Application | Comment | Organism |
---|---|---|
synthesis | development of an efficient process to transform HSP into 2,5-dihydroxypyridine (2,5-DHP) with heterologously expressed HSP hydroxylase and NADH-regenerating system, because 2,5-DHP, the product of the reaction catalyzed by HSP hydroxylase, is a valuable precursor for the chemical synthesis of 5-aminolevulinic acid, which is applied as a plant growth hormone, a herbicide and in cancer therapy | Agrobacterium tumefaciens |
Cloned (Comment) | Organism |
---|---|
DNA and amino acid sequence determination and analysis, sequence comparisons, RT-PCR expression analysis, recombinant expression of His-tagged enzyme in Escherichia coli strain BL21(DE3) | Agrobacterium tumefaciens |
Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|
90000 | - |
gel filtration | Agrobacterium tumefaciens |
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 | Agrobacterium tumefaciens | - |
2,5-dihydroxypyridine + succinate semialdehyde + 2 NAD+ + H2O | - |
? | |
4-(6-hydroxypyridin-3-yl)-4-oxobutanoate + 2 NADH + 2 H+ + O2 | Agrobacterium tumefaciens S33 | - |
2,5-dihydroxypyridine + succinate semialdehyde + 2 NAD+ + H2O | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Agrobacterium tumefaciens | W8E0Y0 | i.e. Rhizobium radiobacter | - |
Agrobacterium tumefaciens S33 | W8E0Y0 | i.e. Rhizobium radiobacter | - |
Purification (Comment) | Organism |
---|---|
native enzyme 64.4fold from Agrobacterium tumefaciens strain S33 cells by ammonium sulfate fractionation, hydrophobic interaction chromatography, desalting gel filtration, and two different steps of anion exchange chromatography, followed by gel filtration. Recombinant His-tagged enzyme from Escherichia coli strain BL21(DE3) by nickel affinity chromatography, anion exchange chromatography, and ultrafiltration. During purification, all the buffers are supplemented with 0.005 mM FAD. When FAD is omitted, the purification yield is very low | Agrobacterium tumefaciens |
Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
---|---|---|---|
10.3 | - |
purified native enzyme, pH 8.0, 30°C | Agrobacterium tumefaciens |
18.8 | - |
purified recombinant enzyme, pH 8.0, 30°C | Agrobacterium tumefaciens |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
4-(6-hydroxypyridin-3-yl)-4-oxobutanoate + 2 NADH + 2 H+ + O2 | - |
Agrobacterium tumefaciens | 2,5-dihydroxypyridine + succinate semialdehyde + 2 NAD+ + H2O | - |
? | |
4-(6-hydroxypyridin-3-yl)-4-oxobutanoate + 2 NADH + 2 H+ + O2 | oxidative decarboxylation of HSP to 2,5-dihydroxypyridine in presence of NADH and FAD | Agrobacterium tumefaciens | 2,5-dihydroxypyridine + succinate semialdehyde + 2 NAD+ + H2O | - |
? | |
4-(6-hydroxypyridin-3-yl)-4-oxobutanoate + 2 NADH + 2 H+ + O2 | - |
Agrobacterium tumefaciens S33 | 2,5-dihydroxypyridine + succinate semialdehyde + 2 NAD+ + H2O | - |
? | |
4-(6-hydroxypyridin-3-yl)-4-oxobutanoate + 2 NADH + 2 H+ + O2 | oxidative decarboxylation of HSP to 2,5-dihydroxypyridine in presence of NADH and FAD | Agrobacterium tumefaciens S33 | 2,5-dihydroxypyridine + succinate semialdehyde + 2 NAD+ + H2O | - |
? | |
additional information | no activity with p-nitrophenol or 6-hydroxynicotinate as substrates | Agrobacterium tumefaciens | ? | - |
? | |
additional information | no activity with p-nitrophenol or 6-hydroxynicotinate as substrates | Agrobacterium tumefaciens S33 | ? | - |
? |
Subunits | Comment | Organism |
---|---|---|
homodimer | 2 * 45000, SDS-PAGE | Agrobacterium tumefaciens |
Synonyms | Comment | Organism |
---|---|---|
6-hydroxy-3-succinoylpyridine hydroxylase | - |
Agrobacterium tumefaciens |
AWN88_01205 | gene name, UniProt | Agrobacterium tumefaciens |
HSP hydroxylase | - |
Agrobacterium tumefaciens |
NADH-dependent HSP hydroxylase | - |
Agrobacterium tumefaciens |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
30 | - |
assay at | Agrobacterium tumefaciens |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
8 | - |
- |
Agrobacterium tumefaciens |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
FAD | required for activity | Agrobacterium tumefaciens | |
NADH | dependent on | Agrobacterium tumefaciens |
General Information | Comment | Organism |
---|---|---|
metabolism | 6-hydroxy-3-succinoylpyridine hydroxylase catalyzes a central step of nicotine degradation. 6-Hydroxy-3-succinoylpyridine (HSP) is a key intermediate connecting the two pathways, pyridine pathway and pyrrolidine pathway, overview | Agrobacterium tumefaciens |
physiological function | the key enzyme HSP hydroxylase is involved in the fused nicotine degradation pathway of the pyridine and pyrrolidine pathways | Agrobacterium tumefaciens |