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Literature summary for 3.7.1.8 extracted from
- Evidence for a gem-diol reaction intermediate in bacterial C-C hydrolase enzymes BphD and MhpC from 13C NMR spectroscopy (2006), Biochemistry, 45, 12461-12469.
Cloned(Commentary)
| Cloned (Comment) | Organism |
|---|---|
- |
Escherichia coli |
| into the pET-14b vector, overexpression in Escherichia coli BL21(DE3) cells | Paraburkholderia xenovorans |
Protein Variants
| Protein Variants | Comment | Organism |
|---|---|---|
| D237N | reduced activity | Paraburkholderia xenovorans |
| H114A | reduced activity, is able to accept the 6-phenyl-containing substrate, on a shorter time scale | Escherichia coli |
| H265A | reduced activity | Paraburkholderia xenovorans |
| S112A | reduced activity | Paraburkholderia xenovorans |
KM Value [mM]
| KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| 0.0017 | - |
ethyl-2-acetoxy-6-keto-6-phenylhexa-2,4-dienoate | mutant D237N | Paraburkholderia xenovorans |  |
| 0.002 | - |
ethyl-2-acetoxy-6-keto-6-phenylhexa-2,4-dienoate | His6-BphD | Paraburkholderia xenovorans | |
| 0.0238 | - |
ethyl-2-acetoxy-6-keto-6-phenylhexa-2,4-dienoate | mutant H114A | Escherichia coli | |
| 0.037 | - |
ethyl-2-acetoxy-6-keto-6-phenylhexa-2,4-dienoate | mutant H265A | Paraburkholderia xenovorans | |
| 0.0373 | - |
ethyl-2-acetoxy-6-keto-6-phenylhexa-2,4-dienoate | mutant S112A | Paraburkholderia xenovorans | |
Organism
| Organism | UniProt | Comment | Textmining |
|---|---|---|---|
| Escherichia coli | - |
- |
- |
| Paraburkholderia xenovorans | - |
LB400 | - |
Purification (Commentary)
| Purification (Comment) | Organism |
|---|---|
- |
Escherichia coli |
| by metal affinity chromatography, to more than 90% homogeneity | Paraburkholderia xenovorans |
Substrates and Products (Substrate)
| Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| ethyl-2-acetoxy-6-keto-6-phenylhexa-2,4-dienoate + H2O | - |
Escherichia coli | benzoate + ethyl 2-(acetyloxy)penta-2,4-dienoate | - |
? | |
| ethyl-2-acetoxy-6-keto-6-phenylhexa-2,4-dienoate + H2O | - |
Paraburkholderia xenovorans | benzoate + ethyl-2-(acetyloxy)penta-2,4-dienoate | - |
? | |
Synonyms
| Synonyms | Comment | Organism |
|---|---|---|
| BphD | - |
Paraburkholderia xenovorans |
| MhpC | - |
Escherichia coli |
Turnover Number [1/s]
| Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| 0.0009 | - |
ethyl-2-acetoxy-6-keto-6-phenylhexa-2,4-dienoate | mutant S112A | Paraburkholderia xenovorans | |
| 0.0016 | - |
ethyl-2-acetoxy-6-keto-6-phenylhexa-2,4-dienoate | mutant H265A | Paraburkholderia xenovorans | |
| 0.028 | - |
ethyl-2-acetoxy-6-keto-6-phenylhexa-2,4-dienoate | mutant D237N | Paraburkholderia xenovorans | |
| 0.036 | - |
ethyl-2-acetoxy-6-keto-6-phenylhexa-2,4-dienoate | mutant H114A | Escherichia coli | |
| 6.5 | - |
ethyl-2-acetoxy-6-keto-6-phenylhexa-2,4-dienoate | His6-BphD | Paraburkholderia xenovorans | |
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