Literature summary for 1.14.12.10 extracted from

Wolfe, M.D.; Altier, D.J.; Stubna, A.; Popescu, C.V.; Muenck, E.; Lipscomb, J.D.
Benzoate 1,2-dioxygenase from Pseudomonas putida: single turnover kinetics and regulation of a two-component Rieske dioxygenase (2002), Biochemistry, 41, 9611-9626.

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Metals/Ions

Metals/Ions	Comment	Organism	Structure
Iron	oxygenase component: 2.7 mol/mol of alphabeta subunit, each alpha subunit contains a Rieske [2Fe-2S] cluster and a mononuclear iron site, reductase component: 2.02 mol/monomer	Pseudomonas putida
Iron	each alpha-subunit contains a Rieske [2Fe-2S] cluster and a mononuclear iron site	Pseudomonas putida

Molecular Weight [Da]

Molecular Weight [Da]	Molecular Weight Maximum [Da]	Comment	Organism
19000	-	alpha3,beta3, 3 * 19000 + 3 * 49000, oxygenase component, SDS-PAGE	Pseudomonas putida
38000	-	reductase component	Pseudomonas putida
49000	-	alpha3,beta3, 3 * 19000 + 3 * 49000, oxygenase component, SDS-PAGE	Pseudomonas putida
195000	-	sedimentation analysis	Pseudomonas putida
195000	-	oxygenase component, sedimentation analysis	Pseudomonas putida
204000	-	SDS-PAGE	Pseudomonas putida

Organism

Organism	UniProt	Comment	Textmining
Pseudomonas putida	-	-	-
Pseudomonas putida mt-2 / ATCC 33015 / DSM 3931 / NCIB 12182 / NCIMB 12182	-	-	-

Purification (Commentary)

Purification (Comment)	Organism
benzoate oxygenase and reductase component	Pseudomonas putida
DEAE-Sepharose Fast Flow column chromatography and Ultrogel AcA 34 gel filtration	Pseudomonas putida

Reaction

Reaction	Comment	Organism	Reaction ID
benzoate + NADH + H+ + O2 = (1R,6S)-1,6-dihydroxycyclohexa-2,4-diene-1-carboxylate + NAD+	proposed catalytic cycle	Pseudomonas putida	a

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg]	Specific Activity Maximum [µmol/min/mg]	Comment	Organism
1.36	-	benzoate oxygenase	Pseudomonas putida
790	-	benzoate reductase, reduction of K3Fe(CN)6	Pseudomonas putida

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.	Reac.
3-chlorobenzoate + O2 + NADH + H+	-	Pseudomonas putida	3-chloro-2-hydro-1,2-dihydroxybenzoate + NAD+	-	?
3-chlorobenzoate + O2 + NADH + H+	-	Pseudomonas putida mt-2 / ATCC 33015 / DSM 3931 / NCIB 12182 / NCIMB 12182	3-chloro-2-hydro-1,2-dihydroxybenzoate + NAD+	-	?
3-fluorobenzoate + O2 + NADH + H+	-	Pseudomonas putida	3-fluoro-2-hydro-1,2-dihydroxybenzoate + NAD+	-	?
3-fluorobenzoate + O2 + NADH + H+	-	Pseudomonas putida mt-2 / ATCC 33015 / DSM 3931 / NCIB 12182 / NCIMB 12182	3-fluoro-2-hydro-1,2-dihydroxybenzoate + NAD+	-	?
3-methylbenzoate + O2 + NADH + H+	-	Pseudomonas putida	3-methyl-2-hydro-1,2-dihydroxybenzoate + NAD+	-	?
3-methylbenzoate + O2 + NADH + H+	-	Pseudomonas putida mt-2 / ATCC 33015 / DSM 3931 / NCIB 12182 / NCIMB 12182	3-methyl-2-hydro-1,2-dihydroxybenzoate + NAD+	-	?
benzoate + O2 + NADH	-	Pseudomonas putida	2-hydro-1,2-dihydroxybenzoate + NAD+	-	?
benzoate + O2 + NADH	-	Pseudomonas putida mt-2 / ATCC 33015 / DSM 3931 / NCIB 12182 / NCIMB 12182	2-hydro-1,2-dihydroxybenzoate + NAD+	-	?

Subunits

Subunits	Comment	Organism
hexamer	alpha3,beta3, 3 * 19000 + 3 * 49000, oxygenase component, SDS-PAGE	Pseudomonas putida
monomer	1 * 38000, reductase component, sedimentation analysis, SDS-PAGE	Pseudomonas putida

Synonyms

Synonyms	Comment	Organism
BZDO	-	Pseudomonas putida

Cofactor

Cofactor	Comment	Organism	Structure
FAD	1 FAD per enzyme monomer	Pseudomonas putida
iron-sulfur center	each alpha-subunit contains a Rieske [2Fe-2S] cluster and a mononuclear iron site	Pseudomonas putida
NADH	-	Pseudomonas putida

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Release 2023.1 (January 2023)