Literature summary extracted from
Ruzzini, A.C.; Ghosh, S.; Horsman, G.P.; Foster, L.J.; Bolin, J.T.; Eltis, L.D.
Identification of an acyl-enzyme intermediate in a meta-cleavage product hydrolase reveals the versatility of the catalytic triad (2012), J. Am. Chem. Soc., 134, 4615-4624.
Cloned(Commentary)
EC Number |
Cloned (Comment) |
Organism |
---|
3.7.1.8 |
gene bphD, expression of wild-type and mutant BphDs |
Paraburkholderia xenovorans |
Crystallization (Commentary)
EC Number |
Crystallization (Comment) |
Organism |
---|
3.7.1.8 |
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
EC Number |
Protein Variants |
Comment |
Organism |
---|
3.7.1.8 |
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 |
3.7.1.8 |
S112A/H265Q |
site-directed mutagenesis, the mutant shows highly reduced activity compared to the wild-type enzyme |
Paraburkholderia xenovorans |
KM Value [mM]
EC Number |
KM Value [mM] |
KM Value Maximum [mM] |
Substrate |
Comment |
Organism |
Structure |
---|
3.7.1.8 |
additional information |
- |
additional information |
pre-steady-state kinetic burst of BphD |
Paraburkholderia xenovorans |
|
Natural Substrates/ Products (Substrates)
EC Number |
Natural Substrates |
Organism |
Comment (Nat. Sub.) |
Natural Products |
Comment (Nat. Pro.) |
Rev. |
Reac. |
---|
3.7.1.8 |
2-hydroxy-6-oxo-6-phenylhexa-2,4-dienoic acid + H2O |
Paraburkholderia xenovorans |
i.e. HOPDA |
2-hydroxypenta-2,4-dienoic acid + benzoate |
- |
? |
|
Organism
EC Number |
Organism |
UniProt |
Comment |
Textmining |
---|
3.7.1.8 |
Paraburkholderia xenovorans |
P47229 |
gene bphD |
- |
Reaction
EC Number |
Reaction |
Comment |
Organism |
Reaction ID |
---|
3.7.1.8 |
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 and Products (Substrate)
EC Number |
Substrates |
Comment Substrates |
Organism |
Products |
Comment (Products) |
Rev. |
Reac. |
---|
3.7.1.8 |
2-hydroxy-6-oxo-6-phenylhexa-2,4-dienoic acid + H2O |
i.e. HOPDA |
Paraburkholderia xenovorans |
2-hydroxypenta-2,4-dienoic acid + benzoate |
- |
? |
|
Synonyms
EC Number |
Synonyms |
Comment |
Organism |
---|
3.7.1.8 |
BphD |
- |
Paraburkholderia xenovorans |
3.7.1.8 |
MCP hydrolase |
- |
Paraburkholderia xenovorans |
3.7.1.8 |
meta-cleavage product hydrolase |
- |
Paraburkholderia xenovorans |
General Information
EC Number |
General Information |
Comment |
Organism |
---|
3.7.1.8 |
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 |
3.7.1.8 |
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 |