Cloned (Comment) | Organism |
---|---|
BphD mutants expressed in a pET-14b vector as an N-terminal His6 fusion protein | Paraburkholderia xenovorans |
Crystallization (Comment) | Organism |
---|---|
H263 mutant by hanging drop vapor diffusion, to 2 A resolution | Escherichia coli |
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 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 | UniProt | Comment | Textmining |
---|---|---|---|
Escherichia coli | - |
- |
- |
Paraburkholderia xenovorans | - |
LB400 | - |
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 | 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 | Comment | Organism |
---|---|---|
BphD | - |
Paraburkholderia xenovorans |
MhpC | - |
Escherichia coli |
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 |