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Literature summary for 3.7.1.8 extracted from
- Catalytic role for arginine 188 in the C-C hydrolase catalytic mechanism for Escherichia coli MhpC and Burkholderia xenovorans LB400 BphD (2006), Biochemistry, 45, 12470-12479.
Cloned(Commentary)
| Cloned (Comment) | Organism |
|---|---|
| BphD mutants expressed in a pET-14b vector as an N-terminal His6 fusion protein | Paraburkholderia xenovorans |
Crystallization (Commentary)
| Crystallization (Comment) | Organism |
|---|---|
| H263 mutant by hanging drop vapor diffusion, to 2 A resolution | Escherichia coli |
Protein Variants
| Protein Variants | Comment | Organism |
|---|---|---|
| C261A | 2fold decreased turnover rate, Cys-261 seems to be not involved in catalysis | Escherichia coli |
| F173D | 1.5fold increased Km value, 100fold decreased turnover rate | Escherichia coli |
| F173G | 8fold increased Km value, 3.5fold decreased turnover rate | Escherichia coli |
| H263A | overal structure similar, but asymmetry of the enzyme dimer more pronounced than for the native enzyme | Escherichia coli |
| N109A | similar Km value as wild-type, 200fold decreased turnover rate | Escherichia coli |
| N109H | similar Km value as wild-type, 350fold decreased turnover rate | Escherichia coli |
| R188K | 5fold increased Km value,35fold decreased turnover rate | Escherichia coli |
| R188Q | first step of enzyme reaction, keto-enol tautomerization, becomes rate-limiting, 11fold increased Km value, 300fold decreased turnover rate | Escherichia coli |
| R190K | similar Km value as wild-type, 700fold decreased turnover rate | Paraburkholderia xenovorans |
| R190Q | 14fold increased Km value, 400fold decreased turnover rate | Paraburkholderia xenovorans |
| W264G | 16fold increased Km value, 10fold decreased turnover rate | Escherichia coli |
KM Value [mM]
| KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| 0.002 | - |
meta-ring fission intermediate | wild-type | Paraburkholderia xenovorans | |
| 0.002 | - |
meta-ring fission intermediate | R188K | Paraburkholderia xenovorans | |
| 0.004 | - |
meta-ring fission intermediate | N109A | Escherichia coli | |
| 0.004 | - |
meta-ring fission intermediate | N109H | Escherichia coli | |
| 0.0042 | - |
meta-ring fission intermediate | C261A | Escherichia coli | |
| 0.0068 | - |
meta-ring fission intermediate | wild-type | Escherichia coli | |
| 0.01 | - |
meta-ring fission intermediate | F173D | Escherichia coli | |
| 0.0282 | - |
meta-ring fission intermediate | R188Q | Paraburkholderia xenovorans | |
| 0.038 | - |
meta-ring fission intermediate | R188K | Escherichia coli | |
| 0.058 | - |
meta-ring fission intermediate | F173G | Escherichia coli | |
| 0.077 | - |
meta-ring fission intermediate | R188Q | Escherichia coli |
Organism
| Organism | UniProt | Comment | Textmining |
|---|---|---|---|
| Escherichia coli | - |
- |
- |
| Paraburkholderia xenovorans | - |
LB400 | - |
Purification (Commentary)
| Purification (Comment) | Organism |
|---|---|
| BphD mutants purified by affinity chromatography to more than 95% homogeneity | Paraburkholderia xenovorans |
| MhpC mutants purified by anion exchange chromatography to more than 95% homogeneity | Escherichia coli |
Substrates and Products (Substrate)
| Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| meta-ring fission intermediate + H2O | catalysis of the hydrolytic C-C cleavage on biphenyl degradation pathway | Paraburkholderia xenovorans | ? | - |
? |  |
| meta-ring fission intermediate + H2O | catalysis of the hydrolytic C-C cleavage on phenylpropionic acid pathway | Escherichia coli | ? | - |
? | |
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 |
|---|---|---|---|---|---|
| 0.009 | - |
meta-ring fission intermediate | R188K | Paraburkholderia xenovorans | |
| 0.016 | - |
meta-ring fission intermediate | R188Q | Paraburkholderia xenovorans | |
| 0.08 | - |
meta-ring fission intermediate | N109H | Escherichia coli | |
| 0.1 | - |
meta-ring fission intermediate | R188Q | Escherichia coli | |
| 0.13 | - |
meta-ring fission intermediate | N109A | Escherichia coli | |
| 0.26 | - |
meta-ring fission intermediate | F173D | Escherichia coli | |
| 0.74 | - |
meta-ring fission intermediate | R188K | Escherichia coli | |
| 2 | 8 | meta-ring fission intermediate | wild-type | Escherichia coli | |
| 6.5 | - |
meta-ring fission intermediate | wild-type | Paraburkholderia xenovorans | |
| 7.5 | - |
meta-ring fission intermediate | F173G | Escherichia coli | |
| 12 | - |
meta-ring fission intermediate | C261A | Escherichia coli |
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