Literature summary for 1.13.11.9 extracted from

Jimenez, J.I.; Acebron, I.; Garcia, J.L.; Diaz, E.; Mancheno, J.M.
A preliminary crystallographic study of recombinant NicX, an Fe2+-dependent 2,5-dihydroxypyridine dioxygenase from Pseudomonas putida KT2440 (2010), Acta Crystallogr. Sect. F, 66, 549-553.

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Activating Compound

Activating Compound	Comment	Organism	Structure
DTT	-	Pseudomonas putida

Cloned(Commentary)

Cloned (Comment)	Organism
functional overexpression in Escherichia coli strain BL21 (DE3)	Pseudomonas putida

Crystallization (Commentary)

Crystallization (Comment)	Organism
purified recombinant NicX, hanging drop vapour diffusion method, 0.001 ml of 14 mg/ml protein in 20 mM HEPES pH 7.5, 0.1 M NaCl, and 2 mM DTT, is mixed with 0.001 ml of reservoir solution, containing 22% w/v PEG 4000, 0.1 M sodium acetate and 0.1 M HEPES pH 7.5, 2 mM DTT, and with 1 ml 50 mM EDTA, 18°C, method optimization, X-ray diffraction structure determination and analysis at 2.0 A resolution	Pseudomonas putida

Crystallization (Comment)

Organism

purified recombinant NicX, hanging drop vapour diffusion method, 0.001 ml of 14 mg/ml protein in 20 mM HEPES pH 7.5, 0.1 M NaCl, and 2 mM DTT, is mixed with 0.001 ml of reservoir solution, containing 22% w/v PEG 4000, 0.1 M sodium acetate and 0.1 M HEPES pH 7.5, 2 mM DTT, and with 1 ml 50 mM EDTA, 18°C, method optimization, X-ray diffraction structure determination and analysis at 2.0 A resolution

Pseudomonas putida

Metals/Ions

Metals/Ions	Comment	Organism	Structure
Fe2+	dependent on	Pseudomonas putida

Molecular Weight [Da]

Molecular Weight [Da]	Molecular Weight Maximum [Da]	Comment	Organism
143200	-	gel filtration and mass spectrometry	Pseudomonas putida

Natural Substrates/ Products (Substrates)

Natural Substrates	Organism	Comment (Nat. Sub.)	Natural Products	Comment (Nat. Pro.)	Rev.	Reac.
2,5-dihydroxypyridine + O2	Pseudomonas putida	-	N-formylmaleamic acid	-	?
2,5-dihydroxypyridine + O2	Pseudomonas putida KT 2240	-	N-formylmaleamic acid	-	?

Organism

Organism	UniProt	Comment	Textmining
Pseudomonas putida	-	-	-
Pseudomonas putida KT 2240	-	-	-

Purification (Commentary)

Purification (Comment)	Organism
recombinant enzyme from Escherichia coli strain BL21(DE3) by anion exchange and hydroxyapatite chromatography	Pseudomonas putida

Storage Stability

Storage Stability	Organism
4°C, purified recombinant enzyme, 20 mM HEPES pH 7.5, 0.1 M NaCl, and 2 mM DTT, maintains its activity for several days	Pseudomonas putida

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.	Reac.
2,5-dihydroxypyridine + O2	-	Pseudomonas putida	N-formylmaleamic acid	-	?
2,5-dihydroxypyridine + O2	-	Pseudomonas putida KT 2240	N-formylmaleamic acid	-	?

Subunits

Subunits	Comment	Organism
hexamer	6 * 38 883, a hexameric assembly composed of two cyclic trimers, gel filtration and mass spectrometry	Pseudomonas putida

Synonyms

Synonyms	Comment	Organism
2,5-DHP dioxygenase	-	Pseudomonas putida
2,5-dihydroxypyridine dioxygenase	-	Pseudomonas putida
NicX	-	Pseudomonas putida

Temperature Optimum [°C]

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

pH Optimum

pH Optimum Minimum	pH Optimum Maximum	Comment	Organism
8	-	assay at	Pseudomonas putida