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Literature summary for 1.14.13.114 extracted from

  • Nakano, H.; Wieser, M.; Hurh, B.; Kawai, T.; Yoshida, T.; Yamane, T.; Nagasawa, T.
    Purification, characterization and gene cloning of 6-hydroxynicotinate 3-monooxygenase from Pseudomonas fluorescens TN5 (1999), Eur. J. Biochem., 260, 120-126.
    View publication on PubMed

Application

Application Comment Organism
synthesis the isolated enzyme is used for the synthesis of 2,5-dihydroxypyridine, a precursor for the chemical synthesis of 5-aminolevulinic acid, which is applied as a plant growth hormone, a herbicide and in cancer therapy Pseudomonas fluorescens

Cloned(Commentary)

Cloned (Comment) Organism
expression in Escherichia coli Pseudomonas fluorescens

Inhibitors

Inhibitors Comment Organism Structure
5,5'-dithiobis(2-nitrobenzoate) 1 mM, complete inhibition Pseudomonas fluorescens
Ag2SO4 1 mM, complete inhibition Pseudomonas fluorescens
CuCl2 1 mM, complete inhibition Pseudomonas fluorescens
HgCl2 1 mM, complete inhibition Pseudomonas fluorescens
additional information no significant effect on enzyme activity is found with metal-chelating agents such o-phenanthroline, 8-hydroxyquinoline, EDTA, disodium 4,5-dihydroxy-m-benzenedisulfonate, fluoride and azide, and other compounds such as KCl, LiCl, NaCl, BaCl2, CaCl2, MnCl2, MgCl2, PbCl2, ZnCl2, CoCl2, SnCl2, FeSO4, FeCl3, NiCl2, CdCl2, AlCl3, iodoacetic acid, hydroxylamine, phenylhydrazine, semicarbazide, cysteamine, alpha,alpha'-dipyridyl and urea Pseudomonas fluorescens
N-ethylmaleimide 1 mM, 69% inhibition Pseudomonas fluorescens
nicotinate potent competitive inhibitor Pseudomonas fluorescens
p-chloromercuribenzoate 1 mM, complete inhibition Pseudomonas fluorescens

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.098
-
6-Hydroxynicotinate pH 7.0, 30°C Pseudomonas fluorescens
0.15
-
4-hydroxybenzoate pH 7.0, 30°C Pseudomonas fluorescens

Localization

Localization Comment Organism GeneOntology No. Textmining
membrane bound to Pseudomonas fluorescens 16020
-

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
42886
-
1 * 42886, including the starting methionine, calculated from sequence Pseudomonas fluorescens

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
6-hydroxynicotinate + NADH + H+ + O2 Pseudomonas fluorescens aerobic catabolism of nicotinic acid. NADH is 5times more effective than NADPH 2,5-dihydroxypyridine + NAD+ + H2O + CO2
-
?
6-hydroxynicotinate + NADH + H+ + O2 Pseudomonas fluorescens TN5 aerobic catabolism of nicotinic acid. NADH is 5times more effective than NADPH 2,5-dihydroxypyridine + NAD+ + H2O + CO2
-
?

Organism

Organism UniProt Comment Textmining
Pseudomonas fluorescens
-
-
-
Pseudomonas fluorescens TN5
-
-
-

Purification (Commentary)

Purification (Comment) Organism
-
Pseudomonas fluorescens

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
6.67
-
-
Pseudomonas fluorescens

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
4-hydroxybenzoate + NADH + H+ + O2 6.0% relative activity compared with 6-hydroxynicotinate Pseudomonas fluorescens hydroquinone + NAD+ + H2O + CO2
-
?
4-hydroxybenzoate + NADH + H+ + O2 6.0% relative activity compared with 6-hydroxynicotinate Pseudomonas fluorescens TN5 hydroquinone + NAD+ + H2O + CO2
-
?
6-hydroxynicotinate + NADH + H+ + O2 aerobic catabolism of nicotinic acid. NADH is 5times more effective than NADPH Pseudomonas fluorescens 2,5-dihydroxypyridine + NAD+ + H2O + CO2
-
?
6-hydroxynicotinate + NADH + H+ + O2 NADH is 5times more effective than NADPH Pseudomonas fluorescens 2,5-dihydroxypyridine + NAD+ + H2O + CO2
-
?
6-hydroxynicotinate + NADH + H+ + O2 aerobic catabolism of nicotinic acid. NADH is 5times more effective than NADPH Pseudomonas fluorescens TN5 2,5-dihydroxypyridine + NAD+ + H2O + CO2
-
?
6-hydroxynicotinate + NADH + H+ + O2 NADH is 5times more effective than NADPH Pseudomonas fluorescens TN5 2,5-dihydroxypyridine + NAD+ + H2O + CO2
-
?
additional information 3-hydroxybenzoate (0.49% relative activity compared with 6-hydroxynicotinate), 2-hydroxybenzoate (0.18% compared with 6-hydroxynicotinate), 2-hydroxynicotinate (0.31% relative activity compared with 6-hydroxynicotinate) and 6-hydroxypyrazine carboxylate (0.19% relative activity compared with 6-hydroxynicotinate) are less effecive substrates or, in the case of nicotinate, 6-methylnicotinate and benzoate, not substrates at all Pseudomonas fluorescens ?
-
?
additional information 3-hydroxybenzoate (0.49% relative activity compared with 6-hydroxynicotinate), 2-hydroxybenzoate (0.18% compared with 6-hydroxynicotinate), 2-hydroxynicotinate (0.31% relative activity compared with 6-hydroxynicotinate) and 6-hydroxypyrazine carboxylate (0.19% relative activity compared with 6-hydroxynicotinate) are less effecive substrates or, in the case of nicotinate, 6-methylnicotinate and benzoate, not substrates at all Pseudomonas fluorescens TN5 ?
-
?

Subunits

Subunits Comment Organism
monomer 1 * 40000-42000, SDS-PAGE Pseudomonas fluorescens
monomer 1 * 42886, including the starting methionine, calculated from sequence Pseudomonas fluorescens

Synonyms

Synonyms Comment Organism
6-hydroxynicotinic acid 3-monooxygenase
-
Pseudomonas fluorescens

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
35
-
-
Pseudomonas fluorescens

Temperature Stability [°C]

Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
35
-
pH 7.0, 10 min, in the absence of FAD, apoenzyme is stable below Pseudomonas fluorescens
40
-
pH 7.0, 10 min, in the presence of 0.5 mM FAD, holoenzyme is stable below Pseudomonas fluorescens

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
7.6
-
-
Pseudomonas fluorescens

pH Stability

pH Stability pH Stability Maximum Comment Organism
6 9 35°C, 10 min, stable between pH 6.0 and pH 9.0 Pseudomonas fluorescens

Cofactor

Cofactor Comment Organism Structure
FAD the enzyme activity is NADH-dependent and FAD-dependent. The holoenzyme contains 1 M of FAD per 1 M of enzyme. FAD gradually dissociates from the enzyme during purification. Without FAD, no pure enzyme activity is observed, but after the addition of FAD, the apoenzyme is activated immediately Pseudomonas fluorescens
additional information riboflavin or FMN do not serve as enzyme cofactors Pseudomonas fluorescens
NADH the enzyme activity is NADH-dependent and FAD-dependent. NADH is 5times more effective than NADPH Pseudomonas fluorescens

Ki Value [mM]

Ki Value [mM] Ki Value maximum [mM] Inhibitor Comment Organism Structure
0.49
-
nicotinate
-
Pseudomonas fluorescens