Activating Compound | Comment | Organism | Structure |
---|---|---|---|
BphJ | 15fold increased activity | Paraburkholderia xenovorans |
Cloned (Comment) | Organism |
---|---|
coexpression of bphI and bphJ in Escherichia coli using two compatible plasmids (pBTL4 and pET28a) yielded soluble proteins | Paraburkholderia xenovorans |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
Cd2+ | 56.0% activity at 0.1 mM and 5.4% activity at 1 mM chloride salt | Paraburkholderia xenovorans | |
Co2+ | 19.3% activity at 0.1 mM and 18.3% activity at 1 mM chloride salt | Paraburkholderia xenovorans | |
Mg2+ | 3.4% activity at 0.1 mM and 14.8% activity at 1 mM chloride salt | Paraburkholderia xenovorans | |
Mn2+ | 100% activity at 0.1 mM and 1 mM chloride salt | Paraburkholderia xenovorans | |
Ni2+ | 2.6% activity at 0.1 mM and 17.5% activity at 1 mM chloride salt | Paraburkholderia xenovorans | |
oxalate | - |
Paraburkholderia xenovorans |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
additional information | - |
additional information | presence of NAD+, NADP+, NADH or CoA leads to an at least 5fold reduction in the Km for 4-hydroxy-2-oxopentanoate | Paraburkholderia xenovorans | |
0.18 | - |
4-hydroxy-2-oxohexanoate | pH 8.0, 25°C | Paraburkholderia xenovorans | |
0.22 | - |
4-hydroxy-2-oxopentanoate | pH 8.0, 25°C | Paraburkholderia xenovorans | |
0.35 | - |
4-hydroxy-2-oxoheptanoate | pH 8.0, 25°C | Paraburkholderia xenovorans | |
7.5 | - |
oxaloacetate | pH 8.0, 25°C | Paraburkholderia xenovorans |
Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|
32000 | - |
BphI-BphJ-complex, alpha2beta2, 2 * 32000 BphJ, 2 * 37000 BphI | Paraburkholderia xenovorans |
37000 | - |
SDS-PAGE, in agreement with the predicted molecular mass calculated from amino acid sequence | Paraburkholderia xenovorans |
140000 | - |
native molecular mass of the purified BphI-BphJ-complex, estimated by gel filtration | Paraburkholderia xenovorans |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
(S)-4-hydroxy-2-oxohexanoate | Paraburkholderia xenovorans | - |
propanal + pyruvate | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Paraburkholderia xenovorans | P51015 | - |
- |
Purification (Comment) | Organism |
---|---|
BphI and BphJ form a stable complex as they bind and coelute from Ni2+-NTA column, although only BphJ has the histidine tag. After purification, the N-terminal histidine tag of BphJ is proteolytically cleaved by thrombin digestion | Paraburkholderia xenovorans |
Storage Stability | Organism |
---|---|
purified enzyme can be stored at -80°C, without loss of activity for at least 12 months | Paraburkholderia xenovorans |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
(S)-4-hydroxy-2-oxohexanoate | - |
Paraburkholderia xenovorans | propanal + pyruvate | - |
? | |
4-hydroxy-2-oxoheptanoate | - |
Paraburkholderia xenovorans | pyruvate + butanal | - |
? | |
4-hydroxy-2-oxohexanoate | - |
Paraburkholderia xenovorans | pyruvate + propionaldehyde | - |
? | |
4-hydroxy-2-oxopentanoate | - |
Paraburkholderia xenovorans | pyruvate + acetaldehyde | - |
? | |
Oxaloacetate | - |
Paraburkholderia xenovorans | Pyruvate + CO2 | - |
? |
Subunits | Comment | Organism |
---|---|---|
heterotetramer | BphI-BphJ-complex, alpha2beta2, 2 * 32000 BphJ, 2 * 37000 BphI | Paraburkholderia xenovorans |
Synonyms | Comment | Organism |
---|---|---|
BphI | - |
Paraburkholderia xenovorans |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
additional information | - |
additional information | 2 and 5fold increases in kcat observed when BphI reaction is conducted in the presence of NAD+ and NADH | Paraburkholderia xenovorans | |
0.63 | - |
4-hydroxy-2-oxoheptanoate | pH 8.0, 25°C | Paraburkholderia xenovorans | |
3.8 | - |
4-hydroxy-2-oxohexanoate | pH 8.0, 25°C | Paraburkholderia xenovorans | |
4.1 | - |
4-hydroxy-2-oxopentanoate | pH 8.0, 25°C | Paraburkholderia xenovorans | |
16.5 | - |
oxaloacetate | pH 8.0, 25°C | Paraburkholderia xenovorans |
Ki Value [mM] | Ki Value maximum [mM] | Inhibitor | Comment | Organism | Structure |
---|---|---|---|---|---|
0.00093 | - |
oxalate | pH 8.0, 25°C | Paraburkholderia xenovorans |
General Information | Comment | Organism |
---|---|---|
metabolism | enzyme forms a bifunctional complex with EC 1.2.1.87 (BphJ), propanal dehydrogenase (CoA-propanoylating) and is part of the polychlorinated biphenyl degradation pathway | Paraburkholderia xenovorans |
kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
additional information | - |
additional information | highest catalytic efficiency of BphI occurs when NADH is present, which is 25fold higher than the value obtained without nucleotides | Paraburkholderia xenovorans | |
1.8 | - |
4-hydroxy-2-oxoheptanoate | pH 8.0, 25°C | Paraburkholderia xenovorans | |
2.2 | - |
oxaloacetate | pH 8.0, 25°C | Paraburkholderia xenovorans | |
18.6 | - |
4-hydroxy-2-oxopentanoate | pH 8.0, 25°C | Paraburkholderia xenovorans | |
21.1 | - |
4-hydroxy-2-oxohexanoate | pH 8.0, 25°C | Paraburkholderia xenovorans |