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Information on EC 3.1.8.1 - aryldialkylphosphatase and Organism(s) Brevundimonas diminuta and UniProt Accession P0A434
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3.1.8.1 aryldialkylphosphataseIUBMB 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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Word Map
- 3.1.8.1
- arylesterase
- paraoxonase-1
- cholesterol
- nerve
- atherosclerosis
- cardiovascular
-
low-density
-
insecticide
- triglyceride
- coronary
-
apolipoproteins
- acetylcholinesterase
- malondialdehyde
- diminuta
-
poison
- soman
- p-nitrophenol
-
warfare
- sarin
-
bioremediation
- chlorpyrifos
- vx
-
antiatherogenic
-
decontamination
-
bioscavenger
- atherogenesis
- cholinesterase
- phenylacetate
- medicine
-
binuclear
-
phosphotriesters
-
atherogenic
- flavobacterium
- carboxylesterase
-
hdl-cholesterol
- apoa-i
- butyrylcholinesterase
- fluorophosphate
-
surface-expressed
- diazoxon
- tabun
-
thiolactone
-
g-type
-
salt-stimulated
-
triesters
- malathion
- ochrobactrum
- sphingobium
- radiobacter
- cyclosarin
-
loligo
- synthesis
- environmental protection
- drug development
- degradation
- biotechnology
- diagnostics
- agriculture
- analysis
- nutrition
The taxonomic range for the selected organisms is: Brevundimonas diminuta
The enzyme appears in selected viruses and cellular organisms
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
topSYNONYM 
ORGANISM
UNIPROT
COMMENTARY 
LITERATURE
phosphotriesterase
-
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
-
-
-
-
PTE
OPH
-
-
-
-
organophosphate hydrolase
-
-
-
-
organophosphorus hydrolase
-
-
-
-
phosphotriesterase
-
-
-
-
PTE
-
-
-
-
REACTION
REACTION DIAGRAM
COMMENTARY
ORGANISM
UNIPROT
LITERATURE
an aryl dialkyl phosphate + H2O = dialkyl phosphate + an aryl alcohol
catalytic mechanism and active site structure, substrate docking mechanism
an aryl dialkyl phosphate + H2O = dialkyl phosphate + an aryl alcohol
mechanism of hydrolysis of organophosphates by the bacterial phosphotriesterase, reaction modelling, kinetic model
an aryl dialkyl phosphate + H2O = dialkyl phosphate + an aryl alcohol
mechanism
-
an aryl dialkyl phosphate + H2O = dialkyl phosphate + an aryl alcohol
kinetics and mechanism
-
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
hydrolysis of phosphoric triester
-
-
-
-
PATHWAY SOURCE
PATHWAYS
-
-, -, -, -
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
117698-12-1
-
SUBSTRATE
PRODUCT
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate)
(Substrate)
LITERATURE
(Substrate)
(Substrate)
COMMENTARY
(Product)
(Product)
LITERATURE
(Product)
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
r=reversible
ir=irreversible
?=not specified
(RS)-propan-2-yl methylphosphonofluoridate + H2O
isopropyl phosphate methylphosphonate + fluoride
i.e. sarin
-
-
?
(S)-[2-(diethylamino)ethyl] O,O-diethyl phosphorothioate + H2O
diethyl phosphate + 2-(diethylamino)ethane-1-thiol
-
-
-
?
(S)-[2-(diethylamino)ethyl] O,O-dimethyl phosphorothioate + H2O
dimethyl phosphate + 2-(diethylamino)ethane-1-thiol
-
-
-
?
(S)-[2-(diethylamino)ethyl] O-(2-methylpropyl) methylphosphonothioate + H2O
2-methylpropyl methylphosphonate + 2-(diethylamino)ethane-1-thiol
-
-
-
?
(S)-[2-[di(propan-2-yl)amino]ethyl] O,O-diethyl phosphorothioate + H2O
diethyl phosphate + 2-[di(propan-2-yl)amino]ethane-1-thiol
-
-
-
?
(S)-[2-[di(propan-2-yl)amino]ethyl] O,O-dimethyl phosphorothioate + H2O
dimethyl phosphate + 2-[di(propan-2-yl)amino]ethane-1-thiol
-
-
-
?
(S)-[2-[di(propan-2-yl)amino]ethyl] O-ethyl methylphosphonothioate + H2O
ethyl methylphosphonate + [di(propan-2-yl)amino]ethane-1-thiol
-
-
-
?
2-methoxycarbonyl-1-methylvinyl dimethyl phosphate + H2O
?
i.e. mevinphos
-
-
?
3-chloro-4-methyl-2-oxo-2H-chromen-7-yl ethyl methylphosphonate + H2O
3-chloro-7-hydroxy-4-methyl-2H-chromen-2-one + ethyl methylphosphonate
-
-
-
?
3-[fluoro(methyl)phosphoryl]oxy-2,2-dimethylbutane + H2O
Pinacolyl methylphosphonate + fluoride
i.e. soman
-
-
?
4-acetylphenyl (R)-2-methylpropyl methylphosphonate + H2O
(R)-2-methylpropyl methylphosphonate + 4-acetylphenol
-
-
-
?
4-acetylphenyl (S)-2-methylpropyl methylphosphonate + H2O
(S)-2-methylpropyl methylphosphonate + 4-acetylphenol
-
-
-
?
cyclohexylmethylphosphonofluoridate + H2O
?
i.e. cyclosarin, activity of 3.1.8.2
-
-
?
cyclohexylmethylphosphonofluoridate + H2O
cyclohexyl methylphosphonate + fluoride
i.e. cyclosarin
-
-
?
demeton-S + H2O
2-ethylsulfanyl-ethanethiol + diethyl phosphate
-
-
-
?
diethyl (3,5,6-trichloropyridin-2-yl) phosphate + H2O
?
i.e. chlorpyrifos-oxon
-
-
?
diethyl 2-(dimethoxyphosphorylsulfanyl)butanedioate + H2O
?
i.e. malaoxon
-
-
?
diethyl 4-chlorophenyl phosphate + H2O
4-chlorophenol + diethyl phosphate
-
-
-
?
diethyl 4-chlorophenyl thiophosphate + H2O
4-chlorophenol + diethyl thiophosphate
-
-
-
?
diethyl 4-nitrophenyl phosphate + H2O
4-nitrophenol + diethyl phosphate
-
-
-
?
diethyl-paraoxon + H2O
diethyl phosphate + 4-nitrophenol
-
-
-
?
dimethyl-parathion + H2O
dimethyl thiophosphate + 4-nitrophenol
-
-
-
?
N,N-diethyl-2-(methyl-(2-methylpropoxy)phosphoryl)sulfanylethanamine + H2O
?
i.e. VR
-
-
?
O,O-diethyl-S-[2-diethylaminoethyl]thiophosphate + H2O
?
i.e. amiton or VG
-
-
?
O,O-diethyl-S-[2-diisopropylaminoethyl]thiophosphate + H2O
?
i.e. diisopropyl-amiton
-
-
?
O,O-diethyl-S-[2-dimethylaminoethyl]thiophosphate + H2O
?
i.e. dimethyl-amiton
-
-
?
O-ethyl S-(2-diisopropylaminoethyl)methylphosphonothioate + H2O
?
also called VX
-
-
?
O-ethyl-S-(2-di-n-propylaminoethyl)methylphosphonothiolate + H2O
?
i.e. n-propyl-VX
-
-
?
O-ethyl-S-(2-diethylaminoethyl)ethylphosphonothiolate + H2O
?
i.e. VE
-
-
?
O-ethyl-S-(2-diethylaminoethyl)methylphosphonothiolate + H2O
?
i.e. VM
-
-
?
O-ethyl-S-(2-diisopropylaminoethyl)ethylphosphonothiolate + H2O
?
i.e. isopropyl-VE
-
-
?
O-ethyl-S-(2-diisopropylaminoethyl)methylphosphonothiolate + H2O
?
i.e. VX
-
-
?
O-ethyl-S-(2-dimethylaminoethyl)ethylphosphonothiolate + H2O
?
i.e. methyl-VE
-
-
?
O-ethyl-S-(2-dimethylaminoethyl)methylphosphonothiolate + H2O
?
i.e. dimethyl-VX
-
-
?
O-isobutyl S-(2-N,N-diethylaminoethyl)methylphosphonothioate + H2O
?
also called VR
-
-
?
O-isobutyl-S-(2-diethylaminoethyl)methylphosphonothiolate + H2O
?
i.e. VR or Russian VX
-
-
?
O-methyl-S-(2-diethylaminoethyl)methylphosphonothiolate + H2O
?
i.e. methyl-VM
-
-
?
O-n-butyl-S-(2-diethylaminoethyl)methylphosphonothiolate + H2O
?
i.e. CVX or Chinese VX
-
-
?
1,2,2-trimethylpropyl methylphosphonofluoridate + H2O
3,3-dimethylbutan-2-yl methylphosphonate + HF
-
-
-
-
?
CVX + H2O
?
-
i.e. diethylamino-ethyl-O-butyl methylphosphonothioate
-
-
?
cyclosarin + H2O
methyl-phosphonic acid monofluoride + cyclohexanol
-
-
-
-
?
isopropyl methylphosphonofluoridate + H2O
propan-2-yl methylphosphonate + HF
-
-
-
-
?
methyl parathion + H2O
4-nitrophenol + dimethyl thiophosphate
-
-
-
-
?
methyl parathion + H2O
dimethyl thiophosphate + 4-nitrophenol
-
-
-
-
?
parathion + H2O
diethylthiophosphate + 4-nitrophenol
-
i.e. diethyl-4-nitrophenyl thiophosphate
-
-
?
russian VX + H2O
methyl-phosphonothioic acid S-(2-diethylamino-ethyl) ester + 2-methylpropanol
-
-
-
-
?
sarin + H2O
methyl-phosphonic acid monofluoride + isopropyl alcohol
-
-
-
-
?
soman + H2O
methyl-phosphonic acid monofluoride + 1,2,2-trimethylpropanol
-
-
-
-
?
VR + H2O
?
-
i.e. diethylamino-ethyl-O-isobutyl methylphosphonothioate
-
-
?
VX + H2O
?
-
i.e. diisopropylamino-ethyl-O-ethyl methylphosphonothioate
-
-
?
VX + H2O
S-[2-(diisopropylamino)ethyl]-methylphosphonothioic acid + ethanol
-
-
-
-
?
coroxon + H2O
diethyl phosphate + chlorferon
substrate is used for in vitro activity detection
-
-
?
coroxon + H2O
diethyl phosphate + chlorferon
-
substrate is used for in vitro activity detection
-
-
?
dimethyl-paraoxon + H2O
dimethyl phosphate + 4-nitrophenol
-
-
-
?
dimethyl-paraoxon + H2O
dimethyl phosphate + 4-nitrophenol
fluorimetric detection on solid medium
-
-
?
dimethyl-paraoxon + H2O
dimethyl phosphate + 4-nitrophenol
-
fluorimetric detection on solid medium
-
-
?
paraoxon + H2O
4-nitrophenol + diethyl phosphate
-
-
-
?
paraoxon + H2O
4-nitrophenol + diethyl phosphate
-
the enzyme prevents, independent of compound aging, the inhibition of acetylcholinesterase by paraoxon, and also by VX, sarin and soman, the latter are phosphyloxime products of acetlycholinesterase reaction with organophosphate inhibitors, overview
-
-
?
paraoxon + H2O
diethylphosphate + 4-nitrophenol
-
i.e. diethyl-4-nitrophenyl phosphate
-
-
?
additional information
?
-
the enzyme is involved in detoxification of a broad range of organophosphorus pesticides and chemical warfare agents sarin and VX
-
-
?
additional information
?
-
the enzyme also exhibits activity of EC 3.1.8.2, diisopropyl-fluorophosphatase. Enzyme-substrate docking study, overview
-
-
?
additional information
?
-
-
the enzyme also exhibits activity of EC 3.1.8.2, diisopropyl-fluorophosphatase. Enzyme-substrate docking study, overview
-
-
?
additional information
?
-
wild-type PTE exhibits little stereoselectivity against the relatively small phosphonate center of O-ethyl-S-(2-diisopropylaminoethyl)methylphosphonothiolate, i.e. VX, but the elevated pKa of the thiol-leaving group provides a significant challenge for enzyme-catalyzed hydrolysis. Wild-type PTE has enzymatic activity for the hydrolysis of racemic VR similar to VX, but the larger isobutyl group attached to the phosphorus center results in a 25fold preference for the RP-enantiomer. Computational docking study of wild-type and mutant enzymes with the substrates
-
-
?
additional information
?
-
-
OPH catalyzes hydrolysis of organophosphorus compounds, OPC15-20, to give phosphoric or alkylphosphonic acid derivatives, which is accompanied by a decrease in the pH of the reaction medium through release of two protons
-
-
?
NATURAL SUBSTRATE
NATURAL PRODUCT
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate)
(Substrate)
LITERATURE
(Substrate)
(Substrate)
COMMENTARY
(Product)
(Product)
LITERATURE
(Product)
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
r=reversible
ir=irreversible
?=not specified
diethyl-paraoxon + H2O
diethyl phosphate + 4-nitrophenol
-
-
-
?
dimethyl-parathion + H2O
dimethyl thiophosphate + 4-nitrophenol
-
-
-
?
O-ethyl-S-(2-diisopropylaminoethyl)methylphosphonothiolate + H2O
?
i.e. VX
-
-
?
additional information
?
-
the enzyme is involved in detoxification of a broad range of organophosphorus pesticides and chemical warfare agents sarin and VX
-
-
?
dimethyl-paraoxon + H2O
dimethyl phosphate + 4-nitrophenol
fluorimetric detection on solid medium
-
-
?
dimethyl-paraoxon + H2O
dimethyl phosphate + 4-nitrophenol
-
fluorimetric detection on solid medium
-
-
?
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
Co2+
Zn2+
Co2+
Zn2+
additional information
-
requirement for divalent metal ions, the activity of recombinant His6-OPH decreases in the series: Co2+, Ni2+, Mn2+, Cu2+, Zn2+
Co2+
the Co2+ form of OPH is the most active of the metal-liganded enzyme forms
Co2+
Co-PTE has Zn2+ and Co2+ in equimolar amount, the later being located most likely in the labile beta-site. A single Co2+ in the active site already provides a significant enhancement of the catalytic efficiency compared to Zn-PTE
Zn2+
the Zn2+ form of OPH is one of the most stable dimeric proteins ever identified
Zn2+
recombinant Zn-PTE contains only Zn2+ whereas Co-PTE has Zn2+ and Co2+ in equimolar amount, the later being located most likely in the labile beta-site. 0.05 mM ZnCl2 has a modest stabilizing effect, as Zn-PTE and Co-PTE retain respectively 38% and 27% of their activity 2 min after dilution. There are neither additives nor synergistic effect of ZnCl2 and beta-lactoglobulin
Zn2+
required, a metalloenzyme, the active site contains two divalent metal ions essential for its catalytic activity and also 6 amino acids, including four histidine, one aspartic acid and one lysine
Co2+
-
activates, preferred divalent metal ion by recombinant His6-OPH
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
diethyl 4-methylbenzylphosphonate
the inhibitor does not bind at the active site, but binds exclusively into a well-defined surface pocket 12 A away from the active site, docking, overview
ZnCl2
induces a non-competitive partial inhibition of Zn2+-PTE and Co2+-PTE at pH 8.5. Inhibition of hydrolysis of O,O-(3-chloro-4-methyl-2-oxo-2H-chromen-7-yl)ethylmethylphosphonate is one order of magnitude higher than for paraoxon. Inhibition results from interactions with colloidal Zn(OH)2 formed in alkaline buffer that alters the catalytic machinery. ZnCl2 has an inhibitory effect when a protein environment stabilizes PTE. Adding ZnCl2 to the enzyme stabilized by beta-lactoglobulin results in 10% and 20% decrease in activity for Co-PTE and Zn-PTE, respectively
1-hexynyl diethyl phosphate
-
rapid inactivation, reactiviation by increasing pH
1-hexynyl dimethyl phosphate
-
rapid inactivation, reactiviation by increasing pH
1-hexynyl diphenyl phosphate
-
rapid inactivation, reactiviation by increasing pH
1-propynyl diethyl phosphate
-
rapid inactivation, reactiviation by increasing pH
ethynyl diethyl phosphate
-
rapid inactivation, reactiviation by increasing pH
additional information
without additives, both Zn2+- and Co-PTE dramatically loose about 90% of their activity upon dilution. Activity of both Zn2+- and Co-PTE remains unchanged upon addition of CoCl2, CdCl2, FeSO4 and MgCl2 up to 2 mM
-
additional information
-
without additives, both Zn2+- and Co-PTE dramatically loose about 90% of their activity upon dilution. Activity of both Zn2+- and Co-PTE remains unchanged upon addition of CoCl2, CdCl2, FeSO4 and MgCl2 up to 2 mM
-
ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
beta-Lactoglobulin
maximum activity in the presence of 0.1% (w/v) beta-lactoglobulin. There are neither additives nor synergistic effect of ZnCl2 and beta-lactoglobulin
-
additional information
-
efficient enzyme-catalyzed hydrolysis of OPC requires essential stabilization of the enzyme in the acidic pH region
-
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
0.35
(S)-[2-[di(propan-2-yl)amino]ethyl] O,O-diethyl phosphorothioate
pH 9.0, 30°C, recombinant mutant A80V/F132V/K185R/H254Q/H257Y/I274N
0.6
(S)-[2-[di(propan-2-yl)amino]ethyl] O,O-diethyl phosphorothioate
pH 9.0, 30°C, recombinant mutant I106C/F132V/H254Q/H257Y/A270V/L272M/I274N/S308L
0.65
(S)-[2-[di(propan-2-yl)amino]ethyl] O,O-diethyl phosphorothioate
pH 9.0, 30°C, recombinant mutant F132V/H254Q/H257F/S308L
0.73
(S)-[2-[di(propan-2-yl)amino]ethyl] O,O-diethyl phosphorothioate
pH 9.0, 30°C, recombinant mutant A80V/F132V/K185R/H254Q/H257Y/I274N/S308L
0.76
(S)-[2-[di(propan-2-yl)amino]ethyl] O,O-diethyl phosphorothioate
pH 9.0, 30°C, recombinant mutant I106C/F132V/H254Q/H257F/S308L
0.8
(S)-[2-[di(propan-2-yl)amino]ethyl] O,O-diethyl phosphorothioate
pH 9.0, 30°C, recombinant mutant A80V/I106C/F132V/K185R/H254Q/H257Y/I274N/S308L
0.87
(S)-[2-[di(propan-2-yl)amino]ethyl] O,O-diethyl phosphorothioate
pH 9.0, 30°C, recombinant wild-type enzyme
1
(S)-[2-[di(propan-2-yl)amino]ethyl] O,O-diethyl phosphorothioate
pH 9.0, 30°C, recombinant mutant F132V/H254Q/H257F
1.2
(S)-[2-[di(propan-2-yl)amino]ethyl] O,O-diethyl phosphorothioate
pH 9.0, 30°C, recombinant mutant I106C/F132V/H254Q/H257Y/A270V/L272M/I274N/S308L/Y309F
1.4
(S)-[2-[di(propan-2-yl)amino]ethyl] O,O-diethyl phosphorothioate
pH 9.0, 30°C, recombinant mutant H254Q/H257F
1.6
(S)-[2-[di(propan-2-yl)amino]ethyl] O,O-diethyl phosphorothioate
pH 9.0, 30°C, recombinant mutant A80V/I106C/F132V/K185R/H254Q/H257Y/I274N
2.8
(S)-[2-[di(propan-2-yl)amino]ethyl] O,O-diethyl phosphorothioate
pH 9.0, 30°C, recombinant mutant I106G/F132V/H254Q/H257F/S308L
3.2
(S)-[2-[di(propan-2-yl)amino]ethyl] O,O-diethyl phosphorothioate
pH 9.0, 30°C, recombinant mutant I106C/F132V/H254Q/H257F
0.15
4-acetylphenyl (R)-2-methylpropyl methylphosphonate
pH 9.0, 30°C, recombinant mutant F132V/H254Q/H257F/S308L
0.16
4-acetylphenyl (R)-2-methylpropyl methylphosphonate
pH 9.0, 30°C, recombinant mutant A80V/F132V/K185R/H254Q/H257Y/I274N/S308L
0.18
4-acetylphenyl (R)-2-methylpropyl methylphosphonate
pH 9.0, 30°C, recombinant mutant I106C/F132V/H254Q/H257F/S308L
0.23
4-acetylphenyl (R)-2-methylpropyl methylphosphonate
pH 9.0, 30°C, recombinant mutant A80V/I106C/F132V/K185R/H254Q/H257Y/I274N/S308L
0.36
4-acetylphenyl (R)-2-methylpropyl methylphosphonate
pH 9.0, 30°C, recombinant mutant H254Q/H257F
0.41
4-acetylphenyl (R)-2-methylpropyl methylphosphonate
pH 9.0, 30°C, recombinant mutant A80V/I106G/F132V/K185R/H254Q/H257Y/I274N/S308L
0.62
4-acetylphenyl (R)-2-methylpropyl methylphosphonate
pH 9.0, 30°C, recombinant mutant I106C/F132V/H254Q/H257Y/A270V/L272M/I274N/S308L
0.71
4-acetylphenyl (R)-2-methylpropyl methylphosphonate
pH 9.0, 30°C, recombinant mutant I106G/F132V/H254Q/H257Y/A270V/L272M/I274N/S308L
1.4
4-acetylphenyl (R)-2-methylpropyl methylphosphonate
pH 9.0, 30°C, recombinant mutant I106G/F132V/H254Q/H257F/S308L
1.7
4-acetylphenyl (R)-2-methylpropyl methylphosphonate
pH 9.0, 30°C, recombinant wild-type enzyme
1.7
4-acetylphenyl (R)-2-methylpropyl methylphosphonate
pH 9.0, 30°C, recombinant mutant A80V/F132V/K185R/H254Q/H257Y/I274N
2
4-acetylphenyl (R)-2-methylpropyl methylphosphonate
pH 9.0, 30°C, recombinant mutant I106C/F132V/H254Q/H257F
2.1
4-acetylphenyl (R)-2-methylpropyl methylphosphonate
pH 9.0, 30°C, recombinant mutant A80V/I106C/F132V/K185R/H254Q/H257Y/I274N
4
4-acetylphenyl (R)-2-methylpropyl methylphosphonate
pH 9.0, 30°C, recombinant mutant I106G/F132V/H254Q/H257F
0.03
4-acetylphenyl (S)-2-methylpropyl methylphosphonate
pH 9.0, 30°C, recombinant mutant H254Q/H257F
0.13
4-acetylphenyl (S)-2-methylpropyl methylphosphonate
pH 9.0, 30°C, recombinant mutant A80V/I106C/F132V/K185R/H254Q/H257Y/I274N
0.18
4-acetylphenyl (S)-2-methylpropyl methylphosphonate
pH 9.0, 30°C, recombinant mutant A80V/I106C/F132V/K185R/H254Q/H257Y/I274N
0.18
4-acetylphenyl (S)-2-methylpropyl methylphosphonate
pH 9.0, 30°C, recombinant mutant A80V/I106G/F132V/K185R/H254Q/H257Y/I274N/S308L
0.19
4-acetylphenyl (S)-2-methylpropyl methylphosphonate
pH 9.0, 30°C, recombinant mutant I106C/F132V/H254Q/H257F
0.19
4-acetylphenyl (S)-2-methylpropyl methylphosphonate
pH 9.0, 30°C, recombinant mutant I106C/F132V/H254Q/H257F/S308L
0.2
4-acetylphenyl (S)-2-methylpropyl methylphosphonate
pH 9.0, 30°C, recombinant mutant I106G/F132V/H254Q/H257F/S308L
0.23
4-acetylphenyl (S)-2-methylpropyl methylphosphonate
pH 9.0, 30°C, recombinant mutant A80V/I106C/F132V/K185R/H254Q/H257Y/I274N/S308L
0.24
4-acetylphenyl (S)-2-methylpropyl methylphosphonate
pH 9.0, 30°C, recombinant mutant I106G/F132V/H254Q/H257F
0.31
4-acetylphenyl (S)-2-methylpropyl methylphosphonate
pH 9.0, 30°C, recombinant mutant I106G/F132V/H254Q/H257Y/A270V/L272M/I274N/S308L
0.5
4-acetylphenyl (S)-2-methylpropyl methylphosphonate
pH 9.0, 30°C, recombinant mutant I106C/F132V/H254Q/H257Y/A270V/L272M/I274N/S308L
1.5
4-acetylphenyl (S)-2-methylpropyl methylphosphonate
pH 9.0, 30°C, recombinant mutant A80V/F132V/K185R/H254Q/H257Y/I274N
1.5
4-acetylphenyl (S)-2-methylpropyl methylphosphonate
pH 9.0, 30°C, recombinant mutant F132V/H254Q/H257F/S308L
1.7
4-acetylphenyl (S)-2-methylpropyl methylphosphonate
pH 9.0, 30°C, recombinant mutant A80V/F132V/K185R/H254Q/H257Y/I274N/S308L
4.5
4-acetylphenyl (S)-2-methylpropyl methylphosphonate
pH 9.0, 30°C, recombinant wild-type enzyme
6.2
demeton-S
mutant enzyme H254R/H257L, in tripart buffer (pH 8.0)
7.6
demeton-S
mutant enzyme H254R/H257F, in tripart buffer (pH 8.0)
1.6
diethyl 4-chlorophenyl phosphate
recombinant wild-type enzyme bound to Zn2+, pH 9.0, 30°C
3.3
diethyl 4-chlorophenyl phosphate
recombinant wild-type enzyme bound to Cd2+ and Zn2+, pH 9.0, 30°C
7
diethyl 4-chlorophenyl phosphate
recombinant wild-type enzyme bound to Cd2+, pH 9.0, 30°C
0.3
diethyl 4-chlorophenyl thiophosphate
recombinant wild-type enzyme bound to Cd2+, pH 9.0, 30°C
0.5
diethyl 4-chlorophenyl thiophosphate
recombinant wild-type enzyme bound to Zn2+, pH 9.0, 30°C
0.6
diethyl 4-chlorophenyl thiophosphate
recombinant wild-type enzyme bound to Cd2+ and Zn2+, pH 9.0, 30°C
0.34
diethyl 4-nitrophenyl phosphate
recombinant wild-type enzyme bound to Zn2+, pH 9.0, 30°C
0.74
diethyl 4-nitrophenyl phosphate
recombinant wild-type enzyme bound to Cd2+ and Zn2+, pH 9.0, 30°C
1.9
diethyl 4-nitrophenyl phosphate
recombinant wild-type enzyme bound to Cd2+, pH 9.0, 30°C
0.0017
diethyl-paraoxon
pH 9.0, 30°C, recombinant mutant A80V/F132V/K185R/H254Q/H257Y/I274N
0.0036
diethyl-paraoxon
pH 9.0, 30°C, recombinant mutant A80V/I106C/F132V/K185R/H254Q/H257Y/I274N
0.0053
diethyl-paraoxon
pH 9.0, 30°C, recombinant mutant A80V/I106C/F132V/K185R/H254Q/H257Y/I274N/S308L
0.0053
diethyl-paraoxon
pH 9.0, 30°C, recombinant mutant H254Q/H257F
0.0054
diethyl-paraoxon
pH 9.0, 30°C, recombinant mutant F132V/H254Q/H257F/S308L
0.0056
diethyl-paraoxon
pH 9.0, 30°C, recombinant mutant I106C/F132V/H254Q/H257F/S308L
0.0067
diethyl-paraoxon
pH 9.0, 30°C, recombinant mutant I106C/F132V/H254Q/H257F
0.0072
diethyl-paraoxon
pH 9.0, 30°C, recombinant mutant I106C/F132V/H254Q/H257Y/A270V/L272M/I274N/S308L
0.008
diethyl-paraoxon
pH 9.0, 30°C, recombinant mutant A80V/F132V/K185R/H254Q/H257Y/I274N/S308L
0.0092
diethyl-paraoxon
pH 9.0, 30°C, recombinant mutant A80V/I106G/F132V/K185R/H254Q/H257Y/I274N
0.0092
diethyl-paraoxon
pH 9.0, 30°C, recombinant mutant I106C/F132V/H254Q/H257Y/A270V/L272M/I274N/S308L/Y309F
0.011
diethyl-paraoxon
pH 9.0, 30°C, recombinant mutant F132V/H254Q/H257F
0.021
diethyl-paraoxon
pH 9.0, 30°C, recombinant mutant A80V/I106G/F132V/K185R/H254Q/H257Y/I274N/S308L
0.022
diethyl-paraoxon
pH 9.0, 30°C, recombinant mutant I106G/F132V/H254Q/H257F/S308L
0.025
diethyl-paraoxon
pH 9.0, 30°C, recombinant mutant I106G/F132V/H254Q/H257F
0.033
diethyl-paraoxon
pH 9.0, 30°C, recombinant mutant I106G/F132V/H254Q/H257Y/A270V/L272M/I274N/S308L
0.081
diethyl-paraoxon
pH 9.0, 30°C, recombinant wild-type enzyme
0.05
paraoxon
mutant enzyme H254R/H257F, in 20 mM CHES buffer (pH 9.0) at 25°C
0.07
paraoxon
mutant enzyme H254R/H257L, in 20 mM CHES buffer (pH 9.0) at 25°C
0.09
paraoxon
mutant enzyme H254R, in 20 mM CHES buffer (pH 9.0) at 25°C
0.1
paraoxon
wild type enzyme, in 20 mM CHES buffer (pH 9.0) at 25°C
0.3
paraoxon
mutant enzyme H257L, in 20 mM CHES buffer (pH 9.0) at 25°C
0.6
parathion
recombinant wild-type enzyme bound to Cd2+ and Zn2+, pH 9.0, 30°C
1.5
parathion
recombinant wild-type enzyme bound to Zn2+, pH 9.0, 30°C
1.7
parathion
recombinant wild-type enzyme bound to Cd2+, pH 9.0, 30°C
0.104
paraoxon
-
pH 10.5, 25°C, 3-fluorotyrosine-containing recombinant His6-tagged OPH
0.29
paraoxon
-
native enzyme, in 50 mM borate buffer (pH 8.5, 0.1 mM CoCl2), at 25°C
additional information
additional information
kinetic evaluation, mutant H254R
-
additional information
additional information
kinetics of the Zn-enzyme and the Cd-enzyme, wild-type and mutant, kinetic model, overview
-
additional information
additional information
-
kinetics of the Zn-enzyme and the Cd-enzyme, wild-type and mutant, kinetic model, overview
-
additional information
additional information
classical Michaelis-Menten kinetics
-
additional information
additional information
reaction kinetics of wild-type and mutant enzymes, overview
-
additional information
additional information
-
reaction kinetics of wild-type and mutant enzymes, overview
-
additional information
additional information
-
Km values of paraoxon hydrolysis with different metal ions in the active site
-
additional information
additional information
-
kinetics of recombinant His6-tagged wild-type OPH and 3-fluorotyrosine-containing recombinant His6-tagged OPH
-
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
1.1
(S)-[2-[di(propan-2-yl)amino]ethyl] O,O-diethyl phosphorothioate
pH 9.0, 30°C, recombinant wild-type enzyme
3 - 6
(S)-[2-[di(propan-2-yl)amino]ethyl] O,O-diethyl phosphorothioate
pH 9.0, 30°C, recombinant mutant I106C/F132V/H254Q/H257Y/A270V/L272M/I274N/S308L/Y309F
5.1
(S)-[2-[di(propan-2-yl)amino]ethyl] O,O-diethyl phosphorothioate
pH 9.0, 30°C, recombinant mutant A80V/F132V/K185R/H254Q/H257Y/I274N
6.1
(S)-[2-[di(propan-2-yl)amino]ethyl] O,O-diethyl phosphorothioate
pH 9.0, 30°C, recombinant mutant H254Q/H257F
8.6
(S)-[2-[di(propan-2-yl)amino]ethyl] O,O-diethyl phosphorothioate
pH 9.0, 30°C, recombinant mutant A80V/I106C/F132V/K185R/H254Q/H257Y/I274N
13
(S)-[2-[di(propan-2-yl)amino]ethyl] O,O-diethyl phosphorothioate
pH 9.0, 30°C, recombinant mutant I106C/F132V/H254Q/H257F
14
(S)-[2-[di(propan-2-yl)amino]ethyl] O,O-diethyl phosphorothioate
pH 9.0, 30°C, recombinant mutant F132V/H254Q/H257F/S308L
16
(S)-[2-[di(propan-2-yl)amino]ethyl] O,O-diethyl phosphorothioate
pH 9.0, 30°C, recombinant mutant I106C/F132V/H254Q/H257F/S308L
19
(S)-[2-[di(propan-2-yl)amino]ethyl] O,O-diethyl phosphorothioate
pH 9.0, 30°C, recombinant mutant F132V/H254Q/H257F
22
(S)-[2-[di(propan-2-yl)amino]ethyl] O,O-diethyl phosphorothioate
pH 9.0, 30°C, recombinant mutant A80V/F132V/K185R/H254Q/H257Y/I274N/S308L
23
(S)-[2-[di(propan-2-yl)amino]ethyl] O,O-diethyl phosphorothioate
pH 9.0, 30°C, recombinant mutant A80V/I106C/F132V/K185R/H254Q/H257Y/I274N/S308L
31
(S)-[2-[di(propan-2-yl)amino]ethyl] O,O-diethyl phosphorothioate
pH 9.0, 30°C, recombinant mutant I106G/F132V/H254Q/H257F/S308L
51
(S)-[2-[di(propan-2-yl)amino]ethyl] O,O-diethyl phosphorothioate
pH 9.0, 30°C, recombinant mutant I106C/F132V/H254Q/H257Y/A270V/L272M/I274N/S308L
17
4-acetylphenyl (R)-2-methylpropyl methylphosphonate
pH 9.0, 30°C, recombinant mutant A80V/I106G/F132V/K185R/H254Q/H257Y/I274N/S308L
45
4-acetylphenyl (R)-2-methylpropyl methylphosphonate
pH 9.0, 30°C, recombinant mutant A80V/I106C/F132V/K185R/H254Q/H257Y/I274N
46
4-acetylphenyl (R)-2-methylpropyl methylphosphonate
pH 9.0, 30°C, recombinant mutant I106C/F132V/H254Q/H257F/S308L
50
4-acetylphenyl (R)-2-methylpropyl methylphosphonate
pH 9.0, 30°C, recombinant mutant F132V/H254Q/H257F/S308L
55
4-acetylphenyl (R)-2-methylpropyl methylphosphonate
pH 9.0, 30°C, recombinant mutant A80V/F132V/K185R/H254Q/H257Y/I274N/S308L
57
4-acetylphenyl (R)-2-methylpropyl methylphosphonate
pH 9.0, 30°C, recombinant mutant H254Q/H257F
58
4-acetylphenyl (R)-2-methylpropyl methylphosphonate
pH 9.0, 30°C, recombinant mutant I106G/F132V/H254Q/H257Y/A270V/L272M/I274N/S308L
72
4-acetylphenyl (R)-2-methylpropyl methylphosphonate
pH 9.0, 30°C, recombinant mutant A80V/I106C/F132V/K185R/H254Q/H257Y/I274N/S308L
84
4-acetylphenyl (R)-2-methylpropyl methylphosphonate
pH 9.0, 30°C, recombinant wild-type enzyme
100
4-acetylphenyl (R)-2-methylpropyl methylphosphonate
pH 9.0, 30°C, recombinant mutant I106G/F132V/H254Q/H257F
101
4-acetylphenyl (R)-2-methylpropyl methylphosphonate
pH 9.0, 30°C, recombinant mutant I106C/F132V/H254Q/H257Y/A270V/L272M/I274N/S308L
122
4-acetylphenyl (R)-2-methylpropyl methylphosphonate
pH 9.0, 30°C, recombinant mutant I106C/F132V/H254Q/H257F
160
4-acetylphenyl (R)-2-methylpropyl methylphosphonate
pH 9.0, 30°C, recombinant mutant A80V/F132V/K185R/H254Q/H257Y/I274N
174
4-acetylphenyl (R)-2-methylpropyl methylphosphonate
pH 9.0, 30°C, recombinant mutant I106G/F132V/H254Q/H257F/S308L
3 - 6
4-acetylphenyl (S)-2-methylpropyl methylphosphonate
pH 9.0, 30°C, recombinant mutant I106G/F132V/H254Q/H257F/S308L
5
4-acetylphenyl (S)-2-methylpropyl methylphosphonate
pH 9.0, 30°C, recombinant mutant A80V/I106C/F132V/K185R/H254Q/H257Y/I274N
8.1
4-acetylphenyl (S)-2-methylpropyl methylphosphonate
pH 9.0, 30°C, recombinant mutant I106C/F132V/H254Q/H257F
8.7
4-acetylphenyl (S)-2-methylpropyl methylphosphonate
pH 9.0, 30°C, recombinant mutant H254Q/H257F
14
4-acetylphenyl (S)-2-methylpropyl methylphosphonate
pH 9.0, 30°C, recombinant mutant F132V/H254Q/H257F/S308L
21
4-acetylphenyl (S)-2-methylpropyl methylphosphonate
pH 9.0, 30°C, recombinant mutant I106C/F132V/H254Q/H257F/S308L
25
4-acetylphenyl (S)-2-methylpropyl methylphosphonate
pH 9.0, 30°C, recombinant wild-type enzyme
29
4-acetylphenyl (S)-2-methylpropyl methylphosphonate
pH 9.0, 30°C, recombinant mutant A80V/F132V/K185R/H254Q/H257Y/I274N/S308L
33
4-acetylphenyl (S)-2-methylpropyl methylphosphonate
pH 9.0, 30°C, recombinant mutant A80V/I106C/F132V/K185R/H254Q/H257Y/I274N/S308L
40
4-acetylphenyl (S)-2-methylpropyl methylphosphonate
pH 9.0, 30°C, recombinant mutant I106G/F132V/H254Q/H257F
51
4-acetylphenyl (S)-2-methylpropyl methylphosphonate
pH 9.0, 30°C, recombinant mutant A80V/F132V/K185R/H254Q/H257Y/I274N
53
4-acetylphenyl (S)-2-methylpropyl methylphosphonate
pH 9.0, 30°C, recombinant mutant A80V/I106C/F132V/K185R/H254Q/H257Y/I274N
56
4-acetylphenyl (S)-2-methylpropyl methylphosphonate
pH 9.0, 30°C, recombinant mutant I106C/F132V/H254Q/H257Y/A270V/L272M/I274N/S308L
159
4-acetylphenyl (S)-2-methylpropyl methylphosphonate
pH 9.0, 30°C, recombinant mutant A80V/I106G/F132V/K185R/H254Q/H257Y/I274N/S308L
166
4-acetylphenyl (S)-2-methylpropyl methylphosphonate
pH 9.0, 30°C, recombinant mutant I106G/F132V/H254Q/H257Y/A270V/L272M/I274N/S308L
34
demeton-S
mutant enzyme H254R/H257F, in tripart buffer (pH 8.0)
50
demeton-S
mutant enzyme H254R/H257L, in tripart buffer (pH 8.0)
0.13
diethyl 4-chlorophenyl phosphate
recombinant wild-type enzyme bound to Cd2+ and Zn2+, pH 9.0, 30°C
0.23
diethyl 4-chlorophenyl phosphate
recombinant wild-type enzyme bound to Cd2+, pH 9.0, 30°C
0.36
diethyl 4-chlorophenyl phosphate
recombinant wild-type enzyme bound to Zn2+, pH 9.0, 30°C
1.1
diethyl 4-chlorophenyl thiophosphate
recombinant wild-type enzyme bound to Cd2+, pH 9.0, 30°C
2.2
diethyl 4-chlorophenyl thiophosphate
recombinant wild-type enzyme bound to Zn2+, pH 9.0, 30°C
2.4
diethyl 4-chlorophenyl thiophosphate
recombinant wild-type enzyme bound to Cd2+ and Zn2+, pH 9.0, 30°C
2060
diethyl 4-nitrophenyl phosphate
recombinant wild-type enzyme bound to Cd2+ and Zn2+, pH 9.0, 30°C
2300
diethyl 4-nitrophenyl phosphate
recombinant wild-type enzyme bound to Zn2+, pH 9.0, 30°C
2500
diethyl 4-nitrophenyl phosphate
recombinant wild-type enzyme bound to Cd2+, pH 9.0, 30°C
15
diethyl-paraoxon
pH 9.0, 30°C, recombinant mutant A80V/F132V/K185R/H254Q/H257Y/I274N
33
diethyl-paraoxon
pH 9.0, 30°C, recombinant mutant I106C/F132V/H254Q/H257F
35
diethyl-paraoxon
pH 9.0, 30°C, recombinant mutant A80V/I106C/F132V/K185R/H254Q/H257Y/I274N
38
diethyl-paraoxon
pH 9.0, 30°C, recombinant mutant I106C/F132V/H254Q/H257F/S308L
41
diethyl-paraoxon
pH 9.0, 30°C, recombinant mutant H254Q/H257F
48
diethyl-paraoxon
pH 9.0, 30°C, recombinant mutant I106G/F132V/H254Q/H257F
58
diethyl-paraoxon
pH 9.0, 30°C, recombinant mutant A80V/I106G/F132V/K185R/H254Q/H257Y/I274N
66
diethyl-paraoxon
pH 9.0, 30°C, recombinant mutant F132V/H254Q/H257F/S308L
85
diethyl-paraoxon
pH 9.0, 30°C, recombinant mutant I106C/F132V/H254Q/H257Y/A270V/L272M/I274N/S308L
103
diethyl-paraoxon
pH 9.0, 30°C, recombinant mutant A80V/I106C/F132V/K185R/H254Q/H257Y/I274N/S308L
108
diethyl-paraoxon
pH 9.0, 30°C, recombinant mutant F132V/H254Q/H257F
116
diethyl-paraoxon
pH 9.0, 30°C, recombinant mutant A80V/F132V/K185R/H254Q/H257Y/I274N/S308L
131
diethyl-paraoxon
pH 9.0, 30°C, recombinant mutant I106C/F132V/H254Q/H257Y/A270V/L272M/I274N/S308L/Y309F
446
diethyl-paraoxon
pH 9.0, 30°C, recombinant mutant A80V/I106G/F132V/K185R/H254Q/H257Y/I274N/S308L
456
diethyl-paraoxon
pH 9.0, 30°C, recombinant mutant I106G/F132V/H254Q/H257F/S308L
545
diethyl-paraoxon
pH 9.0, 30°C, recombinant mutant I106G/F132V/H254Q/H257Y/A270V/L272M/I274N/S308L
2230
diethyl-paraoxon
pH 9.0, 30°C, recombinant wild-type enzyme
450
paraoxon
mutant enzyme H254R/H257F, in 20 mM CHES buffer (pH 9.0) at 25°C
640
paraoxon
mutant enzyme H254R/H257L, in 20 mM CHES buffer (pH 9.0) at 25°C
780
paraoxon
pH 8.5, recombinant wild-type enzyme-MBP fusion protein
1200
paraoxon
mutant enzyme H254R, in 20 mM CHES buffer (pH 9.0) at 25°C
1280
paraoxon
pH 8.5, recombinant genetic variants 1.5E-MBP fusion protein
1430
paraoxon
pH 8.5, recombinant genetic variants 1.6F-MBP fusion protein
2150
paraoxon
pH 8.5, recombinant genetic variants 2.S5-MBP fusion protein
6900
paraoxon
wild type enzyme, in 20 mM CHES buffer (pH 9.0) at 25°C
8000
paraoxon
mutant enzyme H257L, in 20 mM CHES buffer (pH 9.0) at 25°C
300
parathion
recombinant wild-type enzyme bound to Cd2+ and Zn2+, pH 9.0, 30°C
600
parathion
recombinant wild-type enzyme bound to Cd2+, pH 9.0, 30°C
720
parathion
recombinant wild-type enzyme bound to Zn2+, pH 9.0, 30°C
125
paraoxon
-
pH 10.5, 25°C, 3-fluorotyrosine-containing recombinant His6-tagged OPH
kcat/KM VALUE [1/mMs-1]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
1.264
(S)-[2-[di(propan-2-yl)amino]ethyl] O,O-diethyl phosphorothioate
pH 9.0, 30°C, recombinant wild-type enzyme
4.0625
(S)-[2-[di(propan-2-yl)amino]ethyl] O,O-diethyl phosphorothioate
pH 9.0, 30°C, recombinant mutant I106C/F132V/H254Q/H257F
4.357
(S)-[2-[di(propan-2-yl)amino]ethyl] O,O-diethyl phosphorothioate
pH 9.0, 30°C, recombinant mutant H254Q/H257F
5.375
(S)-[2-[di(propan-2-yl)amino]ethyl] O,O-diethyl phosphorothioate
pH 9.0, 30°C, recombinant mutant A80V/I106C/F132V/K185R/H254Q/H257Y/I274N
11.071
(S)-[2-[di(propan-2-yl)amino]ethyl] O,O-diethyl phosphorothioate
pH 9.0, 30°C, recombinant mutant I106G/F132V/H254Q/H257F/S308L
14.571
(S)-[2-[di(propan-2-yl)amino]ethyl] O,O-diethyl phosphorothioate
pH 9.0, 30°C, recombinant mutant A80V/F132V/K185R/H254Q/H257Y/I274N
19
(S)-[2-[di(propan-2-yl)amino]ethyl] O,O-diethyl phosphorothioate
pH 9.0, 30°C, recombinant mutant F132V/H254Q/H257F
21.053
(S)-[2-[di(propan-2-yl)amino]ethyl] O,O-diethyl phosphorothioate
pH 9.0, 30°C, recombinant mutant I106C/F132V/H254Q/H257F/S308L
21.539
(S)-[2-[di(propan-2-yl)amino]ethyl] O,O-diethyl phosphorothioate
pH 9.0, 30°C, recombinant mutant F132V/H254Q/H257F/S308L
28.75
(S)-[2-[di(propan-2-yl)amino]ethyl] O,O-diethyl phosphorothioate
pH 9.0, 30°C, recombinant mutant A80V/I106C/F132V/K185R/H254Q/H257Y/I274N/S308L
30
(S)-[2-[di(propan-2-yl)amino]ethyl] O,O-diethyl phosphorothioate
pH 9.0, 30°C, recombinant mutant I106C/F132V/H254Q/H257Y/A270V/L272M/I274N/S308L/Y309F
30.137
(S)-[2-[di(propan-2-yl)amino]ethyl] O,O-diethyl phosphorothioate
pH 9.0, 30°C, recombinant mutant A80V/F132V/K185R/H254Q/H257Y/I274N/S308L
85
(S)-[2-[di(propan-2-yl)amino]ethyl] O,O-diethyl phosphorothioate
pH 9.0, 30°C, recombinant mutant I106C/F132V/H254Q/H257Y/A270V/L272M/I274N/S308L
21.4
4-acetylphenyl (R)-2-methylpropyl methylphosphonate
pH 9.0, 30°C, recombinant mutant A80V/I106C/F132V/K185R/H254Q/H257Y/I274N
25
4-acetylphenyl (R)-2-methylpropyl methylphosphonate
pH 9.0, 30°C, recombinant mutant I106G/F132V/H254Q/H257F
32.7
4-acetylphenyl (R)-2-methylpropyl methylphosphonate
pH 9.0, 30°C, recombinant mutant A80V/I106G/F132V/K185R/H254Q/H257Y/I274N/S308L
34.375
4-acetylphenyl (R)-2-methylpropyl methylphosphonate
pH 9.0, 30°C, recombinant mutant A80V/F132V/K185R/H254Q/H257Y/I274N/S308L
49.4
4-acetylphenyl (R)-2-methylpropyl methylphosphonate
pH 9.0, 30°C, recombinant wild-type enzyme
61
4-acetylphenyl (R)-2-methylpropyl methylphosphonate
pH 9.0, 30°C, recombinant mutant I106C/F132V/H254Q/H257F
81.7
4-acetylphenyl (R)-2-methylpropyl methylphosphonate
pH 9.0, 30°C, recombinant mutant I106G/F132V/H254Q/H257Y/A270V/L272M/I274N/S308L
94.1
4-acetylphenyl (R)-2-methylpropyl methylphosphonate
pH 9.0, 30°C, recombinant mutant A80V/F132V/K185R/H254Q/H257Y/I274N
124.3
4-acetylphenyl (R)-2-methylpropyl methylphosphonate
pH 9.0, 30°C, recombinant mutant I106G/F132V/H254Q/H257F/S308L
158.3
4-acetylphenyl (R)-2-methylpropyl methylphosphonate
pH 9.0, 30°C, recombinant mutant H254Q/H257F
162.9
4-acetylphenyl (R)-2-methylpropyl methylphosphonate
pH 9.0, 30°C, recombinant mutant I106C/F132V/H254Q/H257Y/A270V/L272M/I274N/S308L
255.6
4-acetylphenyl (R)-2-methylpropyl methylphosphonate
pH 9.0, 30°C, recombinant mutant I106C/F132V/H254Q/H257F/S308L
313
4-acetylphenyl (R)-2-methylpropyl methylphosphonate
pH 9.0, 30°C, recombinant mutant A80V/I106C/F132V/K185R/H254Q/H257Y/I274N/S308L
333.3
4-acetylphenyl (R)-2-methylpropyl methylphosphonate
pH 9.0, 30°C, recombinant mutant F132V/H254Q/H257F/S308L
5.55
4-acetylphenyl (S)-2-methylpropyl methylphosphonate
pH 9.0, 30°C, recombinant wild-type enzyme
9.3
4-acetylphenyl (S)-2-methylpropyl methylphosphonate
pH 9.0, 30°C, recombinant mutant F132V/H254Q/H257F/S308L
17.1
4-acetylphenyl (S)-2-methylpropyl methylphosphonate
pH 9.0, 30°C, recombinant mutant A80V/F132V/K185R/H254Q/H257Y/I274N/S308L
34
4-acetylphenyl (S)-2-methylpropyl methylphosphonate
pH 9.0, 30°C, recombinant mutant A80V/F132V/K185R/H254Q/H257Y/I274N
38.5
4-acetylphenyl (S)-2-methylpropyl methylphosphonate
pH 9.0, 30°C, recombinant mutant A80V/I106C/F132V/K185R/H254Q/H257Y/I274N
42.6
4-acetylphenyl (S)-2-methylpropyl methylphosphonate
pH 9.0, 30°C, recombinant mutant I106C/F132V/H254Q/H257F
110.5
4-acetylphenyl (S)-2-methylpropyl methylphosphonate
pH 9.0, 30°C, recombinant mutant I106C/F132V/H254Q/H257F/S308L
112
4-acetylphenyl (S)-2-methylpropyl methylphosphonate
pH 9.0, 30°C, recombinant mutant I106C/F132V/H254Q/H257Y/A270V/L272M/I274N/S308L
143.5
4-acetylphenyl (S)-2-methylpropyl methylphosphonate
pH 9.0, 30°C, recombinant mutant A80V/I106C/F132V/K185R/H254Q/H257Y/I274N/S308L
166.67
4-acetylphenyl (S)-2-methylpropyl methylphosphonate
pH 9.0, 30°C, recombinant mutant I106G/F132V/H254Q/H257F
180
4-acetylphenyl (S)-2-methylpropyl methylphosphonate
pH 9.0, 30°C, recombinant mutant I106G/F132V/H254Q/H257F/S308L
290
4-acetylphenyl (S)-2-methylpropyl methylphosphonate
pH 9.0, 30°C, recombinant mutant H254Q/H257F
294.4
4-acetylphenyl (S)-2-methylpropyl methylphosphonate
pH 9.0, 30°C, recombinant mutant A80V/I106C/F132V/K185R/H254Q/H257Y/I274N
535.5
4-acetylphenyl (S)-2-methylpropyl methylphosphonate
pH 9.0, 30°C, recombinant mutant I106G/F132V/H254Q/H257Y/A270V/L272M/I274N/S308L
883.3
4-acetylphenyl (S)-2-methylpropyl methylphosphonate
pH 9.0, 30°C, recombinant mutant A80V/I106G/F132V/K185R/H254Q/H257Y/I274N/S308L
1920
diethyl-paraoxon
pH 9.0, 30°C, recombinant mutant I106G/F132V/H254Q/H257F
4925
diethyl-paraoxon
pH 9.0, 30°C, recombinant mutant I106C/F132V/H254Q/H257F
6304
diethyl-paraoxon
pH 9.0, 30°C, recombinant mutant A80V/I106G/F132V/K185R/H254Q/H257Y/I274N
6786
diethyl-paraoxon
pH 9.0, 30°C, recombinant mutant I106C/F132V/H254Q/H257F/S308L
7736
diethyl-paraoxon
pH 9.0, 30°C, recombinant mutant H254Q/H257F
8824
diethyl-paraoxon
pH 9.0, 30°C, recombinant mutant A80V/F132V/K185R/H254Q/H257Y/I274N
9722
diethyl-paraoxon
pH 9.0, 30°C, recombinant mutant A80V/I106C/F132V/K185R/H254Q/H257Y/I274N
9818
diethyl-paraoxon
pH 9.0, 30°C, recombinant mutant F132V/H254Q/H257F
11806
diethyl-paraoxon
pH 9.0, 30°C, recombinant mutant I106C/F132V/H254Q/H257Y/A270V/L272M/I274N/S308L
12222
diethyl-paraoxon
pH 9.0, 30°C, recombinant mutant F132V/H254Q/H257F/S308L
14239
diethyl-paraoxon
pH 9.0, 30°C, recombinant mutant I106C/F132V/H254Q/H257Y/A270V/L272M/I274N/S308L/Y309F
14500
diethyl-paraoxon
pH 9.0, 30°C, recombinant mutant A80V/F132V/K185R/H254Q/H257Y/I274N/S308L
16515
diethyl-paraoxon
pH 9.0, 30°C, recombinant mutant I106G/F132V/H254Q/H257Y/A270V/L272M/I274N/S308L
19434
diethyl-paraoxon
pH 9.0, 30°C, recombinant mutant A80V/I106C/F132V/K185R/H254Q/H257Y/I274N/S308L
20727
diethyl-paraoxon
pH 9.0, 30°C, recombinant mutant I106G/F132V/H254Q/H257F/S308L
21239
diethyl-paraoxon
pH 9.0, 30°C, recombinant mutant A80V/I106G/F132V/K185R/H254Q/H257Y/I274N/S308L
27531
diethyl-paraoxon
pH 9.0, 30°C, recombinant wild-type enzyme
Ki VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
3
NiCl2
Co2+-PTE, at pH 8.5 and 30°C, with paraoxon as substrate
0.25
diethyl 4-methylbenzylphosphonate
mutant H254R, pH 9.0, 25°C, versus paraoxon
0.99
diethyl 4-methylbenzylphosphonate
wild-type enzyme, pH 9.0, 25°C, versus paraoxon
0.052
ZnCl2
Co2+-PTE, at pH 8.5 and 30°C, with paraoxon as substrate
0.155
ZnCl2
Zn2+-PTE, at pH 8.5 and 30°C, with paraoxon as substrate
0.23
ZnCl2
Co2+-PTE mutant H123N, at pH 8.5 and 30°C, with paraoxon as substrate
0.3
ZnCl2
Co2+-PTE mutant H123A, at pH 8.5 and 30°C, with paraoxon as substrate
0.63
ZnCl2
Co2+-PTE mutant H123I, at pH 8.5 and 30°C, with paraoxon as substrate
IC50 VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
0.083
substrate O-ethyl-S-(2-diisopropylaminoethyl)methylphosphonothiolate, pH 8.0, 25°C, enzyme mutant C23 administered in guinea pigs in vivo
0.567
substrate cyclosarin, pH 8.0, 25°C, enzyme administered in guinea pigs in vivo
1.4
substrate cyclosarin, pH 8.0, 37°C, enzyme administered in guinea pigs in vivo
additional information
additional information
-
assay method evaluation of analogues with fluorescent leaving groups for screening and selection of the enzyme that efficiently hydrolyzes organophosphorus nerve agents, overview
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
10.5
pH RANGE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
5.5 - 12
-
the fluorinated enzyme exhibits stable catalytic activity in the pH range, 35-50% of maximal activity at pH 5.5 and pH 12.0
additional information
additional information
pH profiles of wild-type and mutant enzymes substituted with different metals, overview
additional information
-
pH profiles of wild-type and mutant enzymes substituted with different metals, overview
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
30
60
TEMPERATURE RANGE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
pI VALUE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
7.6
-
the fluorinated enzyme exhibits stable catalytic activity in the pH range, 35-50% of maximal activity at pH 5.5 and pH 12.0
8.5
ORGANISM
COMMENTARY
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
-
UniProt
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
SOURCE
additional information
additional information
enzyme OPH supports Brevundimonas diminuta growth with methyl-parathion as sole phosphate source
additional information
-
enzyme OPH supports Brevundimonas diminuta growth with methyl-parathion as sole phosphate source
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY
GeneOntology No.
LITERATURE
SOURCE
enzyme OPH is tightly bound to the inner membrane. It is targeted to the inner membrane of Brevundimonas diminuta in a pre-folded conformation by the twin arginine transport (Tat) pathway
additional information
additional information
the OPH signal peptide contains an invariant cysteine residue at the junction of the signal peptidase (Spase) cleavage site along with a well conserved lipobox motif
-
additional information
-
the OPH signal peptide contains an invariant cysteine residue at the junction of the signal peptidase (Spase) cleavage site along with a well conserved lipobox motif
-
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
malfunction
opd null mutants of Brevundimonas diminuta fail to grow using the organophosphate insecticide methyl parathion as sole source of phosphate
physiological function
additional information
physiological function
evaluation of protective effect of the enzyme in plasma of rats against in vitro challenge of intoxication with parathion or paraoxon in rats, effectiveness of the enzyme as bioscavenger in vivo, overview
physiological function
organophosphate hydrolase (OPH) hydrolyzes the triester bond found in a variety of organophosphate insecticides and nerve agents. Membrane-bound OPH interacts with the outer membrane efflux protein TolC and with PstS, the periplasmic component of the ABC transporter complex (PstSACB) involved in phosphate transport. Interaction of enzyme OPH with PstS appears to facilitate transport of phosphate generated from organophosphates due to the combined action of OPH and periplasmically located phosphatases. Enzyme OPH supports Brevundimonas diminuta growth with methyl-parathion as sole phosphate source
additional information
enzyme homology modeling and molecular dynamics simulations, overview. The active site contains two divalent metal ions essential for its catalytic activity and also 6 amino acids, including four histidine, one aspartic acid and one lysine. Disulfide bridge confirmation in the enzyme structure
additional information
identification of the OPH interactome using purified native wild-type enzyme. Protein-protein interactions study using the bacterial two-hybrid system
additional information
-
identification of the OPH interactome using purified native wild-type enzyme. Protein-protein interactions study using the bacterial two-hybrid system
UNIPROT
ENTRY NAME
ORGANISM
NO. OF AA
NO. OF TRANSM. HELICES
MOLECULAR WEIGHT[Da]
SOURCE
SEQUENCE
LOCALIZATION PREDICTION
The reliability class of the localization prediction ranges from 1 (very reliable) to 5 (not reliable).
The reliability class of the localization prediction ranges from 1 (very reliable) to 5 (not reliable). PDB
SCOP
CATH
UNIPROT
ORGANISM
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
36290
SUBUNIT
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
homodimer
monomer
?
-
x * 37691.1, recombinant N-terminally His6-tagged mutant 3-fluorotyrosine-containing OPH, mass spectrometry, x * 37602.9, recombinant N-terminally His6-tagged wild-type OPH, mass spectrometry
additional information
homodimer
each monomer includes eight strands of parallel beta-pleated sheets in a beta-barrel structure. Secondary structure analysis of wild-type and mutant enzymes, overview
additional information
OPH has no C-terminal hydrophobic helical domain
additional information
-
OPH has no C-terminal hydrophobic helical domain
POSTTRANSLATIONAL MODIFICATION
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
lipoprotein
purified enzyme OPH is modified with the fatty acids palmitate and stearate, identification of fatty acids attached to the enzyme. Mature OPH is linked to myristic and oleic fatty acids
proteolytic modification
the N-terminal signal peptide of OPH is cleaved off during biosynthesis and therefore cannot serve as a signal anchor. In silico analysis of the OPH signal peptide predicts the existence of both signal peptidase II (SpaseII) and multiple signal peptidase I (SpaseI) cleavage sites in pre-OPH, with the SpaseII cleavage site predicted with the highest level of confidence
CRYSTALLIZATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
hanging drop vapour diffusion method, in 10% PEG6000, 0.1 M HEPES, pH 7.0, 1% diethyl 4-methylbenzylphosphonate, and 5 mM sodium azide
purified recombinant enzyme mutants A80V/F132V/K185R/H254Q/H257Y/I274N/S308L and I106C/F132V/H254Q/H257Y/A270V/L272M/I274N/S308L, vapor diffusion method, mixing of 0.001 ml of 10 mg/ml protein in 50 mM HEPES, pH 8.5, and 1.0 mM CoCl2, with 0.001 ml of precipitant solution containing 100 mM sodium cacodylate, pH 5.5-7.0, 0.2 M magnesium acetate, and 15-30% PEG 8000, and equilbration against 0.5 ml of the precipitant solution, 7-21 days, 18°C, X-ray diffraction structure determination and analysis, molecular replacement using the coordinates of wild-type PTE, PDB ID 1DPM, as the search model
purified recombinant mutant enzyme H254R bound to inhibitor diethyl 4-methylbenzylphosphonate, 5.8 mg/ml protein in 20 mM HEPES, pH 7.5, hanging drop vapour diffusion method, room temperature, the precipitation solution contains 9% PEG 6000, 100 mM CHES, pH 9.0, 1% diethyl 4-methylbenzylphosphonate, and 5 mM sodium azide, no Co2+ is added, X-ray diffraction structure determination and analysis at 1.9 A resolution
in complex with diisopropyl methyl phosphonate and with triethyl phosphate
-
PROTEIN VARIANTS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
A204C/T234C
site-directed mutagenesis, Ala204 and Thr234 are located on the alpha-helices and gamma turn, respectively, thermostability of the mutant enzyme is increased compared to the wild-type, due to increased rigidity
A80V/F132V/K185R/H254Q/H257Y/I274N/S308L
site-directed mutagenesis, the mutant combines expression-enhancing mutations (A80V/K185R/I274N) with additional changes in the active site to achieve a high kcat/Km for the hydrolysis of the SP-enantiomer of ethyl-S-(2-diisopropylaminoethyl)methylphosphonothiolate, i.e. VX. Three-dimensional structure, overview
A80V/I106C/F132V/K185R/H254Q/H257Y/I274N
site-directed mutagenesis
A80V/I106C/F132V/K185R/H254Q/H257Y/I274N/S308L
site-directed mutagenesis
A80V/I106G/F132V/K185R/H254Q/H257Y/I274N
site-directed mutagenesis
A80V/I106G/F132V/K185R/H254Q/H257Y/I274N/S308L
site-directed mutagenesis
C24S
site-directed mutagenesis, mutation of the invariant cysteine residue in the lipobox motif of the OPH signal peptide
D233A
site-directed mutagenesis, structure and kinetics of the mutant enzyme with different bound metal ions compared to the wild-type enzyme, overview
D233N
site-directed mutagenesis, structure and kinetics of the mutant enzyme with different bound metal ions compared to the wild-type enzyme, overview
D301A
site-directed mutagenesis, structure and kinetics of the mutant enzyme with different bound metal ions compared to the wild-type enzyme, overview
D301N
site-directed mutagenesis, structure and kinetics of the mutant enzyme with different bound metal ions compared to the wild-type enzyme, overview
G74C/A78C
site-directed mutagenesis, Gly74 and Ala78 are located on the gamma turn and beta-turn, respectively, which link gamma turn and beta-turn adjacent together, thermostability of the mutant enzyme is decreased compared to the wild-type, due to increased flexibility
H123A
KM remains unaffected, whereas kcat decreases 2.5fold. Alters the bacterial expression yield of Co2+-PTE and Ki for Zn(OH)2 inhibition
H123I
KM remains unaffected, whereas kcat decreases 10fold. Alters the bacterial expression yield of Co2+-PTE and Ki for Zn(OH)2 inhibition
H123N
KM remains unaffected, whereas kcat decreases 80fold. Alters the bacterial expression yield of Co2+-PTE and Ki for Zn(OH)2 inhibition
H254A
site-directed mutagenesis, structure and kinetics of the mutant enzyme with different bound metal ions compared to the wild-type enzyme, overview
H254N
site-directed mutagenesis, structure and kinetics of the mutant enzyme with different bound metal ions compared to the wild-type enzyme, overview
H254Q/H257F
site-directed mutagenesis, the mutant exhibits a 100fold improvement for the hydrolysis of O-ethyl-S-(2-diisopropylaminoethyl)methylphosphonothiolate, i.e. VX, relative to the wild-type enzyme
H254R
H254R/H257F
the mutant enzyme shows reduced activity towards paraoxon and increased activity towards demeton-S compared to the wild type enzyme
H254R/H257L
the variant has higher turnover numbers for the chemical warfare agents O-ethyl S-diisopropyl aminomethyl methylphosphonothioate and O-isobutyl S-N,N-diethylaminoethyl methylphosphonothioate compared to the wild type enzyme, the demeton-S activity is over 14times higher as that of the wild type enzyme
H257L
the mutant enzyme shows increased activity towards paraoxon and reduced activity toward demeton-S compared to the wild type enzyme
I106C/F132V/H254Q/H257F/S308L
site-directed mutagenesis, the mutant shows a similar catalytic efficiency for the hydrolysis of ethyl-S-(2-diisopropylaminoethyl)methylphosphonothiolate, i.e. VX, as mutant H254Q/H257F, but a 3fold improvement in kcat
I106C/F132V/H254Q/H257Y/A270V/L272M/I274N/S308L
site-directed mutagenesis, the mutant has a kcat value for ethyl-S-(2-diisopropylaminoethyl)methylphosphonothiolate, i.e. VX, that is enhanced by 150fold relative to wild-type PTE. Three-dimensional structure, overview
I106C/F132V/H254Q/H257Y/A270V/L272M/I274N/S308L/Y309F
site-directed mutagenesis
I106G/F132V/H254Q/H257Y/A270V/L272M/I274N/S308L
site-directed mutagenesis
T128C/ E124C
site-directed mutagenesis, thermostability and kcat of the mutant enzyme are increased compared to the wild-type
T128C/ E153C
site-directed mutagenesis, Thr128 and Glu153 are located on the beta-sheet and alpha-helices, respectively, thermostability of the mutant enzyme is increased compared to the wild-type, due to increased rigidity
Y309F
I106A/F132A/H254Y
-
stereoselectivity of enzyme toward chiral phosphorous center of substrate is reversed
additional information
H254R
site-directed mutagenesis, the mutation increases the mutants activity with substrate demeton-S, a VX analogue, 4fold and decreases the activity with paraoxon 14fold as well as activity towards diisopropyl phosphorofluoridate, a sarin analogue, 183fold compared to the wild-type enzyme
H254R
the mutant enzyme is not completely denatured by urea concentrations up to 8.5 M even after incubation for 24 h, the mutant is more stable than the wild type enzyme and shows increased activity with demeton-S 9times over that of the wild type enzyme
Y309F
site-directed mutagenesis, structure and kinetics of the mutant enzyme with different bound metal ions compared to the wild-type enzyme, overview
additional information
a PTE mutant dubbed C23 is engineered, exhibiting reversed stereoselectivity and high catalytic efficiency (kcat/KM) for the hydrolysis of the toxic enantiomers of nerve agents VX, CVX, and VR. The other mutants A53, IV-A1, IV-H3, B141, and RD1-G83 are less effective, activities and mutant-substrate docking, overview. The only exception is IV-A1 with amiton. Most variants are highly efficient when hydrolyzing N,N-diisopropyl substrates and were 2fold less efficient with N,N-di-ethyl substrates and 5fold less efficient with N,N-di-methyl substrates
additional information
-
a PTE mutant dubbed C23 is engineered, exhibiting reversed stereoselectivity and high catalytic efficiency (kcat/KM) for the hydrolysis of the toxic enantiomers of nerve agents VX, CVX, and VR. The other mutants A53, IV-A1, IV-H3, B141, and RD1-G83 are less effective, activities and mutant-substrate docking, overview. The only exception is IV-A1 with amiton. Most variants are highly efficient when hydrolyzing N,N-diisopropyl substrates and were 2fold less efficient with N,N-di-ethyl substrates and 5fold less efficient with N,N-di-methyl substrates
additional information
appropriate amino acid pairs for the introduction of disulfide bridge to improve protein thermostability are selected, engineering of de novo disulfide bridges is explored as a means to increase the thermal stability of enzymes in the rational method of protein engineering. Circular dichroism (CD) spectrometry and intrinsic fluorescence experiments are applied to evaluate the structural changes of mutants compared to the wild-type after introducing disulfide bridge
additional information
computational docking study of wild-type and mutant enzymes with the toxic substrates
additional information
generation of opd null mutants, strain DS010
additional information
-
construction of a 3-fluorotyrosine-containing enzyme by replacement of Tyr residues, Tyr292, Tyr239, and Tyr248 residues are completely replaced by the fluorinated analogue, while the Tyr156 residue is replaced by 90%, overview, the substitution extends substantially the pH range of action of the fluorine-substituted enzyme compared with the unsubstituted one, the mutant shows increased pH and temperature stability, but reduced activity
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
65
60 min, the wild-type enzyme had a residual activity of 31%, the residual activities of enzyme mutants G74C/A78C, A204C/T234C, and T128C/E153C are 28%, 33%, and 46.36%, respectively
37
-
the conjugates display improved thermal stability, with a half-life at 37°C and pH 7.4 increasing 48fold from 0.256 h to 12.3 h for native phosphotriesterase and phosphotriesterase-MPEG 5 kDa, respectively
50
-
15 min, purified recombinant His6-tagged wild-type OPH, 50% remaining activity
60
-
15 min, purified 3-fluorotyrosine-containing recombinant His6-tagged OPH, stable up to
75
-
15 min, purified 3-fluorotyrosine-containing recombinant His6-tagged OPH, loss of 20% activity
85
-
15 min, purified 3-fluorotyrosine-containing recombinant His6-tagged OPH, complete inactivation
GENERAL STABILITY
ORGANISM
UNIPROT
LITERATURE
efficient enzyme-catalyzed hydrolysis of OPC requires essential stabilization of the enzyme in the acidic pH region
-
the hydrophobic poly(propylene oxide) blocks of Pluronic interact with hydrophobic amino acids of the enzyme, resulting in an increased activity and pot life for dilute concentrations, methanol inhibition, enhanced stability against high temperature, and extended shelf life
-
PURIFICATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
by ion exchange chromatography, affinity chromatography and ultrafiltration
membrane preparation from wild-type and mutant cells, solubilization of OPH from the inner membrane using a buffer that contains Triton X-100, Triton X-114, n-dodecyl beta-D-maltoside, and digitonin, followed by dialysis, and immunoaffinity chromatography
recombinant wild-type and mutant enzymes from cell-free enzyme extract by precipitation of nucleic acids by protamine sulfate, ammonium sulfate fractionation, and gel filtration, followed by anion exchange chromatography
recombinant wild-type and mutant enzymes from Escherichia coli strain BL21 by ammonium sulfate fractionation, anion exchange chromatography, and gel filtration
recombinant His6-tagged from Escherichia coli strain SG13009 OPH 29-43fold by agarose-based nickel affinity chromatography, and 60fold by affinity chromatography on divalent metal ion-iminodiacetic acidpolyacrylamide cryogel, usage of Ni2+, Co2+ or Cu2+ as methyl ions, method development and evaluation, effect of sample pretreatment on His6-OPH purification, overview
-
recombinant His6-tagged OPH from Escherichia coli by nickel affinity chromatography
-
recombinant N-terminally His6-tagged OPH from Escherichia coli strain B-2935 by Co2+-iminodiacetic acid-polyacrylamide cryogel affinity chromatography
-
CLONED (Commentary)
ORGANISM
UNIPROT
LITERATURE
directed evolution of phosphotriesterase from Pseudomonas diminuta for heterologous expression in Escherichia coli results in stabilization of the metal-free state
expression of wild-type and mutant enzymes in Escherichia coli strain BL21
gene opd is encoded by the organophosphate degradation (opd) island, recombinant expression of His10-tagged wild-type enzyme in Brevundimonas diminuta enzyme-deficient strain DS010. Protein-protein interactions study using the bacterial two-hybrid system in Escherichia coli strain BTH101
gene opd, recombinant expression of wild-type and mutant MBP-tagged enzymes in Escherichia coli strain BL21(DE3)
PTEm gene without the 29-amino acid leader cloned into pET17b vector, expressed in Escherichia coli HMS 174 (DE3) pLysS. PCR fragments of mutants cloned into the expression plasmid pET17b and expressed in Escherichia coli BL21 (DE3) pLysS
recombinant expression of wild-type and mutant enzymes in Escherichia coli strain BL21(DE3)
expression of His6-tagged OPH in enzyme-deficient Escherichia coli strain SG13009
-
expression of His6-tagged OPH in Escherichia coli strains DH5alpha and SG13009, the His6-tagged enzyme shows altered properties compared to the wild-type enzyme
-
expression of N-terminally His6-tagged wild-type and 3-fluorotyrosine-containing OPH in Escherichia coli strain B-2935 devoid of the metabolic ways of tyrosine biosynthesis, the presence of 3-F-Tyr in the medium inhibits the cell growth
-
APPLICATION
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
biotechnology
future biotechnological developments of PTE as a detoxifying enzyme
environmental protection
the enzyme is involved in detoxification of organophosphorus pesticides and chemical warfare agents sarin and VX
biotechnology
-
chemo-enzymatic procedure for chiral synthesis of soman analogues, use of mutants to reverse stereoselectivity
degradation
-
activity and stability of organophosphorus hydrolase are enhanced by interactions between the hydrophobic poly(propylene oxide) block of amphiphilic Pluronics and the enzyme. The strategy provides an efficient route to new formulations for decontaminating organophosphate neurotoxins
additional information
additional information
the organophosphorus-hydrolyzing enzymes such as Brevundimonas diminuta phosphotriesterase (PTE) are used for development of alternative catalytic bioscavengers to treat organophosphorus agents, e.g. pesticides and warfare agents, intoxications of humans
additional information
-
the organophosphorus-hydrolyzing enzymes such as Brevundimonas diminuta phosphotriesterase (PTE) are used for development of alternative catalytic bioscavengers to treat organophosphorus agents, e.g. pesticides and warfare agents, intoxications of humans
REF.
AUTHORS
TITLE
JOURNAL
VOL.
PAGES
YEAR
ORGANISM (UNIPROT)
PUBMED ID
SOURCE
Dumas, D.P.; Durst, H.D.; Landis, W.G.; Raushel, F.M.; Wild, J.R.
Inactivation of organophosphorus nerve agents by the phosphotriesterase from Pseudomonas diminuta
Arch. Biochem. Biophys.
277
155-159
1990
Brevundimonas diminuta
Efremenko, E.N.; Sergeeva, V.S.
Organophosphate hydrolase - an enzyme catalyzing degradation of phosphorus-containing toxins and pesticides
Russ. Chem. Bull. (Translation of Izvestiya Akademii Nauk, Seriya Khimicheskaya)
50
1826-1832
2001
Brevundimonas diminuta, Flavobacterium sp., Rattus norvegicus
-
Benning, M.M.; Hong, S.B.; Raushel, F.M.; Holden, H.M.
The binding of substrate analogs to phosphotriesterase
J. Biol. Chem.
275
30556-30560
2000
Brevundimonas diminuta
Banzon, J.A.; Kuo, J.M.; Miles, B.W.; Fischer, D.R.; Stang, P.J.; Raushel, F.M.
Mechanism-based inactivation of phosphotriesterase by reaction of a critical histidine with a ketene intermediate
Biochemistry
34
743-749
1995
Brevundimonas diminuta
Li, W.S.; Lum, K.T.; Chen-Goodspeed, M.; Sogorb, M.A.; Raushel, F.M.
Stereoselective detoxification of chiral sarin and soman analogues by phosphotriesterase
Bioorg. Med. Chem.
9
2083-2091
2001
Brevundimonas diminuta
Grimsley, J.K.; Calamini, B.; Wild, J.R.; Mesecar, A.D.
Structural and mutational studies of organophosphorus hydrolase reveal a cryptic and functional allosteric-binding site
Arch. Biochem. Biophys.
442
169-179
2005
Brevundimonas diminuta (P0A434)
Herkenhoff, S.; Szinicz, L.; Rastogi, V.K.; Cheng, T.C.; DeFrank, J.J.; Worek, F.
Effect of organophosphorus hydrolysing enzymes on obidoxime-induced reactivation of organophosphate-inhibited human acetylcholinesterase
Arch. Toxicol.
78
338-343
2004
Brevundimonas diminuta
Aubert, S.D.; Li, Y.; Raushel, F.M.
Mechanism for the hydrolysis of organophosphates by the bacterial phosphotriesterase
Biochemistry
43
5707-5715
2004
Brevundimonas diminuta (P0A434), Brevundimonas diminuta
Briseno-Roa, L.; Hill, J.; Notman, S.; Sellers, D.; Smith, A.P.; Timperley, C.M.; Wetherell, J.; Williams, N.H.; Williams, G.R.; Fersht, A.R.; Griffiths, A.D.
Analogues with fluorescent leaving groups for screening and selection of enzymes that efficiently hydrolyze organophosphorus nerve agents
J. Med. Chem.
49
246-255
2006
Brevundimonas diminuta, Homo sapiens
Roodveldt, C.; Tawfik, D.S.
Directed evolution of phosphotriesterase from Pseudomonas diminuta for heterologous expression in Escherichia coli results in stabilization of the metal-free state
Protein Eng. Des. Sel.
18
51-58
2005
Brevundimonas diminuta (P0A434), Brevundimonas diminuta
Efremenko, E.; Votchitseva, Y.; Plieva, F.; Galaev, I.; Mattiasson, B.
Purification of His6-organophosphate hydrolase using monolithic supermacroporous polyacrylamide cryogels developed for immobilized metal affinity chromatography
Appl. Microbiol. Biotechnol.
70
558-563
2006
Brevundimonas diminuta
Votchitseva, Y.A.; Efremenko, E.N.; Aliev, T.K.; Varfolomeyev, S.D.
Properties of hexahistidine-tagged organophosphate hydrolase
Biochemistry (Moscow)
71
167-172
2006
Brevundimonas diminuta, Brevundimonas diminuta VKM B-1297
Votchitseva, Y.A.; Efremenko, E.N.; Varfolomeyev, S.D.
Insertion of an unnatural amino acid into the protein structure: preparation and properties of 3-fluorotyrosine-containing organophosphate hydrolase
Russ. Chem. Bull.
55
369-374
2006
Brevundimonas diminuta
-
Reeves, T.E.; Wales, M.E.; Grimsley, J.K.; Li, P.; Cerasoli, D.M.; Wild, J.R.
Balancing the stability and the catalytic specificities of OP hydrolases with enhanced V-agent activities
Protein Eng. Des. Sel.
21
405-412
2008
Brevundimonas diminuta (P0A434)
Carletti, E.; Jacquamet, L.; Loiodice, M.; Rochu, D.; Masson, P.; Nachon, F.
Update on biochemical properties of recombinant Pseudomonas diminuta phosphotriesterase
J. Enzyme Inhib. Med. Chem.
24
1045-1055
2009
Brevundimonas diminuta (P0A434), Brevundimonas diminuta
Trovaslet-Leroy, M.; Musilova, L.; Renault, F.; Brazzolotto, X.; Misik, J.; Novotny, L.; Froment, M.T.; Gillon, E.; Loiodice, M.; Verdier, L.; Masson, P.; Rochu, D.; Jun, D.; Nachon, F.
Organophosphate hydrolases as catalytic bioscavengers of organophosphorus nerve agents
Toxicol. Lett.
206
14-23
2011
Brevundimonas diminuta, Homo sapiens
Kim, M.; Gkikas, M.; Huang, A.; Kang, J.W.; Suthiwangcharoen, N.; Nagarajan, R.; Olsen, B.D.
Enhanced activity and stability of organophosphorus hydrolase via interaction with an amphiphilic polymer
Chem. Commun. (Camb. )
50
5345-5348
2014
Brevundimonas diminuta
Goldsmith, M.; Eckstein, S.; Ashani, Y.; Greisen, P.; Leader, H.; Sussman, J.L.; Aggarwal, N.; Ovchinnikov, S.; Tawfik, D.S.; Baker, D.; Thiermann, H.; Worek, F.
Catalytic efficiencies of directly evolved phosphotriesterase variants with structurally different organophosphorus compounds in vitro
Arch. Toxicol.
90
2711-2724
2016
Brevundimonas diminuta (P0A434), Brevundimonas diminuta
Bigley, A.N.; Mabanglo, M.F.; Harvey, S.P.; Raushel, F.M.
Variants of phosphotriesterase for the enhanced detoxification of the chemical warfare agent VR
Biochemistry
54
5502-5512
2015
Brevundimonas diminuta (P0A434)
Ashani, Y.; Leader, H.; Aggarwal, N.; Silman, I.; Worek, F.; Sussman, J.L.; Goldsmith, M.
Invitro evaluation of the catalytic activity of paraoxonases and phosphotriesterases predicts the enzyme circulatory levels required for invivo protection against organophosphate intoxications
Chem. Biol. Interact.
259
252-256
2016
Pseudomonas sp., Brevundimonas diminuta (P0A434), Homo sapiens (P27169), Agrobacterium tumefaciens (Q93LD7)
Santillan, J.Y.; Dettorre, L.A.; Lewkowicz, E.S.; Iribarren, A.M.
New and highly active microbial phosphotriesterase sources
FEMS Microbiol. Lett.
363
fnw276
2016
Streptomyces phaeochromogenes, Pseudarthrobacter oxydans, Nocardia asteroides, Streptomyces griseus, Rhodococcus corynebacterioides, Brevundimonas diminuta (P0A434), Brevundimonas diminuta, Streptomyces phaeochromogenes CCRC 10811, Pseudarthrobacter oxydans ATCC 14358, Pseudarthrobacter oxydans ATCC 14359, Nocardia asteroides ATCC 19296, Rhodococcus corynebacterioides ATCC 14898, Streptomyces griseus ATCC 39116
Parthasarathy, S.; Parapatla, H.; Nandavaram, A.; Palmer, T.; Siddavattam, D.
Organophosphate hydrolase is a lipoprotein and interacts with Pi-specific transport system to facilitate growth of Brevundimonas diminuta using OP insecticide as source of Phosphate
J. Biol. Chem.
291
7774-7785
2016
Brevundimonas diminuta (P0A434), Brevundimonas diminuta
Farnoosh, G.; Khajeh, K.; Latifi, A.M.; Aghamollaei, H.
Engineering and introduction of de novo disulphide bridges in organophosphorus hydrolase enzyme for thermostability improvement
J. Biosci.
41
577-588
2016
Brevundimonas diminuta (P0A434)
Santillan, J.Y.; Dettorre, L.A.; Lewkowicz, E.S.; Iribarren, A.M.
New and highly active microbial phosphotriesterase sources
FEMS Microbiol. Lett.
363
fnw276
2016
Streptomyces phaeochromogenes, Pseudarthrobacter oxydans, Nocardia asteroides, Streptomyces griseus, Rhodococcus corynebacterioides, Brevundimonas diminuta (P0A434), Brevundimonas diminuta, Streptomyces phaeochromogenes CCRC 10811, Pseudarthrobacter oxydans ATCC 14358, Pseudarthrobacter oxydans ATCC 14359, Nocardia asteroides ATCC 19296, Rhodococcus corynebacterioides ATCC 14898, Streptomyces griseus ATCC 39116
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