Cloned (Comment) | Organism |
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co-overexpression of TDO-F, TDO-R, TDO-O and apo-TDO-O in Escherichia coli | Pseudomonas putida |
Crystallization (Comment) | Organism |
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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 | 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 | 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 | ? | - |
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Organism | UniProt | Comment | Textmining |
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Pseudomonas putida | - |
- |
- |
Purification (Comment) | Organism |
---|---|
recombinant TDO-F, TDO-R, TDO-O and apo-TDO-O from Escherichia coli | Pseudomonas putida |
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 |
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 | ? | - |
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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 | Comment | Organism |
---|---|---|
TDO | - |
Pseudomonas putida |
toluene 2,3-dioxygenase | - |
Pseudomonas putida |