Literature summary for 1.14.12.11 extracted from

Friemann, R.; Lee, K.; Brown, E.N.; Gibson, D.T.; Eklund, H.; Ramaswamy, S.
Structures of the multicomponent Rieske non-heme iron toluene 2,3-dioxygenase enzyme system (2009), Acta Crystallogr. Sect. D, 65, 24-33.

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Cloned(Commentary)

Cloned (Comment)	Organism
co-overexpression of TDO-F, TDO-R, TDO-O and apo-TDO-O in Escherichia coli	Pseudomonas putida

Crystallization (Commentary)

Crystallization (Comment)	Organism
purified recombinant enzyme system components, hanging-drop or sitting-drop vapour diffusion method, from 1.4 M ammonium sulfate, 0.1 M HEPES, pH 7.7. TDO-F crystallized in 38% PEG 8000, 0.1 M MES pH 6.1 or 37% PEG 8000, 0.1 M MES, pH 5.8. Apo-TDO-O crystallizes in 40% w/v PEG 600, 0.1 M sodium citrate, pH 5.8. Cocrystallization experiments with Fe(NH4)2(SO4)2 and toluene were performed under strict anaerobic conditions in 40% w/v PEG 600, 0.1 M sodium citrate pH 6.1, 20-50 mM Fe(NH4)2(SO4)2 and 20 mM toluene, X-ray diffraction structure determinhation and analysis at 1.2-3.2 A resolution, molecular replacement, structure modelling, overview	Pseudomonas putida

Crystallization (Comment)

Organism

purified recombinant enzyme system components, hanging-drop or sitting-drop vapour diffusion method, from 1.4 M ammonium sulfate, 0.1 M HEPES, pH 7.7. TDO-F crystallized in 38% PEG 8000, 0.1 M MES pH 6.1 or 37% PEG 8000, 0.1 M MES, pH 5.8. Apo-TDO-O crystallizes in 40% w/v PEG 600, 0.1 M sodium citrate, pH 5.8. Cocrystallization experiments with Fe(NH4)2(SO4)2 and toluene were performed under strict anaerobic conditions in 40% w/v PEG 600, 0.1 M sodium citrate pH 6.1, 20-50 mM Fe(NH4)2(SO4)2 and 20 mM toluene, X-ray diffraction structure determinhation and analysis at 1.2-3.2 A resolution, molecular replacement, structure modelling, overview

Pseudomonas putida

Metals/Ions

Metals/Ions	Comment	Organism	Structure
Iron	the catalytic subunit contains a Rieske [2Fe2S] cluster and mononuclear iron at the active site. The iron is not strongly bound and is easily removed during enzyme purification	Pseudomonas putida

Natural Substrates/ Products (Substrates)

Natural Substrates	Organism	Comment (Nat. Sub.)	Natural Products	Comment (Nat. Pro.)	Rev.	Reac.
additional information	Pseudomonas putida	the enzyme is organized in a multicomponent Rieske non-heme iron toluene 2,3-dioxygenase enzyme system. The TDO system is composed of a reductase, TDO-R, a Rieske [2Fe2S] ferredoxin, TDO-F, and a terminal dioxygenase, TDO-O, overview. TDO-F shuttles electrons from NADH via a flavin in TDO-R to TDO-O, which catalyzes the enantioselective addition of dioxygen to the aromatic nucleus to form cis-(1R,2S)-dihydroxy-3-methylcyclohexa-3,5-diene	?	-	?

Organism

Organism	UniProt	Comment	Textmining
Pseudomonas putida	-	-	-

Purification (Commentary)

Purification (Comment)	Organism
recombinant TDO-F, TDO-R, TDO-O and apo-TDO-O from Escherichia coli	Pseudomonas putida

Reaction

Reaction	Comment	Organism	Reaction ID
toluene + NADH + H+ + O2 = (1S,2R)-3-methylcyclohexa-3,5-diene-1,2-diol + NAD+	reaction mechanism, structure-function reklationship of the enzyme organized in a multicomponent Rieske non-heme iron toluene 2,3-dioxygenase enzyme system, overview	Pseudomonas putida	a

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.	Reac.
additional information	the enzyme is organized in a multicomponent Rieske non-heme iron toluene 2,3-dioxygenase enzyme system. The TDO system is composed of a reductase, TDO-R, a Rieske [2Fe2S] ferredoxin, TDO-F, and a terminal dioxygenase, TDO-O, overview. TDO-F shuttles electrons from NADH via a flavin in TDO-R to TDO-O, which catalyzes the enantioselective addition of dioxygen to the aromatic nucleus to form cis-(1R,2S)-dihydroxy-3-methylcyclohexa-3,5-diene	Pseudomonas putida	?	-	?
additional information	enzyme active site structure, the orientation of the toluene substrate in the active site is consistent with the regiospecificity of oxygen incorporation seen in the product formed, overview	Pseudomonas putida	?	-	?

Subunits

Subunits	Comment	Organism
hexamer	enzyme is a heterohexamer containing a catalytic and a structural subunit type, consists of three domains: an FAD-binding domain, an NADH-binding domain and a C-terminal domain	Pseudomonas putida
More	the enzyme is organized in a multicomponent Rieske non-heme iron toluene 2,3-dioxygenase enzyme system, overview	Pseudomonas putida

Synonyms

Synonyms	Comment	Organism
TDO	-	Pseudomonas putida
toluene 2,3-dioxygenase	-	Pseudomonas putida