Literature summary for 3.1.1.1 extracted from

Boonyuen, U.; Promnares, K.; Junkree, S.; Day, N.; Imwong, M.
Efficient in vitro refolding and functional characterization of recombinant human liver carboxylesterase (CES1) expressed in E. coli (2015), Protein Expr. Purif., 107, 68-75 .

Cloned(Commentary)

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

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 [mM]	KM Value Maximum [mM]	Substrate	Comment	Organism
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

Localization	Comment	Organism	GeneOntology No.	Textmining
microsome	-	Homo sapiens	-	-

Organism

Organism	UniProt	Comment	Textmining
Homo sapiens	P23141	-	-

Posttranslational Modification

Posttranslational Modification	Comment	Organism
glycoprotein	the enzyme sequence contains one N-glycosylation site	Homo sapiens

Purification (Commentary)

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 (Commentary)

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

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

Source Tissue	Comment	Organism	Textmining
liver	-	Homo sapiens	-

Specific Activity [micromol/min/mg]

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 and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.	Reac.
4-nitrophenyl acetate + H2O	-	Homo sapiens	4-nitrophenol + acetate	-	?

Synonyms

Synonyms	Comment	Organism
CES1	-	Homo sapiens
human liver carboxylesterase	-	Homo sapiens

Temperature Optimum [°C]

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

Turnover Number [1/s]

Turnover Number Minimum [1/s]	Turnover Number Maximum [1/s]	Substrate	Comment	Organism
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

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

General Information

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