Cloned (Comment) | Organism |
---|---|
gene bphD, expression of wild-type and mutant BphDs | Paraburkholderia xenovorans |
Crystallization (Comment) | Organism |
---|---|
BphD H265Q and S112A/H265Q mutants in complex with substrate 2-hydroxy-6-oxo-6-phenylhexa-2,4-dienoic acid, sitting drop vapour diffusion method, 0.001 ml of protein solution containing 4 mg/ml protein in 20 mM HEPES, pH 7.5, 20°C, 9 days, is mixed with 0.001 ml of reservoir solution containing 2.4 M malonate at pH 6.0-7.0, X-ray diffraction structure determination and analysis at 1.3 A and 1.9 A resolutions, respectively | Paraburkholderia xenovorans |
Protein Variants | Comment | Organism |
---|---|---|
H265Q | site-directed mutagenesis, the mutant shows highly reduced activity compared to the wild-type enzyme and does not show formation of the carbanion catalytic intermediate in contrast to the wild-type enzyme | Paraburkholderia xenovorans |
S112A/H265Q | site-directed mutagenesis, the mutant shows highly reduced activity compared to the wild-type enzyme | Paraburkholderia xenovorans |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
additional information | - |
additional information | pre-steady-state kinetic burst of BphD | Paraburkholderia xenovorans |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
2-hydroxy-6-oxo-6-phenylhexa-2,4-dienoic acid + H2O | Paraburkholderia xenovorans | i.e. HOPDA | 2-hydroxypenta-2,4-dienoic acid + benzoate | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Paraburkholderia xenovorans | P47229 | gene bphD | - |
Reaction | Comment | Organism | Reaction ID |
---|---|---|---|
2,6-dioxo-6-phenylhexa-3-enoate + H2O = benzoate + 2-oxopent-4-enoate | formation of a catalytic intermediate carbanion during hydrolysis, the carbanion abstracts a proton from Ser112, thereby completing tautomerization and generating a serinate for nucleophilic attack on the C6-carbonyl, catalytic mechanism, overview. BphD is a half-site reactive enzyme with versatility of the Ser-His-Asp triad, role of the catalytic His in acylation and deacylation | 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 | 2-hydroxypenta-2,4-dienoic acid + benzoate | - |
? |
Synonyms | Comment | Organism |
---|---|---|
BphD | - |
Paraburkholderia xenovorans |
MCP hydrolase | - |
Paraburkholderia xenovorans |
meta-cleavage product hydrolase | - |
Paraburkholderia xenovorans |
General Information | Comment | Organism |
---|---|---|
evolution | meta-cleavage product (MCP) hydrolases are members of the alpha/beta-hydrolase superfamily that utilize a Ser-His-Asp triad to catalyze the hydrolysis of a C?C bond | Paraburkholderia xenovorans |
metabolism | BphD is the MCP hydrolase from the biphenyl degradation pathway and hydrolyzes 2-hydroxy-6-oxo-6-phenylhexa-2,4-dienoic acid (HOPDA) to 2-hydroxypenta-2,4-dienoic acid (HPD) and benzoate | Paraburkholderia xenovorans |