Literature summary for 3.1.8.2 extracted from

Yun, H.; Yu, J.; Kim, S.; Lee, N.; Lee, J.; Lee, S.; Kim, N.D.; Yu, C.; Rho, J.
Expression and purification of biologically active recombinant human paraoxonase 1 from a Drosophila S2 stable cell line (2017), Protein Expr. Purif., 131, 34-41 .

Cloned(Commentary)

Cloned (Comment)	Organism
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	Homo sapiens

Cloned (Comment)

Organism

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

Homo sapiens

Protein Variants

Protein Variants	Comment	Organism
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	Homo sapiens

KM Value [mM]

KM Value [mM]	KM Value Maximum [mM]	Substrate	Comment	Organism	Structure
3.63	-	diisopropyl fluorophosphate	pH 10.5, 25°C, recombinant PON1 wild-type enzyme	Homo sapiens
4.7	-	diisopropyl fluorophosphate	pH 10.5, 25°C, recombinant PON1-hFc fusion enzyme	Homo sapiens

Localization

Localization	Comment	Organism	GeneOntology No.	Textmining
extracellular	the enzyme circulates in the blood bound to high-density lipoprotein (HDL)	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.
diisopropyl fluorophosphate + H2O	Homo sapiens	-	diisopropyl phosphate + fluoride	-	?

Organism

Organism	UniProt	Comment	Textmining
Homo sapiens	P27169	-	-

Purification (Commentary)

Purification (Comment)	Organism
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	Homo sapiens

Source Tissue

Source Tissue	Comment	Organism	Textmining
liver	primarily expressed in liver	Homo sapiens	-

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg]	Specific Activity Maximum [µmol/min/mg]	Comment	Organism
13.8	-	purified recombinant PON1 wild-type enzyme, pH 10.5, 25°C, substrate diisopropyl fluorophosphate	Homo sapiens
17.95	-	purified recombinant PON1-hFc fusion enzyme, pH 10.5, 25°C, substrate diisopropyl fluorophosphate	Homo sapiens

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.
diisopropyl fluorophosphate + H2O	-	Homo sapiens	diisopropyl phosphate + fluoride	-	?
diisopropyl fluorophosphate + H2O	spectrophotometric determination with phenol red	Homo sapiens	diisopropyl phosphate + fluoride	-	?
additional information	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	Homo sapiens	?	-	?

Synonyms

Synonyms	Comment	Organism
DFPase	-	Homo sapiens
More	cf. EC 3.1.8.1	Homo sapiens
PON1	-	Homo sapiens

Temperature Optimum [°C]

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

Temperature Stability [°C]

Temperature Stability Minimum [°C]	Temperature Stability Maximum [°C]	Comment	Organism
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	Homo sapiens

Turnover Number [1/s]

Turnover Number Minimum [1/s]	Turnover Number Maximum [1/s]	Substrate	Comment	Organism	Structure
9.69	-	diisopropyl fluorophosphate	pH 10.5, 25°C, recombinant PON1 wild-type enzyme	Homo sapiens
20.35	-	diisopropyl fluorophosphate	pH 10.5, 25°C, recombinant PON1-hFc fusion enzyme	Homo sapiens

pH Optimum

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

pH Stability

pH Stability	pH Stability Maximum	Comment	Organism
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	Homo sapiens

kcat/KM [mM/s]

kcat/KM Value [1/mMs^-1]	kcat/KM Value Maximum [1/mMs^-1]	Substrate	Comment	Organism	Structure
2.67	-	diisopropyl fluorophosphate	pH 10.5, 25°C, recombinant PON1 wild-type enzyme	Homo sapiens
4.33	-	diisopropyl fluorophosphate	pH 10.5, 25°C, recombinant PON1-hFc fusion enzyme	Homo sapiens