Cloned (Comment) | Organism |
---|---|
recombinant overexpression of His-tagged enzyme in Escherichia coli strain BL21(DE3), quantitative RT-PCR enzyme expression analysis | Entamoeba histolytica |
Protein Variants | Comment | Organism |
---|---|---|
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] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
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 | |
0.13 | - |
acetyl-CoA | at pH 7.0 and 37°C | Entamoeba histolytica | |
0.14 | - |
acetyl-CoA | pH 7.0, 37°C, recombinant enzyme | Entamoeba histolytica | |
0.14 | - |
acetyl-CoA | at pH 6.0 and 37°C | Entamoeba histolytica | |
0.2 | - |
CoA | pH 6.0, 37°C, recombinant enzyme | Entamoeba histolytica | |
0.2 | - |
CoA | at pH 7.0 and 37°C | Entamoeba histolytica | |
0.3 | - |
ATP | pH 6.0, 37°C, recombinant enzyme | Entamoeba histolytica | |
0.3 | - |
ATP | at pH 7.0 and 37°C | Entamoeba histolytica | |
0.4 | - |
CoA | at pH 6.0 and 37°C | Entamoeba histolytica | |
0.6 | - |
ATP | at pH 6.0 and 37°C | Entamoeba histolytica | |
0.8 | - |
acetate | pH 6.0, 37°C, recombinant enzyme | Entamoeba histolytica | |
0.8 | - |
acetate | at pH 7.0 and 37°C | Entamoeba histolytica | |
0.9 | - |
ADP | at pH 7.0 and 37°C | Entamoeba histolytica | |
1.3 | - |
acetate | at pH 6.0 and 37°C | Entamoeba histolytica | |
1.8 | - |
phosphate | at pH 7.0 and 37°C | Entamoeba histolytica | |
1.9 | - |
ADP | pH 7.0, 37°C, recombinant enzyme | Entamoeba histolytica | |
1.9 | - |
ADP | at pH 6.0 and 37°C | Entamoeba histolytica | |
2.7 | - |
phosphate | pH 7.0, 37°C, recombinant enzyme | Entamoeba histolytica | |
2.7 | - |
phosphate | at pH 6.0 and 37°C | Entamoeba histolytica |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Mg2+ | required | Entamoeba histolytica |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
ADP + phosphate + acetyl-CoA | Entamoeba histolytica | - |
ATP + acetate + CoA | - |
r | |
ATP + acetate + CoA | Entamoeba histolytica | - |
ADP + phosphate + acetyl-CoA | - |
r |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Entamoeba histolytica | Q9NAT4 | - |
- |
Purification (Comment) | Organism |
---|---|
Co2+ affinity column chromatography | Entamoeba histolytica |
recombinant His-tagged enzyme from Escherichia coli strain BL21(DE3) by cobalt affinity chromatography | Entamoeba histolytica |
Source Tissue | Comment | Organism | Textmining |
---|---|---|---|
trophozoite | - |
Entamoeba histolytica | - |
Storage Stability | Organism |
---|---|
-20°C, 50% (v/v) glycerol, 4 weeks, no loss of activity | Entamoeba histolytica |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
ADP + phosphate + acetyl-CoA | - |
Entamoeba histolytica | ATP + acetate + CoA | - |
r | |
ATP + acetate + CoA | - |
Entamoeba histolytica | ADP + phosphate + acetyl-CoA | - |
r | |
additional information | metabolites of both reaction directions are determined by gas chromatography | Entamoeba histolytica | ? | - |
? |
Subunits | Comment | Organism |
---|---|---|
homodimer | 2 * 79000, SDS-PAGE | Entamoeba histolytica |
Synonyms | Comment | Organism |
---|---|---|
Acetyl-CoA synthetase (ADP-forming) | - |
Entamoeba histolytica |
ACS | - |
Entamoeba histolytica |
ADP-forming acetyl-CoA synthetase | - |
Entamoeba histolytica |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
37 | - |
assay at | Entamoeba histolytica |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
34 | - |
ADP | pH 7.0, 37°C, recombinant enzyme | Entamoeba histolytica | |
34 | - |
phosphate | pH 7.0, 37°C, recombinant enzyme | Entamoeba histolytica | |
34 | - |
acetyl-CoA | pH 7.0, 37°C, recombinant enzyme | Entamoeba histolytica | |
34 | - |
acetyl-CoA | at pH 6.0 and 37°C | Entamoeba histolytica | |
35 | - |
acetate | at pH 6.0 and 37°C | Entamoeba histolytica | |
46 | - |
ATP | pH 6.0, 37°C, recombinant enzyme | Entamoeba histolytica | |
46 | - |
CoA | pH 6.0, 37°C, recombinant enzyme | Entamoeba histolytica | |
46 | - |
acetate | pH 6.0, 37°C, recombinant enzyme | Entamoeba histolytica | |
46 | - |
acetate | at pH 7.0 and 37°C | Entamoeba histolytica | |
55 | - |
acetyl-CoA | at pH 7.0 and 37°C | Entamoeba histolytica |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
6 | - |
forward reaction | Entamoeba histolytica |
7 | - |
reverse reaction | Entamoeba histolytica |
8 | - |
- |
Entamoeba histolytica |
pH Minimum | pH Maximum | Comment | Organism |
---|---|---|---|
6 | 7 | at pH 6.0-7.0, the recombinant enzyme shows 1560% of the maximum activity | Entamoeba histolytica |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
ATP | - |
Entamoeba histolytica |
General Information | Comment | Organism |
---|---|---|
malfunction | amoebae lacking enzyme activity are unable to reestablish the acetyl-CoA/CoA ratio after an oxidative stress challenge | Entamoeba histolytica |
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 |
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 |
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 Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
12 | - |
phosphate | at pH 6.0 and 37°C | Entamoeba histolytica | |
12 | - |
phosphate | pH 7.0, 37°C, recombinant enzyme | Entamoeba histolytica | |
17 | - |
ADP | at pH 6.0 and 37°C | Entamoeba histolytica | |
17 | - |
ADP | pH 7.0, 37°C, recombinant enzyme | Entamoeba histolytica | |
26 | - |
acetate | at pH 6.0 and 37°C | Entamoeba histolytica | |
32 | - |
phosphate | at pH 7.0 and 37°C | Entamoeba histolytica | |
58 | - |
acetate | at pH 7.0 and 37°C | Entamoeba histolytica | |
58 | - |
acetate | pH 6.0, 37°C, recombinant enzyme | Entamoeba histolytica | |
59 | - |
ATP | at pH 6.0 and 37°C | Entamoeba histolytica | |
76 | - |
ADP | at pH 7.0 and 37°C | Entamoeba histolytica | |
90 | - |
CoA | at pH 6.0 and 37°C | Entamoeba histolytica | |
144 | - |
ATP | at pH 7.0 and 37°C | Entamoeba histolytica | |
144 | - |
ATP | pH 6.0, 37°C, recombinant enzyme | Entamoeba histolytica | |
213 | - |
CoA | at pH 7.0 and 37°C | Entamoeba histolytica | |
213 | - |
CoA | pH 6.0, 37°C, recombinant enzyme | Entamoeba histolytica | |
239 | - |
acetyl-CoA | at pH 6.0 and 37°C | Entamoeba histolytica | |
239 | - |
acetyl-CoA | pH 7.0, 37°C, recombinant enzyme | Entamoeba histolytica | |
458 | - |
acetyl-CoA | at pH 7.0 and 37°C | Entamoeba histolytica |