General Stability | Organism |
---|---|
1-hydroxy-2-naphthoic acid hydroxylase in the cell-free extract is stabilized by 0.1 mM 1-hydroxy-2-naphthoic acid, 5 mM FAD, 2 mM DTT, and 5% glycerol | Alcaligenes sp. |
repeated freezing and thawing led to inactivation of the enzyme | Alcaligenes sp. |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.0716 | - |
1-hydroxy-2-naphthoate | pH 7.5, 30°C, with NADH | Alcaligenes sp. | |
0.0755 | - |
1-hydroxy-2-naphthoate | pH 7.5, 30°C, with NADPH | Alcaligenes sp. | |
0.0846 | - |
NADPH | pH 7.5, 30°C | Alcaligenes sp. | |
0.087 | - |
NADH | pH 7.5, 30°C | Alcaligenes sp. |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
additional information | no effect by 1 mM of Fe2+, Fe3+, Mg2+, Mn2+, Ca2+, Zn2+, and Cu2+, and by metal chelators, such as EDTA, 2,2-dipyridyl and 1',10'-phenanthroline, at 1 mM | Alcaligenes sp. |
Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|
34000 | - |
2 * 34000, SDS-PAGE | Alcaligenes sp. |
66000 | - |
gel filtration | Alcaligenes sp. |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
1-hydroxy-2-naphthoate + NADH + O2 | Alcaligenes sp. | a flavoprotein monooxygenase specific for 1-hydroxy-2-naphthoic acid, no activity with any other hydroxynaphthoic acid analogues, o-phthalic acid, 3,4-dihydroxybenzoic acid, or salicylic acid. In the naphthalene route, 1-hydroxy-2-naphthoic acid is metabolized via 1,2-dihydroxynaphthalene and salicylic acid to catechol by a series of enzymatic steps similar to naphthalene metabolic pathway | 1,2-dihydroxynaphthalene + NAD+ + H2O + CO2 | - |
? | |
1-hydroxy-2-naphthoate + NADPH + O2 | Alcaligenes sp. | a flavoprotein monooxygenase specific for 1-hydroxy-2-naphthoic acid, no activity with any other hydroxynaphthoic acid analogues, o-phthalic acid, 3,4-dihydroxybenzoic acid, or salicylic acid | 1,2-dihydroxynaphthalene + NADP+ + H2O + CO2 | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Alcaligenes sp. | - |
phenanthrene-degrading | - |
no activity in Pseudomonas putida | - |
- |
- |
no activity in Pseudomonas putida CSV86 | - |
- |
- |
Purification (Comment) | Organism |
---|---|
native enzyme 81fold by heat treatment at 60°C, ammonium sulfate fractionation and anion exchange chromatography. Additional purification steps, such as hydrophobic interaction chromatography or gel filtration, lead to the total or a significant, 70%, loss of activity, respectively, without achieving any further purification | Alcaligenes sp. |
Source Tissue | Comment | Organism | Textmining |
---|---|---|---|
culture condition:phenanthrene-grown cell | - |
Alcaligenes sp. | - |
culture condition:salicylate-grown cell | significantly lower activity compared to phenanthrene-grown cells | Alcaligenes sp. | - |
Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
---|---|---|---|
0.0031 | - |
salicylate-grown cells, pH 7.5, 30°C | Alcaligenes sp. |
0.0877 | - |
phenanthrene-grown cells, pH 7.5, 30°C | Alcaligenes sp. |
25.3 | - |
purified enzyme, cofactor NADPH, pH 7.5, 30°C | Alcaligenes sp. |
Storage Stability | Organism |
---|---|
-20°C, purified enzyme, stabilized by 0.1 mM 1-hydroxy-2-naphthoic acid, 5 mM FAD, 2 mM DTT, and 5% glycerol, retains 100% activity, after 60 days | Alcaligenes sp. |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
1-hydroxy-2-naphthoate + NADH + O2 | a flavoprotein monooxygenase specific for 1-hydroxy-2-naphthoic acid, no activity with any other hydroxynaphthoic acid analogues, o-phthalic acid, 3,4-dihydroxybenzoic acid, or salicylic acid | Alcaligenes sp. | 1,2-dihydroxynaphthalene + NAD+ + H2O + CO2 | - |
? | |
1-hydroxy-2-naphthoate + NADH + O2 | a flavoprotein monooxygenase specific for 1-hydroxy-2-naphthoic acid, no activity with any other hydroxynaphthoic acid analogues, o-phthalic acid, 3,4-dihydroxybenzoic acid, or salicylic acid. In the naphthalene route, 1-hydroxy-2-naphthoic acid is metabolized via 1,2-dihydroxynaphthalene and salicylic acid to catechol by a series of enzymatic steps similar to naphthalene metabolic pathway | Alcaligenes sp. | 1,2-dihydroxynaphthalene + NAD+ + H2O + CO2 | - |
? | |
1-hydroxy-2-naphthoate + NADPH + O2 | a flavoprotein monooxygenase specific for 1-hydroxy-2-naphthoic acid, no activity with any other hydroxynaphthoic acid analogues, o-phthalic acid, 3,4-dihydroxybenzoic acid, or salicylic acid | Alcaligenes sp. | 1,2-dihydroxynaphthalene + NADP+ + H2O + CO2 | - |
? | |
additional information | substrate specificity, overview. No activity on salicylic acid, 3-hydroxy-2-naphthoic acid, 2-hydroxy-1-naphthoic acid, 3-hydroxybenzoic acid, 4-hydroxybenzoic acid, 3,4-dihydroxybenzoic acid, 1-naphthol, 2-naphthol, 1-naphthoic acid, 2-naphthoic acid, gentisic acid or catechol | Alcaligenes sp. | ? | - |
? |
Subunits | Comment | Organism |
---|---|---|
homodimer | 2 * 34000, SDS-PAGE | Alcaligenes sp. |
Synonyms | Comment | Organism |
---|---|---|
1-hydroxy-2-naphthoic acid hydroxylase | - |
Alcaligenes sp. |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
30 | - |
assay at | Alcaligenes sp. |
Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
---|---|---|---|
60 | - |
5 min, cell-free extract, the enzyme is stable in presence of 1-hydroxy-2-naphthoic acid | Alcaligenes sp. |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7.5 | - |
- |
Alcaligenes sp. |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
FAD | holoenzyme contains FAD. Km is 0.0047 mM. Preparation of the apoenzyme by the acid-ammonium sulfate, at 2 M, dialysis method, the apoenzyme is colorless, inactive and loses the characteristic flavin absorption spectra but regains about 90% of maximal activity when reconstituted with FAD | Alcaligenes sp. | |
additional information | FMN is a poor substitute for FAD | Alcaligenes sp. | |
NADH | Vmax/Km is similar for NADPH and NADH | Alcaligenes sp. | |
NADPH | Vmax/Km is similar for NADPH and NADH | Alcaligenes sp. |
Organism | Comment | Expression |
---|---|---|
Alcaligenes sp. | the enzyme is induced by growth on phenanthrene | up |
General Information | Comment | Organism |
---|---|---|
metabolism | in the naphthalene route, 1-hydroxy-2-naphthoic acid is metabolized via 1,2-dihydroxynaphthalene and salicylic acid to catechol by a series of enzymatic steps similar to naphthalene metabolic pathway | Alcaligenes sp. |