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Literature summary for 6.4.1.8 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
Search for more literature for 6.4.1.8

Cloned(Commentary)

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

Inhibitors

Inhibitors Comment Organism Structure
beta,gamma-Imido-ATP 2 mM, complete inhibition of acetophenone carboxylase activity, non-competitive inhibition Aromatoleum aromaticum
Carbamoyl phosphate 2 mM: 50% inhibition of ATPase activity. 8 mM: 50% inhibition of carboxylating activity Aromatoleum aromaticum
Mg2+ 20 mM: 50% inhibition Aromatoleum aromaticum
Mn2+ 10 mM: 50% inhibition Aromatoleum aromaticum
additional information acetophenone carboxylation activity in cell extracts is not sensitive to avidin, indicating that the enzyme is independent of biotin Aromatoleum aromaticum
Ni2+ 1 mM: 50% inhibition of ATPase activity. 2.5 mM: 50% inhibition of carboxylation activity Aromatoleum aromaticum
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]

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

Metals/Ions

Metals/Ions Comment Organism Structure
K+ 20 mM, required for optimal activity Aromatoleum aromaticum
Mg2+ the enzyme is dependent on the presence of Mg2+ or Mn2+ Aromatoleum aromaticum
Mn2+ the enzyme is dependent on the presence of Mg2+ or Mn2+ Aromatoleum aromaticum
NH4+ 40 mM, required for optimal activity Aromatoleum aromaticum
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]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
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
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
40000
-
 apcE gene product, gel filtration Aromatoleum aromaticum
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
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
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
485000
-
 apcABCD gene products, gel filtration Aromatoleum aromaticum

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
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
-
 ?
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

Organism UniProt Comment Textmining
Aromatoleum aromaticum
-
-
-
Aromatoleum aromaticum EbN1
-
-
-

Purification (Commentary)

Purification (Comment) Organism
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]

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

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
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
-
 ?
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
-
 ?
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
-
 ?
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
-
 ?
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
-
 ?
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
-
 ?
2 ATP + propiophenone + HCO3- + H2O + H+ 108% of the activity with acetophenone Aromatoleum aromaticum 2 ADP + 2 phosphate + 4-oxo-4-phenylbutanoate
-
 ?
2 ATP + propiophenone + HCO3- + H2O + H+ 108% of the activity with acetophenone Aromatoleum aromaticum EbN1 2 ADP + 2 phosphate + 4-oxo-4-phenylbutanoate
-
 ?
additional information no activity with acetone, butanone, 4-hydroxy-acetophenone or 4-amino-acetophenone Aromatoleum aromaticum ?
-
 ?
additional information no activity with acetone, butanone, 4-hydroxy-acetophenone or 4-amino-acetophenone Aromatoleum aromaticum EbN1 ?
-
 ?

Subunits

Subunits Comment Organism
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]

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

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
6.5
-
-
 Aromatoleum aromaticum

Expression

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