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Literature summary for 2.7.2.12 extracted from

  • Pineda, E.; Vazquez, C.; Encalada, R.; Nozaki, T.; Sato, E.; Hanadate, Y.; Nequiz, M.; Olivos-Garcia, A.; Moreno-Sanchez, R.; Saavedra, E.
    Roles of acetyl-CoA synthetase (ADP-forming) and acetate kinase (PPi-forming) in ATP and PPi supply in Entamoeba histolytica (2016), Biochim. Biophys. Acta, 1860, 1163-1172.
    View publication on PubMed

Cloned(Commentary)

Cloned (Comment) Organism
gene CL6EHI_170010, DNA and amino acid sequence determination and analysis, quantitative expression analysis by qRT-PCR, recombinant overexpression of His6-tagged enzyme in Escherichia coli strain BL21(DE3) Entamoeba histolytica

Protein Variants

Protein Variants Comment Organism
additional information constructio of an acetate kinase antisense knockout mutant strain Entamoeba histolytica

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.02
-
acetyl phosphate pH 7.0, 37°C, recombinant enzyme Entamoeba histolytica
0.07
-
acetyl phosphate pH 6.0, 37°C, recombinant enzyme Entamoeba histolytica
2.2
-
phosphate pH 7.0, 37°C, recombinant enzyme Entamoeba histolytica
2.6
-
phosphate pH 6.0, 37°C, recombinant enzyme Entamoeba histolytica

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
45000
-
-
Entamoeba histolytica
77000
-
-
Entamoeba histolytica

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
phosphate + acetyl phosphate Entamoeba histolytica
-
diphosphate + acetate
-
r

Organism

Organism UniProt Comment Textmining
Entamoeba histolytica C4M1C3
-
-

Purification (Commentary)

Purification (Comment) Organism
recombinant His6-tagged enzyme from Escherichia coli strain BL21(DE3) by cobalt affinity chromatography Entamoeba histolytica

Source Tissue

Source Tissue Comment Organism Textmining
amoeba
-
Entamoeba histolytica
-
trophozoite
-
Entamoeba histolytica
-

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
diphosphate + acetate
-
Entamoeba histolytica phosphate + acetyl phosphate
-
r
phosphate + acetyl phosphate
-
Entamoeba histolytica diphosphate + acetate
-
r

Subunits

Subunits Comment Organism
monomer 1 * 45000, about, sequence calculation, x * 77000, about, recombinant His6-tagged enzyme, SDS-PAGE Entamoeba histolytica

Synonyms

Synonyms Comment Organism
ACK
-
Entamoeba histolytica
PPi-forming acetate kinase
-
Entamoeba histolytica

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
37
-
assay at Entamoeba histolytica

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
21
-
acetyl phosphate pH 6.0, 37°C, recombinant enzyme Entamoeba histolytica
30
-
acetyl phosphate pH 7.0, 37°C, recombinant enzyme Entamoeba histolytica

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
6
-
diphosphate and acetate formation reaction, assay at Entamoeba histolytica
7
-
acetyl phosphate formation reaction, assay at Entamoeba histolytica

General Information

General Information Comment Organism
malfunction AcK gene silencing does not affect acetate production in the parasites but promotes a slight decrease (10-20%) in the hexose phosphates and phosphate levels. Amoebae lacking ADP-forming acetyl-CoA synthetase activity are unable to reestablish the acetyl-CoA/CoA ratio after an oxidative stress challenge Entamoeba histolytica
physiological function The enzyme does not importantly contribute to ATP and phospate supply in the amoebae. Recombinant enzyme AcK shows activity only in the acetate formation direction. Enzyme AcK does not contribute to acetate production but might be marginally involved in phosphate and hexosephosphate homeostasis Entamoeba histolytica

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
9
-
phosphate pH 6.0, 37°C, recombinant enzyme Entamoeba histolytica
12
-
phosphate pH 7.0, 37°C, recombinant enzyme Entamoeba histolytica
300
-
acetyl phosphate pH 6.0, 37°C, recombinant enzyme Entamoeba histolytica
1500
-
acetyl phosphate pH 7.0, 37°C, recombinant enzyme Entamoeba histolytica