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Literature summary extracted from

  • Jobst, B.; Schühle, K.; Linne, U.; Heider, J.
    ATP-dependent carboxylation of acetophenone by a novel type of carboxylase (2010), J. Bacteriol., 192, 1387-1394.
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

EC Number Cloned (Comment) Organism
6.4.1.8 heterologous expression of the apcEC-His6 subunit in Escherichia coli Aromatoleum aromaticum

Inhibitors

EC Number Inhibitors Comment Organism Structure
6.4.1.8 beta,gamma-Imido-ATP 2 mM, complete inhibition of acetophenone carboxylase activity, non-competitive inhibition Aromatoleum aromaticum
6.4.1.8 Carbamoyl phosphate 2 mM: 50% inhibition of ATPase activity. 8 mM: 50% inhibition of carboxylating activity Aromatoleum aromaticum
6.4.1.8 Mg2+ 20 mM: 50% inhibition Aromatoleum aromaticum
6.4.1.8 Mn2+ 10 mM: 50% inhibition Aromatoleum aromaticum
6.4.1.8 additional information acetophenone carboxylation activity in cell extracts is not sensitive to avidin, indicating that the enzyme is independent of biotin Aromatoleum aromaticum
6.4.1.8 Ni2+ 1 mM: 50% inhibition of ATPase activity. 2.5 mM: 50% inhibition of carboxylation activity Aromatoleum aromaticum
6.4.1.8 Zn2+ the enzyme contains 1 Zn2+ ion per alphabetagammadelta core complex. In spite of the presence of Zn in the enzyme, it is strongly inhibited by Zn2+ ions. 0.2 mM ZnCl2: 50% inhibition of ATPase activity and carboxylation activity Aromatoleum aromaticum

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
6.4.1.8 0.033
-
 acetophenone pH 6.5, 30°C Aromatoleum aromaticum
6.4.1.8 0.5
-
 ATP pH 6.5, 30°C Aromatoleum aromaticum
6.4.1.8 0.54
-
 HCO3- pH 6.5, 30°C Aromatoleum aromaticum

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
6.4.1.8 K+ 20 mM, required for optimal activity Aromatoleum aromaticum
6.4.1.8 Mg2+ the enzyme is dependent on the presence of Mg2+ or Mn2+ Aromatoleum aromaticum
6.4.1.8 Mn2+ the enzyme is dependent on the presence of Mg2+ or Mn2+ Aromatoleum aromaticum
6.4.1.8 NH4+ 40 mM, required for optimal activity Aromatoleum aromaticum
6.4.1.8 Zn2+ the enzyme contains 1 Zn2+ ion per alphabetagammadelta core complex. In spite of the presence of Zn in the enzyme, it is strongly inhibited by Zn2+ ions Aromatoleum aromaticum

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
6.4.1.8 15000
-
 1 * 70000 (alpha) + 1 * 15000 (beta) + 1 * 87000 (gamma) + 1 * 75000 (delta) + 1 * 34000 (epsilon), the genes (apcABCDE) of the five subunits form an apparent operon Aromatoleum aromaticum
6.4.1.8 34000
-
 1 * 70000 (alpha) + 1 * 15000 (beta) + 1 * 87000 (gamma) + 1 * 75000 (delta) + 1 * 34000 (epsilon), the genes (apcABCDE) of the five subunits form an apparent operon Aromatoleum aromaticum
6.4.1.8 40000
-
 apcE gene product, gel filtration Aromatoleum aromaticum
6.4.1.8 70000
-
 1 * 70000 (alpha) + 1 * 15000 (beta) + 1 * 87000 (gamma) + 1 * 75000 (delta) + 1 * 34000 (epsilon), the genes (apcABCDE) of the five subunits form an apparent operon Aromatoleum aromaticum
6.4.1.8 75000
-
 1 * 70000 (alpha) + 1 * 15000 (beta) + 1 * 87000 (gamma) + 1 * 75000 (delta) + 1 * 34000 (epsilon), the genes (apcABCDE) of the five subunits form an apparent operon Aromatoleum aromaticum
6.4.1.8 87000
-
 1 * 70000 (alpha) + 1 * 15000 (beta) + 1 * 87000 (gamma) + 1 * 75000 (delta) + 1 * 34000 (epsilon), the genes (apcABCDE) of the five subunits form an apparent operon Aromatoleum aromaticum
6.4.1.8 485000
-
 apcABCD gene products, gel filtration Aromatoleum aromaticum

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
6.4.1.8 2 ATP + acetophenone + HCO3- + H2O + H+ Aromatoleum aromaticum anaerobic ethylbenzene metabolism in the betaproteobacterium Aromatoleum aromaticum is initiated by anaerobic oxidation to acetophenone via (S)-1-phenylethanol. The subsequent carboxylation of acetophenone to benzoylacetate is catalyzed by an acetophenone-induced enzyme 2 ADP + 2 phosphate + 3-oxo-3-phenylpropanoate
-
 ?
6.4.1.8 2 ATP + acetophenone + HCO3- + H2O + H+ Aromatoleum aromaticum EbN1 anaerobic ethylbenzene metabolism in the betaproteobacterium Aromatoleum aromaticum is initiated by anaerobic oxidation to acetophenone via (S)-1-phenylethanol. The subsequent carboxylation of acetophenone to benzoylacetate is catalyzed by an acetophenone-induced enzyme 2 ADP + 2 phosphate + 3-oxo-3-phenylpropanoate
-
 ?

Organism

EC Number Organism UniProt Comment Textmining
6.4.1.8 Aromatoleum aromaticum
-
-
-
6.4.1.8 Aromatoleum aromaticum EbN1
-
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
6.4.1.8 during purification, acetophenone carboxylase dissociates into inactive subcomplexes consisting of the 70-, 15-, 87-, and 75-kDa subunits (apcABCD gene products) and the 34-kDa subunit (apcE gene product), respectively. Acetophenone carboxylase activity is restored by mixing the purified subcomplexes Aromatoleum aromaticum

Specific Activity [micromol/min/mg]

EC Number Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
6.4.1.8 0.051
-
 pH 6.5, 30°C Aromatoleum aromaticum

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
6.4.1.8 2 ATP + 4-acetylpyridine + HCO3- + H2O + H+ 22% of the activity with acetophenone Aromatoleum aromaticum 2 ADP + 2 phosphate + 2-oxo-3-(pyridin-4-yl)propanoate
-
 ?
6.4.1.8 2 ATP + 4-acetylpyridine + HCO3- + H2O + H+ 22% of the activity with acetophenone Aromatoleum aromaticum EbN1 2 ADP + 2 phosphate + 2-oxo-3-(pyridin-4-yl)propanoate
-
 ?
6.4.1.8 2 ATP + acetophenone + HCO3- + H2O + H+ anaerobic ethylbenzene metabolism in the betaproteobacterium Aromatoleum aromaticum is initiated by anaerobic oxidation to acetophenone via (S)-1-phenylethanol. The subsequent carboxylation of acetophenone to benzoylacetate is catalyzed by an acetophenone-induced enzyme Aromatoleum aromaticum 2 ADP + 2 phosphate + 3-oxo-3-phenylpropanoate
-
 ?
6.4.1.8 2 ATP + acetophenone + HCO3- + H2O + H+ the enzyme shows uncoupled ATPase activity with either bicarbonate or acetophenone in the absence of the second substrate. These observations indicate that both substrates may be phosphorylated. Purified enzyme shows no activity with GTP, CTP, UTP or ITP Aromatoleum aromaticum 2 ADP + 2 phosphate + 3-oxo-3-phenylpropanoate
-
 ?
6.4.1.8 2 ATP + acetophenone + HCO3- + H2O + H+ anaerobic ethylbenzene metabolism in the betaproteobacterium Aromatoleum aromaticum is initiated by anaerobic oxidation to acetophenone via (S)-1-phenylethanol. The subsequent carboxylation of acetophenone to benzoylacetate is catalyzed by an acetophenone-induced enzyme Aromatoleum aromaticum EbN1 2 ADP + 2 phosphate + 3-oxo-3-phenylpropanoate
-
 ?
6.4.1.8 2 ATP + acetophenone + HCO3- + H2O + H+ the enzyme shows uncoupled ATPase activity with either bicarbonate or acetophenone in the absence of the second substrate. These observations indicate that both substrates may be phosphorylated. Purified enzyme shows no activity with GTP, CTP, UTP or ITP Aromatoleum aromaticum EbN1 2 ADP + 2 phosphate + 3-oxo-3-phenylpropanoate
-
 ?
6.4.1.8 2 ATP + propiophenone + HCO3- + H2O + H+ 108% of the activity with acetophenone Aromatoleum aromaticum 2 ADP + 2 phosphate + 4-oxo-4-phenylbutanoate
-
 ?
6.4.1.8 2 ATP + propiophenone + HCO3- + H2O + H+ 108% of the activity with acetophenone Aromatoleum aromaticum EbN1 2 ADP + 2 phosphate + 4-oxo-4-phenylbutanoate
-
 ?
6.4.1.8 additional information no activity with acetone, butanone, 4-hydroxy-acetophenone or 4-amino-acetophenone Aromatoleum aromaticum ?
-
 ?
6.4.1.8 additional information no activity with acetone, butanone, 4-hydroxy-acetophenone or 4-amino-acetophenone Aromatoleum aromaticum EbN1 ?
-
 ?

Subunits

EC Number Subunits Comment Organism
6.4.1.8 pentamer 1 * 70000 (alpha) + 1 * 15000 (beta) + 1 * 87000 (gamma) + 1 * 75000 (delta) + 1 * 34000 (epsilon), the genes (apcABCDE) of the five subunits form an apparent operon Aromatoleum aromaticum

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
6.4.1.8 30
-
 assay at Aromatoleum aromaticum

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
6.4.1.8 6.5
-
-
 Aromatoleum aromaticum

Expression

EC Number Organism Comment Expression
6.4.1.8 Aromatoleum aromaticum the enzyme is synthesized at high levels in cells grown on ethylbenzene or acetophenone, but not in cells grown on benzoate up