Cloned (Comment) | Organism |
---|---|
recombinant expression of wild-type and mutant His-tagged enzyme CES1 in Escherichia coli strain BL21(DE3) in inclusion bodies, the recombinant CES1 protein is expressed differently when grown at different temperatures, expression of CES1 grown at 25°C was much higher than in cells grown at 37°C | Homo sapiens |
Protein Variants | Comment | Organism |
---|---|---|
A270S | site-directed mutagenesis, recombinant mutant shows increased activity compared to the recombinant wild-type | Homo sapiens |
D204E | site-directed mutagenesis, recombinant mutant shows increased activity compared to the recombinant wild-type | Homo sapiens |
S76N | site-directed mutagenesis, the recombinant mutant shows increased activity compared to the recombinant wild-type | Homo sapiens |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
additional information | - |
additional information | Michaelis-Menten steady-state kinetic | Homo sapiens | |
0.263 | - |
4-nitrophenyl acetate | recombinant refolded His-taggged mutant A270S, pH 7.5, 37°C | Homo sapiens | |
0.389 | - |
4-nitrophenyl acetate | recombinant refolded His-taggged mutant S76N, pH 7.5, 37°C | Homo sapiens | |
0.417 | - |
4-nitrophenyl acetate | recombinant refolded His-taggged mutant D204E, pH 7.5, 37°C | Homo sapiens | |
0.579 | - |
4-nitrophenyl acetate | recombinant refolded His-taggged wild-type enzyme, pH 7.5, 37°C | Homo sapiens |
Localization | Comment | Organism | GeneOntology No. | Textmining |
---|---|---|---|---|
microsome | - |
Homo sapiens | - |
- |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Homo sapiens | P23141 | - |
- |
Posttranslational Modification | Comment | Organism |
---|---|---|
glycoprotein | the enzyme sequence contains one N-glycosylation site | Homo sapiens |
Purification (Comment) | Organism |
---|---|
recombinant refolded His-taged enzyme from Escherichia coli strain BL21(DE3) by cobalt affinity chromatography using an elution buffer containing 20 mM sodium phosphate, pH 7.4, 300 mM NaCl, 8 M urea, 250 mM imidazole, and 2 mM 2-mercaptoethanol | Homo sapiens |
Renatured (Comment) | Organism |
---|---|
refolding of recombinant His-tagged CES1 expressed in Escherichia coli inclusion bodies is successful in Tris-HCl at pH 7.5 containing a combination of 1% glycerol and 2 mM 2-mercaptoethanol, whereas a mixture of other additives (trehalose, sorbitol and sucrose) and 2-mercaptoethanol fails to recover a functional protein. His-tagged recombinant CES1 retains its biological activity after refolding and can be used directly without removing the fusion tag | Homo sapiens |
Source Tissue | Comment | Organism | Textmining |
---|---|---|---|
liver | - |
Homo sapiens | - |
Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
---|---|---|---|
73 | - |
purified recombinant refolded His-taggged wild-type enzyme, pH 7.5, 37°C, substrate 4-nitrophenyl acetate | Homo sapiens |
109 | - |
purified recombinant refolded His-taggged mutant A270S, pH 7.5, 37°C, substrate 4-nitrophenyl acetate | Homo sapiens |
128 | - |
purified recombinant refolded His-taggged mutant S76N, pH 7.5, 37°C, substrate 4-nitrophenyl acetate | Homo sapiens |
193 | - |
purified recombinant refolded His-taggged mutant D204E, pH 7.5, 37°C, substrate 4-nitrophenyl acetate | Homo sapiens |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
4-nitrophenyl acetate + H2O | - |
Homo sapiens | 4-nitrophenol + acetate | - |
? |
Synonyms | Comment | Organism |
---|---|---|
CES1 | - |
Homo sapiens |
human liver carboxylesterase | - |
Homo sapiens |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
37 | - |
assay at | Homo sapiens |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
7.6 | - |
4-nitrophenyl acetate | recombinant refolded His-taggged wild-type enzyme, pH 7.5, 37°C | Homo sapiens | |
11 | - |
4-nitrophenyl acetate | recombinant refolded His-taggged mutant A270S, pH 7.5, 37°C | Homo sapiens | |
13 | - |
4-nitrophenyl acetate | recombinant refolded His-taggged mutant S76N, pH 7.5, 37°C | Homo sapiens | |
20 | - |
4-nitrophenyl acetate | recombinant refolded His-taggged mutant D204E, pH 7.5, 37°C | Homo sapiens |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7.5 | - |
assay at | Homo sapiens |
General Information | Comment | Organism |
---|---|---|
physiological function | human liver carboxylesterase 1 (CES1) plays a critical role in the hydrolysis of various ester- and amide-containing molecules, including active metabolites, drugs and prodrugs | Homo sapiens |