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Literature summary for 1.17.1.5 extracted from

  • Hurh, B.; Yamane, T.; Nagasawa, T.
    Purification and characterization of nicotinic acid dehydrogenase from Pseudomonas fluorescens TN5 (1994), J. Ferment. Bioeng., 78, 19-26.
No PubMed abstract available

General Stability

General Stability Organism
membrane-bound enzyme is stable, but solubilized enzyme is very labile, 20 mM nicotinate stabilizes Pseudomonas fluorescens

Inhibitors

Inhibitors Comment Organism Structure
2,2'-dipyridyl 1 mM, 10 min at 35°C, 21% loss of activity Pseudomonas fluorescens
Acriflavin 1 mM, 10 min at 35°C, 22% loss of activity Pseudomonas fluorescens
AgNO3 1 mM, 10 min at 35°C, complete loss of activity Pseudomonas fluorescens
CuCl2 1 mM, 10 min at 35°C, complete loss of activity Pseudomonas fluorescens
HgCl2 1 mM, 10 min at 35°C, complete loss of activity Pseudomonas fluorescens
KCN strong inhibition, partially restored by addition of phenazine methosulfate Pseudomonas fluorescens
N-ethylmaleimide 1 mM, 10 min at 35°C, complete loss of activity Pseudomonas fluorescens
NaN3 1 mM, 10 min at 35°C, 17% loss of activity Pseudomonas fluorescens
p-chloromercuribenzoate 1 mM, 10 min at 35°C, complete loss of activity Pseudomonas fluorescens
phenylhydrazine 1 mM, 10 min at 35°C, 15% loss of activity Pseudomonas fluorescens

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.11
-
nicotinate
-
Pseudomonas fluorescens
0.7
-
3-pyridinesulfonate
-
Pseudomonas fluorescens
2.1
-
2-Hydroxynicotinate
-
Pseudomonas fluorescens
9.8
-
2-pyrazinecarboxylate
-
Pseudomonas fluorescens
150
-
3-Cyanopyridine
-
Pseudomonas fluorescens

Localization

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

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
80000
-
1 * 80000, SDS-PAGE, in presence of 50 mM 2-mercaptoethanol Pseudomonas fluorescens
100000
-
gel filtration Pseudomonas fluorescens

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
nicotinate + H2O + electron acceptor Pseudomonas fluorescens first step of nicotinate catabolism 6-hydroxynicotinate + reduced electron acceptor
-
?
nicotinate + H2O + electron acceptor Pseudomonas fluorescens enzyme is linked to the cytochrome respiratory chain, phenazine methosulfate may accelerate the electron transfer from the electron respiratory chain to oxygen 6-hydroxynicotinate + reduced electron acceptor
-
?
nicotinate + H2O + electron acceptor Pseudomonas fluorescens TN5 first step of nicotinate catabolism 6-hydroxynicotinate + reduced electron acceptor
-
?
nicotinate + H2O + electron acceptor Pseudomonas fluorescens TN5 enzyme is linked to the cytochrome respiratory chain, phenazine methosulfate may accelerate the electron transfer from the electron respiratory chain to oxygen 6-hydroxynicotinate + reduced electron acceptor
-
?

Organism

Organism UniProt Comment Textmining
Delftia acidovorans
-
-
-
Pseudomonas fluorescens
-
-
-
Pseudomonas fluorescens TN5
-
-
-
Serratia marcescens
-
-
-
Serratia marcescens IFO 12648
-
-
-

Purification (Commentary)

Purification (Comment) Organism
126fold purification Pseudomonas fluorescens

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
1
-
2-hydroxynicotinate Pseudomonas fluorescens
12
-
3-cyanopyridine Pseudomonas fluorescens
104
-
3-pyridinesulfonate Pseudomonas fluorescens
170
-
2-pyrazinecarboxylate Pseudomonas fluorescens
672
-
nicotinate Pseudomonas fluorescens

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2-hydroxynicotinate + H2O + electron acceptor hydroxylation at a low rate Pseudomonas fluorescens ?
-
?
2-pyrazinecarboxylate + H2O + electron acceptor
-
Pseudomonas fluorescens ?
-
?
2-pyrazinecarboxylate + H2O + electron acceptor
-
Pseudomonas fluorescens TN5 ?
-
?
3-cyanopyridine + H2O + electron acceptor hydroxylation at a low rate Pseudomonas fluorescens ?
-
?
3-pyridinesulfonate + H2O + electron acceptor
-
Pseudomonas fluorescens ?
-
?
additional information not as substrates: nicotinamide, methyl nicotinate, beta-picoline, 3-pyridylcarbinol, quinolinic acid, 3-pyridylacetic acid, 3-hydroxypyridine, 3-chloropyridine, not as electron acceptors: methylene blue, FAD, FMN, NAD+, NADP+, horse heart cytochrome c, 2,3,5-triphenyl-2H-tetrazolium, neutral red, safranin O, menadione Pseudomonas fluorescens ?
-
?
additional information not as substrates: nicotinamide, methyl nicotinate, beta-picoline, 3-pyridylcarbinol, quinolinic acid, 3-pyridylacetic acid, 3-hydroxypyridine, 3-chloropyridine, not as electron acceptors: methylene blue, FAD, FMN, NAD+, NADP+, horse heart cytochrome c, 2,3,5-triphenyl-2H-tetrazolium, neutral red, safranin O, menadione Pseudomonas fluorescens TN5 ?
-
?
nicotinate + H2O + electron acceptor
-
Serratia marcescens 6-hydroxynicotinate + reduced electron acceptor
-
?
nicotinate + H2O + electron acceptor
-
Delftia acidovorans 6-hydroxynicotinate + reduced electron acceptor
-
?
nicotinate + H2O + electron acceptor purified enzyme probably loses its own native electron acceptor during the course of solubilization and purification, electron acceptors: phenazine methosulfate, potassium ferricyanide and with lower activity nitro blue tetrazolium, negative polarity at the 3-position on the pyridine ring of the substrate is important for reaction Pseudomonas fluorescens 6-hydroxynicotinate + reduced electron acceptor
-
?
nicotinate + H2O + electron acceptor first step of nicotinate catabolism Pseudomonas fluorescens 6-hydroxynicotinate + reduced electron acceptor
-
?
nicotinate + H2O + electron acceptor enzyme is linked to the cytochrome respiratory chain, phenazine methosulfate may accelerate the electron transfer from the electron respiratory chain to oxygen Pseudomonas fluorescens 6-hydroxynicotinate + reduced electron acceptor
-
?
nicotinate + H2O + electron acceptor
-
Serratia marcescens IFO 12648 6-hydroxynicotinate + reduced electron acceptor
-
?
nicotinate + H2O + electron acceptor purified enzyme probably loses its own native electron acceptor during the course of solubilization and purification, electron acceptors: phenazine methosulfate, potassium ferricyanide and with lower activity nitro blue tetrazolium, negative polarity at the 3-position on the pyridine ring of the substrate is important for reaction Pseudomonas fluorescens TN5 6-hydroxynicotinate + reduced electron acceptor
-
?
nicotinate + H2O + electron acceptor first step of nicotinate catabolism Pseudomonas fluorescens TN5 6-hydroxynicotinate + reduced electron acceptor
-
?
nicotinate + H2O + electron acceptor enzyme is linked to the cytochrome respiratory chain, phenazine methosulfate may accelerate the electron transfer from the electron respiratory chain to oxygen Pseudomonas fluorescens TN5 6-hydroxynicotinate + reduced electron acceptor
-
?

Subunits

Subunits Comment Organism
monomer 1 * 80000, SDS-PAGE, in presence of 50 mM 2-mercaptoethanol Pseudomonas fluorescens

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
35
-
assay at Pseudomonas fluorescens
50
-
-
Pseudomonas fluorescens

Temperature Stability [°C]

Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
50
-
10 min in 0.1 M potassium phosphate buffer, pH 7, stable below Pseudomonas fluorescens

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
8.3
-
-
Pseudomonas fluorescens

pH Range

pH Minimum pH Maximum Comment Organism
4.5 9 broad pH-range with maximum at pH 8.3 Pseudomonas fluorescens

pH Stability

pH Stability pH Stability Maximum Comment Organism
5 8.8 10 min at 50°C in various buffers, stable Pseudomonas fluorescens

Cofactor

Cofactor Comment Organism Structure
additional information no cytochromes or flavoprotein components Pseudomonas fluorescens
phenazine methosulfate phenazine methosulfate-requiring enzyme Pseudomonas fluorescens