Cloned (Comment) | Organism |
---|---|
mutations confirmed by sequence analysis | Paraburkholderia xenovorans |
Crystallization (Comment) | Organism |
---|---|
E257Q-NADP+ mutant at 1.4 A resolution, C296A-NADP+ at 1.4 A resolution, E167A mutant at 1.6 A resolution, E496A mutant at 1.65 A resolution | Paraburkholderia xenovorans |
Protein Variants | Comment | Organism |
---|---|---|
C296A | same purification behavior as the native enzyme indicating that the mutation does not alter basic enzyme structure, but no catalytic activity detected consistent with a critical nucleophilic role for C296 | Paraburkholderia xenovorans |
E167A | significant reduction in enzyme activity but not perturbing the overall structure, significant reduction in activity of 77%, likely candidate to present the catalytic water to the general base E257 that leads to deacylation and product release | Paraburkholderia xenovorans |
E257Q | tolerated with respect to overall protein stability, but dramatic reduction in kcat value and no activity detectable, E257 serves probably as the primary general base to deprotonate the nucleophilic C296 | Paraburkholderia xenovorans |
E400A | 91% reduction in kcat value, E400 plays an essential role in facilitating the acquisition of the hydride conformation | Paraburkholderia xenovorans |
E496A | significant reduction in enzyme activity but not perturbing the overall structure, significant reduction in activity of 90% | Paraburkholderia xenovorans |
H485A | dramatic reduction in enzyme solubility | Paraburkholderia xenovorans |
H485Q | completely recalcitrant to purification, H485 may be important in stabilizing the nicotinamide amide moiety of NADP+ | Paraburkholderia xenovorans |
K168A | significant reduction in enzyme activity but not perturbing the overall structure, significant reduction in activity of 92.5%, K168 likely serves to stabilize charges and tune pKa of the active site glutamate side-chains | Paraburkholderia xenovorans |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
cis-3,4-dehydroadipyl-CoA semialdehyde + NADP+ + H2O | Paraburkholderia xenovorans | catalytic mechanism proposed in detail | cis-3,4-dehydroadipyl-CoA + NADPH + H+ | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Paraburkholderia xenovorans | Q13WK4 | - |
- |
Purification (Comment) | Organism |
---|---|
by nickel affinity column and gel filtration | Paraburkholderia xenovorans |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
cis-3,4-dehydroadipyl-CoA semialdehyde + NADP+ + H2O | catalytic mechanism proposed in detail | Paraburkholderia xenovorans | cis-3,4-dehydroadipyl-CoA + NADPH + H+ | - |
? | |
propionaldehyde + NADP+ + H2O | propionaldehyde is used as the aldehydic substrate in enzyme assay because of commercial unavailability of the native substrate | Paraburkholderia xenovorans | propionate + NADPH + H+ | - |
? |
Synonyms | Comment | Organism |
---|---|---|
ALDH | - |
Paraburkholderia xenovorans |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
additional information | - |
additional information | no activities detectable for C296A and E257Q mutant enzymes | Paraburkholderia xenovorans | |
5.75 | - |
propionaldehyde | H485A mutant enzyme, pH 7.5, temperature not specified in the publication | Paraburkholderia xenovorans | |
12.1 | - |
propionaldehyde | K168A mutant enzyme, pH 7.5, temperature not specified in the publication | Paraburkholderia xenovorans | |
14.6 | - |
propionaldehyde | E400A mutant enzyme, pH 7.5, temperature not specified in the publication | Paraburkholderia xenovorans | |
16.2 | - |
propionaldehyde | E496A mutant enzyme, pH 7.5, temperature not specified in the publication | Paraburkholderia xenovorans | |
36.8 | - |
propionaldehyde | E167A mutant enzyme, pH 7.5, temperature not specified in the publication | Paraburkholderia xenovorans | |
162 | - |
propionaldehyde | wild type enzyme, pH 7.5, temperature not specified in the publication | Paraburkholderia xenovorans |
General Information | Comment | Organism |
---|---|---|
metabolism | enzyme catalyzes an essential step in the metabolism of benzoate in Burkholderia xenovorans LB400 | Paraburkholderia xenovorans |