Saccharolobus solfataricus (Q97VT7), Saccharolobus solfataricus, Saccharolobus solfataricus P2 (Q97VT7)
Manually annotated by BRENDA team
Restaino, O.F.; Borzacchiello, M.G.; Scognamiglio, I.; Porzio, E.; Manco, G., Fedele, L., Donatiello, C., De Rosa, M.; Schiraldi, C.
Boosted large-scale production and purification of a thermostable archaeal phosphotriesterase-like lactonase for organophosphate decontamination
J. Ind. Microbiol. Biotechnol.
44
363-375
2017
Saccharolobus solfataricus (Q97VT7), Saccharolobus solfataricus, Saccharolobus solfataricus DSM 1617 (Q97VT7)
Manually annotated by BRENDA team
Del Giudice, I.; Coppolecchia, R.; Merone, L.; Porzio, E.; Carusone, T.M.; Mandrich, L.; Worek, F.; Manco, G.
An efficient thermostable organophosphate hydrolase and its application in pesticide decontamination
Biotechnol. Bioeng.
113
724-734
2016
Saccharolobus solfataricus (Q97VT7)
Manually annotated by BRENDA team
Restaino, O.F.; Borzacchiello, M.G.; Scognamiglio, I.; Fedele, L.; Alfano, A.; Porzio, E.; Manco, G.; De Rosa, M.; Schiraldi, C.
High yield production and purification of two recombinant thermostable phosphotriesterase-like lactonases from Sulfolobus acidocaldarius and Sulfolobus solfataricus useful as bioremediation tools and bioscavengers
BMC Biotechnol.
18
18
2018
Sulfolobus acidocaldarius, Saccharolobus solfataricus (Q97VT7)
Manually annotated by BRENDA team
Remy, B.; Plener, L.; Poirier, L.; Elias, M.; Daude, D.; Chabriere, E.
Harnessing hyperthermostable lactonase from Sulfolobus solfataricus for biotechnological applications
Sci. Rep.
6
37780
2016
Saccharolobus solfataricus
Manually annotated by BRENDA team
Jacquet, P.; Hiblot, J.; Daude, D.; Bergonzi, C.; Gotthard, G.; Armstrong, N.; Chabriere, E.; Elias, M.
Rational engineering of a native hyperthermostable lactonase into a broad spectrum phosphotriesterase
Sci. Rep.
7
16745
2017
Saccharolobus solfataricus (Q97VT7), Saccharolobus solfataricus
Manually annotated by BRENDA team