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Literature summary for 3.5.1.4 extracted from
- Identification of active sites in amidase: Evolutionary relationship between amide bond- and peptide bond-cleaving enzymes (1997), Proc. Natl. Acad. Sci. USA, 94, 11987-11991.
Application
| Application | Comment | Organism |
|---|---|---|
| synthesis | - |
Rhodococcus rhodochrous |
KM Value [mM]
| KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| 0.15 | - |
Benzamide | purified wild-type | Escherichia coli |  |
| 0.16 | - |
Benzamide | purified C203A mutant | Escherichia coli | |
| 0.68 | - |
Benzamide | purified D191E mutant | Escherichia coli | |
Organism
| Organism | UniProt | Comment | Textmining |
|---|---|---|---|
| Escherichia coli | - |
harboring pALJ30 and pALJ20 | - |
| Escherichia coli JM109 | - |
harboring pALJ30 and pALJ20 | - |
| Rhodococcus rhodochrous | - |
- |
- |
| Rhodococcus rhodochrous J1 | - |
- |
- |
Purification (Commentary)
| Purification (Comment) | Organism |
|---|---|
| a. transformant : sonication, lysis, dialysis against urea, b. wild-type amidase: dialysis, Mono-Q HR 5/5 column, ammonium sulfate precipitation, dialysis | Escherichia coli |
| dialysis, column chromatography and ammonium sulfate fractionation | Rhodococcus rhodochrous |
Specific Activity [micromol/min/mg]
| Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
|---|---|---|---|
| 0.0021 | - |
D191E mutant, supernatant of cell-free extract, 0.75% of wild-type activity | Escherichia coli |
| 0.0306 | - |
C203A mutant, supernatant of cell-free extract, 10.8% of wild-type activity | Escherichia coli |
| 0.162 | - |
D191E mutant, purified, 1.33% of purified wild-type amidase | Escherichia coli |
| 0.283 | - |
wild-type amidase | Escherichia coli |
| 1.4 | - |
C203Amutant, purified, 11.5% of purified wild-type amidase | Escherichia coli |
| 11.6 | - |
wild-type amidase after purification | Escherichia coli |
| 12.2 | - |
purified from a soluble fraction | Rhodococcus rhodochrous |
| 12.2 | - |
wild-type amidase, purified | Rhodococcus rhodochrous |
Substrates and Products (Substrate)
| Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| benzamide + H2O | - |
Escherichia coli | benzoic acid + NH3 | - |
? | |
| benzamide + H2O | - |
Escherichia coli JM109 | benzoic acid + NH3 | - |
? | |
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