Cloned (Comment) | Organism |
---|---|
overexpression of wild-type and mutant BphDs in Escherichia coli strain Rosetta(DE3)pLysS | Paraburkholderia xenovorans |
Protein Variants | Comment | Organism |
---|---|---|
H265A | site-directed mutagenesis, the mutant shows highly reduced activity compared to the wild-type enzyme | Paraburkholderia xenovorans |
H265Q | site-directed mutagenesis, the mutant shows highly reduced activity compared to the wild-type enzyme, rapid acylation of the variant during C-C bond cleavage suggesting that the serinate forms via a substrate-assisted mechanism in the reaction | Paraburkholderia xenovorans |
S112A | site-directed mutagenesis, the mutant shows highly reduced activity compared to the wild-type enzyme | Paraburkholderia xenovorans |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
ethanol | - |
Paraburkholderia xenovorans | |
methanol | - |
Paraburkholderia xenovorans | |
additional information | no effects by n-propanol and 2-propanol | Paraburkholderia xenovorans |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
additional information | - |
additional information | in the steady-state hydrolysis of HOPDA, kcat/Km values are independent of methanol concentration, while both kcat and Km values increase with methanol concentration, stopped-flow spectrophotometry and steady-state kinetics, overview | Paraburkholderia xenovorans | |
0.0002 | - |
2-Hydroxy-6-oxo-6-phenylhexa-2,4-dienoic acid | pH 7.5, 25°C, recombinant wild-type BphD, without methanol | Paraburkholderia xenovorans | |
0.00085 | - |
2-Hydroxy-6-oxo-6-phenylhexa-2,4-dienoic acid | pH 7.5, 25°C, recombinant wild-type BphD, with 494 mM methanol | Paraburkholderia xenovorans | |
0.002 | - |
2-Hydroxy-6-oxo-6-phenylhexa-2,4-dienoic acid | pH 7.5, 25°C, recombinant wild-type BphD, with 12940 mM methanol | Paraburkholderia xenovorans |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Paraburkholderia xenovorans | - |
gene bphD | - |
Reaction | Comment | Organism | Reaction ID |
---|---|---|---|
2,6-dioxo-6-phenylhexa-3-enoate + H2O = benzoate + 2-oxopent-4-enoate | general base and nucleophilic catalytic reaction mechanisms, overview | Paraburkholderia xenovorans |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
2-hydroxy-6-oxo-6-phenylhexa-2,4-dienoic acid + H2O | i.e. HOPDA | Paraburkholderia xenovorans | ? | - |
? | |
4-nitrophenyl benzoate + H2O | the serine nucleophile is activated by the His-Asp dyad | Paraburkholderia xenovorans | 4-nitrophenol + benzoate | - |
? |
Synonyms | Comment | Organism |
---|---|---|
BphD | - |
Paraburkholderia xenovorans |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
25 | - |
assay at | Paraburkholderia xenovorans |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
additional information | - |
additional information | in the steady-state hydrolysis of HOPDA, kcat/Km values are independent of methanol concentration, while both kcat and Km values increase with methanol concentration, stopped-flow spectrophotometry and steady-state kinetics, overview | Paraburkholderia xenovorans | |
2 | 8 | 2-Hydroxy-6-oxo-6-phenylhexa-2,4-dienoic acid | pH 7.5, 25°C, recombinant wild-type BphD, with 12940 mM methanol | Paraburkholderia xenovorans | |
6.3 | - |
4-nitrophenyl benzoate | pH 7.5, 25°C | Paraburkholderia xenovorans | |
6.5 | - |
2-Hydroxy-6-oxo-6-phenylhexa-2,4-dienoic acid | pH 7.5, 25°C, recombinant wild-type BphD, without methanol | Paraburkholderia xenovorans | |
26 | - |
2-Hydroxy-6-oxo-6-phenylhexa-2,4-dienoic acid | pH 7.5, 25°C, recombinant wild-type BphD, with 494 mM methanol | Paraburkholderia xenovorans |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7.5 | - |
assay at | Paraburkholderia xenovorans |
General Information | Comment | Organism |
---|---|---|
malfunction | rapid acylation of the H265Q variant during C-C bond cleavage suggests that the serinate forms via a substrate-assisted mechanism in the reaction | Paraburkholderia xenovorans |
metabolism | BphD is a meta-cleavage product (MCP) hydrolase which catalyzes C-C bond fission in the aerobic catabolism of aromatic compounds by bacteria utilizing a Ser-His-Asp triad to catalyze hydrolysis via an acyl-enzyme intermediate | Paraburkholderia xenovorans |
kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
14 | - |
2-Hydroxy-6-oxo-6-phenylhexa-2,4-dienoic acid | pH 7.5, 25°C, recombinant wild-type BphD, with 12940 mM methanol | Paraburkholderia xenovorans | |
31 | - |
2-Hydroxy-6-oxo-6-phenylhexa-2,4-dienoic acid | pH 7.5, 25°C, recombinant wild-type BphD, with 494 mM methanol | Paraburkholderia xenovorans | |
32 | - |
2-Hydroxy-6-oxo-6-phenylhexa-2,4-dienoic acid | pH 7.5, 25°C, recombinant wild-type BphD, without methanol | Paraburkholderia xenovorans |