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Literature summary for 2.8.3.12 extracted from

  • Mack, M.; Buckel, W
    Conversion of glutaconate CoA-transferase from Acidaminococcus fermentans into an acyl-CoA hydrolase by site-directed mutagenesis (1997), FEBS Lett., 405, 209-212.
    View publication on PubMed

Cloned(Commentary)

Cloned (Comment) Organism
overexpression of betaE54N and betaE54A mutant enzyme in Escherichia coli, fusion of the genes gctA and gctB, encoding the 2 subunits of Gct, yields GctF, which is expressed in Escherichia coli as a 65 kDa protein with 30% of wild-type activity Acidaminococcus fermentans
overexpression of wild-type and betaE54D mutant enzyme in Escherichia coli Acidaminococcus fermentans

Crystallization (Commentary)

Crystallization (Comment) Organism
betaE54D mutant enzyme, expressed in Escherichia coli Acidaminococcus fermentans

Protein Variants

Protein Variants Comment Organism
E54A mutation in the subunit GctB, inactive mutant Acidaminococcus fermentans
E54A mutant without CoA-transferase or acyl-CoA hydrolase activity Acidaminococcus fermentans
E54D mutation in the subunit GctB Acidaminococcus fermentans
E54D mutation decreases CoA-transferase and increases acyl-CoA hydrolase activity, mechanism, mutant catalyses the hydrolysis of glutaryl-CoA, acetyl-CoA and 3-butenoyl-CoA, mutant Escherichia coli strain shows reduced growth rate after induction Acidaminococcus fermentans
E54N mutation in the subunit GctB, inactive mutant Acidaminococcus fermentans
E54N mutant without CoA-transferase or acyl-CoA hydrolase activity Acidaminococcus fermentans

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.028
-
glutaryl-CoA pH 7, cosubstrate acetate, wild-type enzyme Acidaminococcus fermentans
0.064
-
glutaryl-CoA pH 7, cosubstrate acetate, 65 kDa GctF fusion protein Acidaminococcus fermentans

Organism

Organism UniProt Comment Textmining
Acidaminococcus fermentans
-
-
-

Purification (Commentary)

Purification (Comment) Organism
betaE54D mutant enzyme and 65 kDa GctF fusion protein, expressed in Escherichia coli Acidaminococcus fermentans

Reaction

Reaction Comment Organism Reaction ID
acetyl-CoA + (E)-glutaconate = acetate + glutaconyl-1-CoA reaction mechanism Acidaminococcus fermentans

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
additional information
-
-
Acidaminococcus fermentans

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
acetyl-CoA + (E)-glutaconate
-
Acidaminococcus fermentans acetate + glutaconyl-1-CoA
-
?
glutaryl-CoA + acetate
-
Acidaminococcus fermentans glutarate + acetyl-CoA
-
?
additional information catalytic residue is E-54 of subunit B, catalytic mechanism Acidaminococcus fermentans ?
-
?

Subunits

Subunits Comment Organism
octamer (alphabeta)4 Acidaminococcus fermentans

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
additional information
-
additional information
-
Acidaminococcus fermentans
39
-
glutaryl-CoA pH 7, cosubstrate acetate, 65 kDa GctF fusion protein Acidaminococcus fermentans
140
-
glutaryl-CoA pH 7, cosubstrate acetate, wild-type enzyme Acidaminococcus fermentans

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
7
-
assay at Acidaminococcus fermentans