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Literature summary extracted from
- 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)
| EC Number | Cloned (Comment) | Organism |
|---|---|---|
| 3.1.8.1 | 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 |
| 3.1.8.2 | 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
| EC Number | Protein Variants | Comment | Organism |
|---|---|---|---|
| 3.1.8.1 | 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 |
| 3.1.8.2 | 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]
| EC Number | KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|---|
| 3.1.8.1 | 0.53 | - |
diethyl-paraoxon | pH 10.5, 25°C, recombinant PON1 wild-type enzyme | Homo sapiens |  |
| 3.1.8.1 | 0.81 | - |
diethyl-paraoxon | pH 10.5, 25°C, recombinant PON1-hFc fusion enzyme | Homo sapiens | |
| 3.1.8.2 | 3.63 | - |
diisopropyl fluorophosphate | pH 10.5, 25°C, recombinant PON1 wild-type enzyme | Homo sapiens | |
| 3.1.8.2 | 4.7 | - |
diisopropyl fluorophosphate | pH 10.5, 25°C, recombinant PON1-hFc fusion enzyme | Homo sapiens | |
Localization
| EC Number | Localization | Comment | Organism | GeneOntology No. | Textmining |
|---|---|---|---|---|---|
| 3.1.8.1 | extracellular | the enzyme circulates in the blood bound to high-density lipoprotein (HDL) | Homo sapiens | - |
- |
| 3.1.8.2 | extracellular | the enzyme circulates in the blood bound to high-density lipoprotein (HDL) | Homo sapiens | - |
- |
Metals/Ions
| EC Number | Metals/Ions | Comment | Organism | Structure |
|---|---|---|---|---|
| 3.1.8.1 | Ca2+ | dependent on | Homo sapiens | |
| 3.1.8.2 | Ca2+ | dependent on | Homo sapiens | |
Natural Substrates/ Products (Substrates)
| EC Number | Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
|---|---|---|---|---|---|---|---|
| 3.1.8.1 | diethyl-paraoxon + H2O | Homo sapiens | - |
diethyl phosphate + 4-nitrophenol | - |
? | |
| 3.1.8.2 | diisopropyl fluorophosphate + H2O | Homo sapiens | - |
diisopropyl phosphate + fluoride | - |
? | |
Organism
| EC Number | Organism | UniProt | Comment | Textmining |
|---|---|---|---|---|
| 3.1.8.1 | Homo sapiens | P27169 | - |
- |
| 3.1.8.2 | Homo sapiens | P27169 | - |
- |
Purification (Commentary)
| EC Number | Purification (Comment) | Organism |
|---|---|---|
| 3.1.8.1 | 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 |
| 3.1.8.2 | 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
| EC Number | Source Tissue | Comment | Organism | Textmining |
|---|---|---|---|---|
| 3.1.8.1 | liver | primarily expressed in liver | Homo sapiens | - |
| 3.1.8.2 | liver | primarily expressed in liver | Homo sapiens | - |
Specific Activity [micromol/min/mg]
| EC Number | Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
|---|---|---|---|---|
| 3.1.8.1 | 0.15 | - |
purified recombinant PON1 wild-type enzyme, pH 10.5, 25°C, substrate diethyl-paraoxon | Homo sapiens |
| 3.1.8.1 | 0.18 | - |
purified recombinant PON1-hFc fusion enzyme, pH 10.5, 25°C, substrate diethyl-paraoxon | Homo sapiens |
| 3.1.8.2 | 13.8 | - |
purified recombinant PON1 wild-type enzyme, pH 10.5, 25°C, substrate diisopropyl fluorophosphate | Homo sapiens |
| 3.1.8.2 | 17.95 | - |
purified recombinant PON1-hFc fusion enzyme, pH 10.5, 25°C, substrate diisopropyl fluorophosphate | Homo sapiens |
Substrates and Products (Substrate)
| EC Number | Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
|---|---|---|---|---|---|---|---|
| 3.1.8.1 | diethyl-paraoxon + H2O | - |
Homo sapiens | diethyl phosphate + 4-nitrophenol | - |
? | |
| 3.1.8.1 | 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 | ? | - |
? | |
| 3.1.8.2 | diisopropyl fluorophosphate + H2O | - |
Homo sapiens | diisopropyl phosphate + fluoride | - |
? | |
| 3.1.8.2 | diisopropyl fluorophosphate + H2O | spectrophotometric determination with phenol red | Homo sapiens | diisopropyl phosphate + fluoride | - |
? | |
| 3.1.8.2 | 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
| EC Number | Synonyms | Comment | Organism |
|---|---|---|---|
| 3.1.8.1 | h-PON1 | - |
Homo sapiens |
| 3.1.8.1 | human paraoxonase 1 | - |
Homo sapiens |
| 3.1.8.1 | paraoxonase 1 | - |
Homo sapiens |
| 3.1.8.1 | PON1 | - |
Homo sapiens |
| 3.1.8.2 | DFPase | - |
Homo sapiens |
| 3.1.8.2 | More | cf. EC 3.1.8.1 | Homo sapiens |
| 3.1.8.2 | PON1 | - |
Homo sapiens |
Temperature Optimum [°C]
| EC Number | Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
|---|---|---|---|---|
| 3.1.8.1 | 25 | 30 | paraoxonase activity of recombinant PON1-hFc or PON1 | Homo sapiens |
| 3.1.8.2 | 25 | - |
assay at | Homo sapiens |
Temperature Range [°C]
| EC Number | Temperature Minimum [°C] | Temperature Maximum [°C] | Comment | Organism |
|---|---|---|---|---|
| 3.1.8.1 | 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]
| EC Number | Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
|---|---|---|---|---|
| 3.1.8.1 | 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 |
| 3.1.8.2 | 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]
| EC Number | Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|---|
| 3.1.8.1 | 0.104 | - |
diethyl-paraoxon | pH 10.5, 25°C, recombinant PON1 wild-type enzyme | Homo sapiens | |
| 3.1.8.1 | 0.202 | - |
diethyl-paraoxon | pH 10.5, 25°C, recombinant PON1-hFc fusion enzyme | Homo sapiens | |
| 3.1.8.2 | 9.69 | - |
diisopropyl fluorophosphate | pH 10.5, 25°C, recombinant PON1 wild-type enzyme | Homo sapiens | |
| 3.1.8.2 | 20.35 | - |
diisopropyl fluorophosphate | pH 10.5, 25°C, recombinant PON1-hFc fusion enzyme | Homo sapiens | |
pH Optimum
| EC Number | pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
|---|---|---|---|---|
| 3.1.8.1 | 10.5 | - |
paraoxonase activity of recombinant PON1-hFc or PON1 | Homo sapiens |
| 3.1.8.2 | 8 | - |
assay at | Homo sapiens |
pH Range
| EC Number | pH Minimum | pH Maximum | Comment | Organism |
|---|---|---|---|---|
| 3.1.8.1 | 7.5 | 10.5 | optimal activity at pH 10.5, inactivation at pH 7.5 | Homo sapiens |
pH Stability
| EC Number | pH Stability | pH Stability Maximum | Comment | Organism |
|---|---|---|---|---|
| 3.1.8.1 | 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 |
| 3.1.8.2 | 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]
| EC Number | kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|---|
| 3.1.8.1 | 0.2 | - |
diethyl-paraoxon | pH 10.5, 25°C, recombinant PON1 wild-type enzyme | Homo sapiens | |
| 3.1.8.1 | 0.25 | - |
diethyl-paraoxon | pH 10.5, 25°C, recombinant PON1-hFc fusion enzyme | Homo sapiens | |
| 3.1.8.2 | 2.67 | - |
diisopropyl fluorophosphate | pH 10.5, 25°C, recombinant PON1 wild-type enzyme | Homo sapiens | |
| 3.1.8.2 | 4.33 | - |
diisopropyl fluorophosphate | pH 10.5, 25°C, recombinant PON1-hFc fusion enzyme | Homo sapiens | |
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