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Literature summary extracted from

  • Gencic, S.; Duin, E.C.; Grahame, D.A.
    Tight coupling of partial reactions in the cc from Methanosarcina thermophila: acetyl C-C bond fragmentation at the a cluster promoted by protein conformational changes (2010), J. Biol. Chem., 285, 15450-15463.
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

EC Number Cloned (Comment) Organism
2.3.1.169 expression of ACSD beta in Escherichia coli strain NM522 Methanosarcina thermophila
2.3.1.169 gene ascB, expression of full-length protein, designated ACSCh732 amino acids, and as a form lacking the 317-amino acid N-terminal domain, designated ACSChDELTAN, 415 amino acids, in Escherichia coli strain NM522 Carboxydothermus hydrogenoformans

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
2.3.1.169 additional information
-
 additional information kinetics of recombinant ACSCh732 and ACSChDELTAN for acetyl-CoA synthase activity, acetyltransferase activity, and acetyl-CoA/CO exchange activity, overview Carboxydothermus hydrogenoformans
2.3.1.169 additional information
-
 additional information kinetics of recombinant ACSD beta for acetyl-CoA synthase activity, acetyltransferase activity, and acetyl-CoA/CO exchange activity, overview Methanosarcina thermophila
2.3.1.169 0.066
-
 methylcorrinoid protein pH 7.2, 25°C, recombinant ACSChDELTAN Carboxydothermus hydrogenoformans
2.3.1.169 0.087
-
 CO pH 7.2, 25°C, recombinant ACSD beta Methanosarcina thermophila
2.3.1.169 0.36
-
 CO pH 7.2, 25°C, recombinant ACSChDELTAN Carboxydothermus hydrogenoformans
2.3.1.169 0.46
-
 methylcorrinoid protein pH 7.2, 25°C, recombinant ACSCh732 Carboxydothermus hydrogenoformans
2.3.1.169 0.53
-
 methylcorrinoid protein pH 7.2, 25°C, recombinant ACSD beta Methanosarcina thermophila

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
2.3.1.169 Fe2+ a [Fe4S4] cluster Methanosarcina thermophila
2.3.1.169 Fe2+ Fe/S-containing active site metal center, the A cluster Carboxydothermus hydrogenoformans
2.3.1.169 Ni2+ nickel-containing active site metal center, the A cluster, a binuclear Ni-Ni center bridged by a cysteine thiolate to an [Fe4S4] cluster. Ni2+-CO equatorial coordination environment in closed buried hydrophobic and open solvent-exposed states Methanosarcina thermophila

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
2.3.1.169 acetyl-CoA + corrinoid protein Methanosarcina thermophila
-
 CoA + CO + methylcorrinoid protein
-
 r
2.3.1.169 acetyl-CoA + corrinoid protein Carboxydothermus hydrogenoformans
-
 CoA + CO + methylcorrinoid protein
-
 r
2.3.1.169 acetyl-CoA + corrinoid protein Carboxydothermus hydrogenoformans DSM 6008
-
 CoA + CO + methylcorrinoid protein
-
 r
2.3.1.169 acetyl-CoA + corrinoid protein Methanosarcina thermophila TM-1
-
 CoA + CO + methylcorrinoid protein
-
 r

Organism

EC Number Organism UniProt Comment Textmining
2.3.1.169 Carboxydothermus hydrogenoformans
-
 gene acsB
-
2.3.1.169 Carboxydothermus hydrogenoformans DSM 6008
-
 gene acsB
-
2.3.1.169 Methanosarcina thermophila
-
-
-
2.3.1.169 Methanosarcina thermophila TM-1
-
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
2.3.1.169 recombinant ACSCh732 and ACSChDELTAN from Escherichia coli strain NM522 by anion exchange and hydroxyapatite chromatography, followed by another and different step of anion exchange chromatography and by hydrophobic interaction chromatography to 94-95% purity forACSCh and around 98% purity for ACSChDELTAN Carboxydothermus hydrogenoformans
2.3.1.169 recombinant ACSD beta from Escherichia coli strain NM522 by anion exchange chromatography, followed by hydrophobic interaction chromatography and another and different step of ion exchange of anion exchange chromatography to around 98% purity for ACDS beta Methanosarcina thermophila

Reaction

EC Number Reaction Comment Organism Reaction ID
2.3.1.169 acetyl-CoA + a [Co(I) corrinoid Fe-S protein] = CO + CoA + a [methyl-Co(III) corrinoid Fe-S protein] chemical steps and conformational changes in the mechanism of acetyl-CoA synthase, overview Methanosarcina thermophila
a
2.3.1.169 acetyl-CoA + a [Co(I) corrinoid Fe-S protein] = CO + CoA + a [methyl-Co(III) corrinoid Fe-S protein] chemical steps and conformational changes in the mechanism of acetyl-CoA synthase, overview Carboxydothermus hydrogenoformans
a

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2.3.1.169 acetyl-CoA + corrinoid protein
-
 Methanosarcina thermophila CoA + CO + methylcorrinoid protein
-
 r
2.3.1.169 acetyl-CoA + corrinoid protein
-
 Carboxydothermus hydrogenoformans CoA + CO + methylcorrinoid protein
-
 r
2.3.1.169 acetyl-CoA + corrinoid protein
-
 Carboxydothermus hydrogenoformans DSM 6008 CoA + CO + methylcorrinoid protein
-
 r
2.3.1.169 acetyl-CoA + corrinoid protein
-
 Methanosarcina thermophila TM-1 CoA + CO + methylcorrinoid protein
-
 r
2.3.1.169 additional information an Fe/S-containing active site metal center, the A cluster, catalyzes acetyl C–C bond formation/breakdown. Carbonyl group exchange of acetyl-CoA with CO is a hallmark of CODH/ACS, coupling analysis of the recombinant A cluster protein of acetyl-CoA synthase of Carboxydothermus hydrogenoformans, ACSCh, and truncated ACSCh lacking its 317-amino acid N-terminal domain, overview Carboxydothermus hydrogenoformans ?
-
 ?
2.3.1.169 additional information an nickel-containing active site metal center, the A cluster, catalyzes acetyl C-C bond formation/breakdown. Carbonyl group exchange of acetyl-CoA with CO is weakly active in ACDS, and exchange with CO2 is up to 350 times faster, indicating tight coupling of CO release at the A cluster to CO oxidation to CO2 at the C cluster in CO dehydrogenase, coupling analysis of the recombinant A cluster protein of ACDS. Direct role of the ACS N-terminal domain in promoting acetyl C-C bond fragmentation. Protein conformational changes, related to open/closed states have direct effects on the coordination geometry and stability of the A cluster Ni2+-acetyl intermediate, controlling Ni2-acetyl fragmentation and Ni2(CO)(CH3) condensation. Involvement of subunit-subunit interactions in ACDS, versus interdomain contacts in ACS, ensures that CO is not released from the ACDS beta-subunit in the absence of appropriate interactions with the alpha2epsilon2 CO dehydrogenase component, ACDS complex partial reactions in the overall synthesis and cleavage of acetyl-CoA, overview Methanosarcina thermophila ?
-
 ?
2.3.1.169 additional information an Fe/S-containing active site metal center, the A cluster, catalyzes acetyl C–C bond formation/breakdown. Carbonyl group exchange of acetyl-CoA with CO is a hallmark of CODH/ACS, coupling analysis of the recombinant A cluster protein of acetyl-CoA synthase of Carboxydothermus hydrogenoformans, ACSCh, and truncated ACSCh lacking its 317-amino acid N-terminal domain, overview Carboxydothermus hydrogenoformans DSM 6008 ?
-
 ?
2.3.1.169 additional information an nickel-containing active site metal center, the A cluster, catalyzes acetyl C-C bond formation/breakdown. Carbonyl group exchange of acetyl-CoA with CO is weakly active in ACDS, and exchange with CO2 is up to 350 times faster, indicating tight coupling of CO release at the A cluster to CO oxidation to CO2 at the C cluster in CO dehydrogenase, coupling analysis of the recombinant A cluster protein of ACDS. Direct role of the ACS N-terminal domain in promoting acetyl C-C bond fragmentation. Protein conformational changes, related to open/closed states have direct effects on the coordination geometry and stability of the A cluster Ni2+-acetyl intermediate, controlling Ni2-acetyl fragmentation and Ni2(CO)(CH3) condensation. Involvement of subunit-subunit interactions in ACDS, versus interdomain contacts in ACS, ensures that CO is not released from the ACDS beta-subunit in the absence of appropriate interactions with the alpha2epsilon2 CO dehydrogenase component, ACDS complex partial reactions in the overall synthesis and cleavage of acetyl-CoA, overview Methanosarcina thermophila TM-1 ?
-
 ?

Subunits

EC Number Subunits Comment Organism
2.3.1.169 More open and closed conformations of ACS, overview Methanosarcina thermophila

Synonyms

EC Number Synonyms Comment Organism
2.3.1.169 ACDS multienzyme complex
-
 Methanosarcina thermophila
2.3.1.169 acetyl-CoA decarbonylase/synthase multienzyme complex
-
 Methanosarcina thermophila
2.3.1.169 CO dehydrogenase/acetyl-CoA synthase
-
 Carboxydothermus hydrogenoformans
2.3.1.169 CODH/ACS
-
 Carboxydothermus hydrogenoformans

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
2.3.1.169 25
-
 assay at Methanosarcina thermophila
2.3.1.169 25
-
 assay at Carboxydothermus hydrogenoformans

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
2.3.1.169 0.00012
-
 acetyl-CoA pH 7.2, 25°C, recombinant ACSChDELTAN, acetyl-CoA/CO exchange activity Carboxydothermus hydrogenoformans
2.3.1.169 0.0002
-
 acetyl-CoA pH 7.2, 25°C, recombinant ACSD beta, acetyl-CoA/CO exchange activity Methanosarcina thermophila
2.3.1.169 0.142
-
 acetyl-CoA pH 7.2, 25°C, recombinant ACSCh732, acetyl-CoA/CO exchange activity Carboxydothermus hydrogenoformans

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
2.3.1.169 6.7
-
 acetyltransferase assay at Carboxydothermus hydrogenoformans
2.3.1.169 6.7 7.2 assay at Methanosarcina thermophila
2.3.1.169 7.2
-
 acetyl-CoA synthase assay at Carboxydothermus hydrogenoformans

General Information

EC Number General Information Comment Organism
2.3.1.169 evolution comparison of bifunctional CO dehydrogenase/acetyl-CoA synthase enzyme from anaerobic bacteria and of the acetyl-CoA decarbonylase/synthase (ACDS) multienzyme complex from Archaea, and of the role of the ACS N-terminal domain in promoting acetyl C-C bond fragmentation at the A cluster, overview Methanosarcina thermophila
2.3.1.169 evolution comparison of bifunctional CO dehydrogenase/acetyl-CoA synthase enzyme from anaerobic bacteria and of the acetyl-CoA decarbonylase/synthase (ACDS) multienzyme complex from Archaea, and of the role of the ACS N-terminal domain in promoting acetyl C-C bond fragmentation at the A cluster, overview Carboxydothermus hydrogenoformans
2.3.1.169 malfunction in the ACSChDLETAN truncation mutant, the Km value is decreased to about one-seventh of its value in the full-length protein, and the Vmax value is increased by a factor of around 4.4. Overall, the Vmax/Km ratio increases by around 30fold, indicating an apparent unmasking of the intrinsic catalytic efficiency for overall synthesis of acetyl-CoA. Changes in the kinetics of acetyl-CoA synthesis are possibly due to differences in CO accessibility to the A cluster in different forms of the enzyme Carboxydothermus hydrogenoformans
2.3.1.169 additional information open and closed conformations of ACS, overview Methanosarcina thermophila
2.3.1.169 physiological function direct synthesis and cleavage of acetyl-CoA are carried out by the acetyl-CoA decarbonylase/synthase, ACDS, multienzyme complex in Archaea Methanosarcina thermophila
2.3.1.169 physiological function direct synthesis and cleavage of acetyl-CoA are carried out by the bifunctional CO dehydrogenase/acetyl-CoA synthase enzyme in anaerobic bacteria Carboxydothermus hydrogenoformans