Literature summary for 1.14.13.114 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 .

Search for more literature for 1.14.13.114

Cloned(Commentary)

Cloned (Comment)	Organism
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
gene nicC, DNA and amino acid sequence determination and analysis, sequence comparisons	Neobacillus niacini
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)

Crystallization (Comment)	Organism
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

Crystallization (Comment)

Organism

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

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

KM Value [mM]

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

Natural Substrates/ Products (Substrates)

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

Organism

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

Purification (Commentary)

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

Reaction

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

Substrates and Products (Substrate)

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

Subunits

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

Synonyms

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

Temperature Optimum [°C]

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

Turnover Number [1/s]

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

pH Optimum

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

pH Range

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

Cofactor

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

Ki Value [mM]

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

General Information

General Information	Comment	Organism
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
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
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
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
physiological function	6-hydroxynicotinic acid 3-monooxygenase is a decarboxylative hydroxylase involved in aerobic nicotinate degradation	Neobacillus niacini
physiological function	6-hydroxynicotinic acid 3-monooxygenase is a decarboxylative hydroxylase involved in aerobic nicotinate degradation	Pseudomonas putida
physiological function	6-hydroxynicotinic acid 3-monooxygenase is a decarboxylative hydroxylase involved in aerobic nicotinate degradation	Bordetella bronchiseptica

kcat/KM [mM/s]

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