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Literature summary extracted from

  • Hicks, K.; Yuen, M.; Zhen, W.; Gerwig, T.; Story, R.; Kopp, M.; Snider, M.
    Structural and biochemical characterization of 6-hydroxynicotinic acid 3-monooxygenase, a novel decarboxylative hydroxylase involved in aerobic nicotinate degradation (2016), Biochemistry, 55, 3432-3446 .
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
1.14.13.114 gene Bb1770, DNA and amino acid sequence determination and analysis, sequence comparisons, recombinant expression of His-tagged enzyme in Escherichia coli strains MachI and BL21(DE3) Bordetella bronchiseptica
1.14.13.114 gene nicC, DNA and amino acid sequence determination and analysis, sequence comparisons Neobacillus niacini
1.14.13.114 gene nicC, DNA and amino acid sequence determination and analysis, sequence comparisons, recombinant expression of His-tagged enzyme in Escherichia coli strains MachI and BL21(DE3) Pseudomonas putida

Crystallization (Commentary)

EC Number Crystallization (Comment) Organism
1.14.13.114 purified recombinant His-tagged selenomethionine-labeled enzyme, hanging drop vapor diffusion method, mixing of 0.0015 ml of protein solution with 0.0015 ml of reservoir solution containing 0.1 M Tris-HCl, pH 7.7-8.3, 27-30% PEG 4000, and 0.2 M magnesium chloride hexahydrate, 18°C, 2-3 days, method optimization, X-ray diffraction structure determination and analysis using SAD phasing at 2.1 A resolution Pseudomonas putida

Inhibitors

EC Number Inhibitors Comment Organism Structure
1.14.13.114 6-hydroxynicotinaldehyde competitive inhibition with respect to 6-hydroxynicotinate is weak Bordetella bronchiseptica
1.14.13.114 iodoacetamide
-
 Bordetella bronchiseptica
1.14.13.114 methylmethanethiosulfonate
-
 Bordetella bronchiseptica

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
1.14.13.114 additional information
-
 additional information kinetic analysis Pseudomonas putida
1.14.13.114 additional information
-
 additional information kinetic analysis Bordetella bronchiseptica
1.14.13.114 0.003
-
 NADH pH 7.5, 25°C Pseudomonas putida
1.14.13.114 0.006
-
 NADH pH 7.5, 25°C Bordetella bronchiseptica
1.14.13.114 0.085
-
 6-Hydroxynicotinate pH 7.5, 25°C Bordetella bronchiseptica
1.14.13.114 0.097
-
 6-Hydroxynicotinate pH 7.5, 25°C Pseudomonas putida

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
1.14.13.114 6-hydroxynicotinate + NADH + H+ + O2 Neobacillus niacini
-
 2,5-dihydroxypyridine + NAD+ + H2O + CO2
-
 ?
1.14.13.114 6-hydroxynicotinate + NADH + H+ + O2 Pseudomonas putida
-
 2,5-dihydroxypyridine + NAD+ + H2O + CO2
-
 ?
1.14.13.114 6-hydroxynicotinate + NADH + H+ + O2 Bordetella bronchiseptica
-
 2,5-dihydroxypyridine + NAD+ + H2O + CO2
-
 ?
1.14.13.114 6-hydroxynicotinate + NADH + H+ + O2 Bordetella bronchiseptica ATCC BAA-588 / NCTC 13252 / RB50
-
 2,5-dihydroxypyridine + NAD+ + H2O + CO2
-
 ?

Organism

EC Number Organism UniProt Comment Textmining
1.14.13.114 Bordetella bronchiseptica A0A0H3LLR0
-
-
1.14.13.114 Bordetella bronchiseptica ATCC BAA-588 / NCTC 13252 / RB50 A0A0H3LLR0
-
-
1.14.13.114 Neobacillus niacini
-
-
-
1.14.13.114 Pseudomonas putida Q88FY2
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
1.14.13.114 recombinant His-tagged enzyme from Escherichia coli strain BL21(DE3) by nickel affinity chromatography Pseudomonas putida
1.14.13.114 recombinant His-tagged enzyme from Escherichia coli strain BL21(DE3) by nickel affinity chromatography Bordetella bronchiseptica

Reaction

EC Number Reaction Comment Organism Reaction ID
1.14.13.114 6-hydroxynicotinate + NADH + H+ + O2 = 2,5-dihydroxypyridine + NAD+ + H2O + CO2 proposal of two catalytic mechanisms, overview Pseudomonas putida
a
1.14.13.114 6-hydroxynicotinate + NADH + H+ + O2 = 2,5-dihydroxypyridine + NAD+ + H2O + CO2 proposal of two catalytic mechanisms, overview Bordetella bronchiseptica
a

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
1.14.13.114 6-hydroxynicotinate + NADH + H+ + O2
-
 Neobacillus niacini 2,5-dihydroxypyridine + NAD+ + H2O + CO2
-
 ?
1.14.13.114 6-hydroxynicotinate + NADH + H+ + O2
-
 Pseudomonas putida 2,5-dihydroxypyridine + NAD+ + H2O + CO2
-
 ?
1.14.13.114 6-hydroxynicotinate + NADH + H+ + O2
-
 Bordetella bronchiseptica 2,5-dihydroxypyridine + NAD+ + H2O + CO2
-
 ?
1.14.13.114 6-hydroxynicotinate + NADH + H+ + O2
-
 Bordetella bronchiseptica ATCC BAA-588 / NCTC 13252 / RB50 2,5-dihydroxypyridine + NAD+ + H2O + CO2
-
 ?

Subunits

EC Number Subunits Comment Organism
1.14.13.114 More structure of the PpNicC monomer, modeling, overview Pseudomonas putida

Synonyms

EC Number Synonyms Comment Organism
1.14.13.114 6-hydroxynicotinic acid 3-monooxygenase
-
 Neobacillus niacini
1.14.13.114 6-hydroxynicotinic acid 3-monooxygenase
-
 Pseudomonas putida
1.14.13.114 6-hydroxynicotinic acid 3-monooxygenase
-
 Bordetella bronchiseptica
1.14.13.114 BB1770
-
 Bordetella bronchiseptica
1.14.13.114 BbNicC
-
 Bordetella bronchiseptica
1.14.13.114 NicC
-
 Pseudomonas putida
1.14.13.114 PpNicC
-
 Pseudomonas putida

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
1.14.13.114 25
-
 assay at Pseudomonas putida
1.14.13.114 25
-
 assay at Bordetella bronchiseptica

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
1.14.13.114 2.2
-
 NADH pH 7.5, 25°C Pseudomonas putida
1.14.13.114 2.2
-
 6-Hydroxynicotinate pH 7.5, 25°C Pseudomonas putida
1.14.13.114 4.2
-
 NADH pH 7.5, 25°C Bordetella bronchiseptica
1.14.13.114 4.2
-
 6-Hydroxynicotinate pH 7.5, 25°C Bordetella bronchiseptica

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
1.14.13.114 7.5
-
 assay at Pseudomonas putida
1.14.13.114 7.5
-
 assay at Bordetella bronchiseptica

pH Range

EC Number pH Minimum pH Maximum Comment Organism
1.14.13.114 additional information
-
 pH-dependence of the enzyme fitting to a double-bell curve pH-rate profile, overview Bordetella bronchiseptica

Cofactor

EC Number Cofactor Comment Organism Structure
1.14.13.114 FAD
-
 Bordetella bronchiseptica
1.14.13.114 FAD NicC FAD binding site structure, overview Pseudomonas putida
1.14.13.114 NADH
-
 Neobacillus niacini
1.14.13.114 NADH
-
 Pseudomonas putida
1.14.13.114 NADH
-
 Bordetella bronchiseptica

Ki Value [mM]

EC Number Ki Value [mM] Ki Value maximum [mM] Inhibitor Comment Organism Structure
1.14.13.114 additional information
-
 additional information inhibition kinetics Bordetella bronchiseptica
1.14.13.114 2.98
-
 6-hydroxynicotinaldehyde pH 7.5, 25°C, recombinant enzyme Bordetella bronchiseptica

General Information

EC Number General Information Comment Organism
1.14.13.114 metabolism the enzyme is involved in the nicotinate degradation pathway catalyzing the the second of three oxidations of nicotinate that activate the pyridine toward ring cleavage by aerobic bacteria, overview Neobacillus niacini
1.14.13.114 metabolism the enzyme is involved in the nicotinate degradation pathway catalyzing the the second of three oxidations of nicotinate that activate the pyridine toward ring cleavage by aerobic bacteria, overview Pseudomonas putida
1.14.13.114 metabolism the enzyme is involved in the nicotinate degradation pathway catalyzing the the second of three oxidations of nicotinate that activate the pyridine toward ring cleavage by aerobic bacteria, overview Bordetella bronchiseptica
1.14.13.114 additional information modeling of substrate 6-hydroxynicotinate into the active site, substrate binding site structure, overview. The nicC gene in Pseudomonas putida strain KT2440 has been misidentified as salicylate hydroxylase, nahG Pseudomonas putida
1.14.13.114 physiological function 6-hydroxynicotinic acid 3-monooxygenase is a decarboxylative hydroxylase involved in aerobic nicotinate degradation Neobacillus niacini
1.14.13.114 physiological function 6-hydroxynicotinic acid 3-monooxygenase is a decarboxylative hydroxylase involved in aerobic nicotinate degradation Pseudomonas putida
1.14.13.114 physiological function 6-hydroxynicotinic acid 3-monooxygenase is a decarboxylative hydroxylase involved in aerobic nicotinate degradation Bordetella bronchiseptica

kcat/KM [mM/s]

EC Number kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
1.14.13.114 22.6
-
 6-Hydroxynicotinate pH 7.5, 25°C Pseudomonas putida
1.14.13.114 49.4
-
 6-Hydroxynicotinate pH 7.5, 25°C Bordetella bronchiseptica
1.14.13.114 700
-
 NADH pH 7.5, 25°C Bordetella bronchiseptica
1.14.13.114 733.3
-
 NADH pH 7.5, 25°C Pseudomonas putida