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Literature summary 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)

EC Number Cloned (Comment) Organism
6.2.1.13 recombinant overexpression of His-tagged enzyme in Escherichia coli strain BL21(DE3), quantitative RT-PCR enzyme expression analysis Entamoeba histolytica

Protein Variants

EC Number Protein Variants Comment Organism
6.2.1.13 additional information gradual ACS gene silencing (49-93%) significantly decreases the acetate flux without affecting the levels of glycolytic metabolites and ATP in trophozoites. Amoebae lacking ACS activity are unable to reestablish the acetyl-CoA/CoA ratio after an oxidative stress challenge Entamoeba histolytica

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
2.7.2.12 0.02
-
 acetyl phosphate at pH 6.0 and 37°C Entamoeba histolytica
2.7.2.12 0.07
-
 acetyl phosphate at pH 6.0 and 37°C Entamoeba histolytica
2.7.2.12 2.2
-
 phosphate at pH 6.0 and 37°C Entamoeba histolytica
2.7.2.12 2.6
-
 phosphate at pH 6.0 and 37°C Entamoeba histolytica
6.2.1.13 additional information
-
 additional information Michaelis-Menten kinetics, the amoebal ACS displays hyperbolic kinetics for its substrates acetyl-CoA, Pi and ADP and its products acetate, ATP and CoA Entamoeba histolytica
6.2.1.13 0.13
-
 acetyl-CoA at pH 7.0 and 37°C Entamoeba histolytica
6.2.1.13 0.14
-
 acetyl-CoA pH 7.0, 37°C, recombinant enzyme Entamoeba histolytica
6.2.1.13 0.14
-
 acetyl-CoA at pH 6.0 and 37°C Entamoeba histolytica
6.2.1.13 0.2
-
 CoA pH 6.0, 37°C, recombinant enzyme Entamoeba histolytica
6.2.1.13 0.2
-
 CoA at pH 7.0 and 37°C Entamoeba histolytica
6.2.1.13 0.3
-
 ATP pH 6.0, 37°C, recombinant enzyme Entamoeba histolytica
6.2.1.13 0.3
-
 ATP at pH 7.0 and 37°C Entamoeba histolytica
6.2.1.13 0.4
-
 CoA at pH 6.0 and 37°C Entamoeba histolytica
6.2.1.13 0.6
-
 ATP at pH 6.0 and 37°C Entamoeba histolytica
6.2.1.13 0.8
-
 acetate pH 6.0, 37°C, recombinant enzyme Entamoeba histolytica
6.2.1.13 0.8
-
 acetate at pH 7.0 and 37°C Entamoeba histolytica
6.2.1.13 0.9
-
 ADP at pH 7.0 and 37°C Entamoeba histolytica
6.2.1.13 1.3
-
 acetate at pH 6.0 and 37°C Entamoeba histolytica
6.2.1.13 1.8
-
 phosphate at pH 7.0 and 37°C Entamoeba histolytica
6.2.1.13 1.9
-
 ADP pH 7.0, 37°C, recombinant enzyme Entamoeba histolytica
6.2.1.13 1.9
-
 ADP at pH 6.0 and 37°C Entamoeba histolytica
6.2.1.13 2.7
-
 phosphate pH 7.0, 37°C, recombinant enzyme Entamoeba histolytica
6.2.1.13 2.7
-
 phosphate at pH 6.0 and 37°C Entamoeba histolytica

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
6.2.1.13 Mg2+ required Entamoeba histolytica

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
6.2.1.13 ADP + phosphate + acetyl-CoA Entamoeba histolytica
-
 ATP + acetate + CoA
-
 r
6.2.1.13 ATP + acetate + CoA Entamoeba histolytica
-
 ADP + phosphate + acetyl-CoA
-
 r

Organism

EC Number Organism UniProt Comment Textmining
2.7.2.12 Entamoeba histolytica C4M1C3
-
-
6.2.1.13 Entamoeba histolytica Q9NAT4
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
2.7.2.12 Co2+ affinity column chromatography Entamoeba histolytica
6.2.1.13 Co2+ affinity column chromatography Entamoeba histolytica
6.2.1.13 recombinant His-tagged enzyme from Escherichia coli strain BL21(DE3) by cobalt affinity chromatography Entamoeba histolytica

Source Tissue

EC Number Source Tissue Comment Organism Textmining
2.7.2.12 trophozoite
-
 Entamoeba histolytica
-
6.2.1.13 trophozoite
-
 Entamoeba histolytica
-

Storage Stability

EC Number Storage Stability Organism
6.2.1.13 -20°C, 50% (v/v) glycerol, 4 weeks, no loss of activity Entamoeba histolytica

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2.7.2.12 additional information D-glucose 6-phosphate, D-fructose 6-phosphate, D-fructose 1,6-bisphophate, ADP, ATP and AMP used at the physiological concentrations and 1 mM each of 3-phosphoglycerate, 2-phosphoglycerate, phosphoenolpyruvate, UDP-glucose, UTP, O-phosphoserine and phosphocreatine are no substrates for the enzyme. The enzyme has no activity in the acetyl phosphate-forming direction Entamoeba histolytica ?
-
-
2.7.2.12 phosphate + acetyl phosphate
-
 Entamoeba histolytica diphosphate + acetate
-
 ir
6.2.1.13 ADP + phosphate + acetyl-CoA
-
 Entamoeba histolytica ATP + acetate + CoA
-
 r
6.2.1.13 ATP + acetate + CoA
-
 Entamoeba histolytica ADP + phosphate + acetyl-CoA
-
 r
6.2.1.13 additional information metabolites of both reaction directions are determined by gas chromatography Entamoeba histolytica ?
-
 ?

Subunits

EC Number Subunits Comment Organism
2.7.2.12 homodimer 2 * 45000, His-tagged enzyme, SDS-PAGE Entamoeba histolytica
6.2.1.13 homodimer 2 * 79000, SDS-PAGE Entamoeba histolytica

Synonyms

EC Number Synonyms Comment Organism
2.7.2.12 acetate kinase (PPi-forming)
-
 Entamoeba histolytica
2.7.2.12 ACK
-
 Entamoeba histolytica
2.7.2.12 PPi-forming acetate kinase
-
 Entamoeba histolytica
6.2.1.13 Acetyl-CoA synthetase (ADP-forming)
-
 Entamoeba histolytica
6.2.1.13 ACS
-
 Entamoeba histolytica
6.2.1.13 ADP-forming acetyl-CoA synthetase
-
 Entamoeba histolytica

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
6.2.1.13 37
-
 assay at Entamoeba histolytica

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
2.7.2.12 21
-
 phosphate at pH 6.0 and 37°C Entamoeba histolytica
2.7.2.12 21
-
 acetyl phosphate at pH 6.0 and 37°C Entamoeba histolytica
2.7.2.12 30
-
 phosphate at pH 7.0 and 37°C Entamoeba histolytica
2.7.2.12 30
-
 acetyl phosphate at pH 7.0 and 37°C Entamoeba histolytica
6.2.1.13 34
-
 ADP pH 7.0, 37°C, recombinant enzyme Entamoeba histolytica
6.2.1.13 34
-
 phosphate pH 7.0, 37°C, recombinant enzyme Entamoeba histolytica
6.2.1.13 34
-
 acetyl-CoA pH 7.0, 37°C, recombinant enzyme Entamoeba histolytica
6.2.1.13 34
-
 acetyl-CoA at pH 6.0 and 37°C Entamoeba histolytica
6.2.1.13 35
-
 acetate at pH 6.0 and 37°C Entamoeba histolytica
6.2.1.13 46
-
 ATP pH 6.0, 37°C, recombinant enzyme Entamoeba histolytica
6.2.1.13 46
-
 CoA pH 6.0, 37°C, recombinant enzyme Entamoeba histolytica
6.2.1.13 46
-
 acetate pH 6.0, 37°C, recombinant enzyme Entamoeba histolytica
6.2.1.13 46
-
 acetate at pH 7.0 and 37°C Entamoeba histolytica
6.2.1.13 55
-
 acetyl-CoA at pH 7.0 and 37°C Entamoeba histolytica

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
2.7.2.12 7.5
-
-
 Entamoeba histolytica
6.2.1.13 6
-
 forward reaction Entamoeba histolytica
6.2.1.13 7
-
 reverse reaction Entamoeba histolytica
6.2.1.13 8
-
-
 Entamoeba histolytica

pH Range

EC Number pH Minimum pH Maximum Comment Organism
6.2.1.13 6 7 at pH 6.0-7.0, the recombinant enzyme shows 15–60% of the maximum activity Entamoeba histolytica

Cofactor

EC Number Cofactor Comment Organism Structure
6.2.1.13 ATP
-
 Entamoeba histolytica

General Information

EC Number General Information Comment Organism
2.7.2.12 metabolism the enzyme does not contribute to acetate production but might be marginally involved in diphosphate and hexosephosphate homeostasis Entamoeba histolytica
6.2.1.13 malfunction amoebae lacking enzyme activity are unable to reestablish the acetyl-CoA/CoA ratio after an oxidative stress challenge Entamoeba histolytica
6.2.1.13 malfunction gradual ACS gene silencing (49-93%) significantly decreases the acetate flux without affecting the levels of glycolytic metabolites and ATP in trophozoites. Amoebae lacking ACS activity are unable to reestablish the acetyl-CoA/CoA ratio after an oxidative stress challenge Entamoeba histolytica
6.2.1.13 metabolism the enzyme is related to the phosphate-dependent fermentative glycolysis in Entamoeba histolytica. The end-products of carbohydrate catabolism in the parasite are ethanol and acetate, produced by the bifunctional aldehyde-alcohol dehydrogenase (ADHE) and ACS, respectively, overview Entamoeba histolytica
6.2.1.13 metabolism the main role of the enzyme in the parasite energy metabolism is to recycle CoA for glycolysis to proceed under aerobic conditions Entamoeba histolytica

kcat/KM [mM/s]

EC Number kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
2.7.2.12 9
-
 phosphate at pH 6.0 and 37°C Entamoeba histolytica
2.7.2.12 12
-
 phosphate at pH 6.0 and 37°C Entamoeba histolytica
2.7.2.12 355
-
 acetyl phosphate at pH 6.0 and 37°C Entamoeba histolytica
2.7.2.12 1400
-
 acetyl phosphate at pH 6.0 and 37°C Entamoeba histolytica
6.2.1.13 12
-
 phosphate at pH 6.0 and 37°C Entamoeba histolytica
6.2.1.13 12
-
 phosphate pH 7.0, 37°C, recombinant enzyme Entamoeba histolytica
6.2.1.13 17
-
 ADP at pH 6.0 and 37°C Entamoeba histolytica
6.2.1.13 17
-
 ADP pH 7.0, 37°C, recombinant enzyme Entamoeba histolytica
6.2.1.13 26
-
 acetate at pH 6.0 and 37°C Entamoeba histolytica
6.2.1.13 32
-
 phosphate at pH 7.0 and 37°C Entamoeba histolytica
6.2.1.13 58
-
 acetate at pH 7.0 and 37°C Entamoeba histolytica
6.2.1.13 58
-
 acetate pH 6.0, 37°C, recombinant enzyme Entamoeba histolytica
6.2.1.13 59
-
 ATP at pH 6.0 and 37°C Entamoeba histolytica
6.2.1.13 76
-
 ADP at pH 7.0 and 37°C Entamoeba histolytica
6.2.1.13 90
-
 CoA at pH 6.0 and 37°C Entamoeba histolytica
6.2.1.13 144
-
 ATP at pH 7.0 and 37°C Entamoeba histolytica
6.2.1.13 144
-
 ATP pH 6.0, 37°C, recombinant enzyme Entamoeba histolytica
6.2.1.13 213
-
 CoA at pH 7.0 and 37°C Entamoeba histolytica
6.2.1.13 213
-
 CoA pH 6.0, 37°C, recombinant enzyme Entamoeba histolytica
6.2.1.13 239
-
 acetyl-CoA at pH 6.0 and 37°C Entamoeba histolytica
6.2.1.13 239
-
 acetyl-CoA pH 7.0, 37°C, recombinant enzyme Entamoeba histolytica
6.2.1.13 458
-
 acetyl-CoA at pH 7.0 and 37°C Entamoeba histolytica