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Information on EC 3.1.8.2 - diisopropyl-fluorophosphatase and Organism(s) Homo sapiens and UniProt Accession P27169
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3.1.8.2 diisopropyl-fluorophosphataseIUBMB Comments
Acts on phosphorus anhydride bonds (such as phosphorus-halide and phosphorus-cyanide) in organophosphorus compounds (including 'nerve gases'). Inhibited by chelating agents; requires divalent cations. Related to EC 3.1.8.1 aryldialkylphosphatase.
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Word Map
- 3.1.8.2
- dfpase
-
organophosphorus
- dfp
- degradation
- squid
- paraoxonase
- sarin
- phosphotriesterases
- paraoxon
- environmental protection
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loligo
-
opaas
- arylesterase
-
bioscavenger
- vx
- methylphosphonofluoridate
- medicine
- cyclosarin
-
six-bladed
-
carbanilate
- udp-glucosyltransferase
- diisopropylphosphorofluoridate
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2.8.2.1
- arylsulfotransferase
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udp-glucuronyltransferase
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2.3.1.5
- analysis
The taxonomic range for the selected organisms is: Homo sapiens
The expected taxonomic range for this enzyme is: Bacteria, Eukaryota
The expected taxonomic range for this enzyme is: Bacteria, Eukaryota
Synonyms
diisopropylfluorophosphatase, somanase, opa anhydrolase, ptes5, diisopropyl-fluorophosphatase, 
more
topSYNONYM 
ORGANISM
UNIPROT
COMMENTARY 
LITERATURE
DFPase
di-isopropylphosphorofluoridate fluorohydrolase
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dialkylfluorophosphatase
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diisopropyl phosphorofluoridate hydrolase
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diisopropylfluorophosphatase
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diisopropylfluorophosphonate dehalogenase
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diisopropylphosphofluoridase
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isopropylphosphorofluoridase
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OPA anhydrase
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OPA anhydrolase
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OPAA
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OPAA-2
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organophosphate acid anhydrase
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organophosphorus acid anhydrolase
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somanase
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tabunase
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additional information
DFPase
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-
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REACTION
REACTION DIAGRAM
COMMENTARY
ORGANISM
UNIPROT
LITERATURE
diisopropyl fluorophosphate + H2O = diisopropyl phosphate + fluoride
molecular details of the catalytic mechanism of h-PON1
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
hydrolysis of phosphoric triester
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SYSTEMATIC NAME
IUBMB Comments
diisopropyl-fluorophosphate fluorohydrolase
Acts on phosphorus anhydride bonds (such as phosphorus-halide and phosphorus-cyanide) in organophosphorus compounds (including 'nerve gases'). Inhibited by chelating agents; requires divalent cations. Related to EC 3.1.8.1 aryldialkylphosphatase.
CAS REGISTRY NUMBER
COMMENTARY
9032-18-2
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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
diethyl-paraoxon + H2O
diethyl phosphate + 4-nitrophenol
reaction of EC 3.1.8.1
-
-
?
chlorpyrifos oxon + H2O
diethyl phosphate + 3,5,6-trichloropyridin-2-ol
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-
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?
chlorpyrifos oxon + H2O
diethyl phosphate + 3,5,6-trichloropyridin-2-ol
i.e. CPO, a metabolite of chlorpyrifos that is used as a pesticide in agriculture industry
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-
?
diisopropyl fluorophosphate + H2O
diisopropyl phosphate + fluoride
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?
diisopropyl fluorophosphate + H2O
diisopropyl phosphate + fluoride
highly toxic structural analogue of G-class type of nerve agents
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-
?
diisopropyl fluorophosphate + H2O
diisopropyl phosphate + fluoride
spectrophotometric determination with phenol red
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?
additional information
?
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human PON1 is a calcium-dependent promiscuous enzyme (phosphotriesterase, arylesterase and lactonase) with a wide range of substrates. Human PON1 is capable of hydrolyzing a broad range of organophosphorus compounds, including paraoxon, diisopropylfluorophosphate (DFP) and nerve agents such as sarin, soman and VX
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-
?
additional information
?
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the enzyme is also active with substrates of EC 3.1.1.81, quorum-quenching N-acyl-homoserine lactonase, and EC 3.1.8.1, paraoxonase, hydrolyzing organophosphorus compounds. Measurement of N-oxodecanoyl-DL-homoserine lactone (3O-C10AHL)-hydrolyzing activity (EC 3.1.1.81) of recombinant h-PON1 enzymes is determined by using a recombinant quorum-sensing reporter Escherichia coli strain
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-
?
additional information
?
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additional substrate: dipeptide Gly-Pro, prolidase activity of enzyme
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?
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
chlorpyrifos oxon + H2O
diethyl phosphate + 3,5,6-trichloropyridin-2-ol
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-
-
?
diisopropyl fluorophosphate + H2O
diisopropyl phosphate + fluoride
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?
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
2-hydroxyquinoline
a specific reversible competitive inhibitor of h-PON1 that is known to bind in the active site of the enzyme and inhibit the hydrolytic activities of the enzyme
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
3.63
diisopropyl fluorophosphate
pH 10.5, 25°C, recombinant PON1 wild-type enzyme
4.7
diisopropyl fluorophosphate
pH 10.5, 25°C, recombinant PON1-hFc fusion enzyme
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
9.69
diisopropyl fluorophosphate
pH 10.5, 25°C, recombinant PON1 wild-type enzyme
20.35
diisopropyl fluorophosphate
pH 10.5, 25°C, recombinant PON1-hFc fusion enzyme
kcat/KM VALUE [1/mMs-1]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
2.67
diisopropyl fluorophosphate
pH 10.5, 25°C, recombinant PON1 wild-type enzyme
4.33
diisopropyl fluorophosphate
pH 10.5, 25°C, recombinant PON1-hFc fusion enzyme
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
13.8
purified recombinant PON1 wild-type enzyme, pH 10.5, 25°C, substrate diisopropyl fluorophosphate
17.95
purified recombinant PON1-hFc fusion enzyme, pH 10.5, 25°C, substrate diisopropyl fluorophosphate
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
ORGANISM
COMMENTARY
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
SOURCE
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY
GeneOntology No.
LITERATURE
SOURCE
the enzyme circulates in the blood bound to high-density lipoprotein (HDL)
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GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
malfunction
insufficient organophosphate-hydrolyzing activity of native enzyme affirms the urgent need to develop improved variant(s) having enhanced organophosphate-hydrolyzing activity. Enzyme mutants show altered substrate specificity with increased activity against paraoxon and lactone substrates, overview
physiological function
human paraoxonase 1 (h-PON1) is a serum enzyme that can hydrolyze a variety of substrates, including organophosphate (OP) compounds. PON1 can hydrolyze and inactivate a variety of organophosphate (OP) compounds, including certain OP pesticides and nerve agents (NAs). It is a potential candidate for the development of antidote against OP poisoning in humans. The enzyme possesses anti-inflammatory, anti-oxidative, anti-diabetic and quorum sensor-hydrolyzing activities, it is proposed that the lactonase activity of the enzyme is important for these defensive roles, cf. EC 3.1.1.81
additional information
h-PON1 is a polymorphic enzyme. Molecular docking analysis, homology modelling, overview
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). MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
SUBUNIT
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
additional information
features observed in the structure of PON1, i.e. the six-bladed beta-propeller scaffold, the three alpha-helices at the top of the propeller and the putative calcium-binding residues, are well conserved in the modelled structures
POSTTRANSLATIONAL MODIFICATION
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
PROTEIN VARIANTS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
H115W/R192K
site-directed mutagenesis, the mutant shows altered substrate specificity compared to wild-type
H115W/R192K/A137T
site-directed mutagenesis, the mutant shows altered substrate specificity compared to wild-type
H115W/R192K/A137T/D94H/S211T
site-directed mutagenesis, the mutant shows altered substrate specificity compared to wild-type
H115W/R192K/A137T/L130F
site-directed mutagenesis, the mutant shows altered substrate specificity compared to wild-type
H115W/R192K/A137T/M127I/D263H
site-directed mutagenesis, the mutant shows altered substrate specificity compared to wild-type
H115W/R192K/A137T/S81R/P165A
site-directed mutagenesis, the mutant shows altered substrate specificity compared to wild-type
L55M
natural polymorphism, the polymorphism at the 55th position of h-PON1 does not affect the catalytic properties of the enzyme
R192E
natural polymorphism, polymorphism at position 192 plays an important role in determining the substrate specificity and catalytic efficiency of the enzyme
additional information
additional information
h-PON1 is a polymorphic enzyme. A random mutagenesis approach is used to increase the organophosphate (OP)-hydrolyzing activity of recombinant enzyme h-PON1. The mutants not only show a 10-340fold increased OP-hydrolyzing activity against different OP substrates but also exhibit differential lactonase and arylesterase activities, molecular docking studies, overview. Random mutagenesis using Escherichia coli XL-1 Red mutator strain. All mutations result in a considerable decrease in the delta-valerolactone-hydrolyzing activity of the enzyme
additional information
the catalytic efficiency of the recombinant human paraoxonase 1 fused to human immunoglobulin Fc domain, i.e. recombinant PON1-hFc, towards diisopropyl fluorophosphate (DFP) and paraoxon hydrolysis is 1.63 and 1.24fold higher, respectively, than the recombinant human wild-type PON1
pH STABILITY
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
5 - 10
the pH stability of activity of recombinant PON1-hFc is similar compared to recombinant wild-type PON1, highest stability at pH 7.0-8.0, profiles overview
752102
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
30 - 60
the thermostability of hydrolytic activity of recombinant PON1-hFc is slightly increased compared to recombinant wild-type PON1, inactivation of PON1 at 55°C, inactivation of PON1-hFc at 60°C, profiles overview
PURIFICATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
recombinant human paraoxonase 1 fused to human immunoglobulin Fc domain from Drosophila melanogaster S2 cells by protein A affinity chromatography and dialysis. Recombinant His-tagged enzyme from Escherichia coli strain BL21(DE3) by nickel affinity chromatography and dialysis
recombinant wild-type and mutant enzymes refolded from Escherichia coli strain BL21(DE3) inclusion bodies by ion exchange chromatography
CLONED (Commentary)
ORGANISM
UNIPROT
LITERATURE
gene PON1, recombinant expression of the human paraoxonase 1 in Drosophila melanogaster S2 stable cell line with induction by CuSO4 for 14 days. The recombinant human PON1 is fused with the human immunoglobulin Fc domain (PON1-hFc) to improve protein stability and purification efficiency, method optimization, overview. Recombinant expression of His-tagged enzyme in Escherichia coli strain BL21(DE3) harboring the pGTf2 plasmid expressing GroEL-GroES and TF chaperones
gene PON1, recombinant expression of wild-type and mutant enzymes in Escherichia coli strain BL21(DE3) in inclusion bodies, subcloning in Escherichia coli strain DH5alpha
EXPRESSION
ORGANISM
UNIPROT
LITERATURE
the coexpression of recombinant human SMP30 with GroES/GroEL/Tf at 15°C, combined with the addition of a membrane fluidizer, increases osmolytes, and a two-step expression results in the highest enhancement of solubility and DFPase activity
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RENATURED/Commentary
ORGANISM
UNIPROT
LITERATURE
refolding of recombinant wild-type and mutant enzymes from Escherichia coli strain BL21(DE3) inclusion bodies. The recombinant proteins are refolded to their active form by in vitro refolding, and the active protein present in the refolding reaction mixture is further purified
APPLICATION
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
medicine
human paraoxonase 1 (h-PON1) is a potential candidate for the development of antidote against organophosphate (OP) compounds poisoning in humans. Insufficient organophosphate-hydrolyzing activity of native enzyme affirms the urgent need to develop improved variant(s) having enhanced organophosphate-hydrolyzing activity
REF.
AUTHORS
TITLE
JOURNAL
VOL.
PAGES
YEAR
ORGANISM (UNIPROT)
PUBMED ID
SOURCE
Wang, S.H.; Zhi, Q.W.; Sun, M.J.
Dual activities of human prolidase
Toxicol. In Vitro
20
71-77
2006
Homo sapiens
Choi, M.S.; Saxena, A.; Chilukuri, N.
A strategy for the production of soluble human senescence marker protein-30 in Escherichia coli
Biochem. Biophys. Res. Commun.
393
509-513
2010
Homo sapiens
Tripathy, R.K.; Aggarwal, G.; Bajaj, P.; Kathuria, D.; Bharatam, P.V.; Pande, A.H.
Towards understanding the catalytic mechanism of human paraoxonase 1 experimental and in silico mutagenesis studies
Appl. Biochem. Biotechnol.
182
1642-1662
2017
Homo sapiens (P27169)
EXTERNAL LINKS (specific for EC-Number 3.1.8.2)
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