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 (Comment) | Organism |
---|---|
betaE54D mutant enzyme, expressed in Escherichia coli | Acidaminococcus fermentans |
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 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 | UniProt | Comment | Textmining |
---|---|---|---|
Acidaminococcus fermentans | - |
- |
- |
Purification (Comment) | Organism |
---|---|
betaE54D mutant enzyme and 65 kDa GctF fusion protein, expressed in Escherichia coli | Acidaminococcus fermentans |
Reaction | Comment | Organism | Reaction ID |
---|---|---|---|
acetyl-CoA + (E)-glutaconate = acetate + glutaconyl-1-CoA | reaction mechanism | Acidaminococcus fermentans |
Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
---|---|---|---|
additional information | - |
- |
Acidaminococcus fermentans |
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 | Comment | Organism |
---|---|---|
octamer | (alphabeta)4 | Acidaminococcus fermentans |
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 Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7 | - |
assay at | Acidaminococcus fermentans |