Any feedback?
Please rate this page
(literature.php)
(0/150)

BRENDA support

Literature summary for 3.1.8.1 extracted from

  • 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 (2017), Appl. Biochem. Biotechnol., 182, 1642-1662 .
    View publication on PubMed

Application

Application Comment Organism
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 Homo sapiens

Cloned(Commentary)

Cloned (Comment) Organism
expressed in Escherichia coli BL21(DE3) cells Homo sapiens
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 Homo sapiens

Protein Variants

Protein Variants Comment Organism
H115W/R192K site-directed mutagenesis, the mutant shows altered substrate specificity compared to wild-type Homo sapiens
H115W/R192K the mutant enzyme exhibits considerably increased organophosphate-hydrolyzing activity compared to the wild type enzyme Homo sapiens
H115W/R192K/A137T site-directed mutagenesis, the mutant shows altered substrate specificity compared to wild-type Homo sapiens
H115W/R192K/A137T/D94H/S211T site-directed mutagenesis, the mutant shows altered substrate specificity compared to wild-type Homo sapiens
H115W/R192K/A137T/L130F site-directed mutagenesis, the mutant shows altered substrate specificity compared to wild-type Homo sapiens
H115W/R192K/A137T/M127I/D263H site-directed mutagenesis, the mutant shows altered substrate specificity compared to wild-type Homo sapiens
H115W/R192K/A137T/S81R/P165A site-directed mutagenesis, the mutant shows altered substrate specificity compared to wild-type Homo sapiens
L55M natural polymorphism, the polymorphism at the 55th position of h-PON1 does not affect the catalytic properties of the enzyme Homo sapiens
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 Homo sapiens
R180T the mutant enzyme exhibits 180fold increased ethyl paraoxon-hydrolyzing activity compared to the wild type enzyme Homo sapiens
R192E natural polymorphism, polymorphism at position 192 plays an important role in determining the substrate specificity and catalytic efficiency of the enzyme Homo sapiens
R460T the mutant enzyme exhibits 23fold increased ethyl paraoxon-hydrolyzing activity and 340fold increased diisopropylfluorophosphate-hydrolyzing activity compared to the wild type enzyme Homo sapiens
R478T the mutant enzyme exhibits 8fold increased ethyl paraoxon-hydrolyzing activity compared to the wild type enzyme Homo sapiens
R784T the mutant enzyme exhibits 3fold increased ethyl paraoxon-hydrolyzing activity compared to the wild type enzyme Homo sapiens
R789T the mutant enzyme exhibits 15fold increased ethyl paraoxon-hydrolyzing activity compared to the wild type enzyme Homo sapiens

Inhibitors

Inhibitors Comment Organism Structure
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; specific inhibitor Homo sapiens

Metals/Ions

Metals/Ions Comment Organism Structure
Ca2+ dependent on Homo sapiens

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
chlorpyrifos oxon + H2O Homo sapiens
-
diethyl phosphate + 3,5,6-trichloropyridin-2-ol
-
?
diethyl-paraoxon + H2O Homo sapiens
-
diethyl phosphate + 4-nitrophenol
-
?

Organism

Organism UniProt Comment Textmining
Homo sapiens
-
-
-
Homo sapiens P27169
-
-

Purification (Commentary)

Purification (Comment) Organism
Q-Sepharose column chromatography Homo sapiens
recombinant wild-type and mutant enzymes refolded from Escherichia coli strain BL21(DE3) inclusion bodies by ion exchange chromatography Homo sapiens

Reaction

Reaction Comment Organism Reaction ID
an aryl dialkyl phosphate + H2O = dialkyl phosphate + an aryl alcohol molecular details of the catalytic mechanism of h-PON1 Homo sapiens

Renatured (Commentary)

Renatured (Comment) Organism
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 Homo sapiens

Source Tissue

Source Tissue Comment Organism Textmining
blood serum
-
Homo sapiens
-
serum
-
Homo sapiens
-

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
chlorpyrifos oxon + H2O
-
Homo sapiens diethyl phosphate + 3,5,6-trichloropyridin-2-ol
-
?
chlorpyrifos oxon + H2O i.e. CPO, a metabolite of chlorpyrifos that is used as a pesticide in agriculture industry Homo sapiens diethyl phosphate + 3,5,6-trichloropyridin-2-ol
-
?
chlorpyrifosoxon + H2O
-
Homo sapiens 3,5,6-trichloro-pyridin-2-ol + diethyl phosphate
-
?
diethyl-paraoxon + H2O
-
Homo sapiens diethyl phosphate + 4-nitrophenol
-
?
diisopropyl fluorophosphate + H2O reaction of EC 3.1.8.2, highly toxic structural analogue of G-class type of nerve agents Homo sapiens diisopropyl phosphate + fluoride
-
?
diisopropylfluorophosphate + H2O
-
Homo sapiens ?
-
?
ethyl paraoxon + H2O
-
Homo sapiens 4-nitrophenol + diethyl phosphate
-
?
additional information the enzyme is also active with substrates of EC 3.1.1.81, quorum-quenching N-acyl-homoserine lactonase, and EC 3.1.8.2, diisopropyl-fluorophosphatase, hydrolyzing diisopropyl-fluorophosphates and phosphorus-halide and phosphorus-cyanide bonds in 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 Homo sapiens ?
-
?
phenyl acetate + H2O
-
Homo sapiens phenol + acetate
-
?

Subunits

Subunits Comment Organism
? x * 45000, SDS-PAGE Homo sapiens
? x * 45000, about, SDS-PAGE Homo sapiens
More 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 Homo sapiens

Synonyms

Synonyms Comment Organism
h-PON1
-
Homo sapiens
More cf. EC 3.1.8.2 and EC 3.1.1.81 Homo sapiens
paraoxonase 1
-
Homo sapiens

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
25
-
assay at Homo sapiens

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
8
-
assay at Homo sapiens

General Information

General Information Comment Organism
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 Homo sapiens
additional information h-PON1 is a polymorphic enzyme. Molecular docking analysis, homology modelling, overview Homo sapiens
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 Homo sapiens