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Literature summary 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.
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

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

Crystallization (Commentary)

EC Number Crystallization (Comment) Organism
1.14.12.11 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

EC Number Metals/Ions Comment Organism Structure
1.14.12.11 Iron the catalytic subunit contains a Rieske [2Fe–2S] 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)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
1.14.12.11 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 [2Fe–2S] 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 ?
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Organism

EC Number Organism UniProt Comment Textmining
1.14.12.11 Pseudomonas putida
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Purification (Commentary)

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

Reaction

EC Number Reaction Comment Organism Reaction ID
1.14.12.11 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)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
1.14.12.11 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 [2Fe–2S] 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 ?
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 ?
1.14.12.11 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 ?
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 ?

Subunits

EC Number Subunits Comment Organism
1.14.12.11 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
1.14.12.11 More the enzyme is organized in a multicomponent Rieske non-heme iron toluene 2,3-dioxygenase enzyme system, overview Pseudomonas putida

Synonyms

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