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 | 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 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 | Comment | Organism | GeneOntology No. | Textmining |
---|---|---|---|---|
extracellular | the enzyme circulates in the blood bound to high-density lipoprotein (HDL) | Homo sapiens | - |
- |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Ca2+ | dependent on | Homo sapiens |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
diethyl-paraoxon + H2O | Homo sapiens | - |
diethyl phosphate + 4-nitrophenol | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Homo sapiens | P27169 | - |
- |
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 | Comment | Organism | Textmining |
---|---|---|---|
liver | primarily expressed in liver | Homo sapiens | - |
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 | 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 | Comment | Organism |
---|---|---|
h-PON1 | - |
Homo sapiens |
human paraoxonase 1 | - |
Homo sapiens |
paraoxonase 1 | - |
Homo sapiens |
PON1 | - |
Homo sapiens |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
25 | 30 | paraoxonase activity of recombinant PON1-hFc or PON1 | Homo sapiens |
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 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 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 Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
10.5 | - |
paraoxonase activity of recombinant PON1-hFc or PON1 | Homo sapiens |
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 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 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 |