Cloned (Comment) | Organism |
---|---|
gene hspB, phylogenetic analysis and tree, recombinant expression of His6-tagged enzyme in Escherichia coli strain BL21(DE3) | Pseudomonas putida |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
additional information | - |
additional information | stopped-flow spectroscopy and kinetic analysis | Pseudomonas putida | |
0.173 | - |
4-(6-hydroxypyridin-3-yl)-4-oxobutanoate | pH 8.0, 25°C, recombinant His-tagged enzyme | Pseudomonas putida | |
0.0294 | - |
NADH | pH 8.0, 25°C, recombinant His-tagged enzyme | Pseudomonas putida |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
4-(6-hydroxypyridin-3-yl)-4-oxobutanoate + 2 NADH + 2 H+ + O2 | Pseudomonas putida | - |
2,5-dihydroxypyridine + succinate semialdehyde + 2 NAD+ + H2O | - |
? | |
4-(6-hydroxypyridin-3-yl)-4-oxobutanoate + 2 NADH + 2 H+ + O2 | Pseudomonas putida S16 | - |
2,5-dihydroxypyridine + succinate semialdehyde + 2 NAD+ + H2O | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Pseudomonas putida | - |
- |
- |
Pseudomonas putida S16 | - |
- |
- |
Purification (Comment) | Organism |
---|---|
recombinant His6-tagged enzyme from Escherichia coli strain BL21(DE3) by nickel affinity chromatography | Pseudomonas putida |
Storage Stability | Organism |
---|---|
-80°C, purified recombinant His-tagged enzyme HspB, as a frozen solution or dry powder, no change in activity for at least 3 months, pH 8.0 | Pseudomonas putida |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
additional information | mass spectrometric analysis of substrates and products, with recombinant enzyme | Pseudomonas putida | ? | - |
? | |
additional information | mass spectrometric analysis of substrates and products, with recombinant enzyme | Pseudomonas putida S16 | ? | - |
? | |
4-(6-hydroxypyridin-3-yl)-4-oxobutanoate + 2 NADH + 2 H+ + O2 | - |
Pseudomonas putida | 2,5-dihydroxypyridine + succinate semialdehyde + 2 NAD+ + H2O | - |
? | |
4-(6-hydroxypyridin-3-yl)-4-oxobutanoate + 2 NADH + 2 H+ + O2 | - |
Pseudomonas putida S16 | 2,5-dihydroxypyridine + succinate semialdehyde + 2 NAD+ + H2O | - |
? |
Synonyms | Comment | Organism |
---|---|---|
hspB | - |
Pseudomonas putida |
6-hydroxy-3-succinoyl-pyridine 3-monooxygenase | - |
Pseudomonas putida |
HSP 3-monooxygenase | - |
Pseudomonas putida |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
25 | - |
assay at | Pseudomonas putida |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
5.49 | - |
NADH | pH 8.0, 25°C, recombinant His-tagged enzyme | Pseudomonas putida | |
7.74 | - |
4-(6-hydroxypyridin-3-yl)-4-oxobutanoate | pH 8.0, 25°C, recombinant His-tagged enzyme | Pseudomonas putida |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
8 | - |
assay at | Pseudomonas putida |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
NADH | - |
Pseudomonas putida | |
FAD | - |
Pseudomonas putida |
General Information | Comment | Organism |
---|---|---|
physiological function | 6-hydroxy-3-succinoyl-pyridine (HSP) 3-monooxygenase (HspB) is a flavoprotein essential to the pyrrolidine pathway of nicotine degradation, it catalyzes pyridine-ring beta-hydroxylation, resulting in carbon-carbon cleavage and production of 2,5-dihydroxypyridine | Pseudomonas putida |
evolution | phylogenetic analysis reveals that HspB is the most closely related to two p-nitrophenol 4-monooxygenases, and the experimental results exhibit that p-nitrophenol is a substrate of HspB | Pseudomonas putida |
additional information | free H2O2 does not catalyze the HspB enzyme reaction | Pseudomonas putida |