Literature summary for 3.1.8.1 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
0.53	-	diethyl-paraoxon	pH 10.5, 25°C, recombinant PON1 wild-type enzyme	Homo sapiens
0.81	-	diethyl-paraoxon	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.
diethyl-paraoxon + H2O	Homo sapiens	-	diethyl phosphate + 4-nitrophenol	-	?

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
0.15	-	purified recombinant PON1 wild-type enzyme, pH 10.5, 25°C, substrate diethyl-paraoxon	Homo sapiens
0.18	-	purified recombinant PON1-hFc fusion enzyme, pH 10.5, 25°C, substrate diethyl-paraoxon	Homo sapiens

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.	Reac.
diethyl-paraoxon + H2O	-	Homo sapiens	diethyl phosphate + 4-nitrophenol	-	?
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
h-PON1	-	Homo sapiens
human paraoxonase 1	-	Homo sapiens
paraoxonase 1	-	Homo sapiens
PON1	-	Homo sapiens

Temperature Optimum [°C]

Temperature Optimum [°C]	Temperature Optimum Maximum [°C]	Comment	Organism
25	30	paraoxonase activity of recombinant PON1-hFc or PON1	Homo sapiens

Temperature Range [°C]

Temperature Minimum [°C]	Temperature Maximum [°C]	Comment	Organism
25	50	optimal activity at 25°C, inactivation at 50°C, the paraoxonase activity of recombinant PON1-hFc or PON1 rapidly decreases when the temperature is over 30°C	Homo sapiens

Temperature Stability [°C]

Temperature Stability Minimum [°C]	Temperature Stability Maximum [°C]	Comment	Organism
30	60	the thermostability of hydrolyric 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
0.104	-	diethyl-paraoxon	pH 10.5, 25°C, recombinant PON1 wild-type enzyme	Homo sapiens
0.202	-	diethyl-paraoxon	pH 10.5, 25°C, recombinant PON1-hFc fusion enzyme	Homo sapiens

pH Optimum

pH Optimum Minimum	pH Optimum Maximum	Comment	Organism
10.5	-	paraoxonase activity of recombinant PON1-hFc or PON1	Homo sapiens

pH Range

pH Minimum	pH Maximum	Comment	Organism
7.5	10.5	optimal activity at pH 10.5, inactivation at pH 7.5	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
0.2	-	diethyl-paraoxon	pH 10.5, 25°C, recombinant PON1 wild-type enzyme	Homo sapiens
0.25	-	diethyl-paraoxon	pH 10.5, 25°C, recombinant PON1-hFc fusion enzyme	Homo sapiens