Cloned (Comment) | Organism |
---|---|
coexpression of bphI and bphJ in Escherichia coli using two compatible plasmids (pBTL4 and pET28a) yield soluble proteins | Paraburkholderia xenovorans |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.0348 | - |
NAD+ | app. Km-value with acetaldehyde as substrate, pH 8.0 and 25°C | Paraburkholderia xenovorans | |
0.561 | - |
NADP+ | app. Km-value with acetaldehyde as substrate, pH 8.0 and 25°C | Paraburkholderia xenovorans | |
7.7 | - |
Isobutyraldehyde | app. Km-value, pH 8.0 and 25°C, NAD+ | Paraburkholderia xenovorans | |
23.1 | - |
propionaldehyde | app. Km-value, pH 8.0 and 25°C, NAD+ | Paraburkholderia xenovorans | |
23.6 | - |
acetaldehyde | app. Km-value, pH 8.0 and 25°C, NAD+ | Paraburkholderia xenovorans | |
31.7 | - |
Butyraldehyde | app. Km-value, pH 8.0 and 25°C, NAD+ | Paraburkholderia xenovorans |
Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|
32000 | - |
SDS-PAGE, in agreement with the predicted molecular mass calculated from amino acid sequence | Paraburkholderia xenovorans |
37000 | - |
BphI-BphJ-complex, alpha2beta2, 2 * 32000 BphJ, 2 * 37000 BphI, estimated by gel filtration | 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. |
---|---|---|---|---|---|---|
propanal + CoA + NAD+ | Paraburkholderia xenovorans | - |
propanoyl-CoA + NADH + H+ | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Paraburkholderia xenovorans | Q79AF6 | - |
- |
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. |
---|---|---|---|---|---|---|
acetaldehyde + CoA + NAD+ | - |
Paraburkholderia xenovorans | acetyl-CoA + NADH + H+ | - |
? | |
acetaldehyde + CoA + NADP+ | BphJ is able to utilize NAD+ and NADP+ with comparable kcat values, but the apparent Km-value for NAD+ is 16fold lower than for NADP+ | Paraburkholderia xenovorans | acetyl-CoA + NADPH + H+ | - |
? | |
butyraldehyde + CoA + NAD+ | - |
Paraburkholderia xenovorans | butyryl-CoA + NADH + H+ | - |
? | |
isobutyraldehyde + CoA + NAD+ | - |
Paraburkholderia xenovorans | isobutyryl-CoA + NADH + H+ | - |
? | |
additional information | no activity with succinic semialdehyde and picolinaldehyde | Paraburkholderia xenovorans | ? | - |
? | |
pentaldehyde + CoA + NAD+ | - |
Paraburkholderia xenovorans | pentanoyl-CoA + NADH + H+ | - |
? | |
propanal + CoA + NAD+ | - |
Paraburkholderia xenovorans | propanoyl-CoA + NADH + H+ | - |
? | |
propionaldehyde + CoA + NAD+ | - |
Paraburkholderia xenovorans | propanoyl-CoA + NADH + H+ | - |
? |
Subunits | Comment | Organism |
---|---|---|
heterotetramer | BphI-BphJ-complex, alpha2beta2, 2 * 32000 BphJ, 2 * 37000 BphI, estimated by gel filtration | Paraburkholderia xenovorans |
Synonyms | Comment | Organism |
---|---|---|
BphJ | - |
Paraburkholderia xenovorans |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
9.5 | - |
Butyraldehyde | pH 8.0 and 25°C, NAD+ | Paraburkholderia xenovorans | |
11.2 | - |
Isobutyraldehyde | pH 8.0 and 25°C, NAD+ | Paraburkholderia xenovorans | |
12.9 | - |
NADP+ | with acetaldehyde as substrate, pH 8.0 and 25°C | Paraburkholderia xenovorans | |
14.1 | - |
NAD+ | with acetaldehyde as substrate, pH 8.0 and 25°C | Paraburkholderia xenovorans | |
16.3 | - |
propionaldehyde | pH 8.0 and 25°C, NAD+ | Paraburkholderia xenovorans | |
17.2 | - |
acetaldehyde | pH 8.0 and 25°C, NAD+ | Paraburkholderia xenovorans |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
NAD+ | BphJ is able to utilize NAD+ and NADP+ with comparable kcat values, but the apparent Km-value for NAD+ is 16fold lower than for NADP+ | Paraburkholderia xenovorans | |
NADP+ | BphJ is able to utilize NAD+ and NADP+ with comparable kcat values, but the apparent Km-value for NAD+ is 16fold lower than for NADP+ | Paraburkholderia xenovorans |
General Information | Comment | Organism |
---|---|---|
metabolism | enzyme forms a bifunctional complex with EC 4.1.3.43 (BphI), 4-hydroxy-2-oxohexanoate aldolase 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 |
---|---|---|---|---|---|
0.076 | - |
pentaldehyde | app. Km-value, pH 8.0 and 25°C, NAD+ | Paraburkholderia xenovorans | |
0.3 | - |
Butyraldehyde | app. Km-value, pH 8.0 and 25°C, NAD+ | Paraburkholderia xenovorans | |
0.7 | - |
propionaldehyde | app. Km-value, pH 8.0 and 25°C, NAD+ | Paraburkholderia xenovorans | |
0.73 | - |
acetaldehyde | app. Km-value, pH 8.0 and 25°C, NAD+ | Paraburkholderia xenovorans | |
1.4 | - |
Isobutyraldehyde | app. Km-value, pH 8.0 and 25°C, NAD+ | Paraburkholderia xenovorans | |
22.99 | - |
NADP+ | app. Km-value with acetaldehyde as substrate, pH 8.0 and 25°C | Paraburkholderia xenovorans | |
405.2 | - |
NAD+ | app. Km-value with acetaldehyde as substrate, pH 8.0 and 25°C | Paraburkholderia xenovorans |