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Literature summary for 3.1.1.7 extracted from

  • Sato, R.; Matsumoto, T.; Hidaka, N.; Imai, Y.; Abe, K.; Takahashi, S.; Yamada, R.H.; Kera, Y.
    Cloning and expression of carp acetylcholinesterase gene in Pichia pastoris and characterization of the recombinant enzyme (2009), Protein Expr. Purif., 64, 205-212.
    View publication on PubMed

Cloned(Commentary)

Cloned (Comment) Organism
expression analysis by semi-quantitative PCR, sequence comparisons, subcloning of carp AChE in Escherichia coli strain DH5alpha, and expression of wild-type and mutant enzymes in Pichia pastoris strain X-33 Cyprinus carpio

Protein Variants

Protein Variants Comment Organism
additional information construction of two mutant enzymes, one lacking the T peptide and the second lacking the T peptide and containing the alpha-factor prepro signal sequence from Saccharomyces cerevisiae in place of the original signal sequence Cyprinus carpio

Inhibitors

Inhibitors Comment Organism Structure
acetylthiocholine the recombinant carp AChE shows substrate inhibition at high substrate concentrations of acetyl- and propionylthiocholine Cyprinus carpio
acetylthiocholine iodide
-
Cyprinus carpio
butyrylthiocholine
-
Cyprinus carpio
butyrylthiocholine iodide
-
Cyprinus carpio
BW284c51
-
Cyprinus carpio
diisopropylfluorophosphate
-
Cyprinus carpio
edrophonium
-
Cyprinus carpio
ethopropazine low inhibition Cyprinus carpio
Phenylmethylsulfonylfluoride
-
Cyprinus carpio
procainamide low inhibition Cyprinus carpio
propionylthiocholine the recombinant carp AChE shows substrate inhibition at high substrate concentrations of acetyl- and propionylthiocholine Cyprinus carpio

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.000232
-
acetylthiocholine iodide pH 7.6, 40°C, native enzyme Cyprinus carpio
0.00041
-
butyrylthiocholine iodide pH 7.6, 40°C, native enzyme Cyprinus carpio
0.000526
-
propionylthiocholine iodide pH 7.6, 40°C, native enzyme Cyprinus carpio
0.00205
-
acetylthiocholine iodide pH 7.6, 40°C, recombinant enzyme Cyprinus carpio
0.00605
-
propionylthiocholine iodide pH 7.6, 40°C, recombinant enzyme Cyprinus carpio
0.109
-
butyrylthiocholine iodide pH 7.6, 40°C, recombinant enzyme Cyprinus carpio

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
22000
-
x * 73000 + x * 54000 + x * 22000, recombinant enzyme, SDS-PAGE Cyprinus carpio
54000
-
x * 73000 + x * 54000 + x * 22000, recombinant enzyme, SDS-PAGE Cyprinus carpio
73000
-
x * 73000 + x * 54000 + x * 22000, recombinant enzyme, SDS-PAGE Cyprinus carpio

Organism

Organism UniProt Comment Textmining
Cyprinus carpio B7X755
-
-

Posttranslational Modification

Posttranslational Modification Comment Organism
glycoprotein the enzyme is N-glycosylated, and the sequence contains N-glycosylation consensus sites Cyprinus carpio

Purification (Commentary)

Purification (Comment) Organism
recombinant wild-type and mutant enzymes 276fold from Pichia pastoris strain X-33 culture supernatant by anion exchange and edrophonium adsorption chromatography, followed by gel filtration Cyprinus carpio

Source Tissue

Source Tissue Comment Organism Textmining
muscle
-
Cyprinus carpio
-

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
42
-
mutant enzyme lacking the T peptide and possessing a Saccharomyces cerevisiae alpha-factor prepro signal sequence in recombinant Pichia pastoris cells Cyprinus carpio
176
-
wild-type enzyme in recombinant Pichia pastoris cells Cyprinus carpio
181
-
mutant enzyme lacking the T peptide in recombinant Pichia pastoris cells Cyprinus carpio
4970
-
purified recombinant wild-type enzyme Cyprinus carpio

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
butyrylthiocholine iodide + H2O
-
Cyprinus carpio thiocholine iodide + butyrate
-
?
propionylthiocholine iodide + H2O
-
Cyprinus carpio thiocholine iodide + propionate
-
?
S-acetylthiocholine iodide + H2O detection with 5,5'-dithio-bis-2-nitrobenzoic acid, i.e. DTNB, the enzyme contains a Ser, Glu, and His catalytic triad, overview Cyprinus carpio acetate + thiocholine iodide
-
?

Subunits

Subunits Comment Organism
? x * 73000 + x * 54000 + x * 22000, recombinant enzyme, SDS-PAGE Cyprinus carpio

Synonyms

Synonyms Comment Organism
AChE
-
Cyprinus carpio

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
40
-
-
Cyprinus carpio

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
7.6
-
-
Cyprinus carpio

Ki Value [mM]

Ki Value [mM] Ki Value maximum [mM] Inhibitor Comment Organism Structure
0.0056
-
acetylthiocholine iodide pH 7.6, 40°C, native enzyme Cyprinus carpio
0.175
-
acetylthiocholine iodide pH 7.6, 40°C, recombinant enzyme Cyprinus carpio
0.418
-
butyrylthiocholine iodide pH 7.6, 40°C, native enzyme Cyprinus carpio

IC50 Value

IC50 Value IC50 Value Maximum Comment Organism Inhibitor Structure
0.00217
-
pH 7.6, 40°C, recombinant enzyme Cyprinus carpio ethopropazine
0.00294
-
pH 7.6, 40°C, recombinant enzyme Cyprinus carpio Phenylmethylsulfonylfluoride
0.00305
-
pH 7.6, 40°C, native enzyme Cyprinus carpio Phenylmethylsulfonylfluoride
0.00335
-
pH 7.6, 40°C, native enzyme Cyprinus carpio ethopropazine
0.00347
-
pH 7.6, 40°C, native enzyme Cyprinus carpio procainamide
0.00356
-
pH 7.6, 40°C, recombinant enzyme Cyprinus carpio procainamide
0.00439
-
pH 7.6, 40°C, recombinant enzyme Cyprinus carpio diisopropylfluorophosphate
0.00441
-
pH 7.6, 40°C, native enzyme Cyprinus carpio diisopropylfluorophosphate
0.00522
-
pH 7.6, 40°C, recombinant enzyme Cyprinus carpio edrophonium
0.0066
-
pH 7.6, 40°C, native enzyme Cyprinus carpio edrophonium
0.00682
-
pH 7.6, 40°C, recombinant enzyme Cyprinus carpio BW284c51
0.00722
-
pH 7.6, 40°C, native enzyme Cyprinus carpio BW284c51