Cloned (Comment) | Organism |
---|---|
overexpression of wild-type and betaE54D mutant enzyme in Escherichia coli | Acidaminococcus fermentans |
Protein Variants | Comment | Organism |
---|---|---|
E54A | mutation in the subunit GctB, inactive mutant | Acidaminococcus fermentans |
E54D | mutation in the subunit GctB | Acidaminococcus fermentans |
E54D | replacement of the catalytic glutamate by aspartate converts the mutant enzyme to a thiol ester hydrolase | Acidaminococcus fermentans |
E54N | mutation in the subunit GctB, inactive mutant | Acidaminococcus fermentans |
E54Q | mutation in the subunit GctB | Acidaminococcus fermentans |
E54Q | 1% as active as wild-type enzyme, by incubating with both substrates for 20 h at room temperature, glutamine is completely converted to glutamate yielding a fully active CoA-transferase | Acidaminococcus fermentans |
Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|
29018 | - |
alpha4beta4, 4 * 35573 + 4 * 29018, MALDI-TOF mass spectrometry | Acidaminococcus fermentans |
35573 | - |
alpha4beta4, 4 * 35573 + 4 * 29018, MALDI-TOF mass spectrometry | Acidaminococcus fermentans |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
acetyl-CoA + (R)-2-hydroxyglutarate | Acidaminococcus fermentans | in the course of glutamate fermentation | acetate + (R)-2-hydroxyglutaryl-CoA | - |
r |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Acidaminococcus fermentans | - |
- |
- |
Purification (Comment) | Organism |
---|---|
recombinant wild-type and betaE54D mutant enzyme, expressed in Escherichia coli | Acidaminococcus fermentans |
Reaction | Comment | Organism | Reaction ID |
---|---|---|---|
acetyl-CoA + (E)-glutaconate = acetate + glutaconyl-1-CoA | reaction mechanism | Acidaminococcus fermentans |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
acetyl-CoA + (R)-2-hydroxyglutarate | - |
Acidaminococcus fermentans | acetate + (R)-2-hydroxyglutaryl-1-CoA | - |
r | |
acetyl-CoA + (R)-2-hydroxyglutarate | in the course of glutamate fermentation | Acidaminococcus fermentans | acetate + (R)-2-hydroxyglutaryl-CoA | - |
r | |
glutaryl-CoA + acetate | exchange of oxygen atoms between the substrates and the catalytic residue betaE-54, exchange is site-specific, reversible and requires both substrates, catalytic mechanism | Acidaminococcus fermentans | glutarate + acetyl-CoA | - |
r | |
additional information | wild-type enzyme has significant hydrolase activity using acetyl-CoA as substrate, but no hydrolysis of glutaryl-CoA | Acidaminococcus fermentans | ? | - |
? | |
additional information | catalytic residue is E-54 of subunit B, catalytic mechanism | Acidaminococcus fermentans | ? | - |
? |
Subunits | Comment | Organism |
---|---|---|
octamer | alpha4beta4, 4 * 35573 + 4 * 29018, MALDI-TOF mass spectrometry | Acidaminococcus fermentans |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
25 | - |
assay at, hydrolase activity | Acidaminococcus fermentans |
37 | - |
assay at, CoA-transferase activity | Acidaminococcus fermentans |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7 | - |
assay at | Acidaminococcus fermentans |